165 results on '"Ottenhoff THM"'
Search Results
2. Discovery of HLA-E-Presented Epitopes: MHC-E/Peptide Binding and T-Cell Recognition
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Ruibal, P, Franken, KLMC, van Meijgaarden, KE, Walters, LC, McMichael, AJ, Gillespie, GM, Joosten, SA, and Ottenhoff, THM
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Epitopes ,T-Lymphocytes ,Histocompatibility Antigens Class I ,Humans ,Peptides - Abstract
Understanding the interactions involved during the immunological synapse between peptide, HLA-E molecules, and TCR is crucial to effectively target protective HLA-E-restricted T-cell responses in humans. Here we describe three techniques based on the generation of MHC-E/peptide complexes (MHC-E generically includes HLA-E-like molecules in human and nonhuman species, while HLA-E specifically refers to human molecules), which allow to investigate MHC-E/peptide binding at the molecular level through binding assays and by using peptide loaded HLA-E tetramers, to detect, isolate, and study peptide-specific HLA-E-restricted human T-cells.
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- 2022
3. Immunoglobulin G1 Fc glycosylation as an early hallmark of severe COVID-19
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Pongracz, T, Nouta, J, Wang, Wenjun, van, Meijgaarden KE, Linty, F, Vidarsson, G, Joosten, SA, Ottenhoff, THM, Hokke, CH, de, Vries JJC, Arbous, SM, Roukens, AHE, groups, COVID-19, Afd Biomol.Mass Spect. and Proteomics, Biomolecular Mass Spectrometry and Proteomics, Afd Biomol.Mass Spect. and Proteomics, Biomolecular Mass Spectrometry and Proteomics, Landsteiner Laboratory, and AII - Inflammatory diseases
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Glycosylation ,IgG glycosylation ,Inflammation ,Disease ,Biochemistry ,General Biochemistry, Genetics and Molecular Biology ,Cohort Studies ,chemistry.chemical_compound ,Immune system ,medicine ,Humans ,Prospective Studies ,Fucosylation ,business.industry ,Biochemistry, Genetics and Molecular Biology(all) ,Anti-spike IgG ,SARS-CoV-2 ,Confounding ,COVID-19 ,General Medicine ,Immunoglobulin Fc Fragments ,carbohydrates (lipids) ,Coronavirus ,chemistry ,Immunoglobulin G ,Immunology ,Biomarker (medicine) ,medicine.symptom ,business ,Biomarkers ,Cohort study ,Genetics and Molecular Biology(all) - Abstract
BackgroundImmunoglobulin G1 (IgG1) effector functions are impacted by the structure of fragment crystallizable (Fc) tail-linked N-glycans. Low fucosylation levels on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) spike protein specific (anti-S) IgG1 has been described as a hallmark of severe coronavirus disease 2019 (COVID-19) and may lead to activation of macrophages via immune complexes thereby promoting inflammatory responses, altogether suggesting involvement of IgG1 Fc glycosylation modulated immune mechanisms in COVID-19.MethodsIn this prospective, observational single center cohort study, IgG1 Fc glycosylation was analyzed by liquid chromatography – mass spectrometry following affinity capturing from serial plasma samples of 159 SARS-CoV-2 infected patients.FindingsAt baseline close to disease onset, anti-S IgG1 glycosylation was highly skewed when compared to total plasma IgG1. A rapid, general reduction in glycosylation skewing was observed during the disease course. Low anti-S IgG1 galactosylation and sialylation as well as high bisection were early hallmarks of disease severity, whilst high galactosylation and sialylation and low bisection were found in patients with low disease severity. In line with these observations, anti-S IgG1 glycosylation correlated with various inflammatory markers.InterpretationAssociation of low galactosylation, sialylation as well as high bisection with disease severity suggests that Fc-glycan modulated interactions contribute to disease mechanism. Further studies are needed to understand how anti-S IgG1 glycosylation may contributes to disease mechanism and to evaluate its biomarker