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1. Comparison of preclinical efficacy of immunotherapies against HPV-induced cancers

Author

Anastasia Demidova, Laëtitia Douguet, Ingrid Fert, Yu Wei, Pierre Charneau, and Laleh Majlessi

Subjects
Antitumor immunity, cervix cancers, early E6 and E7 oncoproteins, HPV-induced cancers, immuno-oncotherapy, oropharyngeal cancers, Internal medicine, RC31-1245
Abstract

Introduction Persistent infections with the human papilloma viruses, HPV16 and HPV18, are associated with multiple cancers. Although prophylactic vaccines that induce HPV-neutralizing antibodies are effective against primary infections, they have no effect on HPV-mediated malignancies against which there is no approved immuno-therapy. Active research is ongoing in the immunotherapy of these cancers.Areas covered In this review, we compared the preclinical efficacy of vaccine platforms used to treat HPV-induced tumors in the standard model of mice grafted with TC-1 cells, which express the HPV16 E6 and E7 oncoproteins. We searched for the key words, ‘HPV,’ ‘vaccine,’ ‘therapy,’ ‘E7,’ ‘tumor,’ ‘T cells’, and ‘mice’ for the period from 2005 to 2023 in PubMed and found 330 publications. Among them, we selected the most relevant to extract preclinical antitumor results to enable cross-sectional comparison of their efficacy.Expert opinion section We compared these studies for HPV antigen design, immunization regimen, immunogenicity, and antitumor effect, considering their drawbacks and advantages. Among all strategies used in murine models, certain adjuvanted proteins and viral vectors showed the strongest antitumor effects, with the use of lentiviral vectors being the only approach to result in complete tumor eradication in 100% of experimental individuals while providing the longest-lasting memory.

Published
2024
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2. T-cell immunity induced and reshaped by an anti-HPV immuno-oncotherapeutic lentiviral vector

Author

Ingrid Fert, Laëtitia Douguet, Benjamin Vesin, Fanny Moncoq, Amandine Noirat, Pierre Authié, Sylvain Ciret, Fabien Le Chevalier, Catherine Blanc, Yakov Vitrenko, Pierre Charneau, Laleh Majlessi, and François Anna

Subjects
Immunologic diseases. Allergy, RC581-607, Neoplasms. Tumors. Oncology. Including cancer and carcinogens, RC254-282
Abstract

Abstract We recently developed an immuno-oncotherapy against human papillomavirus (HPV)-induced tumors based on a lentiviral vector encoding the Early E6 and E7 oncoproteins of HPV16 and HPV18 genotypes, namely “Lenti-HPV-07”. The robust and long-lasting anti-tumor efficacy of Lenti-HPV-07 is dependent on CD8+ T-cell induction and remodeling of the tumor microenvironment. Here, we first established that anti-vector immunity induced by Lenti-HPV-07 prime has no impact on the efficacy of a homologous boost to amplify anti-HPV T-cell immunity. To longitudinally monitor the evolution of the T-cell repertoire generated after the prime, homologous or heterologous boost with Lenti-HPV-07, we tracked T-cell clonotypes by deep sequencing of T-Cell Receptor (TCR) variable β and α chain mRNA, applied to whole peripheral blood cells (PBL) and a T cell population specific of an immunodominant E7HPV16 epitope. We observed a hyper-expansion of clonotypes post prime, accompanied by increased frequencies of HPV-07-specific T cells. Additionally, there was a notable diversification of clonotypes post boost in whole PBL, but not in the E7HPV16-specific T cells. We then demonstrated that the effector functions of such Lenti-HPV-07-induced T cells synergize with anti-checkpoint inhibitory treatments by systemic administration of anti-TIM3 or anti-NKG2A monoclonal antibodies. While Lenti-HPV-07 is about to enter a Phase I/IIa clinical trial, these results will help better elucidate its mode of action in immunotherapy against established HPV-mediated malignancies.

Published
2024
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3. Full eradication of pre‐clinical human papilloma virus‐induced tumors by a lentiviral vaccine

Author

Laëtitia Douguet, Ingrid Fert, Jodie Lopez, Benjamin Vesin, Fabien Le Chevalier, Fanny Moncoq, Pierre Authié, Trang‐My Nguyen, Amandine Noirat, Fabien Névo, Catherine Blanc, Maryline Bourgine, David Hardy, François Anna, Laleh Majlessi, and Pierre Charneau

Subjects
early E6‐E7 oncoproteins, immuno‐oncotherapy, intra‐tumoral immune cells, lentiviral vector, tumor microenvironment, Medicine (General), R5-920, Genetics, QH426-470
Abstract

Abstract Human papillomavirus (HPV) infections are the cause of all cervical and numerous oropharyngeal and anogenital cancers. The currently available HPV vaccines, which induce neutralizing antibodies, have no therapeutic effect on established tumors. Here, we developed an immuno‐oncotherapy against HPV‐induced tumors based on a non‐integrative lentiviral vector encoding detoxified forms of the Early E6 and E7 oncoproteins of HPV16 and 18 genotypes, namely, “Lenti‐HPV‐07”. A single intramuscular injection of Lenti‐HPV‐07 into mice bearing established HPV‐induced tumors resulted in complete tumor eradication in 100% of the animals and was also effective against lung metastases. This effect correlated with CD8+ T‐cell induction and profound remodeling of the tumor microenvironment. In the intra‐tumoral infiltrates of vaccinated mice, the presence of large amounts of activated effector, resident memory, and transcription factor T cell factor‐1 (TCF‐1)+ “stem‐like” CD8+ T cells was associated with full tumor eradication. The Lenti‐HPV‐07‐induced immunity was long‐lasting and prevented tumor growth after a late re‐challenge, mimicking tumor relapse. Lenti‐HPV‐07 therapy synergizes with an anti‐checkpoint inhibitory treatment and therefore shows promise as an immuno‐oncotherapy against established HPV‐mediated malignancies.

Published
2023
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4. Preclinical proof of concept of a tetravalent lentiviral T-cell vaccine against dengue viruses

Author

Kirill Nemirov, Pierre Authié, Philippe Souque, Fanny Moncoq, Amandine Noirat, Catherine Blanc, Maryline Bourgine, Laleh Majlessi, and Pierre Charneau

Subjects
dengue, DENV, flavivirus, lentiviral vector, ifnar-/-mice, tetravalent vaccine, Immunologic diseases. Allergy, RC581-607
Abstract

Dengue virus (DENV) is responsible for approximately 100 million cases of dengue fever annually, including severe forms such as hemorrhagic dengue and dengue shock syndrome. Despite intensive vaccine research and development spanning several decades, a universally accepted and approved vaccine against dengue fever has not yet been developed. The major challenge associated with the development of such a vaccine is that it should induce simultaneous and equal protection against the four DENV serotypes, because past infection with one serotype may greatly increase the severity of secondary infection with a distinct serotype, a phenomenon known as antibody-dependent enhancement (ADE). Using a lentiviral vector platform that is particularly suitable for the induction of cellular immune responses, we designed a tetravalent T-cell vaccine candidate against DENV (“LV-DEN”). This vaccine candidate has a strong CD8+ T-cell immunogenicity against the targeted non-structural DENV proteins, without inducing antibody response against surface antigens. Evaluation of its protective potential in the preclinical flavivirus infection model, i.e., mice knockout for the receptor to the type I IFN, demonstrated its significant protective effect against four distinct DENV serotypes, based on reduced weight loss, viremia, and viral loads in peripheral organs of the challenged mice. These results provide proof of concept for the use of lentiviral vectors for the development of efficient polyvalent T-cell vaccine candidates against all DENV serotypes.

Published
2023
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5. Lentiviral vector induces high-quality memory T cells via dendritic cells transduction

Author

Min Wen Ku, Pierre Authié, Fabien Nevo, Philippe Souque, Maryline Bourgine, Marta Romano, Pierre Charneau, and Laleh Majlessi

Subjects
Biology (General), QH301-705.5
Abstract

Ku et al. report a lentiviral (LV) vector system in which expression of vaccine antigens is driven by β2-microglobulin (β2m) promoter. Conducting comparative studies in mice and rats, they find that LV-based vaccine outperforms the ‘gold-standard’ Ad5 by inducing a polyfunctional and long-lived immune response.

Published
2021
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6. Prevalence of SARS-CoV-2 antibodies in France: results from nationwide serological surveillance

Author

Stéphane Le Vu, Gabrielle Jones, François Anna, Thierry Rose, Jean-Baptiste Richard, Sibylle Bernard-Stoecklin, Sophie Goyard, Caroline Demeret, Olivier Helynck, Nicolas Escriou, Marion Gransagne, Stéphane Petres, Corinne Robin, Virgile Monnet, Louise Perrin de Facci, Marie-Noelle Ungeheuer, Lucie Léon, Yvonnick Guillois, Laurent Filleul, Pierre Charneau, Daniel Lévy-Bruhl, Sylvie van der Werf, and Harold Noel

Subjects
Science
Abstract

The percentage of national populations infected during the first stages of the COVID-19 pandemic are unclear owing to limited early testing. Here the authors provide a nation-wide prevalence study of SARS-CoV-2 antibodies in France from the first wave of COVID-19 in 2020, including stratification based on age, sex and region.

Published
2021
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7. A lentiviral vector encoding fusion of light invariant chain and mycobacterial antigens induces protective CD4+ T cell immunity

Author

Jodie Lopez, François Anna, Pierre Authié, Alexandre Pawlik, Min-Wen Ku, Catherine Blanc, Philippe Souque, Fanny Moncoq, Amandine Noirat, David Hardy, Wladimir Sougakoff, Roland Brosch, Françoise Guinet, Pierre Charneau, and Laleh Majlessi

Subjects
CP: Immunology, Biology (General), QH301-705.5
Abstract

Summary: Lentiviral vectors (LVs) are highly efficient at inducing CD8+ T cell responses. However, LV-encoded antigens are processed inside the cytosol of antigen-presenting cells, which does not directly communicate with the endosomal major histocompatibility complex class II (MHC-II) presentation pathway. LVs are thus poor at inducing CD4+ T cell response. To overcome this limitation, we devised a strategy whereby LV-encoded antigens are extended at their N-terminal end with the MHC-II-associated light invariant chain (li), which contains an endosome-targeting signal sequence. When evaluated with an LV-encoded polyantigen composed of CD4+ T cell targets from Mycobacterium tuberculosis, intranasal vaccination in mice triggers pulmonary polyfunctional CD4+ and CD8+ T cell responses. Adjuvantation of these LVs extends the mucosal immunity to Th17 and Tc17 responses. A systemic prime and an intranasal boost with one of these LV induces protection against M. tuberculosis. This strategy improves the protective power of LVs against infections and cancers, where CD4+ T cell immunity plays an important role.

Published
2022
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8. Lentiviral Vectors as a Vaccine Platform against Infectious Diseases

Author

Kirill Nemirov, Maryline Bourgine, François Anna, Yu Wei, Pierre Charneau, and Laleh Majlessi

Subjects
lentiviral vaccine vectors, T-cell vaccines, tropism for dendritic cells, antigen presentation, major histocompatibility complex, inflammation, Pharmacy and materia medica, RS1-441
Abstract

Lentiviral vectors are among the most effective viral vectors for vaccination. In clear contrast to the reference adenoviral vectors, lentiviral vectors have a high potential for transducing dendritic cells in vivo. Within these cells, which are the most efficient at activating naive T cells, lentiviral vectors induce endogenous expression of transgenic antigens that directly access antigen presentation pathways without the need for external antigen capture or cross-presentation. Lentiviral vectors induce strong, robust, and long-lasting humoral, CD8+ T-cell immunity and effective protection against several infectious diseases. There is no pre-existing immunity to lentiviral vectors in the human population and the very low pro-inflammatory properties of these vectors pave the way for their use in mucosal vaccination. In this review, we have mainly summarized the immunological aspects of lentiviral vectors, their recent optimization to induce CD4+ T cells, and our recent data on lentiviral vector-based vaccination in preclinical models, including prophylaxis against flaviviruses, SARS-CoV-2, and Mycobacterium tuberculosis.

Published
2023
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9. Pre-COVID-19 Immunity to Common Cold Human Coronaviruses Induces a Recall-Type IgG Response to SARS-CoV-2 Antigens Without Cross-Neutralisation

Author

Makoto Miyara, Melissa Saichi, Delphine Sterlin, François Anna, Stéphane Marot, Alexis Mathian, Mo Atif, Paul Quentric, Audrey Mohr, Laetitia Claër, Christophe Parizot, Karim Dorgham, Hans Yssel, Jehane Fadlallah, Thibaut Chazal, Julien Haroche, Charles-Edouard Luyt, Julien Mayaux, Alexandra Beurton, Neila Benameur, David Boutolleau, Sonia Burrel, Sophia de Alba, Sasi Mudumba, Rick Hockett, Cary Gunn, Pierre Charneau, Vincent Calvez, Anne-Geneviève Marcelin, Alain Combes, Alexandre Demoule, Zahir Amoura, and Guy Gorochov

Subjects
COVID-19, humoral immune response, coronaviral infection, IgG, IgA, Immunologic diseases. Allergy, RC581-607
Abstract

The capacity of pre-existing immunity to human common coronaviruses (HCoV) to cross-protect against de novo COVID-19is yet unknown. In this work, we studied the sera of 175 COVID-19 patients, 76 healthy donors and 3 intravenous immunoglobulins (IVIG) batches. We found that most COVID-19 patients developed anti-SARS-CoV-2 IgG antibodies before IgM. Moreover, the capacity of their IgGs to react to beta-HCoV, was present in the early sera of most patients before the appearance of anti-SARS-CoV-2 IgG. This implied that a recall-type antibody response was generated. In comparison, the patients that mounted an anti-SARS-COV2 IgM response, prior to IgG responses had lower titres of anti-beta-HCoV IgG antibodies. This indicated that pre-existing immunity to beta-HCoV was conducive to the generation of memory type responses to SARS-COV-2. Finally, we also found that pre-COVID-19-era sera and IVIG cross-reacted with SARS-CoV-2 antigens without neutralising SARS-CoV-2 infectivity in vitro. Put together, these results indicate that whilst pre-existing immunity to HCoV is responsible for recall-type IgG responses to SARS-CoV-2, it does not lead to cross-protection against COVID-19.

Published
2022
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10. Brain cross‐protection against SARS‐CoV‐2 variants by a lentiviral vaccine in new transgenic mice

Author

Min‐Wen Ku, Pierre Authié, Maryline Bourgine, François Anna, Amandine Noirat, Fanny Moncoq, Benjamin Vesin, Fabien Nevo, Jodie Lopez, Philippe Souque, Catherine Blanc, Ingrid Fert, Sébastien Chardenoux, llta Lafosse, Delphine Cussigh, David Hardy, Kirill Nemirov, Françoise Guinet, Francina Langa Vives, Laleh Majlessi, and Pierre Charneau

Subjects
central nervous system, hACE2 transgenic mice, intranasal vaccination, olfactory bulb, SARS‐CoV‐2 emerging variants of concern, Medicine (General), R5-920, Genetics, QH426-470
Abstract

Abstract COVID‐19 vaccines already in use or in clinical development may have reduced efficacy against emerging SARS‐CoV‐2 variants. In addition, although the neurotropism of SARS‐CoV‐2 is well established, the vaccine strategies currently developed have not taken into account protection of the central nervous system. Here, we generated a transgenic mouse strain expressing the human angiotensin‐converting enzyme 2, and displaying unprecedented brain permissiveness to SARS‐CoV‐2 replication, in addition to high permissiveness levels in the lung. Using this stringent transgenic model, we demonstrated that a non‐integrative lentiviral vector, encoding for the spike glycoprotein of the ancestral SARS‐CoV‐2, used in intramuscular prime and intranasal boost elicits sterilizing protection of lung and brain against both the ancestral virus, and the Gamma (P.1) variant of concern, which carries multiple vaccine escape mutations. Beyond induction of strong neutralizing antibodies, the mechanism underlying this broad protection spectrum involves a robust protective T‐cell immunity, unaffected by the recent mutations accumulated in the emerging SARS‐CoV‐2 variants.