potential.FundingThis project received funding from the European Commission’s Horizon2020 research and innovation program for H2020-MSCA-ITN IMforFUTURE, under grant agreement number 721815.Research in contextEvidence before this studyAntibody glycosylation against the spike (S) protein of patients infected with severe acute respiratory syndrome SARS-CoV-2 has been reported as a potentially important determinant of COVID-19 disease severity. Studies have hitherto focused on afucosylation, a modification on immunoglobulin G1 (IgG) Fc-tail-linked N-glycans that enhances effector functions. Most of these studies featured limited sample numbers or were imperfectly matched with respect to demographic and other important confounding factors. Our lab has contributed to some of these studies, and we additionally searched for research articles on PubMed and Google Scholar from January 2020 to October 2021. To date, only two groups studied anti-S IgG1 glycosylation, which resulted in overall three publications found. However, none of these groups found a severity marker between hospitalized non-ICU and ICU patients or studied dynamic changes. Instead, exclusively fucosylation at the first available timepoint has been associated with disease severity between severely ill inpatients and mild outpatients.Added value of this studyIn this prospective, observational single center cohort study, we investigated the severity marker potential of anti-S IgG1 glycosylation in severe and mild hospitalized COVID-19 cases, and correlated these findings with numerous inflammation and clinical markers. Our study reveals low galactosylation and sialylation as well as high bisection on anti-S IgG1 as early hallmarks of severe COVID-19, after correction for age and sex effects. In line with these observations, anti-S IgG1 glycosylation correlated with many inflammatory markers. As days since onset is one of the major confounders of anti-S IgG1 glycosylation due to its highly dynamic nature, we additionally confirmed our findings in time-matched patient subgroups. We believe anti-S IgG1 glycosylation may be applicable for patient stratification upon hospitalization.Implications of all the available evidenceDemographic factors as well as temporal differences should be taken into consideration when analyzing IgG1 glycosylation of COVID-19 patients. Anti-S IgG1 glycosylation is highly dynamic, but is a promising early severity marker in COVID-19.
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- 2022
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4. Repurposing diphenylbutylpiperidine-class antipsychotic drugs for host-directed therapy ofMycobacterium tuberculosisandSalmonella entericainfections
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Heemskerk, MT, primary, Korbee, CJ, additional, Esselink, J, additional, Carvalho dos Santos, C, additional, van Veen, S, additional, Gordijn, IF, additional, Vrieling, F, additional, Walburg, KV, additional, Engele, CG, additional, Dijkman, K, additional, Wilson, L, additional, Verreck, FAW, additional, Ottenhoff, THM, additional, and Haks, MC, additional
- Published
- 2021
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5. RISK6, a 6-gene transcriptomic signature of TB disease risk, diagnosis and treatment response
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Penn-Nicholson, A, Mbandi, SK, Thompson, E, Mendelsohn, SC, Suliman, S, Chegou, NN, Malherbe, ST, Darboe, F, Erasmus, M, Hanekom, WA, Bilek, N, Fisher, M, Kaufmann, SHE, Winter, J, Murphy, M, Wood, R, Morrow, C, Van Rhijn, I, Moody, B, Murray, M, Andrade, BB, Sterling, TR, Sutherland, J, Naidoo, K, Padayatchi, N, Walzl, G, Hatherill, M, Zak, D, Scriba, TJ, Kafaar, F, Workman, L, Mulenga, H, Hughes, EJ, Xasa, O, Veldsman, A, Cloete, Y, Abrahams, D, Moyo, S, Gelderbloem, S, Tameris, M, Geldenhuys, H, Ehrlich, R, Verver, S, Geiter, L, Black, GF, van der Spuy, G, Stanley, K, Kriel, M, Du