Published
2021
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11. Full-Lung Prophylaxis against SARS-CoV-2 by One-Shot or Booster Intranasal Lentiviral Vaccination in Syrian Golden Hamsters

Author

Benjamin Vesin, Pierre Authié, Catherine Blanc, Ingrid Fert, Amandine Noirat, Fabien Le Chevalier, Yu Wei, Min-Wen Ku, Kirill Nemirov, François Anna, David Hardy, Cyril Planchais, Hugo Mouquet, Françoise Guinet, Pierre Charneau, Laleh Majlessi, and Maryline Bourgine

Subjects
lentiviral vectors, SARS-CoV-2 variants, intranasal vaccination, mucosal immunity, lung inflammation, cross-neutralization, Medicine
Abstract

Following the breakthrough of numerous severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants in recent months and the incomplete efficiency of the currently available vaccines, development of more effective vaccines is desirable. Non-integrative, non-cytopathic and non-inflammatory lentiviral vectors elicit sterilizing prophylaxis against SARS-CoV-2 in preclinical animal models and are particularly suitable for mucosal vaccination, which is acknowledged as the most effective in reducing viral transmission. Here, we demonstrate that a single intranasal administration of a vaccinal lentiviral vector encoding a stabilized form of the original SARS-CoV-2 Spike glycoprotein induces full-lung protection of respiratory tracts and strongly reduces pulmonary inflammation in the susceptible Syrian golden hamster model against the prototype SARS-CoV-2. In addition, we show that a lentiviral vector encoding stabilized Spike of SARS-CoV-2 Beta variant (LV::SBeta-2P) prevents pathology and reduces infectious viral loads in lungs and nasal turbinates following inoculation with the SARS-CoV-2 Omicron variant. Importantly, an intranasal boost with LV::SBeta-2P improves cross-seroneutralization much better in LV::SBeta-2P-primed hamsters than in their counterparts primed with an LV-encoding Spike from the ancestral SARS-CoV-2. These results strongly suggest that an immune imprint with the original Spike sequence has a negative impact on cross-protection against new variants. Our results tackle the issue of vaccine effectiveness in people who have already been vaccinated and have vanished immunity and indicate the efficiency of LV-based intranasal vaccination, either as a single dose or as booster.

Published
2022
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12. A Humanized Mouse Strain That Develops Spontaneously Immune-Mediated Diabetes

Author

Sandrine Luce, Sophie Guinoiseau, Alexis Gadault, Franck Letourneur, Patrick Nitschke, Marc Bras, Michel Vidaud, Pierre Charneau, Etienne Larger, Maikel L. Colli, Decio L. Eizirik, François Lemonnier, and Christian Boitard

Subjects
autoimmunity, type 1 diabetes (T1D), humanized mouse, HLA-DQ8, epitopes, preproinsulin, Immunologic diseases. Allergy, RC581-607
Abstract

To circumvent the limitations of available preclinical models for the study of type 1 diabetes (T1D), we developed a new humanized model, the YES-RIP-hB7.1 mouse. This mouse is deficient of murine major histocompatibility complex class I and class II, the murine insulin genes, and expresses as transgenes the HLA-A*02:01 allele, the diabetes high-susceptibility HLA-DQ8A and B alleles, the human insulin gene, and the human co-stimulatory molecule B7.1 in insulin-secreting cells. It develops spontaneous T1D along with CD4+ and CD8+ T-cell responses to human preproinsulin epitopes. Most of the responses identified in these mice were validated in T1D patients. This model is amenable to characterization of hPPI-specific epitopes involved in T1D and to the identification of factors that may trigger autoimmune response to insulin-secreting cells in human T1D. It will allow evaluating peptide-based immunotherapy that may directly apply to T1D in human and complete preclinical model availability to address the issue of clinical heterogeneity of human disease.

Published
2021
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13. Humoral immunity to SARS-CoV-2 and seasonal coronaviruses in children and adults in north-eastern France

Author

Tom Woudenberg, Stéphane Pelleau, François Anna, Mikael Attia, Françoise Donnadieu, Alain Gravet, Caroline Lohmann, Hélène Seraphin, Raphaël Guiheneuf, Catherine Delamare, Karl Stefic, Julien Marlet, Etienne Brochot, Sandrine Castelain, Olivier Augereau, Jean Sibilia, François Dubos, Damia Meddour, Christèle Gras-Le Guen, Marianne Coste-Burel, Berthe-Marie Imbert-Marcille, Anne Chauvire-Drouard, Cyril Schweitzer, Amélie Gatin, Sandra Lomazzi, Aline Joulié, Hervé HAAS, Aymeric Cantais, Frederique Bertholon, Marie-France Chinazzo-Vigouroux, Mohamed SI Abdallah, Laurence Arowas, Pierre Charneau, Bruno Hoen, Caroline Demeret, Sylvie Van Der Werf, Arnaud Fontanet, and Michael White

Subjects
SARS-CoV-2, COVID-19, seroprevalence, sero-epidemiology, seasonal coronaviruses, antibody response, Medicine, Medicine (General), R5-920
Abstract

Background: Children are underrepresented in the COVID-19 pandemic and often experience milder disease than adolescents and adults. Reduced severity is possibly due to recent and more frequent seasonal human coronaviruses (HCoV) infections. We assessed the seroprevalence of SARS-CoV-2 and seasonal HCoV specific antibodies in a large cohort in north-eastern France. Methods: In this cross-sectional seroprevalence study, serum samples were collected from children and adults requiring hospital admission for non-COVID-19 between February and August 2020. Antibody responses to SARS-CoV-2 and seasonal HCoV (229E, HKU1, NL63, OC43) were assessed using a bead-based multiplex assay, Luciferase-Linked ImmunoSorbent Assay, and a pseudotype neutralisation assay. Findings: In 2,408 individuals, seroprevalence of SARS-CoV-2-specific antibodies was 7-8% with three different immunoassays. Antibody levels to seasonal HCoV increased substantially up to the age of 10. Antibody responses in SARS-CoV-2 seropositive individuals were lowest in adults 18-30 years. In SARS-CoV-2 seronegative individuals, we observed cross-reactivity between antibodies to the four HCoV and SARS-CoV-2 Spike. In contrast to other antibodies to SARS-CoV-2, specific antibodies to sub-unit 2 of Spike (S2) in seronegative samples were highest in children. Upon infection with SARS-CoV-2, antibody levels to Spike of betacoronavirus OC43 increased across the whole age spectrum. No SARS-CoV-2 seropositive individuals with low levels of antibodies to seasonal HCoV were observed. Interpretation: Our findings underline significant cross-reactivity between antibodies to SARS-CoV-2 and seasonal HCoV, but provide no significant evidence for cross-protective immunity to SARS-CoV-2 infection due to a recent seasonal HCoV infection. In particular, across all age groups we did not observe SARS-CoV-2 infected individuals with low levels of antibodies to seasonal HCoV. Funding: This work was supported by the « URGENCE COVID-19 » fundraising campaign of Institut Pasteur, by the French Government's Investissement d'Avenir program, Laboratoire d'Excellence Integrative Biology of Emerging Infectious Diseases (Grant No. ANR-10-LABX-62-IBEID), and by the REACTing (Research & Action Emerging Infectious Diseases), and by the RECOVER project funded by the European Union's Horizon 2020 research and innovation programme under grant agreement No. 101003589, and by a grant from LabEx IBEID (ANR-10-LABX-62-IBEID).

Published
2021
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14. Asymptomatic and symptomatic SARS-CoV-2 infections elicit polyfunctional antibodies

Author

Jérémy Dufloo, Ludivine Grzelak, Isabelle Staropoli, Yoann Madec, Laura Tondeur, François Anna, Stéphane Pelleau, Aurélie Wiedemann, Cyril Planchais, Julian Buchrieser, Rémy Robinot, Marie-Noelle Ungeheuer, Hugo Mouquet, Pierre Charneau, Michael White, Yves Lévy, Bruno Hoen, Arnaud Fontanet, Olivier Schwartz, and Timothée Bruel

Subjects
SARS-CoV-2, ADCC, complement, asymptomatic, antibody, Medicine (General), R5-920
Abstract

Summary: Many SARS-CoV-2-infected individuals remain asymptomatic. Little is known about the extent and quality of their antiviral humoral response. Here, we analyze antibody functions in 52 asymptomatic infected individuals, 119 mildly symptomatic, and 21 hospitalized patients with COVID-19. We measure anti-spike immunoglobulin G (IgG), IgA, and IgM levels with the S-Flow assay and map IgG-targeted epitopes with a Luminex assay. We also evaluate neutralization, complement deposition, and antibody-dependent cellular cytotoxicity (ADCC) using replication-competent SARS-CoV-2 or reporter cell systems. We show that COVID-19 sera mediate complement deposition and kill infected cells by ADCC. Sera from asymptomatic individuals neutralize the virus, activate ADCC, and trigger complement deposition. Antibody levels and functions are lower in asymptomatic individuals than they are in symptomatic cases. Antibody functions are correlated, regardless of disease severity. Longitudinal samplings show that antibody functions follow similar kinetics of induction and contraction. Overall, asymptomatic SARS-CoV-2 infection elicits polyfunctional antibodies neutralizing the virus and targeting infected cells.

Published
2021
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15. Multiplexed Quantitation of Intraphagocyte Mycobacterium tuberculosis Secreted Protein Effectors

Author

Fadel Sayes, Catherine Blanc, Louis S. Ates, Nathalie Deboosere, Mickael Orgeur, Fabien Le Chevalier, Matthias I. Gröschel, Wafa Frigui, Ok-Ryul Song, Richard Lo-Man, Florence Brossier, Wladimir Sougakoff, Daria Bottai, Priscille Brodin, Pierre Charneau, Roland Brosch, and Laleh Majlessi

Subjects
Biology (General), QH301-705.5
Abstract

Summary: The pathogenic potential of Mycobacterium tuberculosis largely depends on ESX secretion systems exporting members of the multigenic Esx, Esp, and PE/PPE protein families. To study the secretion and regulation patterns of these proteins while circumventing immune cross-reactions due to their extensive sequence homologies, we developed an approach that relies on the recognition of their MHC class II epitopes by highly discriminative T cell receptors (TCRs) of a panel of T cell hybridomas. The latter were engineered so that each expresses a unique fluorescent reporter linked to specific antigen recognition. The resulting polychromatic and multiplexed imaging assay enabled us to measure the secretion of mycobacterial effectors inside infected host cells. We applied this novel technology to a large panel of mutants, clinical isolates, and host-cell types to explore the host-mycobacteria interplay and its impact on the intracellular bacterial secretome, which also revealed the unexpected capacity of phagocytes from lung granuloma to present mycobacterial antigens via MHC class II. : Sayes et al. develop an approach to express distinct fluorescent reporters that is based on the recognition of specific Mycobacterium tuberculosis MHC class II epitopes by highly discriminative T cell hybridomas. This multiplexed technology allows the study of secretion, subcellular location, and regulation patterns of these instrumental protein members. Keywords: mycobacterium tuberculosis, type VII secretion systems, intracellular bacteria, T-cell hybridomas, mycobacterial virulence factors, bacterial antigen presentation, lentiviral vectors, reporter T cells, in vivo antigen presentation, protein localization

Published
2018
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16. TBVAC2020: Advancing Tuberculosis Vaccines from Discovery to Clinical Development

Author

Stefan H. E. Kaufmann, Hazel M. Dockrell, Nick Drager, Mei Mei Ho, Helen McShane, Olivier Neyrolles, Tom H. M. Ottenhoff, Brij Patel, Danielle Roordink, François Spertini, Steffen Stenger, Jelle Thole, Frank A. W. Verreck, Ann Williams, TBVAC2020 Consortium, Warwick Britton, Jamie Triccas, Claudio Counoupas, Johan Grooten, Marie-Ange Demoitie, Marta Romano, Francoise Mascart, Peter Andersen, Claus Aagaard, Dennis Christensen, Morten Ruhwald, Thomas Lindenstrom, Pierre Charneau, Christophe Guilhot, Antonio Peixoto, Martine Gilleron, Camille Locht, Roland Brosch, Genevieve Inchauspe, Stephane Leung Theung Long, Stefan Kaufmann, January Weiner, Jeroen Maertzdorf, Natalie Neuwenhuizen, Max Bastian, Nadia Caccamo, Delia Goletti, Roberto Nisini, Sung Jae Shin, Hyejon Lee, Alex Sigal, Thomas Scriba, Gerhard Walzl, Andre Loxton, Robert Wilkinson, Pere-Joan Cardona, Cris Vilaplana, Carlos Martin, Dessi Marinova, Nacho Aguilo, Ruedi Aebersold, Etienne Caron, Daniel Pinschewer, Gennaro De Libero, Claire Anne Siegrist, Nicolas Collin, Christophe Barnier-Quer, Peter Sander, Frank Verreck, Tom Ottenhoff, Simone Joosten, Krista van Meijgaarden, Mariateresa Coppola, Annemieke Geluk, Yvonne Perrie, Marc Baird, Michael Levin, Hazel Dockrell, Steven Smith, Helen Fletcher, Gregory Bancroft, Ann Rawkins, Simon Clark, Iman Satti, Elena Stylianou, Martin Vordermeier, and Philip Hogarth

Subjects
tuberculosis, bacille Calmette–Guérin, vaccination, biomarker, clinical trial, portfolio management, Immunologic diseases. Allergy, RC581-607
Abstract

TBVAC2020 is a research project supported by the Horizon 2020 program of the European Commission (EC). It aims at the discovery and development of novel tuberculosis (TB) vaccines from preclinical research projects to early clinical assessment. The project builds on previous collaborations from 1998 onwards funded through the EC framework programs FP5, FP6, and FP7. It has succeeded in attracting new partners from outstanding laboratories from all over the world, now totaling 40 institutions. Next to the development of novel vaccines, TB biomarker development is also considered an important asset to facilitate rational vaccine selection and development. In addition, TBVAC2020 offers portfolio management that provides selection criteria for entry, gating, and priority settings of novel vaccines at an early developmental stage. The TBVAC2020 consortium coordinated by TBVI facilitates collaboration and early data sharing between partners with the common aim of working toward the development of an effective TB vaccine. Close links with funders and other consortia with shared interests further contribute to this goal.

Published
2017
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17. Recombinant Zika NS1 Protein Secreted from Vero Cells Is Efficient for Inducing Production of Immune Serum Directed against NS1 Dimer

Author

Wildriss Viranaicken, Alexia Ndebo, Sandra Bos, Philippe Souque, Gilles Gadea, Chaker El-Kalamouni, Pascale Krejbich-Trotot, Pierre Charneau, Philippe Desprès, and Marjolaine Roche

Subjects
emerging disease, arbovirus, Zika virus, NS1 protein, lentiviral vector, recombinant antigen, humoral immunity, NS1 antiserum, Biology (General), QH301-705.5, Chemistry, QD1-999
Abstract

Zika virus (ZIKV) is a mosquito-borne flavivirus that recently emerged in the South Pacific, Americas, and Caribbean islands, where the larger epidemics were documented. ZIKV infection in humans is responsible for neurological disorders and microcephaly. Flavivirus NS1 is a non-structural glycoprotein that is expressed on the cell surface and secreted as a hexameric lipoprotein particle. Intracellular NS1 exists as a dimer that is required for viral replication, whereas the secreted NS1 hexamer interacts with host factors, leading to pathophysiological conditions. In an effort to dispose of specific anti-ZIKV NS1 immune serum, Vero cells were transduced with a lentiviral vector containing the NS1 gene from an epidemic strain of ZIKV. We showed that stably transduced Vero/ZIKV NS1 cell clone was efficient in the secretion of recombinant NS1 oligomer. Immunization of adult rat with purified extracellular NS1 developed anti-ZIKV antibodies that specifically react with the NS1 dimer produced in human cells infected with African and Asian strains of ZIKV. The rat antibody against ZIKV NS1 dimer is a reliable biological tool that enables the immunological detection of secreted NS1 from host-cells infected with ZIKV.

Published
2017
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18. A Lentiviral Vector Expressing Japanese Encephalitis Virus-like Particles Elicits Broad Neutralizing Antibody Response in Pigs.

Author

Mélissanne de Wispelaere, Meret Ricklin, Philippe Souque, Marie-Pascale Frenkiel, Sylvie Paulous, Obdulio Garcìa-Nicolàs, Artur Summerfield, Pierre Charneau, and Philippe Desprès

Subjects
Arctic medicine. Tropical medicine, RC955-962, Public aspects of medicine, RA1-1270
Abstract

BACKGROUND:Japanese encephalitis virus (JEV) is the major cause of viral encephalitis in Southeast Asia. Vaccination of domestic pigs has been suggested as a "one health" strategy to reduce viral disease transmission to humans. The efficiency of two lentiviral TRIP/JEV vectors expressing the JEV envelope prM and E glycoproteins at eliciting protective humoral response was assessed in a mouse model and piglets. METHODOLOGY/PRINCIPAL FINDINGS:A gene encoding the envelope proteins prM and E from a genotype 3 JEV strain was inserted into a lentiviral TRIP vector. Two lentiviral vectors TRIP/JEV were generated, each expressing the prM signal peptide followed by the prM protein and the E glycoprotein, the latter being expressed either in its native form or lacking its two C-terminal transmembrane domains. In vitro transduction of cells with the TRIP/JEV vector expressing the native prM and E resulted in the efficient secretion of virus-like particles of Japanese encephalitis virus. Immunization of BALB/c mice with TRIP/JEV vectors resulted in the production of IgGs against Japanese encephalitis virus, and the injection of a second dose one month after the prime injection greatly boosted antibody titers. The TRIP/JEV vectors elicited neutralizing antibodies against JEV strains belonging to genotypes 1, 3, and 5. Immunization of piglets with two doses of the lentiviral vector expressing JEV virus-like particles led to high titers of anti-JEV antibodies, that had efficient neutralizing activity regardless of the JEV genotype tested. CONCLUSIONS/SIGNIFICANCE:Immunization of pigs with the lentiviral vector expressing JEV virus-like particles is particularly efficient to prime antigen-specific humoral immunity and trigger neutralizing antibody responses against JEV genotypes 1, 3, and 5. The titers of neutralizing antibodies elicited by the TRIP/JEV vector are sufficient to confer protection in domestic pigs against different genotypes of JEV and this could be of a great utility in endemic regions where more than one genotype is circulating.

Published
2015
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19. TOX4 and NOVA1 proteins are partners of the LEDGF PWWP domain and affect HIV-1 replication.

Author

Mehdi Morchikh, Monica Naughtin, Francesca Di Nunzio, Johan Xavier, Pierre Charneau, Yves Jacob, and Marc Lavigne

Subjects
Medicine, Science
Abstract

PWWP domains are involved in the chromatin attachment of several proteins. They bind to both DNA and proteins and their interaction with specific histone methylation marks define them as a new class of histone code readers. The lens epithelium derived growth factor (LEDGF/p75) contains an N-terminal PWWP domain necessary for its interaction with chromatin but also a C-terminal domain which interacts with several proteins, such as lentiviral integrases. These two domains confer a chromatin-tethering function to LEDGF/p75 and in the case of lentiviral integrases, this tethering participates in the efficiency and site selectivity of integration. Although proteins interacting with LEDGF/p75 C-terminal domain have been extensively studied, no data exist about partners of its PWWP domain regulating its interaction with chromatin. In this study, we report the identification by yeast-two-hybrid of thirteen potential partners of the LEDGF PWWP domain. Five of these interactions were confirmed in mammalian cells, using both a protein complementation assay and co-immunoprecipitation approaches. Three of these partners interact with full length LEDGF/p75, they are specific for PWWP domains of the HDGF family and they require PWWP amino acids essential for the interaction with chromatin. Among them, the transcription activator TOX4 and the splicing cofactor NOVA1 were selected for a more extensive study. These two proteins or their PWWP interacting regions (PIR) colocalize with LEDGF/p75 in Hela cells and interact in vitro in the presence of DNA. Finally, single round VSV-G pseudotyped HIV-1 but not MLV infection is inhibited in cells overexpressing these two PIRs. The observed inhibition of infection can be attributed to a defect in the integration step. Our data suggest that a regulation of LEDGF interaction with chromatin by cellular partners of its PWWP domain could be involved in several processes linked to LEDGF tethering properties, such as lentiviral integration, DNA repair or transcriptional regulation.

Published
2013
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21. A nonintegrative lentiviral vector-based vaccine provides long-term sterile protection against malaria.

Author

Frédéric Coutant, Raul Yusef Sanchez David, Tristan Félix, Aude Boulay, Laxmee Caleechurn, Philippe Souque, Catherine Thouvenot, Catherine Bourgouin, Anne-Sophie Beignon, and Pierre Charneau

Subjects
Medicine, Science
Abstract

Trials testing the RTS,S candidate malaria vaccine and radiation-attenuated sporozoites (RAS) have shown that protective immunity against malaria can be induced and that an effective vaccine is not out of reach. However, longer-term protection and higher protection rates are required to eradicate malaria from the endemic regions. It implies that there is still a need to explore new vaccine strategies. Lentiviral vectors are very potent at inducing strong immunological memory. However their integrative status challenges their safety profile. Eliminating the integration step obviates the risk of insertional oncogenesis. Providing they confer sterile immunity, nonintegrative lentiviral vectors (NILV) hold promise as mass pediatric vaccine by meeting high safety standards. In this study, we have assessed the protective efficacy of NILV against malaria in a robust pre-clinical model. Mice were immunized with NILV encoding Plasmodium yoelii Circumsporozoite Protein (Py CSP) and challenged with sporozoites one month later. In two independent protective efficacy studies, 50% (37.5-62.5) of the animals were fully protected (p = 0.0072 and p = 0.0008 respectively when compared to naive mice). The remaining mice with detectable parasitized red blood cells exhibited a prolonged patency and reduced parasitemia. Moreover, protection was long-lasting with 42.8% sterile protection six months after the last immunization (p = 0.0042). Post-challenge CD8+ T cells to CSP, in contrast to anti-CSP antibodies, were associated with protection (r = -0.6615 and p = 0.0004 between the frequency of IFN-g secreting specific T cells in spleen and parasitemia). However, while NILV and RAS immunizations elicited comparable immunity to CSP, only RAS conferred 100% of sterile protection. Given that a better protection can be anticipated from a multi-antigen vaccine and an optimized vector design, NILV appear as a promising malaria vaccine.

Published
2012
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22. Human nucleoporins promote HIV-1 docking at the nuclear pore, nuclear import and integration.

Author

Francesca Di Nunzio, Anne Danckaert, Thomas Fricke, Patricio Perez, Juliette Fernandez, Emmanuelle Perret, Pascal Roux, Spencer Shorte, Pierre Charneau, Felipe Diaz-Griffero, and Nathalie J Arhel

Subjects
Medicine, Science
Abstract

The nuclear pore complex (NPC) mediates nucleo-cytoplasmic transport of macromolecules and is an obligatory point of passage and functional bottleneck in the replication of some viruses. The Human Immunodeficiency Virus (HIV) has evolved the required mechanisms for active nuclear import of its genome through the NPC. However the mechanisms by which the NPC allows or even assists HIV translocation are still unknown. We investigated the involvement of four key nucleoporins in HIV-1 docking, translocation, and integration: Nup358/RanBP2, Nup214/CAN, Nup98 and Nup153. Although all induce defects in infectivity when depleted, only Nup153 actually showed any evidence of participating in HIV-1 translocation through the nuclear pore. We show that Nup358/RanBP2 mediates docking of HIV-1 cores on NPC cytoplasmic filaments by interacting with the cores and that the C-terminus of Nup358/RanBP2 comprising a cyclophilin-homology domain contributes to binding. We also show that Nup214/CAN and Nup98 play no role in HIV-1 nuclear import per se: Nup214/CAN plays an indirect role in infectivity read-outs through its effect on mRNA export, while the reduction of expression of Nup98 shows a slight reduction in proviral integration. Our work shows the involvement of nucleoporins in diverse and functionally separable steps of HIV infection and nuclear import.

Published
2012
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23. Liver cancer-derived hepatitis C virus core proteins shift TGF-beta responses from tumor suppression to epithelial-mesenchymal transition.

Author

Serena Battaglia, Nassima Benzoubir, Soizic Nobilet, Pierre Charneau, Didier Samuel, Anna Linda Zignego, Azeddine Atfi, Christian Bréchot, and Marie-Françoise Bourgeade

Subjects
Medicine, Science
Abstract

BACKGROUND: Chronic hepatitis C virus (HCV) infection and associated liver cirrhosis represent a major risk factor for hepatocellular carcinoma (HCC) development. TGF-beta is an important driver of liver fibrogenesis and cancer; however, its actual impact in human cancer progression is still poorly known. The aim of this study was to investigate the role of HCC-derived HCV core natural variants on cancer progression through their impact on TGF-beta signaling. PRINCIPAL FINDINGS: We provide evidence that HCC-derived core protein expression in primary human or mouse hepatocyte alleviates TGF-beta responses in terms or growth inhibition or apoptosis. Instead, in these hepatocytes TGF-beta was still able to induce an epithelial to mesenchymal transition (EMT), a process that contributes to the promotion of cell invasion and metastasis. Moreover, we demonstrate that different thresholds of Smad3 activation dictate the TGF-beta responses in hepatic cells and that HCV core protein, by decreasing Smad3 activation, may switch TGF-beta growth inhibitory effects to tumor promoting responses. CONCLUSION/SIGNIFICANCE: Our data illustrate the capacity of hepatocytes to develop EMT and plasticity under TGF-beta, emphasize the role of HCV core protein in the dynamic of these effects and provide evidence for a paradigm whereby a viral protein implicated in oncogenesis is capable to shift TGF-beta responses from cytostatic effects to EMT development.

Published
2009
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24. Protective antiviral immunity conferred by a nonintegrative lentiviral vector-based vaccine.

Author

Frédéric Coutant, Marie-Pascale Frenkiel, Philippe Despres, and Pierre Charneau

Subjects
Medicine, Science
Abstract

Lentiviral vectors are under intense scrutiny as unique candidate viral vector vaccines against tumor and aggressive pathogens because of their ability to initiate potent and durable specific immune responses. Strategies that alleviate safety concerns will facilitate the clinical developments involving lentiviral vectors. In this respect, the development of integration deficient lentiviral vectors circumvents the safety concerns relative to insertional mutagenesis and might pave the way for clinical applications in which gene transfer is targeted to non-dividing cells. We thus evaluated the potential use of nonintegrative lentiviral vectors as vaccination tools since the main targeted cell in vaccination procedures is the non-dividing dendritic cell (DC). In this study, we demonstrated that a single administration of nonintegrative vectors encoding a secreted form of the envelope of a virulent strain of West Nile Virus (WNV) induces a robust B cell response. Remarkably, nonintegrative lentiviral vectors fully protected mice from a challenge with a lethal dose of WNV and a single immunization was sufficient to induce early and long-lasting protective immunity. Thus, nonintegrative lentiviral vectors might represent a safe and efficacious vaccination platform for the development of prophylactic vaccines against infectious agents.

Published
2008
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25. A lentiviral vector expressing a dendritic cell-targeting multimer induces mucosal anti-mycobacterial CD4+ T-cell immunity

Author

François Anna, Jodie Lopez, Fanny Moncoq, Catherine Blanc, Pierre Authié, Amandine Noirat, Ingrid Fert, Philippe Souque, Fabien Nevo, Alexandre Pawlik, David Hardy, Sophie Goyard, Denis Hudrisier, Roland Brosch, Françoise Guinet, Olivier Neyrolles, Pierre Charneau, Laleh Majlessi, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), Pathogénomique mycobactérienne intégrée - Integrated Mycobacterial Pathogenomics, Université Paris Cité (UPCité)-Microbiologie Intégrative et Moléculaire (UMR6047), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Plate-Forme d’Histopathologie / Histopathology Platform, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Plateforme d'Innovation et de Développement de Tests Diagnostiques / Diagnostic Test Innovation and Development Core Facility, Institut de pharmacologie et de biologie structurale (IPBS), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS), Lymphocytes et Immunité - Lymphocytes and Immunity, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), This work was also supported by the Programmes Transversaux de Recherche (PTR # 52-17 from Institut Pasteur to L.M. and P.C.), the EU program TBVAC2020 (contract n°643381 to P.C.), CNRS, University of Toulouse, Agence Nationale de la Recherche/Program d’Investissements d’Avenir (ANR-11-EQUIPEX-0003 to O.N.), the Fondation pour la Recherche Médicale (DEQ20160334902 to O.N.), the Bettencourt Schueller Foundation (Grants Coup d’Élan pour la Recherche Française and Explore-TB to O.N.)., European Project: 643381,H2020,H2020-PHC-2014-single-stage,TBVAC2020(2015), Hardy, David, TBVAC2020, Advancing novel and promising TB vaccine candidates from discovery to preclinical and early clinical development - TBVAC2020 - - H20202015-01-01 - 2018-12-31 - 643381 - VALID, and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV] Life Sciences [q-bio], [SDV]Life Sciences [q-bio], Immunology, Immunology and Allergy
Abstract

International audience; Abstract Most viral vectors, including the potently immunogenic lentiviral vectors (LVs), only poorly direct antigens to the MHC-II endosomal pathway and elicit CD4 + T cells. We developed a new generation of LVs encoding antigen-bearing monomers of collectins substituted at their C-terminal domain with the CD40 ligand ectodomain to target and activate antigen-presenting cells. Host cells transduced with such optimized LVs secreted soluble collectin-antigen polymers with the potential to be endocytosed in vivo and reach the MHC-II pathway. In the murine tuberculosis model, such LVs induced efficient MHC-II antigenic presentation and triggered both CD8 + and CD4 + T cells at the systemic and mucosal levels. They also conferred a significant booster effect, consistent with the importance of CD4 + T cells for protection against Mycobacterium tuberculosis . Given the pivotal role of CD4 + T cells in orchestrating innate and adaptive immunity, this strategy could have a broad range of applications in the vaccinology field.

Published
2022

26. Mice Humanized for MHC and hACE2 with High Permissiveness to SARS-CoV-2 Omicron Replication

Author

Fabien Le Chevalier, Pierre Authié, Sébastien Chardenoux, Maryline Bourgine, Benjamin Vesin, Delphine Cussigh, Yohann Sassier, Ingrid Fert, Amandine Noirat, Kirill Nemirov, François Anna, Marion Bérard, Françoise Guinet, David Hardy, Pierre Charneau, François Lemonnier, Francina Langa-Vives, Laleh Majlessi, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Animalerie Centrale (plateforme) - Central Animal Facility (platform) (C2RA), Lymphocytes et Immunité - Lymphocytes and Immunity, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Plate-Forme d’Histopathologie / Histopathology Platform, Institut Cochin (IC UM3 (UMR 8104 / U1016)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), and Institut Pasteur et Theravectys

Subjects
HLA, Infectious Diseases, SARS-CoV-2, [SDV]Life Sciences [q-bio], Immunology, Transgenesis, Omicron variant, Microbiology, COVID-19 animal model
Abstract

International audience; Human Angiotensin-Converting Enzyme 2 (hACE2) is the major receptor enabling host cell invasion by SARS-CoV-2 via interaction with Spike. The murine ACE2 does not interact efficiently with SARS CoV-2 Spike and therefore the laboratory mouse strains are not permissive to SARS-CoV-2 replication. Here, we generated new hACE2 transgenic mice, which harbor the hACE2 gene under the human keratin 18 promoter, in “HHD-DR1” background. HHD-DR1 mice are fully devoid of murine Major Histocompatibility Complex (MHC) molecules of class-I and -II and express only MHC molecules from Human Leukocyte Antigen (HLA) HLA 02.01, DRA01.01, DRB1.01.01 alleles, widely expressed in human populations. We selected three transgenic strains, with various hACE2 mRNA expression levels and distinctive profiles of lung and/or brain permissiveness to SARS-CoV-2 replication. These new hACE2 transgenic strains display high permissiveness to the replication of SARS-CoV-2 Omicron sub-variants, while the previously available B6.K18-ACE22Prlmn/JAX mice have been reported to be poorly susceptible to infection with Omicron. As a first application, one of these MHC- and ACE2-humanized strains was successfully used to show the efficacy of a lentiviral-based COVID-19 vaccine

Published
2023

27. Mice Humanized for Major Histocompatibility Complex and Angiotensin-Converting Enzyme 2 with High Permissiveness to SARS-CoV-2 Omicron Replication

Author

Fabien Le Chevalier, Pierre Authié, Sébastien Chardenoux, Maryline Bourgine, Benjamin Vesin, Delphine Cussigh, Yohann Sassier, Ingrid Fert, Amandine Noirat, Kirill Nemirov, François Anna, Marion Bérard, Françoise Guinet, David Hardy, Pierre Charneau, François Lemonnier, Francina Langa-Vives, and Laleh Majlessi

Abstract

Human Angiotensin-Converting Enzyme 2 (hACE2) is the major receptor enabling host cell invasion by SARS-CoV-2 via interaction with Spike glycoprotein. The murine ACE2 ortholog does not interact efficiently with SARS-CoV-2 Spike and therefore the conventional laboratory mouse strains are not permissive to SARS-CoV-2 replication. Here, we generated newhACE2transgenic mice, which harbor thehACE2gene under the human keratin 18 promoter, in C57BL/6 “HHD-DR1” background. HHD-DR1 mice are fully devoid of murine Major Histocompatibility Complex (MHC) molecules of class-I and -II and express only MHC molecules from Human Leukocyte Antigen (HLA) HLA 02.01, DRA01.01, DRB1.01.01 alleles, widely expressed in human populations. We selected three transgenic strains, with varioushACE2mRNA expression levels and distinctive profiles of lung and/or brain permissiveness to SARS-CoV-2 replication. Compared to the previously available B6.K18-ACE22Prlmn/JAXmice, which have limited permissiveness to SARS-CoV-2 Omicron replication, these three new hACE2 transgenic strains display higher levels ofhACE2mRNA expression, associated with high permissiveness to the replication of SARS-CoV-2 Omicron sub-variants. As a first application, one of these MHC- and ACE2-humanized strains was successfully used to show the efficacy of a lentiviral vector-based COVID-19 vaccine candidate.