Plessis, N, Nene, N, Roberts, T, Kleynhans, L, Gutschmidt, A, Smith, B, Loxton, AG, Tromp, G, Tabb, D, Ottenhoff, THM, Klein, MR, Haks, MC, Franken, KLMC, Geluk, A, van Meijgaarden, KE, Joosten, SA, Boom, WH, Thiel, B, Mayanja-Kizza, H, Joloba, M, Zalwango, S, Nsereko, M, Okwera, B, Kisingo, H, Parida, SK, Golinski, R, Maertzdorf, J, Weiner, J, Jacobson, M, Dockrell, H, Smith, S, Gorak-Stolinska, P, Hur, YG, Lalor, M, Lee, JS, Crampin, AC, French, N, Ngwira, B, Ben-Smith, A, Watkins, K, Ambrose, L, Simukonda, F, Mvula, H, Chilongo, F, Saul, J, Branson, K, Mahomed, H, Downing, K, The Adolescent Cohort Study team, The GC6-74 Consortium, The SATVI Clinical and Laboratory Team, The ScreenTB Consortium, The AE-TBC Consortium, The RePORT Brazil Team, Peruvian Household Contacts Cohort Team, The CAPRISA IMPRESS team, APH - Methodology, APH - Global Health, AII - Amsterdam institute for Infection and Immunity, Global Health, AII - Infectious diseases, and Translational Immunology Groningen (TRIGR)
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0301 basic medicine ,Oncology ,Male ,GC6-74 Consortium ,AE-TBC Consortium ,lcsh:Medicine ,HIV Infections ,Disease ,Biomarkers/metabolism ,Lung/diagnostic imaging ,ScreenTB Consortium ,Cohort Studies ,Prognostic markers ,0302 clinical medicine ,Immunopathology ,Peruvian Household Contacts Cohort Team ,CAPRISA IMPRESS team ,030212 general & internal medicine ,lcsh:Science ,Lung ,Subclinical infection ,screening and diagnosis ,RNA, Bacterial/metabolism ,Multidisciplinary ,Bacterial/metabolism ,Bacterial ,Prognosis ,Tuberculosis/complications ,RNA, Bacterial ,Detection ,Infectious Diseases ,Mycobacterium tuberculosis/genetics ,Area Under Curve ,Cohort ,Biomarker (medicine) ,HIV/AIDS ,Female ,Adolescent Cohort Study team ,Infection ,Cohort study ,4.2 Evaluation of markers and technologies ,RePORT Brazil Team ,medicine.medical_specialty ,Tuberculosis ,Adolescent ,Point-of-Care Systems ,HIV Infections/complications ,Real-Time Polymerase Chain Reaction/methods ,Real-Time Polymerase Chain Reaction ,Sensitivity and Specificity ,Article ,03 medical and health sciences ,Rare Diseases ,Tuberculosis diagnosis ,Clinical Research ,Internal medicine ,medicine ,Humans ,business.industry ,SATVI Clinical and Laboratory Team ,Prevention ,lcsh:R ,Diagnostic markers ,Mycobacterium tuberculosis ,medicine.disease ,4.1 Discovery and preclinical testing of markers and technologies ,030104 developmental biology ,Good Health and Well Being ,ROC Curve ,Positron-Emission Tomography ,RNA ,lcsh:Q ,business ,Biomarkers - Abstract
Improved tuberculosis diagnostics and tools for monitoring treatment response are urgently needed. We developed a robust and simple, PCR-based host-blood transcriptomic signature, RISK6, for multiple applications: identifying individuals at risk of incident disease, as a screening test for subclinical or clinical tuberculosis, and for monitoring tuberculosis treatment. RISK6 utility was validated by blind prediction using quantitative real-time (qRT) PCR in seven independent cohorts. Prognostic performance significantly exceeded that of previous signatures discovered in the same cohort. Performance for diagnosing subclinical and clinical disease in HIV-uninfected and HIV-infected persons, assessed by area under the receiver-operating characteristic curve, exceeded 85%. As a screening test for tuberculosis, the sensitivity at 90% specificity met or approached the benchmarks set out in World Health Organization target product profiles for non-sputum-based tests. RISK6 scores correlated with lung immunopathology activity, measured by positron emission tomography, and tracked treatment response, demonstrating utility as treatment response biomarker, while predicting treatment failure prior to treatment initiation. Performance of the test in capillary blood samples collected by finger-prick was noninferior to venous blood collected in PAXgene tubes. These results support incorporation of RISK6 into rapid, capillary blood-based point-of-care PCR devices for prospective assessment in field studies.