Published
2022

28. Lower disease activity but higher risk of severe COVID-19 and herpes zoster in patients with systemic lupus erythematosus with pre-existing autoantibodies neutralising IFN-α

Author

Alexis Mathian, Paul Breillat, Karim Dorgham, Paul Bastard, Caroline Charre, Raphael Lhote, Paul Quentric, Quentin Moyon, Alice-Andrée Mariaggi, Suzanne Mouries-Martin, Clara Mellot, François Anna, Julien Haroche, Fleur Cohen-Aubart, Delphine Sterlin, Noël Zahr, Adrian Gervais, Tom Le Voyer, Lucy Bizien, Quentin Amiot, Micheline Pha, Miguel Hié, Francois Chasset, Hans Yssel, Makoto Miyara, Pierre Charneau, Pascale Ghillani-Dalbin, Jean-Laurent Casanova, Flore Rozenberg, Zahir Amoura, and Guy Gorochov

Subjects
COVID-19 Vaccines, SARS-CoV-2, Immunology, COVID-19, Interferon-alpha, Interferon-beta, Herpes Zoster, General Biochemistry, Genetics and Molecular Biology, Rheumatology, Immunology and Allergy, Humans, Animals, Lupus Erythematosus, Systemic, Cattle, Autoantibodies, Retrospective Studies
Abstract

ObjectivesType-I interferons (IFNs-I) have potent antiviral effects. IFNs-I are also overproduced in patients with systemic lupus erythematosus (SLE). Autoantibodies (AAbs) neutralising IFN-α, IFN-β and/or IFN-ω subtypes are strong determinants of hypoxemic COVID-19 pneumonia, but their impact on inflammation remains unknown.MethodsWe retrospectively analysed a monocentric longitudinal cohort of 609 patients with SLE. Serum AAbs against IFN-α were quantified by ELISA and functionally assessed by abolishment of Madin-Darby bovine kidney cell protection by IFN-α2 against vesicular stomatitis virus challenge. Serum-neutralising activity against IFN-α2, IFN-β and IFN-ω was also determined with a reporter luciferase activity assay. SARS-CoV-2 antibody responses were measured against wild-type spike antigen, while serum-neutralising activity was assessed against the SARS-CoV-2 historical strain and variants of concerns.ResultsNeutralising and non-neutralising anti-IFN-α antibodies are present at a frequency of 3.3% and 8.4%, respectively, in individuals with SLE. AAbs neutralising IFN-α, unlike non-neutralising AAbs, are associated with reduced IFN-α serum levels and a reduced likelihood to develop active disease. However, they predispose patients to an increased risk of herpes zoster and severe COVID-19 pneumonia. Severe COVID-19 pneumonia in patients with SLE is mostly associated with combined neutralisation of different IFNs-I. Finally, anti-IFN-α AAbs do not interfere with COVID-19 vaccine humoral immunogenicity.ConclusionThe production of non-neutralising and neutralising anti-IFN-I antibodies in SLE is likely to be a consequence of SLE-associated high IFN-I serum levels, with a beneficial effect on disease activity, yet a greater viral risk. This finding reinforces the recommendations for vaccination against SARS-CoV-2 in SLE.

Published
2022

29. An intranasal lentiviral booster broadens immune recognition of SARS-CoV-2 variants and reinforces the waning mRNA vaccine-induced immunity that it targets to lung mucosa

Author

Benjamin Vesin, Jodie Lopez, Amandine Noirat, Pierre Authié, Ingrid Fert, Fabien Le Chevalier, Fanny Moncoq, Kirill Nemirov, Catherine Blanc, Cyril Planchais, Hugo Mouquet, Françoise Guinet, David Hardy, Christiane Gerke, François Anna, Maryline Bourgine, Laleh Majiessi, and Pierre Charneau

Abstract

As the COVID-19 pandemic continues and new SARS-CoV-2 variants of concern emerge, the adaptive immunity initially induced by the first-generation COVID-19 vaccines wains and needs to be strengthened and broadened in specificity. Vaccination by the nasal route induces mucosal humoral and cellular immunity at the entry point of SARS-CoV-2 into the host organism and has been shown to be the most effective for reducing viral transmission. The lentiviral vaccination vector (LV) is particularly suitable for this route of immunization because it is non-cytopathic, non-replicative and scarcely inflammatory. Here, to set up an optimized cross-protective intranasal booster against COVID-19, we generated an LV encoding stabilized Spike of SARS-CoV-2 Beta variant (LV::SBeta-2P). mRNA vaccine–primed and -boosted mice, with waning primary humoral immunity at 4 months post-vaccination, were boosted intranasally with LV::SBeta-2P. Strong boost effect was detected on cross-sero-neutralizing activity and systemic T-cell immunity. In addition, mucosal anti-Spike IgG and IgA, lung resident B cells, and effector memory and resident T cells were efficiently induced, correlating with complete pulmonary protection against the SARS-CoV-2 Delta variant, demonstrating the suitability of the LV::SBeta-2P vaccine candidate as an intranasal booster against COVID-19.

Published
2022

30. An intranasal lentiviral booster reinforces the waning mRNA vaccine-induced SARS-CoV-2 immunity that it targets to lung mucosa

Author

Benjamin Vesin, Jodie Lopez, Amandine Noirat, Pierre Authié, Ingrid Fert, Fabien Le Chevalier, Fanny Moncoq, Kirill Nemirov, Catherine Blanc, Cyril Planchais, Hugo Mouquet, Françoise Guinet, David Hardy, Francina Langa Vives, Christiane Gerke, François Anna, Maryline Bourgine, Laleh Majlessi, Pierre Charneau, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), Immunologie humorale - Humoral Immunology, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Lymphocytes et Immunité - Lymphocytes and Immunity, Plate-Forme d’Histopathologie / Histopathology Platform, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Centre d'Ingénierie génétique murine - Mouse Genetics Engineering Center (CIGM), Service développement de l'innovation - Innovation Office, Direction des Applications de la Recherche et des Relations industrielles - The Technology Transfer and Industrial Partnership Department (DARRI), Institut Pasteur [Paris] (IP)-Institut Pasteur [Paris] (IP), This work was supported by TheraVectys and Institut Pasteur., The authors are grateful to Dr. Marie José Quentin-Millet and Estelle Besson (TheraVectys) for discussion and advice, to Magali Tichit and Sabine Maurin for excellent technical assistance in preparing histological sections, to Sébastien Chardenoux (CIGM, Institut Pasteur) for excellent technical assistance in B6.K18-hACE2IP−THV mouse transgenesis, and to Marie Laure Loriquet and Dr. Paul-Henri Consigny (Institut Pasteur Medical Center) for their gift of the leftover Moderna vaccines, not unusable in humans, so that this study did not deprive any individual of a vaccine dose during the COVID-19 pandemic. The SARS-CoV2 variant Delta/2021/I7.2 200 was supplied by the Virus and Immunity Unit (Institut Pasteur, Paris, France) headed by Olivier Schwartz. The SARS-CoV-2 Omicron BA.1 variant was initially supplied by the Virus and Immunity Unit (Institut Pasteur, Paris, France) headed by Olivier Schwartz, and was provided to our laboratory by Matthieu Prot and Etienne Simon-Lorière (G5 Evolutionary Genomics of RNA Viruses, Institut Pasteur, Paris, France)., and Majlessi, Laleh

Subjects
COVID-19 Vaccines, lentiviral vaccine, Antibodies, Viral, Mice, [SDV.IMM.VAC] Life Sciences [q-bio]/Immunology/Vaccinology, Drug Discovery, Genetics, mucosal booster vaccine, Animals, Humans, SARS-CoV-2 emerging variants of concern, Lung, Molecular Biology, Pharmacology, Vaccines, Synthetic, Mucous Membrane, SARS-CoV-2, Vaccination, COVID-19, Viral Vaccines, Antibodies, Neutralizing, intranasal vaccination, Molecular Medicine, mucosal immunity, waning anti-COVID-19 immunity, mRNA Vaccines, [SDV.IMM.VAC]Life Sciences [q-bio]/Immunology/Vaccinology
Abstract

International audience; As the coronavirus disease 2019 (COVID-19) pandemic continues and new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants of concern emerge, the adaptive immunity initially induced by the first-generation COVID-19 vaccines starts waning and needs to be strengthened and broadened in specificity. Vaccination by the nasal route induces mucosal, humoral, and cellular immunity at the entry point of SARS-CoV-2 into the host organism and has been shown to be the most effective for reducing viral transmission. The lentiviral vaccination vector (LV) is particularly suitable for this route of immunization owing to its non-cytopathic, non-replicative, and scarcely inflammatory properties. Here, to set up an optimized cross-protective intranasal booster against COVID-19, we generated an LV encoding stabilized spike of SARS-CoV-2 Beta variant (LV::SBeta-2P). mRNA vaccine-primed and -boosted mice, with waning primary humoral immunity at 4 months after vaccination, were boosted intranasally with LV::SBeta-2P. A strong boost effect was detected on cross-sero-neutralizing activity and systemic T cell immunity. In addition, mucosal anti-spike IgG and IgA, lung-resident B cells, and effector memory and resident T cells were efficiently induced, correlating with complete pulmonary protection against the SARS-CoV-2 Delta variant, demonstrating the suitability of the LV::SBeta-2P vaccine candidate as an intranasal booster against COVID-19. LV::SBeta-2P vaccination was also fully protective against Omicron infection of the lungs and central nervous system, in the highly susceptible B6.K18-hACE2IP-THV transgenic mice.

Published
2022

32. Un candidat vaccin lentiviral anti-Covid-19 administrable par voie intranasale

Author

Laleh Majlessi, Pierre Charneau, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris]-TheraVectys, and Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité)

Subjects
0303 health sciences, 03 medical and health sciences, 0302 clinical medicine, [SDV]Life Sciences [q-bio], General Medicine, 030217 neurology & neurosurgery, General Biochemistry, Genetics and Molecular Biology, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology
Abstract

International audience; No abstract available

Published
2021

33. Use of lentiviral vectors in vaccination

Author

Min-Wen Ku, Laleh Majlessi, Pierre Charneau, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), and This manuscript was funded by TheraVectys and Institut Pasteur.

Subjects
Genetic Vectors, Immunology, lentiviral vectors, CD8-Positive T-Lymphocytes, Biology, Communicable Diseases, vaccination CONTACT Laleh Majlessi, Viral vector, Transduction (genetics), Immunity, humoral immunity, Vaccine Development, Drug Discovery, Animals, Humans, nonintegrating lentiviral vectors, long-lasting immune memory, Pharmacology, Immunogenicity, Viral Vaccine, Lentivirus, Vaccination, COVID-19, Viral Vaccines, transduction, Virology, t-cell immunity, Antigen expression in vivo, Humoral immunity, Molecular Medicine, [SDV.IMM.VAC]Life Sciences [q-bio]/Immunology/Vaccinology, CD8
Abstract

International audience; Introduction: Lentiviral vectors have emerged as powerful vectors for vaccination, due to their high efficiency to transduce dendritic cells and to induce long-lasting humoral immunity, CD8 + T cells, and effective protection in numerous preclinical animal models of infection and oncology. Areas covered: Here, we reviewed the literature, highlighting the relevance of lentiviral vectors in vaccinology. We recapitulated both their virological and immunological aspects of lentiviral vectors. We compared lentiviral vectors to the gold standard viral vaccine vectors, i.e. adenoviral vectors, and updated the latest results in lentiviral vector-based vaccination in preclinical models. Expert opinion: Lentiviral vectors are non-replicative, negligibly inflammatory, and not targets of preexisting immunity in human populations. These are major characteristics to consider in vaccine development. The potential of lentiviral vectors to transduce non-dividing cells, including dendritic cells, is determinant in their strong immunogenicity. Notably, lentiviral vectors can be engineered to target antigen expression to specific host cells. The very weak inflammatory properties of these vectors allow their use in mucosal vaccination, with particular interest in infectious diseases that affect the lungs or brain, including COVID-19. Recent results in various preclinical models have reinforced the interest of these vectors in prophylaxis against infectious diseases and in onco-immunotherapy.

Published
2021

34. A lentiviral vector expressing a dendritic cell-targeting multimer induces mucosal anti-mycobacterial CD4

Author

François, Anna, Jodie, Lopez, Fanny, Moncoq, Catherine, Blanc, Pierre, Authié, Amandine, Noirat, Ingrid, Fert, Philippe, Souque, Fabien, Nevo, Alexandre, Pawlik, David, Hardy, Sophie, Goyard, Denis, Hudrisier, Roland, Brosch, Françoise, Guinet, Olivier, Neyrolles, Pierre, Charneau, and Laleh, Majlessi

Subjects
CD4-Positive T-Lymphocytes, Mice, Inbred C57BL, Mice, Genetic Vectors, Animals, Dendritic Cells, CD8-Positive T-Lymphocytes
Abstract

Most viral vectors, including the potently immunogenic lentiviral vectors (LVs), only poorly direct antigens to the MHC-II endosomal pathway and elicit CD4

Published
2021

35. sgDI-tector: defective interfering viral genome bioinformatics for detection of coronavirus subgenomic RNAs

Author

Andrea Di Gioacchino, Rachel Legendre, Yannis Rahou, Valérie Najburg, Pierre Charneau, Benjamin D. Greenbaum, Frédéric Tangy, Sylvie van der Werf, Simona Cocco, Anastassia V. Komarova, Physique pour la médecine (PhysMed Paris), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Physique Statistique et Inférence pour la Biologie, Laboratoire de physique de l'ENS - ENS Paris (LPENS), Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)-Département de Physique de l'ENS-PSL, École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)-Département de Physique de l'ENS-PSL, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Laboratoire d’innovation : vaccins – Innovation lab : vaccines, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), Memorial Sloane Kettering Cancer Center [New York], Weill Cornell Medicine [Cornell University], Cornell University [New York], We acknowledge the ANR (Agence Nationale de la Recherche) and FRM (Fondation de la Recherche Médicale) for funding this work through the AAP Flash-Covid 19 project SARS-Cov-2immunRNAs. A.V.K. received support from ANR through the grant ANR-LBX-62 IBEID CoV-2SENSING/COVID 19. RNA NGS has been supported by France Génomique (ANR-10-INBS-09-09)., We would like to thank J. Pipoli da Fonseca, L. Lemée from Biomics Platform, C2RT, Institut Pasteur, Paris, France for RNA NGS, IBISA and the Illumina COVID-19 Projects’ offer. The authors would like to thank members of the Tangy, van der Werf laboratories and the National Reference Center (CNR) for Respiratory Viruses at the Institut Pasteur for support and valuable discussions., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-10-INBS-0009,France-Génomique,Organisation et montée en puissance d'une Infrastructure Nationale de Génomique(2010), Weill Cornell Medicine [New York], Legendre, Rachel, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID, and Organisation et montée en puissance d'une Infrastructure Nationale de Génomique - - France-Génomique2010 - ANR-10-INBS-0009 - INBS - VALID

Subjects
viruses, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Computational biology, Genome, Viral, Biology, ENCODE, medicine.disease_cause, Genome, Open Reading Frames, medicine, MESH: SARS-CoV-2, Viral rna, user-friendly bioinformatics, MESH: High-Throughput Nucleotide Sequencing, Molecular Biology, Polymerase, [INFO.INFO-BI] Computer Science [cs]/Bioinformatics [q-bio.QM], Subgenomic mRNA, Coronavirus, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, subgenomic RNA, SARS-CoV-2, RNA, Computational Biology, High-Throughput Nucleotide Sequencing, MESH: Open Reading Frames, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, MESH: RNA, Viral, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, defective viral genomes, biology.protein, RNA, Viral, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, MESH: Genome, Viral, [INFO.INFO-BI]Computer Science [cs]/Bioinformatics [q-bio.QM], MESH: Computational Biology
Abstract

Coronavirus RNA-dependent RNA polymerases produce subgenomic RNAs (sgRNAs) that encode viral structural and accessory proteins. User-friendly bioinformatic tools to detect and quantify sgRNA production are urgently needed to study the growing number of next-generation sequencing (NGS) data of SARS-CoV-2. We introduced sgDI-tector to identify and quantify sgRNA in SARS-CoV-2 NGS data. sgDI-tector allowed detection of sgRNA without initial knowledge of the transcription-regulatory sequences. We produced NGS data and successfully detected the nested set of sgRNAs with the ranking M>ORF3a>N>ORF6>ORF7a>ORF8>S>E>ORF7b. We also compared the level of sgRNA production with other types of viral RNA products such as defective interfering viral genomes.

Published
2021

36. BNT162b2 vaccine-induced humoral and cellular responses against SARS-CoV-2 variants in Systemic Lupus Erythematosus

Author

Alexis Mathian, Micheline Pha, Q. Moyon, François Anna, Delphine Sterlin, Fleur Cohen-Aubart, Paul Breillat, Pierre Charneau, Hedi Chaieb, Guy Gorochov, Thi Huong Du Boutin, Pedro Macedo Flores, Sasi Mudumba, Makoto Miyara, Sophia de Alba, Pascale Ghillani-Dalbin, Julien Haroche, Raphael Lhote, Zahir Amoura, Centre d'Immunologie et des Maladies Infectieuses (CIMI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Institut E3M [CHU Pitié-Salpêtrière], CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Centre National de Référence du Lupus Systémique, Syndrome des Anticorps Anti-phospholipides et Maladies Auto-immunes Systémiques Rares [CHU Pitié Salpêtrière], Service de Médecine Interne 2, maladies auto-immunes et systémiques [CHU Pitié-Salpêtrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Institut E3M [CHU Pitié-Salpêtrière], Service d'Immunologie [CHU Pitié-Salpétrière], Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys-Université Paris Cité (UPCité), Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Institut Pasteur [Paris] (IP), Genalyte, The study was supported by the Agence Nationale de la Recherche (ANR Flash COVID19 program, MUCOVID, PI: DS) and by the SARS-CoV-2 Program of the Faculty of Medicine from Sorbonne University ICOViD programs, PI: GG)., ANR-21-COVR-0037,MUCOVID,Evaluation de la capacité neutralisante des IgA contre les variants du SARS-CoV-2(2021), HAL-SU, Gestionnaire, Evaluation de la capacité neutralisante des IgA contre les variants du SARS-CoV-2 - - MUCOVID2021 - ANR-21-COVR-0037 - COVID-19 - VALID, Centre d'Immunologie et de Maladies Infectieuses (CIMI), Institut Pasteur [Paris]-TheraVectys, Institut Pasteur [Paris], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], and Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut E3M [CHU Pitié-Salpêtrière]

Subjects
Male, Antibodies, Viral, Severity of Illness Index, Immunogenicity, Vaccine, 0302 clinical medicine, Interferon, Lupus Erythematosus, Systemic, Immunology and Allergy, Prospective Studies, skin and connective tissue diseases, Prospective cohort study, 0303 health sciences, Systemic lupus erythematosus, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Middle Aged, 3. Good health, Vaccination, medicine.anatomical_structure, Antirheumatic Agents, Female, medicine.drug, Adult, T cell, Immunology, Naive B cell, Systemic Lupus Erythematosus, General Biochemistry, Genetics and Molecular Biology, 03 medical and health sciences, Immune system, Rheumatology, Antigen, medicine, Humans, Adverse effect, BNT162 Vaccine, 030304 developmental biology, 030203 arthritis & rheumatology, Lupus erythematosus, SARS-CoV-2, business.industry, COVID-19, Mycophenolic Acid, systemic, medicine.disease, vaccination, Immunity, Humoral, Methotrexate, business, lupus erythematosus, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

ObjectivesOur aims were to evaluate Systemic Lupus Erythematosus (SLE) disease activity and SARS-CoV-2 specific immune responses after BNT162b2 vaccination.MethodsIn this prospective study, disease activity and clinical assessments were recorded from the first dose of vaccine, until day 15 after the second dose in 126 SLE patients. SARS-CoV-2 antibody responses were measured against wild-type spike antigen while serum-neutralizing activity was assessed against the SARS-CoV-2 historical strain and variants of concerns (VOCs). Vaccine-specific T-cell responses were quantified by Interferon (IFN)-γ release assay after the second dose.ResultsBNT162b2 was well tolerated and no statistically significant variations of BILAG and SLEDAI scores were observed throughout the study in SLE patients with active and inactive disease at baseline. Mycophenolate Mofetil (MMF) and Methotrexate (MTX) treatments were associated with drastically reduced BNT162b2 antibody-response (β=-78; p=0.007, β=-122; pConclusionMMF, MTX and poor baseline humoral immune status, particularly: low naïve B cell frequencies, are independently associated with impaired BNT162b2 mRNA antibody response, delineating SLE patients who might need adapted vaccine regimens and follow-up.KEY MESSAGESWhat is already known about this subject?BNT162b2 efficacy and safety has been described in studies mixing different RMDsWhat does this study add?No serious adverse effects, nor SLE flares have been documented after BNT162b2 in SLE patients.Not only MMF and MTX, but also a poor humoral immune status at baseline impair vaccine antibody responseAlbeit decreased, serum neutralizing activity against VOCs is conferred to vaccine-responders.How might this impact on clinical practice or future developments?These parameters could be helpful for physicians to delineate which patients should have antibody measurement after full BNT162b2 vaccination and should be proposed a third injection of BNT162b2 vaccine.

Published
2021

37. A lentiviral vector encoding fusion of light invariant chain and mycobacterial antigens induces protective CD4

Author

Jodie, Lopez, François, Anna, Pierre, Authié, Alexandre, Pawlik, Min-Wen, Ku, Catherine, Blanc, Philippe, Souque, Fanny, Moncoq, Amandine, Noirat, David, Hardy, Wladimir, Sougakoff, Roland, Brosch, Françoise, Guinet, Pierre, Charneau, and Laleh, Majlessi

Subjects
Antigens, Differentiation, B-Lymphocyte, CD4-Positive T-Lymphocytes, Mice, Inbred C57BL, Antigens, Bacterial, Mice, Genetic Vectors, Lentivirus, Histocompatibility Antigens Class II, Animals, Mycobacterium tuberculosis, CD8-Positive T-Lymphocytes, Mycobacteriaceae
Abstract

Lentiviral vectors (LVs) are highly efficient at inducing CD8

Published
2021

38. Prevalence of SARS-CoV-2 antibodies in France: results from nationwide serological surveillance

Author

Louise Perrin de Facci, Virgile Monnet, Pierre Charneau, Caroline Demeret, Jean-Baptiste Richard, Nicolas Escriou, Daniel Lévy-Bruhl, Gabrielle Jones, Thierry Rose, François Anna, Yvonnick Guillois, Marion Gransagne, Corinne Robin, Sophie Goyard, Stéphane Petres, Stéphane Le Vu, Marie-Noëlle Ungeheuer, Lucie Léon, Laurent Filleul, Sylvie van der Werf, Sibylle Bernard-Stoecklin, Olivier Helynck, Harold Noel, Santé publique France - French National Public Health Agency [Saint-Maurice, France], Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Institut Pasteur [Paris], Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris]-TheraVectys, Biologie Cellulaire des Lymphocytes - Lymphocyte Cell Biology, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Chimie et Biocatalyse, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratoire CERBA [Saint Ouen l'Aumône], Biomnis Sample Library (BSL), Eurofins Biomnis, Investigation Clinique et d’Accès aux Ressources Biologiques (Plate-forme) - Clinical Investigation and Access to BioResources (ICAReB), Centre National de Référence des virus des infections respiratoires (dont la grippe) - National Reference Center Virus Influenzae [Paris] (CNR), We thank Christine Larsen, Bruno Coignard and Jean-Claude Desenclos (Santé publiqueFrance) for their helpful contribution at setting up the study, from Institut Pasteur,Isabelle Cailleau for support in the funding process, Hélène Munier-Lehmann for accessto automate and supply management at the Unit of Chemistry and Biocatalysis, Yves L.Janin for providing the luciferase prosubstrate hikarazine 108, Philippe Souque forproduction of the lentiviral pseudo-types, the whole ICAReB team and COVID-19support staff for sample management at Institut Pasteur, Juliette Paireau (Institut Pas-teur, Santé publique France), Rodolphe Thiébaut (Bordeaux Université) and Xavier deLamballerie (Aix-Marseille Université) for valuable comments onfirst results. We alsothank the team from the Eurofins Biomnis Sample Library and from CerbaHealthcareBenedicte Roquebert (Laboratoire Cerba) and Marie Pierre Guerra (CerbaXpert) forcontributing to sample collection. Santé publique France provided funding to the NRCand to the two centralising biobanks to cover sample collection, preparation, transportand analysis costs. The funder had no role in analysis, interpretation of data or writing ofthe report. S.L.V., G.J. and H.N. had full access to all the data and had responsibility tosubmit for publication., Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-TheraVectys, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Centre National de Référence des virus des infections respiratoires (dont la grippe) - National Reference Center Virus Influenzae [Paris] (CNR - laboratoire coordonnateur), Lassailly-Bondaz, Anne, Santé publique France - French National Public Health Agency, Centre National de Référence des virus des infections respiratoires (dont la grippe) [Paris] (CNR), Santé publique France, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], and Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]

Subjects
0301 basic medicine, Male, Epidemiology, [SDV]Life Sciences [q-bio], General Physics and Astronomy, Antibodies, Viral, Serology, 0302 clinical medicine, Seroepidemiologic Studies, Case fatality rate, Prevalence, Cumulative incidence, 030212 general & internal medicine, Child, Aged, 80 and over, 0303 health sciences, education.field_of_study, Multidisciplinary, biology, Transmission (medicine), Middle Aged, 3. Good health, [SDV] Life Sciences [q-bio], [SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology, Child, Preschool, Female, France, medicine.symptom, Antibody, Adult, medicine.medical_specialty, Adolescent, Science, Population, Asymptomatic, General Biochemistry, Genetics and Molecular Biology, Article, 03 medical and health sciences, Young Adult, medicine, Seroprevalence, Humans, education, Epidemics, 030304 developmental biology, Aged, business.industry, SARS-CoV-2, Public health, Infant, Newborn, COVID-19, Infant, General Chemistry, 030104 developmental biology, Viral infection, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, biology.protein, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, business, Demography
Abstract

Assessment of the cumulative incidence of SARS-CoV-2 infections is critical for monitoring the course and extent of the COVID-19 epidemic. Here, we report estimated seroprevalence in the French population and the proportion of infected individuals who developed neutralising antibodies at three points throughout the first epidemic wave. Testing 11,000 residual specimens for anti-SARS-CoV-2 IgG and neutralising antibodies, we find nationwide seroprevalence of 0.41% (95% CI: 0.05–0.88) mid-March, 4.14% (95% CI: 3.31–4.99) mid-April and 4.93% (95% CI: 4.02–5.89) mid-May 2020. Approximately 70% of seropositive individuals have detectable neutralising antibodies. Infection fatality rate is 0.84% (95% CI: 0.70–1.03) and increases exponentially with age. These results confirm that the nationwide lockdown substantially curbed transmission and that the vast majority of the French population remained susceptible to SARS-CoV-2 in May 2020. Our study shows the progression of the first epidemic wave and provides a framework to inform the ongoing public health response as viral transmission continues globally., The percentage of national populations infected during the first stages of the COVID-19 pandemic are unclear owing to limited early testing. Here the authors provide a nation-wide prevalence study of SARS-CoV-2 antibodies in France from the first wave of COVID-19 in 2020, including stratification based on age, sex and region.

Published
2021

39. SARS-CoV-2 infection in schools in a northern French city: a retrospective serological cohort study in an area of high transmission, France, January to April 2020

Author

Ludivine Grzelak, Charlotte Renaudat, Yoann Madec, Lucie Kuhmel, Laurence Arowas, Caroline Demeret, Kuang-Yu Chen, Christèle Huon, Marie Noelle Ungeheuer, Marc Eloit, Olivier Schwartz, Thomas Bigot, Bernadette Crescenzo-Chaigne, Sandrine Fernandes Pellerin, Arnaud Fontanet, Isabelle Staropoli, Bruno Hoen, Nathalie Jolly, Camille Besombes, Sandie Munier, Isabelle Cailleau, Timothée Bruel, Pierre Charneau, Laura Tondeur, Sarah Temmam, Blanca Liliana Perlaza, Rebecca Grant, Epidémiologie des Maladies Emergentes - Emerging Diseases Epidemiology, Pasteur-Cnam Risques infectieux et émergents (PACRI), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), Conservatoire National des Arts et Métiers [CNAM] (CNAM), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), Découverte de pathogènes – Pathogen discovery, Institut Pasteur [Paris], Hub Bioinformatique et Biostatistique - Bioinformatics and Biostatistics HUB, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Virus et Immunité - Virus and immunity, Université de Paris (UP), Direction de la recherche médicale de l'Institut Pasteur, Investigation Clinique et d’Accès aux Ressources Biologiques (Plate-forme) - Clinical Investigation and Access to BioResources (ICAReB), Centre de Recherche Translationnelle - Center for Translational Science (CRT), Centre Médical de l'Institut Pasteur, Vaccine Research Institute (VRI), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Biologie des ARN et virus influenza - RNA Biology of Influenza Virus, Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris]-TheraVectys, Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, École nationale vétérinaire d'Alfort (ENVA), The study was funded by Institut Pasteur, and several laboratories participating in the study receive funding from the Labex IBEID (ANR-10-LABX-62-IBEID), REACTing and the INCEPTION project (PIA/ANR-16-CONV-0005) for studies focusing on emerging viruses. OS lab is funded by Institut Pasteur, ANRS, Sidaction, the Vaccine Research Institute (ANR-10-LABX-77), 'TIMTAMDEN' ANR-14-CE14-0029, 'CHIKV-Viro- Immuno' ANR-14-CE14-0015-01 and the Gilead HIV cure program. LG is supported by the French Ministry of Higher Education, Research and Innovation. ME lab is funded by Institut Pasteur, Labex IBEID (ANR-10-LABX-62-IBEID), REACTing (Research & Action Emerging Infectious Diseases), EU Grant 101003589 RECoVER., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-16-CONV-0005,INCEPTION,Institut Convergences pour l'étude de l'Emergence des Pathologies au Travers des Individus et des populatiONs(2016), ANR-10-LABX-0077,VRI,Initiative for the creation of a Vaccine Research Institute(2010), ANR-14-CE14-0029,TIMTAMDEN,Rôle des récepteurs TIM et TAM dans l'infection des cellules cibles par le virus de la dengue(2014), ANR-14-CE14-0015,CHIKV-Viro-Immuno,Multiplication et Relation avec l'hôte du virus Chikungunya(2014), European Project: 101003589, H2020-SC1-PHE-CORONAVIRUS-2020,RECOVER(2020), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM), Université Paris Cité (UPCité), Centre Médical de l'Institut Pasteur (CMIP), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), École nationale vétérinaire - Alfort (ENVA), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]-Université de Paris (UP), Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Virus et Immunité - Virus and immunity (CNRS-UMR3569), Vaccine Research Institute [Créteil, France] (VRI), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), and Institut Pasteur [Paris] (IP)-TheraVectys

Subjects
0301 basic medicine, Emerging infectious diseases, medicine.medical_specialty, MESH: Schools, Epidemiology, [SDV]Life Sciences [q-bio], Context (language use), Asymptomatic, Serology, Cohort Studies, 03 medical and health sciences, 0302 clinical medicine, Virology, MESH: Child, Humans, Medicine, MESH: COVID-19, MESH: SARS-CoV-2, 030212 general & internal medicine, Child, MESH: Cohort Studies, Retrospective Studies, Schools, MESH: Humans, Transmission (medicine), business.industry, SARS-CoV-2, Research, 4. Education, Public Health, Environmental and Occupational Health, COVID-19, Outbreak, Retrospective cohort study, MESH: Retrospective Studies, 3. Good health, Coronavirus disease 2019 (COVID-19), MESH: France, 030104 developmental biology, France, medicine.symptom, business, Demography, Cohort study
Abstract

Background Children’s role in SARS-CoV-2 epidemiology remains unclear. We investigated an initially unnoticed SARS-CoV-2 outbreak linked to schools in northern France, beginning as early as mid-January 2020. Aims This retrospective observational study documents the extent of SARS-CoV-2 transmission, linked to an affected high school (n = 664 participants) and primary schools (n = 1,340 study participants), in the context of unsuspected SARS-CoV-2 circulation and limited control measures. Methods Between 30 March and 30 April 2020, all school staff, as well as pupils and their parents and relatives were invited for SARS-CoV-2 antibody testing and to complete a questionnaire covering symptom history since 13 January 2020. Results In the high school, infection attack rates were 38.1% (91/239), 43.4% (23/53), and 59.3% (16/27), in pupils, teachers, and non-teaching staff respectively vs 10.1% (23/228) and 12.0% (14/117) in the pupils’ parents and relatives (p Conclusions Viral circulation can occur in high and primary schools so keeping them open requires consideration of appropriate control measures and enhanced surveillance.