- Published
- 2020
6. Interleukin-10 promoter single-nucleotide polymorphisms as markers for disease susceptibility and disease severity in leprosy
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Moraes, MO, Pacheco, AG, Schonkeren, JJM, Vanderborght, PR, Nery, JAC, Santos, AR, Moraes, ME, Moraes, JR, Ottenhoff, THM, Sampaio, EP, Huizinga, TWJ, and Sarno, EN
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- 2004
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7. Heterogeneity in the granulomatous response to mycobacterial infection in patients with defined genetic mutations in the interleukin 12-dependent interferon-gamma production pathway
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LAMMAS, DA, DE HEER, E, EDGAR, JD, NOVELLI, V, BEN-SMITH, A, BARETTO, R, DRYSDALE, P, BINCH, J, MACLENNAN, C, KUMARARATNE, DS, PANCHALINGAM, S, OTTENHOFF, THM, CASANOVA, J-L, and EMILE, JF
- Published
- 2002
8. TBVAC2020: Advancing Tuberculosis Vaccines from Discovery to Clinical Development
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Kaufmann, SHE, Dockrell, H.M., Drager, N., Ho, M.M., McShane, H., Neyrolles, O., Ottenhoff, THM, Patel, B., Roordink, D., Spertini, F., Stenger, S., Thole, J., Verreck, FAW, Williams, A., TBVAC2020 Consortium, Britton, W., Triccas, J., Counoupas, C., Grooten, J., Demoitie, M.A., Romano, M., Mascart, F., Andersen, P., Aagaard, C., Christensen, D., Ruhwald, M., Lindenstrom, T., Neyrolles, O., Charneau, P., Guilhot, C., Peixoto, A., Gilleron, M., Locht, C., Brosch, R., Inchauspe, G., Long, SLT, Kaufmann, S., Weiner, J., Maertzdorf, J., Neuwenhuizen, N., Bastian, M., Stenger, S., Caccamo, N., Goletti, D., Nisini, R., Shin, S.J., Lee, H., Sigal, A., Scriba, T., Walzl, G., Loxton, A., Wilkinson, R., Cardona, P.J., Vilaplana, C., Martin, C., Marinova, D., Aguilo, N., Spertini, F., Aebersold, R., Caron, E., Pinschewer, D., De Libero, G., Siegrist, C.A., Collin, N., Barnier-Quer, C., Sander, P., Verreck, F., Ottenhoff, T., Joosten, S., van Meijgaarden, K., Coppola, M., Geluk, A., Drager, N., Roordink, D., Thole, J., Perrie, Y., Baird, M., Levin, M., Dockrell, H., Smith, S., Fletcher, H., Bancroft, G., Rawkins, A., Clark, S., Ho, M.M., McShane, H., Satti, I., Stylianou, E., Vordermeier, M., and Hogarth, P.
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bacille Calmette–Guérin ,biomarker ,clinical trial ,discovery ,portfolio management ,tuberculosis ,vaccination - 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
9. Frequency, phenotype and function of Mycobacterium tuberculosis-specific CD8 T cells in patients with active tuberculosis and in individuals with latent infection
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CACCAMO, Nadia Rosalia, GUGGINO, Giuliana, MERAVIGLIA, Serena, GELSOMINO, Giuseppe, SALERNO, Alfredo, DIELI, Francesco, NOUTA, J, KLEIN, M, OTTENHOFF, THM, CACCAMO, N, GUGGINO, G, MERAVIGLIA, S, GELSOMINO, G, NOUTA, J, KLEIN, M, SALERNO, A, DIELI, F, and OTTENHOFF, THM
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Mycobacterium tuberculosis, CD8 T cells, phenotype - Published
- 2008
10. Frequency and subset distribution of human CD8 T cells specific for mycobacterial peptides in healthy contact individuals
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CACCAMO, Nadia Rosalia, MERAVIGLIA, Serena, GUGGINO, Giuliana, LA MENDOLA, Carmela, DIELI, Francesco, SALERNO, Alfredo, NOUTA J, KLEIN M, OTTENHOFF THM, CACCAMO N, MERAVIGLIA S, NOUTA J, GUGGINO G, KLEIN M, LA MENDOLA C, DIELI F, SALERNO A, and OTTENHOFF THM
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- 2007
11. Identification of a Conserved Universal Th Epitope in HIV-1 Reverse Transcriptase That Is Processed and Presented to HIV-Specific CD4+ T Cells by at Least Four Unrelated HLA-DR Molecules
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Burg, Sh, Kwappenberg, Kmc, Geluk, A., Kruk, M., Pontesilli, O., Hovenkamp, E., Franken, Klmc, Meijgaarden, Ke, Jan Wouter Drijfhout, Ottenhoff, Thm, Melief, Cjm, and Offringa, R.
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Immunology ,Immunology and Allergy - Abstract
CD4+ Th cells play an important role in the induction and maintenance of specific T cell immunity. Indications for a protective role of CD4+ T cells against HIV-1 infection were found in subjects who were able to control HIV-1 viremia as well as in highly HIV-1-exposed, yet seronegative, individuals. This study describes the identification of an HIV-1-specific Th epitope that exhibits high affinity binding as well as high immunogenicity in the context of at least four different HLA-DR molecules that together cover 50–60% of the Caucasian, Oriental, and Negroid populations. This HIV-1 reverse transcriptase-derived peptide (RT171–190) is highly conserved among different HIV-1 isolates. Importantly, stimulation of PBL cultures from HIV-1 seronegative donors with this peptide resulted in Th1-type lymphocytes capable of efficient recognition of HIV-1-pulsed APCs. Taken together, these data indicate that peptide RT171–190 constitutes an attractive component of vaccines aiming at induction or enhancement of HIV-1-specific T cell immunity.
- Published
- 1999
12. Definition of Natural T Cell Antigens with Mimicry Epitopes Obtained from Dedicated Synthetic Peptide Libraries
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Hiemstra, Hs, Veelen, Pa, Schloot, Nc, Geluk, A., Meijgaarden, Ke, Willemen, Sjm, Leunissen, Jam, Benckhuijsen, We, Amons, R., Vries, Rrp, Roep, Bo, Ottenhoff, Thm, and Jan Wouter Drijfhout
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Immunology ,Immunology and Allergy - Abstract
Progress has recently been made in the use of synthetic peptide libraries for the identification of T cell-stimulating ligands. T cell epitopes identified from synthetic libraries are mimics of natural epitopes. Here we show how the mimicry epitopes obtained from synthetic peptide libraries enable unambiguous identification of natural T cell Ags. Synthetic peptide libraries were screened with Mycobacterium tuberculosis-reactive and -autoreactive T cell clones. In two cases, database homology searches with mimicry epitopes isolated from a dedicated synthetic peptide library allowed immediate identification of the natural antigenic protein. In two other cases, an amino acid pattern that reflected the epitope requirements of the T cell was determined by substitution and omission mixture analysis. Subsequently, the natural Ag was identified from databases using this refined pattern. This approach opens new perspectives for rapid and reliable Ag definition, representing a feasible alternative to the biochemical and genetic approaches described thus far.