Published
2021

40. Brain and Lung Cross-Protection against Ancestral or Emerging SARS-CoV-2 by Intranasal Lentiviral Vaccination in a New hACE2 Transgenic Murine Model

Author

Francina Langa Vives, Jodie Lopez, Catherine Blanc, llta Lafosse, Ingrid Fert, Sébastien Chardenoux, Pierre Authié, Philippe Souque, Benjamin Vesin, Laleh Majlessi, Amandine Noirat, Kirill Nemirov, Fabien Nevo, Min-Wen Ku, Françoise Guinet, Fanny Moncoq, François Anna, David Hardy, Maryline Bourgine, Delphine Cussigh, and Pierre Charneau

Subjects
Vaccination, Lung, medicine.anatomical_structure, Murine model, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Transgene, medicine, Nasal administration, Biology, Virology, respiratory tract diseases
Abstract

COVID-19 vaccines already in use or in clinical development may have safety concerns, limited immunogenicity in high-risk groups or reduced efficacy against emerging SARS-CoV-2 variants. In addition, although the neurotropism of SARS-CoV-2 is well established, the vaccine strategies currently developed have not taken into account the protection of the central nervous system. Here, we generated a transgenic mouse strain expressing the human Angiotensin Converting Enzyme 2, with unprecedented brain as well as lung permissibility to SARS-CoV-2 replication. Using this stringent transgenic model, we demonstrated that a non-integrative lentiviral vector, encoding for the spike glycoprotein of the ancestral Wuhan SARS-CoV-2, used in intramuscular prime and intranasal boost elicits sterilizing protection of lung and brain against both the Wuhan and the most genetically distant Manaus P.1 SARS-CoV-2 variants. Beyond the induction of strong neutralizing antibodies, the mechanism underlying this broad protection spectrum involves a robust protective spike-specific CD8+ T-cell immunity, unaffected by the recent mutations accumulated in the emerging SARS-CoV-2 variants.

Published
2021

41. Full Brain and Lung Prophylaxis against SARS-CoV-2 by Intranasal Lentiviral Vaccination in a New hACE2 Transgenic Mouse Model or Golden Hamsters

Author

Pierre Charneau, Min-Wen Ku, Philippe Souque, Fanny Moncoq, Ilta Lafosse, François Anna, Sébastien Chardenoux, David Hardy, Maryline Bourgine, Catherine Blanc, Pierre Authié, Benjamin Vesin, Jodie Lopez, Kirill Nemirov, Fabien Nevo, Françoise Guinet, Amandine Noirat, Francina Langa Vives, and Laleh Majlessi

Subjects
Vaccination, Genetically modified mouse, Immunization, business.industry, Neurotropism, Hamster, Medicine, Nasal administration, business, Virology, Viral vector, Transgenic Model
Abstract

SummaryNon-integrative, non-cytopathic and non-inflammatory lentiviral vectors are particularly suitable for mucosal vaccination and recently emerge as a promising strategy to elicit sterilizing prophylaxis against SARS-CoV-2 in preclinical animal models. Here, we demonstrate that a single intranasal administration of a lentiviral vector encoding a prefusion form of SARS-CoV-2 spike glycoprotein induces full protection of respiratory tracts and totally avoids pulmonary inflammation in the susceptible hamster model. More importantly, we generated a new transgenic mouse strain, expressing the human Angiotensin Converting Enzyme 2, with unprecedent brain permissibility to SARS-CoV-2 replication and developing a lethal disease in

Published
2021

42. Promoter’s Choice Drive CAR-T Efficiency

Author

Pierre Langlade-Demoyen, Maria Loustau, François Anna, Pierre Charneau, Maxime Fredon, Olivier Adotevi, Philipe Souque, Francine Garnache-Ottou, Julien Caumartin, Jean-Marie Certoux, and Elodie Bole-Richard

Subjects
Transduction (genetics), Cell culture, Chemistry, Transgene, Cytotoxic T cell, Promoter, Receptor, Chimeric antigen receptor, Cell biology, Viral vector
Abstract

The development and the functionality of Chimeric Antigen Receptor (CAR) therapy is highly linked to the stability and expression level of the receptor, for which the choice of the promoter is key. Although EF1α is a reference promoter to induce strong antigen binding and long-term CAR expression, we investigated the impact of this one and other constitutive promoters (MSCV, PGK, β2m and CMV) on a CAR123 construct in the Blastic Plasmocytoid Dendritic Cell Neoplasm (BPDCN). Our results indicate that lentiviral vector titer is affected by transgene nature and promoter activity. Interestingly, by moderating the expression level of CAR123 at the cell surface, its recognition capacity and selectivity were improved. We observed that in early stages after transduction, CAR123 expression level did not affect lymphocyte’s phenotype and its cytotoxic capacity against BPDCN cell line. However, following long-term culture and decrease of cell-surface CAR expression, a hallmark of transition from activation to memory state of T-cells was observed, and only T-cells transduced with CAR-EF1α or -β2m promoters were still functional. We identified β2m as a promising promoter. Though it shows lower antigen-binding CAR-T cell and antigen recognition rates compared to EF1α, β2m promoter still present interesting CAR expression levels, stability, and binding capacity

Published
2021

43. High seroprevalence but short‐lived immune response to SARS‐CoV‐2 infection in Paris

Author

Pierre Charneau, Philippe Souque, Evelyne Dufour, Marion Gransagne, Stéphane Petres, Ana Ines Lalanne, Thierry Rose, Isabelle Turbiez, François Dejardin, François Anna, Odile Richard-Le Goff, Olivier Helynck, Maude Guillot-Delost, Sophie Goyard, Alexia Savignoni, François-Clément Bidard, Zaineb Choucha, Fabien Nevo, Franck Perez, Delphine Louis, Véronique Gillon, Yves L. Janin, A Gobillion, Nicolas Escriou, Olivier Lantz, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris]-TheraVectys, Biologie Cellulaire des Lymphocytes - Lymphocyte Cell Biology, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], Institut Curie [Paris], Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Institut Pasteur [Paris], Laboratoire d’innovation : vaccins – Innovation lab : vaccines, Risques cliniques et sécurité en santé des femmes et en santé périnatale (RISCQ), Université de Versailles Saint-Quentin-en-Yvelines (UVSQ), Immunité et cancer (U932), Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Plateforme technologique Production et purification de protéines recombinantes – Production and Purification of Recombinant Proteins Technological Platform (PPR), Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology, Chimie et Biocatalyse, This work was made possible thanks to the financial support obtained through the URGENCE nouveau coronavirus fundraising campaign of Institut Pasteur and the financial support of the Fondation Total. This study was funded in part by a grant from Fondation de France and by Institut Curie Institutional funding. The luciferin synthesis development has been supported by DARRI (ValoExpress 2016-2018). LuLISA development has been supported by IARP Pasteur-Carnot MI (2019-2020)., Institut Pasteur [Paris] (IP)-TheraVectys, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP), Université Paris-Saclay, Institut Curie [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Lassailly-Bondaz, Anne, Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Curie [Paris], Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), and Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]

Subjects
0301 basic medicine, Male, Time Factors, Antibodies, Viral, SARS‐CoV‐2, MESH: Antibodies, Neutralizing, Serology, 0302 clinical medicine, Seroepidemiologic Studies, Epidemiology, MESH: COVID-19, Immunology and Allergy, 030212 general & internal medicine, Research Articles, MESH: Immunoglobulin G, 0303 health sciences, education.field_of_study, MESH: Paris, 3. Good health, Titer, Research Article|Basic, ELISA, Female, medicine.symptom, Bioluminescence, Adult, MESH: Pandemics, medicine.medical_specialty, Paris, Population, Immunology, Anosmia, Immunity to infection, Biology, Asymptomatic, Virus, 03 medical and health sciences, COVID‐19, Internal medicine, medicine, Humans, Basic, education, Pandemics, 030304 developmental biology, LuLISA, MESH: Seroepidemiologic Studies, MESH: Humans, SARS-CoV-2, business.industry, MESH: Time Factors, COVID-19, MESH: Adult, Ageusia, Antibodies, Neutralizing, MESH: Male, 030104 developmental biology, Immunization, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, Immunoglobulin G, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, business, MESH: Female, MESH: Antibodies, Viral, 030215 immunology
Abstract

International audience; Background Although the COVID-19 pandemic peaked in March/April 2020 in France, the prevalence of infection is barely known. Herein, we assessed the serological response against the SARS-CoV-2 virus in a large population working in one institution of the Paris conurbation. We set up two high-throughput and sensitive methods to assess SARS CoV-2 Nucleoprotein and Spike protein-specific IgG response along with a pseudo-neutralization assay in sera. We studied 1847 participants who also answered a web-based survey on clinical symptoms.Methods and Results In May-July 2020, 11% (95% CI: 9.7-12.6) of serums were positive for IgG against the SARS-CoV-2 N and S protein and 9.5% (CI:8.2-11.0) were pseudo-neutralizer. The prevalence of immunization was 11.6% (CI:10.2-13.2) considering positivity in at least one assays. In 5% (CI:3.9-7.1) of RT-qPCR positive individuals, no systemic IgGs were detected. Among immune individuals, 21% had been asymptomatic.Anosmia and ageusia occurred in 52% of the IgG-positive individuals and in 3% of the negative ones. In contrast, 30% of the anosmia-ageusia cases were seronegative suggesting that the true prevalence of infection may reach 16.6%. In sera obtained 4-8 weeks after the first sampling anti-N and anti-S IgG titers and pseudo-neutralization activity declined by 31%, 17% and 53%, respectively with half-life of 35, 87 and 28 days, respectively.Conclusions The population studied being not particularly exposed to SARS-CoV-2 infection is representative of active workers in the Paris conurbation, suggesting that the current epidemiological models may underestimate the true prevalence of infection. The short lifespan of the serological systemic responses hinders retrospective assessment of the epidemic extent.

Published
2020
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44. Intranasal vaccination with a lentiviral vector protects against SARS-CoV-2 in preclinical animal models

Author

Hugo Mouquet, Benjamin Vesin, Houda Tabbal, Françoise Guinet, Jodie Lopez, Pierre Authié, Laurence Fiette, Marine Le Dudal, Catherine Blanc, François Anna, Maryline Bourgine, Nicolas Escriou, David Hardy, Fanny Moncoq, Kirill Nemirov, Fabien Nevo, Annette Martin, Pierre Charneau, Philippe Souque, Laleh Majlessi, Emeline Simon, Amandine Noirat, Min Wen Ku, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys, Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Institut Pasteur [Paris] (IP), Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Neuropathologie expérimentale / Experimental neuropathology, Institut Pasteur [Paris] (IP)-Université Paris Cité (UPCité), Institut Mutualiste de Montsouris (IMM), Lymphocytes et Immunité - Lymphocytes and Immunity, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Immunologie humorale - Humoral Immunology, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire d’innovation : vaccins – Innovation lab : vaccines, The authors are grateful to Prof. Sylvie van der Werf (National Reference Centre for Respiratory Viruses, Institut Pasteur, France) for providing the BetaCoV/France/IDF0372/2020 SARS-CoV-2 clinical isolate, supplied through the European Virus Archive Global (Evag), funded by the European Union’s Horizon 2020 program under grant agreement no. 653316. This work was also supported by grants from Institut Pasteur, TheraVectys, and Agence Nationale de la Recherche HuMoCID. M.W.K. is part of the Pasteur - Paris University PhD program and received funding from Institut Carnot Pasteur Microbes & Santé and the European Union’s Horizon 2020 program under Marie Sklodowska-Curie grant agreement no. 665807., ANR-20-COVI-0028,HuMoCID,Développement de modèles murins de COVID-19(2020), European Project: 653316,H2020,H2020-INFRAIA-2014-2015,EVAg(2015), European Project: 665807,H2020,H2020-MSCA-COFUND-2014,PASTEURDOC(2015), Institut Pasteur [Paris]-TheraVectys, Institut Pasteur [Paris], Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Institut Pasteur [Paris]-Université de Paris (UP), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Hardy, David, Développement de modèles murins de COVID-19 - - HuMoCID2020 - ANR-20-COVI-0028 - COVID-19 - VALID, European Virus Archive goes global - EVAg - - H20202015-04-01 - 2019-03-31 - 653316 - VALID, and Institut Pasteur International Docotal Program - PASTEURDOC - - H20202015-10-01 - 2020-10-01 - 665807 - VALID

Subjects
Male, [SDV]Life Sciences [q-bio], Respiratory System, lentiviral vectors, immunoglobulin A, Antibodies, Viral, Mice, Transduction (genetics), 0302 clinical medicine, Cricetinae, respiratory tracts, 0303 health sciences, Viral Load, 3. Good health, Vaccination, [SDV] Life Sciences [q-bio], medicine.anatomical_structure, in vivo Ad5 transduction, Models, Animal, Spike Glycoprotein, Coronavirus, Lentivirus, Female, COVID-19 Vaccines, Genetic Vectors, Immunization, Secondary, Biology, Microbiology, Article, Viral vector, 03 medical and health sciences, Immune system, [SDV.IMM.VAC] Life Sciences [q-bio]/Immunology/Vaccinology, Virology, boost-target, medicine, Animals, neutralizing antibodies, Immunity, Mucosal, Administration, Intranasal, 030304 developmental biology, SARS-CoV-2, lung inflammation, COVID-19, beta-coronavirus, Vaccine efficacy, biology.organism_classification, Antibodies, Neutralizing, intranasal vaccination, Immunology, mucosal immunity, Parasitology, Nasal administration, golden hamsters, [SDV.IMM.VAC]Life Sciences [q-bio]/Immunology/Vaccinology, 030217 neurology & neurosurgery, Respiratory tract
Abstract

To develop a vaccine candidate against COVID-19, we generated a Lentiviral Vector (LV), eliciting neutralizing antibodies against the Spike glycoprotein of SARS-CoV-2. Systemic vaccination by this vector in mice, in which the expression of the SARS-CoV-2 receptor hACE2 has been induced by transduction of respiratory tract cells by an adenoviral vector, confers only partial protection, despite high levels of serum neutralizing activity. However, eliciting an immune response in the respiratory tract through an intranasal boost results in > 3 log10 decrease in the lung viral loads and reduces local inflammation. Moreover, both integrative and non-integrative LV platforms display strong vaccine efficacy and inhibit lung deleterious injury in golden hamsters, which are naturally permissive to SARS-CoV-2 replication and closely mirror human COVID-19 physiopathology. Our results provide evidence of marked prophylactic effects of the LV-based vaccination against SARS-CoV-2 and designate intranasal immunization as a powerful approach against COVID-19., Graphical Abstract, Highlights A lentiviral vector encoding for Spike shows preclinical efficacy as a COVID-19 vaccine. Targeting the immune response to the upper respiratory tract provides critical protection. Intranasal vaccination induces protective mucosal immunity against SARS-CoV-2 in rodents. Lung anti-Spike IgA responses correlate with protection and reduced inflammation., Ku and colleagues present a lentiviral vaccination vector that encodes a full-length, membrane anchored form of SARS-CoV-2 Spike glycoprotein that induces neutralizing antibodies and T-cell responses. An intranasal boost strategy triggers a localized immune response in the upper respiratory tract that provides disease protection in mouse and hamster COVID-19 models.