- Published
- 1998
13. Human T cell responses to peptides of the Mycobacterium leprae 45-kD serine-rich antigen
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Brahmbhatt, S, Hussain, R, Zafar, S, Dawood, G, Ottenhoff, THM, Drijfhout, JW, Bothamley, G, Smith, S, Lopez, FV, and Dockrell, HM
- Abstract
In order to identify T cell epitopes within the Mycobacterium leprae 45-kD serine-rich antigen, we analysed responses to overlapping 17-mer peptides encompassing the whole antigen in non-exposed UK controls, Pakistani leprosy patients and tuberculosis patients in both the United Kingdom and Pakistan. This antigen has been described as M. leprae-specific, although it has a hypothetical homologue in M. tuberculosis. Human peripheral blood mononuclear cells were stimulated with peptide for 5 days and IFN-gamma measured in supernatants by ELISA. Some peptides were recognized more frequently by T cells from tuberculoid leprosy patients than those from UK controls, suggesting that such T cell epitopes might have diagnostic potential, while other peptides induced greater responses among UK control subjects. Short-term cell lines confirmed that these assays detected specific T cell recognition of these peptides. However, many tuberculosis patients also recognized these potentially specific peptides suggesting that there could be a true homologue present in M. tuberculosis.
- Published
- 2002
14. Diagnosis and treatment of lung infection with nontuberculous mycobacteria.
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Arend SM, van Soolingen D, and Ottenhoff THM
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- 2009
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15. Mycobacterium bovis BCG-itis and cervical lymphadenitis due to Salmonella enteritidis in a patient with complete interleukin-12/-23 receptor beta1 deficiency.
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Vosse E, Ottenhoff THM, Paus RA, Verhard EM, Boer T, Dissel JT, and Kuijpers TW
- Abstract
Mendelian susceptibility to mycobacterial disease (MSMD) is a rare disorder with predisposition to severe, sometimes lethal, disease caused by otherwise poorly virulent, non-tuberculous environmental mycobacteria and poorly virulent salmonellae. In patients with MSMD, mutations have been identified in five genes that encode for the proteins IL-12/IL-23p40, IL-12/ IL-23R [beta]1, IFN-R1, IFN-[gamma]R2 and STAT1. These proteins play important roles in the type-1 cytokine pathway, which is crucial for human host defence against intracellular pathogens such as mycobacteria and salmonellae. We report a girl with mild Mycobacterium bovis Bacille Calmette-Guérin (BCG) disease and Salmonella enteritidis cervical lymphadenitis. Despite treatment, she has remained a fecal carrier of S. enteritidis for the past 14 years. She was found to have complete IL-12/IL-23R [beta]1 deficiency. A homozygous r.518G>C IL12RB1 mutation was identified, leading to a non-functional R173P substitution in the IL-12/IL-23R [beta]1 protein. This mutation abrogated IL-12/IL-23R [beta]1 cell-surface expression and resulted in complete lack of T cell responsiveness to both IL-12 and IL-23. [ABSTRACT FROM AUTHOR]
- Published
- 2010
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16. Immunization with human papillomavirus type 16 (HPV16) oncoprotein-loaded dendritic cells as well as protein in adjuvant induces MHC class I-restricted protection to HPV16-induced tumor cells
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Bruijn, Mlh, Schuurhuis, Dh, Vierboom, Mpm, Vermeulen, H., Cock, Kaj, Ooms, Me, Ressing, Me, Toebes, M., Kees Franken, Drijfhout, Jw, Ottenhoff, Thm, Offringa, R., and Melief, Cjm
17. Discovery of isoquinoline sulfonamides as allosteric gyrase inhibitors with activity against fluoroquinolone-resistant bacteria.