Published
2020

45. IgA dominates the early neutralizing antibody response to SARS-CoV-2

Author

Pierre Charneau, Jean Marc Lacorte, Laetitia Claër, Olivier Schwartz, Amélie Guihot, Cary Gunn, Rick Hockett, Pascale Ghillani, Hans Yssel, Alexis Mathian, Jehane Fadlallah, Timothée Bruel, Karim Dorgham, Paul Quentric, François Anna, Makoto Miyara, Salma Fourati, Charles Edouard Luyt, Guy Gorochov, Delphine Sterlin, Hervé Devilliers, Sasi Mudumba, Alexandra Beurton, Julien Mayaux, Zahir Amoura, Christophe Parizot, Audrey Mohr, Anticorps en thérapie et pathologie - Antibodies in Therapy and Pathology, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Centre d'Immunologie et des Maladies Infectieuses (CIMI), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Institut Pasteur [Paris], Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris]-TheraVectys, Service de Médecine Interne (SOC 1 et SOC 2) [CHU de Dijon], Centre Hospitalier Universitaire de Dijon - Hôpital François Mitterrand (CHU Dijon), Service d'Immunologie [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Service d'Anesthésie réanimation [CHU Pitié-Salpêtrière], Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Research Unit on Cardiovascular and Metabolic Diseases [IHU ICAN], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU)-Institut de Cardiométabolisme et Nutrition = Institute of Cardiometabolism and Nutrition [CHU Pitié Salpêtrière] (IHU ICAN), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-CHU Pitié-Salpêtrière [AP-HP], Service de Pneumologie et Réanimation Médicale [CHU Pitié-Salpêtrière] (Département ' R3S '), Neurophysiologie Respiratoire Expérimentale et Clinique (UMRS 1158), Institut National de la Santé et de la Recherche Médicale (INSERM)-Sorbonne Université (SU), Service de Biochimie Endocrinienne et Oncologie [CHU Pitié-Salpêtrière], Centre de recherche sur l'Inflammation (CRI (UMR_S_1149 / ERL_8252 / U1149)), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Virus et Immunité - Virus and immunity, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Vaccine Research Institute (VRI), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Service de médecine interne [CHU Pitié-Salpétrière], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-CHU Pitié-Salpêtrière [AP-HP], Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Sorbonne Université (SU), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-TheraVectys, Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Research Unit on Cardiovascular and Metabolic Diseases (ICAN), Virus et Immunité - Virus and immunity (CNRS-UMR3569), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Vaccine Research Institute [Créteil, France] (VRI), ANR-20-COVI-0025,iCovid,Immunopathologie du COVID-19 à l'Assistance Publique Hôpitaux de Paris(2020), Centre d'Immunologie et de Maladies Infectieuses (CIMI), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP), Unité de Recherche sur les Maladies Cardiovasculaires, du Métabolisme et de la Nutrition = Institute of cardiometabolism and nutrition (ICAN), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-CHU Pitié-Salpêtrière [AP-HP], Sorbonne Université (SU)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Sorbonne Université (SU), Neurophysiologie Respiratoire Expérimentale et Clinique, Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), and Gestionnaire, HAL Sorbonne Université 5

Subjects
Saliva, Time Factors, viruses, Antibodies, Viral, Immunoglobulin E, 03 medical and health sciences, 0302 clinical medicine, Immune system, Immunity, Humans, Medicine, Longitudinal Studies, Neutralizing antibody, Research Articles, 030304 developmental biology, [SDV.MP.VIR] Life Sciences [q-bio]/Microbiology and Parasitology/Virology, 0303 health sciences, biology, SARS-CoV-2, business.industry, Case-control study, COVID-19, General Medicine, Antibodies, Neutralizing, Primary and secondary antibodies, STM r-articles, Immunity, Humoral, Immunoglobulin A, 3. Good health, Coronavirus, Immunoglobulin M, Case-Control Studies, Immunoglobulin G, 030220 oncology & carcinogenesis, Host-Pathogen Interactions, Immunology, [SDV.MP.VIR]Life Sciences [q-bio]/Microbiology and Parasitology/Virology, biology.protein, Antibody, business, Bronchoalveolar Lavage Fluid, Biomarkers, Research Article
Abstract

Early specific antibody responses against SARS-CoV-2 include IgG, IgM, and IgA, but IgA may neutralize virus and control infection to a larger extent., Early action of IgA Humoral immune responses play a critical role in protecting individuals against SARS-CoV-2 infection, particularly through the activity of neutralizing antibodies. Sterlin et al. measured humoral immune responses in the serum, saliva, and bronchoalveolar lavage fluid of SARS-CoV-2–infected patients who experienced a range of COVID-19 disease severity. IgA antibodies dominated the early SARS-CoV-2–specific antibody response compared with IgG and IgM concentrations in these fluids and was associated with expansion of IgA plasmablasts with mucosal homing characteristics. IgA serum concentrations peaked 3 weeks after symptom onset but persisted for several more weeks in saliva, and serum IgA was more potent than IgG in neutralizing SARS-CoV-2. These findings highlight the potential role of IgA during early SARS-CoV-2 infection., Humoral immune responses are typically characterized by primary IgM antibody responses followed by secondary antibody responses associated with immune memory and composed of IgG, IgA, and IgE. Here, we measured acute humoral responses to SARS-CoV-2, including the frequency of antibody-secreting cells and the presence of SARS-CoV-2–specific neutralizing antibodies in the serum, saliva, and bronchoalveolar fluid of 159 patients with COVID-19. Early SARS-CoV-2–specific humoral responses were dominated by IgA antibodies. Peripheral expansion of IgA plasmablasts with mucosal homing potential was detected shortly after the onset of symptoms and peaked during the third week of the disease. The virus-specific antibody responses included IgG, IgM, and IgA, but IgA contributed to virus neutralization to a greater extent compared with IgG. Specific IgA serum concentrations decreased notably 1 month after the onset of symptoms, but neutralizing IgA remained detectable in saliva for a longer time (days 49 to 73 post-symptoms). These results represent a critical observation given the emerging information as to the types of antibodies associated with optimal protection against reinfection and whether vaccine regimens should consider targeting a potent but potentially short-lived IgA response.

Published
2020

46. Asymptomatic and symptomatic SARS-CoV-2 infections elicit polyfunctional antibodies

Author

Stéphane Pelleau, François Anna, Michael T. White, Pierre Charneau, Arnaud Fontanet, Timothée Bruel, Hugo Mouquet, Bruno Hoen, Yoann Madec, Jérémy Dufloo, Yves Levy, Ludivine Grzelak, Isabelle Staropoli, Marie-Noëlle Ungeheuer, Laura Tondeur, Rémy Robinot, Julian Buchrieser, Aurélie Wiedemann, Cyril Planchais, Olivier Schwartz, Virus et Immunité - Virus and immunity, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Université Sorbonne Paris Cité (USPC), Epidémiologie des Maladies Emergentes - Emerging Diseases Epidemiology, Pasteur-Cnam Risques infectieux et émergents (PACRI), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris]-TheraVectys, Malaria : parasites et hôtes - Malaria : parasites and hosts, Institut Pasteur [Paris], Vaccine Research Institute (VRI), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Immunologie humorale - Humoral Immunology, Institut Pasteur [Paris]-Institut National de la Santé et de la Recherche Médicale (INSERM), Investigation Clinique et d’Accès aux Ressources Biologiques (Plate-forme) - Clinical Investigation and Access to BioResources (ICAReB), Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Direction de la recherche médicale de l'Institut Pasteur, Conservatoire National des Arts et Métiers [CNAM] (CNAM), This work was supported by the Urgence COVID-19 Fundraising Campaign of Institut Pasteur. O.S. is funded by Institut Pasteur, ANRS, Sidaction, the Vaccine Research Institute (ANR-10-LABX-77), Labex IBEID (ANR-10-LABX-62-IBEID), the 'TIMTAMDEN' project (a collaboration of Inserm, Institut Pasteur, and IRD, ANR-14-CE14-0029), the ANR 'CHIKV-Viro-Immuno' project (ANR-14-CE14-0015-01), and the Gilead HIV cure program. J.D. and L.G. are supported by the French Ministry of Higher Education, Research and Innovation. H.M. is funded by the Institut Pasteur, the Milieu Intérieur Program (ANR-10-LABX-69-01), Inserm, and REACTing and EU RECOVER grants. C.P. is supported by a fellowship from the Agence Nationale de Recherches sur le Sida et les Hépatites Virales (ANRS)., ANR-10-LABX-0077,VRI,Initiative for the creation of a Vaccine Research Institute(2010), ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-14-CE14-0029,TIMTAMDEN,Rôle des récepteurs TIM et TAM dans l'infection des cellules cibles par le virus de la dengue(2014), ANR-14-CE14-0015,CHIKV-Viro-Immuno,Multiplication et Relation avec l'hôte du virus Chikungunya(2014), ANR-10-LABX-0069,MILIEU INTERIEUR,GENETIC & ENVIRONMENTAL CONTROL OF IMMUNE PHENOTYPE VARIANCE: ESTABLISHING A PATH TOWARDS PERSONALIZED MEDICINE(2010), Virus et Immunité - Virus and immunity (CNRS-UMR3569), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM), Institut Pasteur [Paris] (IP)-TheraVectys, Institut Pasteur [Paris] (IP), Vaccine Research Institute [Créteil, France] (VRI), Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), and HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM)

Subjects
Male, Medicine (General), [SDV]Life Sciences [q-bio], Cell, Antibodies, Viral, Severity of Illness Index, Epitope, Immunoglobulin G, Neutralization, Antigen-Antibody Reactions, Epitopes, 0302 clinical medicine, antibody, Medicine, complement, Antibody-dependent cell-mediated cytotoxicity, 0303 health sciences, biology, Middle Aged, 3. Good health, Killer Cells, Natural, medicine.anatomical_structure, Spike Glycoprotein, Coronavirus, Female, medicine.symptom, Antibody, ADCC, Adult, Adolescent, Coronavirus disease 2019 (COVID-19), Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Asymptomatic, General Biochemistry, Genetics and Molecular Biology, Article, Virus, Young Adult, 03 medical and health sciences, R5-920, Neutralization Tests, Humans, asymptomatic, 030304 developmental biology, SARS-CoV-2, business.industry, Antibody-Dependent Cell Cytotoxicity, COVID-19, Complement System Proteins, Asymptomatic Diseases, Immunology, biology.protein, business, 030217 neurology & neurosurgery
Abstract

A large proportion of SARS-CoV-2 infected individuals remains asymptomatic. Little is known about the extent and quality of their antiviral humoral response. Here, we analyze antibody functions in 52 asymptomatic infected individuals, 119 mild and 21 hospitalized COVID-19 patients. We measured anti-Spike IgG, IgA and IgM levels with the S-Flow assay and map IgG-targeted epitopes by Luminex. We also evaluated neutralization, complement deposition and Antibody-Dependent Cellular Cytotoxicity (ADCC) using replication-competent SARS-CoV-2 or reporter cell systems. We show that COVID-19 sera mediate complement deposition and kill infected cells by ADCC. Sera from asymptomatic individuals neutralize the virus, activate ADCC and trigger complement deposition. Antibody levels and functions are lower in asymptomatic individuals than in symptomatic cases. Antibody functions are correlated, regardless of disease severity. Longitudinal samplings show that antibody functions follow similar kinetics of induction and contraction. Overall, asymptomatic SARS-CoV-2 infection elicits polyfunctional antibodies neutralizing the virus and targeting infected cells., Graphical Abstract, Dufloo et al. show that SARS-CoV-2 infection elicits antibodies that neutralize the virus, activate the complement system and kill infected cells by ADCC. These polyfunctional antibodies are slightly more abundant in symptomatic COVID-19 patients than in asymptomatic individuals and correlate with disease severity.

Published
2020

47. High seroprevalence of SARS-CoV-2 antibodies among people living in precarious situations in Ile de France

Author

Thomas Roederer, Bastien Mollo, Charline Vincent, Birgit Nikolay, Augusto Llosa, Robin Nesbitt, Jessica Vanhomwegen, Thierry Rose, François Anna, Corinne Torre, Emilie Fourrey, Erica Simons, Sophie Goyard, Yves Janin, Pierre Charneau, Oxana Vratskikh, Anneliese Coury, Stefan Vanel, Pierre Mendiharat, Klaudia Porten, William Hennequin, Clair Mills, Francisco Luquero, Epicentre [Paris] [Médecins Sans Frontières], Médecins Sans Frontières [Paris] (MSF), Environnement et Risques infectieux - Environment and Infectious Risks (ERI), Institut Pasteur [Paris] (IP), Biologie Cellulaire des Lymphocytes - Lymphocyte Cell Biology, Institut Pasteur [Paris] (IP)-Institut National de la Santé et de la Recherche Médicale (INSERM), Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys, Chimie et Biocatalyse, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Janin, Yves, Institut Pasteur [Paris], Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur [Paris], Institut Pasteur [Paris]-TheraVectys, and Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]

Subjects
0303 health sciences, education.field_of_study, Coronavirus disease 2019 (COVID-19), [CHIM.ORGA]Chemical Sciences/Organic chemistry, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Population, High seroprevalence, Logistic regression, [CHIM.ORGA] Chemical Sciences/Organic chemistry, 3. Good health, 03 medical and health sciences, 0302 clinical medicine, Geography, Environmental health, Food distribution, Pandemic, Seroprevalence, 030212 general & internal medicine, education, 030304 developmental biology
Abstract

BackgroundA nationwide lockdown was implemented in France on 17 March 2020 to control the COVID-19 pandemic. People living in precarious conditions were relocated by the authorities to emergency shelters, hotels and large venues. Médecins sans Frontières (MSF) then intervened to provide medical care in several of these locations in Paris and in Seine-Saint-Denis, one of its suburbs, between March and June 2020. A seroprevalence survey was conducted to assess the level of exposure to COVID-19 among the population living in the sites. To our knowledge, this is the first assessment of the impact of the pandemic on populations living in insecure conditions in Europe.MethodsWe conducted a cross-sectional seroprevalence study in the food distribution sites, emergency shelters and workers residences supported by MSF in Paris and Seine-Saint-Denis, to determine the extent of COVID-19 exposure as determined by SARS-CoV2 antibody seropositivity. The detection of SARS-COV2 antibodies in serum was performed at the Institut Pasteur of Paris using two LuLISA (Luciferase-Linked Immunosorbent Assay) assays and a Pseudo Neutralization Test. A questionnaire covering sociodemographic characteristics, living conditions, adherence to sanitary recommendations and symptom manifestations was also completed. We describe here the seroprevalence site by site and identify the risk factors for seropositivity using a multivariable logistic regression model with site random effects. We also investigated associations between seropositivity and symptoms eventually reported.FindingsOverall, 426/818 individuals tested positive in the 14 sites investigated. Seroprevalence varied significantly with the type of site (chi2 pInterpretationThe results demonstrate rather high exposure to SARS-COV-2 with important variations between study sites. Living in crowded conditions was identified as the most important explanatory factor for differences in levels of exposure. This study describes the key factors which determine the risk of exposure and illustrates the importance of identifying populations at high risk of exposure in order to orient and adapt prevention and control strategies to their specific needs.