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Bakker AT, Kotsogianni I, Avalos M, Punt JM, Liu B, Piermarini D, Gagestein B, Slingerland CJ, Zhang L, Willemse JJ, Ghimire LB, van den Berg RJHBN, Janssen APA, Ottenhoff THM, van Boeckel CAA, van Wezel GP, Ghilarov D, Martin NI, and van der Stelt M
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- Microbial Sensitivity Tests, Structure-Activity Relationship, Drug Discovery, Allosteric Regulation drug effects, Topoisomerase II Inhibitors pharmacology, Topoisomerase II Inhibitors chemistry, Topoisomerase II Inhibitors chemical synthesis, Isoquinolines chemistry, Isoquinolines pharmacology, Isoquinolines chemical synthesis, Sulfonamides pharmacology, Sulfonamides chemistry, Sulfonamides chemical synthesis, Fluoroquinolones pharmacology, Fluoroquinolones chemistry, Fluoroquinolones chemical synthesis, DNA Gyrase metabolism, Anti-Bacterial Agents pharmacology, Anti-Bacterial Agents chemistry, Anti-Bacterial Agents chemical synthesis, Drug Resistance, Bacterial drug effects, Escherichia coli drug effects, Escherichia coli enzymology
- Abstract
Bacteria have evolved resistance to nearly all known antibacterials, emphasizing the need to identify antibiotics that operate via novel mechanisms. Here we report a class of allosteric inhibitors of DNA gyrase with antibacterial activity against fluoroquinolone-resistant clinical isolates of Escherichia coli. Screening of a small-molecule library revealed an initial isoquinoline sulfonamide hit, which was optimized via medicinal chemistry efforts to afford the more potent antibacterial LEI-800. Target identification studies, including whole-genome sequencing of in vitro selected mutants with resistance to isoquinoline sulfonamides, unanimously pointed to the DNA gyrase complex, an essential bacterial topoisomerase and an established antibacterial target. Using single-particle cryogenic electron microscopy, we determined the structure of the gyrase-LEI-800-DNA complex. The compound occupies an allosteric, hydrophobic pocket in the GyrA subunit and has a mode of action that is distinct from the clinically used fluoroquinolones or any other gyrase inhibitor reported to date. LEI-800 provides a chemotype suitable for development to counter the increasingly widespread bacterial resistance to fluoroquinolones., (© 2024. The Author(s).)
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- 2024
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18. Mtb specific HLA-E restricted T cells are induced during Mtb infection but not after BCG administration in non-human primates and humans.
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Voogd L, van Wolfswinkel M, Satti I, White AD, Dijkman K, Gela A, van Meijgaarden KE, Franken KLMC, Marshall JL, Ottenhoff THM, Scriba TJ, McShane H, Sharpe SA, Verreck FAW, and Joosten SA
- Abstract
Novel vaccines targeting the world's deadliest pathogen Mycobacterium tuberculosis ( Mtb ) are urgently needed as the efficacy of the Bacillus Calmette-Guérin (BCG) vaccine in its current use is limited. HLA-E is a virtually monomorphic unconventional antigen presentation molecule and HLA-E restricted Mtb specific CD8
+ T cells can control intracellular Mtb growth, making HLA-E a promising vaccine target for Mtb . In this study, we evaluated the frequency and phenotype of HLA-E restricted Mtb specific CD4+ /CD8+ T cells in the circulation and bronchoalveolar lavage fluid of two independent non-human primate (NHP) studies and from humans receiving BCG either intradermally or mucosally. BCG vaccination followed by Mtb challenge in NHPs did not affect the frequency of circulating and local HLA-E/ Mtb CD4+ and CD8+ T cells, and we saw the same in humans receiving BCG. HLA-E/ Mtb T cell frequencies were significantly increased after Mtb challenge in unvaccinated NHPs, which was correlated with higher TB pathology. Together, HLA-E/ Mtb restricted T cells are minimally induced by BCG in humans and rhesus macaques (RMs) but can be elicited after Mtb infection in unvaccinated RMs. These results give new insights into targeting HLA-E as a potential immune mechanism against TB.- Published
- 2024
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19. Host-directed therapy with amiodarone in preclinical models restricts mycobacterial infection and enhances autophagy.