Published
2020

48. Serologic responses to SARS-CoV-2 infection among hospital staff with mild disease in eastern France

Author

Yoann Madec, François Anna, Sandrine Fernandes-Pellerin, Marie Mielcarek, Olivier Schwartz, Rebecca Grant, Nicolas Collongues, Isabelle Staropoli, David Rey, Alexandre Bolle, Nathalie Jolly, Laura Tondeur, Charlotte Renaudat, Ludivine Grzelak, Nicolas Lefebvre, Marie-Noëlle Ungeheuer, Arnaud Fontanet, Bruno Hoen, Jérôme De Seze, Timothée Bruel, Catherine Schmidt-Mutter, Samira Fafi-Kremer, Aurélie Velay, Pierre Charneau, Philippe Souque, Nicolas Meyer, Immuno-Rhumatologie Moléculaire, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Virus et Immunité - Virus and immunity, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Laboratoire des sciences de l'ingénieur, de l'informatique et de l'imagerie (ICube), Institut National des Sciences Appliquées - Strasbourg (INSA Strasbourg), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-École Nationale du Génie de l'Eau et de l'Environnement de Strasbourg (ENGEES)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Pasteur-Cnam Risques infectieux et émergents (PACRI), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), CHU Strasbourg, Virus et Immunité - Virus and immunity (CNRS-UMR3569), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Vaccine Research Institute (VRI), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), Epidémiologie des Maladies Emergentes - Emerging Diseases Epidemiology, Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM), Université Paris Cité (UPCité), Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris] (IP)-TheraVectys, Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Institut Pasteur [Paris] (IP), Centre de Recherche Translationnelle - Center for Translational Science (CRT), Investigation Clinique et d’Accès aux Ressources Biologiques (Plate-forme) - Clinical Investigation and Access to BioResources (ICAReB), CIC Strasbourg (Centre d’Investigation Clinique Plurithématique (CIC - P) ), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Nouvel Hôpital Civil de Strasbourg-Hôpital de Hautepierre [Strasbourg], Groupe Méthodes en Recherche Clinique [Strasbourg] (GMRC), Nouvel Hôpital Civil [Strasbourg], CHU Strasbourg-CHU Strasbourg, École Nationale du Génie de l'Eau et de l'Environnement de Strasbourg (ENGEES)-Université de Strasbourg (UNISTRA)-Institut National des Sciences Appliquées - Strasbourg (INSA Strasbourg), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de Recherche en Informatique et en Automatique (Inria)-Les Hôpitaux Universitaires de Strasbourg (HUS)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Direction de la recherche médicale de l'Institut Pasteur, Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM), OS lab is funded by Institut Pasteur, ANRS, Sidaction, the Vaccine Research Institute (ANR- 10-LABX-77), Labex IBEID (ANR-10-LABX-62-IBEID), 'TIMTAMDEN' ANR-14-CE14-0029, 'CHIKV-Viro- Immuno' ANR-14-CE14-0015-01 and the Gilead HIV cure program. LG is supported by the French Ministry of Higher Education, Research and Innovation. SFK lab receives funding from Strasbourg University Hospitals (COVID-HUS, CE-2020-34)., ANR-10-LABX-0077,VRI,Initiative for the creation of a Vaccine Research Institute(2010), ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-14-CE14-0029,TIMTAMDEN,Rôle des récepteurs TIM et TAM dans l'infection des cellules cibles par le virus de la dengue(2014), ANR-14-CE14-0015,CHIKV-Viro-Immuno,Multiplication et Relation avec l'hôte du virus Chikungunya(2014), univOAK, Archive ouverte, Laboratoires d'excellence - Initiative for the creation of a Vaccine Research Institute - - VRI2010 - ANR-10-LABX-0077 - LABX - VALID, Integrative Biology of Emerging Infectious Diseases - - IBEID2010 - ANR-10-LABX-0062 - LABX - VALID, Appel à projets générique - Rôle des récepteurs TIM et TAM dans l'infection des cellules cibles par le virus de la dengue - - TIMTAMDEN2014 - ANR-14-CE14-0029 - Appel à projets générique - VALID, Appel à projets générique - Multiplication et Relation avec l'hôte du virus Chikungunya - - CHIKV-Viro-Immuno2014 - ANR-14-CE14-0015 - Appel à projets générique - VALID, Vaccine Research Institute [Créteil, France] (VRI), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), Institut Pasteur [Paris]-TheraVectys, Institut Pasteur [Paris], and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace (FMNGE)

Subjects
Male, 0301 basic medicine, Future studies, MESH: Coronavirus Infections, [SDV]Life Sciences [q-bio], Antibodies, Viral, Severity of Illness Index, Neutralization, Serology, MESH: Antibodies, Neutralizing, 0302 clinical medicine, [SDV.MHEP.MI]Life Sciences [q-bio]/Human health and pathology/Infectious diseases, MESH: Reverse Transcriptase Polymerase Chain Reaction, MESH: COVID-19, 030212 general & internal medicine, Mild disease, [SDV.MHEP.ME] Life Sciences [q-bio]/Human health and pathology/Emerging diseases, 0303 health sciences, [SDV.MHEP.ME]Life Sciences [q-bio]/Human health and pathology/Emerging diseases, MESH: Middle Aged, biology, Reverse Transcriptase Polymerase Chain Reaction, General Medicine, Middle Aged, MESH: Hospitals, Hospitals, 3. Good health, [SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology, [SDV.IMM.IA] Life Sciences [q-bio]/Immunology/Adaptive immunology, 030220 oncology & carcinogenesis, MESH: RNA, Viral, [SDV.MHEP.MI] Life Sciences [q-bio]/Human health and pathology/Infectious diseases, RNA, Viral, MESH: Betacoronavirus, Female, France, Sample collection, Antibody, Coronavirus Infections, Research Paper, Adult, medicine.medical_specialty, MESH: Pandemics, Coronavirus disease 2019 (COVID-19), Health Personnel, Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), Pneumonia, Viral, Article, General Biochemistry, Genetics and Molecular Biology, Antibodies, Betacoronavirus, 03 medical and health sciences, Internal medicine, MESH: Severity of Illness Index, medicine, Humans, Serologic Tests, MESH: SARS-CoV-2, Symptom onset, Pandemics, 030304 developmental biology, MESH: Humans, business.industry, Mild covid-19, SARS-CoV-2, MESH: Serologic Tests, COVID-19, MESH: Adult, Antibodies, Neutralizing, MESH: Male, Personnel, Hospital, MESH: France, 030104 developmental biology, MESH: Pneumonia, Viral, [SDV.SPEE] Life Sciences [q-bio]/Santé publique et épidémiologie, Antibody Formation, biology.protein, MESH: Health Personnel, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, business, MESH: Female, MESH: Antibodies, Viral
Abstract

PMCID: PMC7502660; International audience; BACKGROUND: The serologic response of individuals with mild forms of SARS-CoV-2 infection is poorly characterized.METHODS: Hospital staff who had recovered from mild forms of PCR-confirmed SARS-CoV-2 infection were tested for anti-SARS-CoV-2 antibodies using two assays: a rapid immunodiagnostic test (99.4% specificity) and the S-Flow assay (~99% specificity). The neutralizing activity of the sera was tested with a pseudovirus-based assay.FINDINGS: Of 162 hospital staff who participated in the investigation, 160 reported SARS-CoV-2 infection that had not required hospital admission and were included in these analyses. The median time from symptom onset to blood sample collection was 24 days (IQR: 21-28, range 13-39). The rapid immunodiagnostic test detected antibodies in 153 (95.6%) of the samples and the S-Flow assay in 159 (99.4%), failing to detect antibodies in one sample collected 18 days after symptom onset (the rapid test did not detect antibodies in that patient). Neutralizing antibodies (NAbs) were detected in 79%, 92% and 98% of samples collected 13-20, 21-27 and 28-41 days after symptom onset, respectively (P = 0.02). INTERPRETATION: Antibodies against SARS-CoV-2 were detected in virtually all hospital staff sampled from 13 days after the onset of COVID-19 symptoms. This finding supports the use of serologic testing for the diagnosis of individuals who have recovered from SARS-CoV-2 infection. The neutralizing activity of the antibodies increased overtime. Future studies will help assess the persistence of the humoral response and its associated neutralization capacity in recovered patients.FUNDINGS: The funders had no role in study design, data collection, interpretation, or the decision to submit the work for publication.

Published
2020

49. Intranasal Vaccination with a Lentiviral Vector Strongly Protects against SARS-CoV-2 in Mouse and Golden Hamster Preclinical Models

Author

Amandine Noirat, Fanny Moncoq, Pierre Charneau, Laleh Majlessi, Jodie Lopez, Philippe Souque, Kirill Nemirov, Fabien Nevo, Pierre Authié, Nicolas Escriou, Maryline Bourgine, Min-Wen Ku, Laurence Fiette, François Anna, Emeline Simon, Hugo Mouquet, Houda Tabbal, Catherine Blanc, Benjamin Vesin, Marine Le Dudal, Françoise Guinet, and Annette Martin

Subjects
biology, business.industry, Antibody titer, Vaccine efficacy, Viral vector, Vaccination, Immune system, Immunology, biology.protein, Medicine, Nasal administration, Antibody, business, Viral load
Abstract

We developed a potent vaccination strategy, based on lentiviral vector (LV), capable of inducing neutralizing antibodies specific to the Spike glycoprotein (S) of SARS-CoV-2, the etiologic agent of CoronaVirus Disease 2019 (COVID-19). Among several LV encoding distinct variants of S, a single one encoding the full-length, membrane anchored S (LV::SFL) triggered high antibody titers in mice, with neutralization activities comparable to patients recovered from COVID-19. LV::SFL systemic vaccination in mice, in which the expression of the CoV2 receptor hACE2 was induced by transduction of the respiratory tract cells by an adenoviral type 5 (Ad5) vector, despite an intense serum neutralizing activity, only ≈1 log10 reduction of lung viral loads was observed after SARS-CoV2 challenge. We thus explored the strategy of targeting the immune response to the upper respiratory tract through an intranasal boost administration. Even though, after a prime and target regimen, the systemic neutralizing activity did not increase substantially, ≈5 log10 decrease in lung viral loads was achieved, with the loads in some animals under the limit of detection of a highly sensitive RT-PCR assay. The conferred protection also avoided largely pulmonary inflammation. We confirmed the vaccine efficacy and inhibition of lung inflammation using both integrative and non-integrative LV platforms in golden hamsters, naturally permissive to SARS-CoV2 replication and restituting human COVID-19 physiopathology. Our results provide the proof-of-principle evidence of marked prophylactic effects of an LV-based vaccination strategy against SARS-CoV-2 in two pre-clinical animal models and designate the intranasal LV::SFL-based immunization as a vigorous and promising vaccine approach against COVID-19.

Published
2020

50. SARS-CoV-2 infection in primary schools in northern France: A retrospective cohort study in an area of high transmission

Author

Yoann Madec, Pierre Charneau, Bruno Hoen, François Anna, Isabelle Cailleau, Marie-Noëlle Ungeheuer, Caroline Demeret, Lucie Kuhmel, Ludivine Grzelak, Laura Tondeur, Arnaud Fontanet, Olivier Schwartz, Rebecca Grant, Timothée Bruel, Isabelle Staropoli, Charlotte Renaudat, Sandrine Fernandes Pellerin, Epidémiologie des Maladies Emergentes - Emerging Diseases Epidemiology, Pasteur-Cnam Risques infectieux et émergents (PACRI), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM)-Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), Virus et Immunité - Virus and immunity, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS), Université Sorbonne Paris Cité (USPC), Direction de la recherche médicale de l'Institut Pasteur, Institut Pasteur [Paris], Investigation Clinique et d’Accès aux Ressources Biologiques (Plate-forme) - Clinical Investigation and Access to BioResources (ICAReB), Centre de Recherche Translationnelle - Center for Translational Science (CRT), Centre Médical de l'Institut Pasteur (CMIP), Laboratoire commun Pasteur-TheraVectys, Institut Pasteur [Paris]-TheraVectys, Virologie Moléculaire et Vaccinologie / Molecular Virology and Vaccinology, Génétique Moléculaire des Virus à ARN - Molecular Genetics of RNA Viruses (GMV-ARN (UMR_3569 / U-Pasteur_2)), Centre National de la Recherche Scientifique (CNRS)-Institut Pasteur [Paris]-Université de Paris (UP), Vaccine Research Institute (VRI), Université Paris-Est Créteil Val-de-Marne - Paris 12 (UPEC UP12), The study was funded by Institut Pasteur, and several laboratories participating in the study receive funding from the Labex IBEID (ANR-10-LABX-62-IBEID), REACTing and the INCEPTION project (PIA/ANR-16-CONV-0005) for studies focusing on emerging viruses. OS lab is funded by Institut Pasteur, ANRS, Sidaction, the Vaccine Research Institute (ANR-10-LABX-77), 'TIMTAMDEN' ANR-14-CE14-0029, 'CHIKV-Viro- Immuno' ANR-14-CE14-0015-01 and the Gilead HIV cure program. LG is supported by the French Ministry of Higher Education, Research and Innovation., ANR-10-LABX-0062,IBEID,Integrative Biology of Emerging Infectious Diseases(2010), ANR-16-CONV-0005,INCEPTION,Institut Convergences pour l'étude de l'Emergence des Pathologies au Travers des Individus et des populatiONs(2016), ANR-10-LABX-0077,VRI,Initiative for the creation of a Vaccine Research Institute(2010), ANR-14-CE14-0029,TIMTAMDEN,Rôle des récepteurs TIM et TAM dans l'infection des cellules cibles par le virus de la dengue(2014), ANR-14-CE14-0015,CHIKV-Viro-Immuno,Multiplication et Relation avec l'hôte du virus Chikungunya(2014), Centre Médical de l'Institut Pasteur, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris] (IP)-Conservatoire National des Arts et Métiers [CNAM] (CNAM), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM), Virus et Immunité - Virus and immunity (CNRS-UMR3569), Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Institut Pasteur [Paris] (IP), Institut Pasteur [Paris] (IP)-TheraVectys, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Vaccine Research Institute [Créteil, France] (VRI), HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-HESAM Université - Communauté d'universités et d'établissements Hautes écoles Sorbonne Arts et métiers université (HESAM)-Institut Pasteur [Paris]-Conservatoire National des Arts et Métiers [CNAM] (CNAM), and Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)

Subjects
business.industry, Transmission (medicine), [SDV]Life Sciences [q-bio], Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), education, Attack rate, School setting, Retrospective cohort study, Familial clustering, Asymptomatic, 3. Good health, 03 medical and health sciences, 0302 clinical medicine, 030225 pediatrics, High transmission, Medicine, [SDV.SPEE]Life Sciences [q-bio]/Santé publique et épidémiologie, 030212 general & internal medicine, medicine.symptom, business, Demography
Abstract

BackgroundThe extent of SARS-CoV-2 transmission among pupils in primary schools and their families is unknown.MethodsBetween 28-30 April 2020, a retrospective cohort study was conducted among pupils, their parents and relatives, and staff of primary schools exposed to SARS-CoV-2 in February and March 2020 in a city north of Paris, France. Participants completed a questionnaire that covered sociodemographic information and history of recent symptoms. A blood sample was tested for the presence of anti-SARS-CoV-2 antibodies using a flow-cytometry-based assay.ResultsThe infection attack rate (IAR) was 45/510 (8.8%), 3/42 (7.1%), 1/28 (3.6%), 76/641 (11.9%) and 14/119 (11.8%) among primary school pupils, teachers, non-teaching staff, parents, and relatives, respectively (P = 0.29). Prior to school closure on February 14, three SARS-CoV-2 infected pupils attended three separate schools with no secondary cases in the following 14 days among pupils, teachers and non-teaching staff of the same schools. Familial clustering of cases was documented by the high proportion of antibodies among parents and relatives of infected pupils (36/59 = 61.0% and 4/9 = 44.4%, respectively). In children, disease manifestations were mild, and 24/58 (41.4%) of infected children were asymptomatic.InterpretationIn young children, SARS-CoV-2 infection was largely mild or asymptomatic and there was no evidence of onwards transmission from children in the school setting.

Published
2020

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