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Kilinç G, Boland R, Heemskerk MT, Spaink HP, Haks MC, van der Vaart M, Ottenhoff THM, Meijer AH, and Saris A
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- Animals, Humans, Disease Models, Animal, Mycobacterium avium drug effects, Lysosomes drug effects, Lysosomes metabolism, Lysosomes microbiology, Amiodarone pharmacology, Autophagy drug effects, Zebrafish microbiology, Macrophages microbiology, Macrophages immunology, Macrophages drug effects, Mycobacterium tuberculosis drug effects, Mycobacterium tuberculosis growth & development, Tuberculosis drug therapy, Tuberculosis microbiology
- Abstract
Mycobacterium tuberculosis ( Mtb ) as well as nontuberculous mycobacteria are intracellular pathogens whose treatment is extensive and increasingly impaired due to the rise of mycobacterial drug resistance. The loss of antibiotic efficacy has raised interest in the identification of host-directed therapeutics (HDT) to develop novel treatment strategies for mycobacterial infections. In this study, we identified amiodarone as a potential HDT candidate that inhibited both intracellular Mtb and Mycobacterium avium in primary human macrophages without directly impairing bacterial growth, thereby confirming that amiodarone acts in a host-mediated manner. Moreover, amiodarone induced the formation of (auto)phagosomes and enhanced autophagic targeting of mycobacteria in macrophages. The induction of autophagy by amiodarone is likely due to enhanced transcriptional regulation, as the nuclear intensity of the transcription factor EB, the master regulator of autophagy and lysosomal biogenesis, was strongly increased. Furthermore, blocking lysosomal degradation with bafilomycin impaired the host-beneficial effect of amiodarone. Finally, amiodarone induced autophagy and reduced bacterial burden in a zebrafish embryo model of tuberculosis, thereby confirming the HDT activity of amiodarone in vivo . In conclusion, we have identified amiodarone as an autophagy-inducing antimycobacterial HDT that improves host control of mycobacterial infections., Importance: Due to the global rise in antibiotic resistance, there is a strong need for alternative treatment strategies against intracellular bacterial infections, including Mycobacterium tuberculosis ( Mtb ) and non-tuberculous mycobacteria. Stimulating host defense mechanisms by host-directed therapy (HDT) is a promising approach for treating mycobacterial infections. This study identified amiodarone, an antiarrhythmic agent, as a potential HDT candidate that inhibits the survival of Mtb and Mycobacterium avium in primary human macrophages. The antimycobacterial effect of amiodarone was confirmed in an in vivo tuberculosis model based on Mycobacterium marinum infection of zebrafish embryos. Furthermore, amiodarone induced autophagy and inhibition of the autophagic flux effectively impaired the host-protective effect of amiodarone, supporting that activation of the host (auto)phagolysosomal pathway is essential for the mechanism of action of amiodarone. In conclusion, we have identified amiodarone as an autophagy-inducing HDT that improves host control of a wide range of mycobacteria., Competing Interests: The authors declare no conflict of interest.
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- 2024
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20. Identification of kinase inhibitors as potential host-directed therapies for intracellular bacteria.
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van den Biggelaar RHGA, Walburg KV, van den Eeden SJF, van Doorn CLR, Meiler E, de Ries AS, Fusco MC, Meijer AH, Ottenhoff THM, and Saris A
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- Animals, Humans, Salmonella Infections drug therapy, Salmonella Infections microbiology, Anti-Bacterial Agents pharmacology, Cell Line, Embryo, Nonmammalian drug effects, Tuberculosis drug therapy, Tuberculosis microbiology, Zebrafish, Salmonella typhimurium drug effects, Mycobacterium tuberculosis drug effects, Mycobacterium tuberculosis enzymology, Protein Kinase Inhibitors pharmacology, Macrophages microbiology, Macrophages drug effects, Macrophages metabolism
- Abstract
The emergence of antimicrobial resistance has created an urgent need for alternative treatments against bacterial pathogens. Here, we investigated kinase inhibitors as potential host-directed therapies (HDTs) against intracellular bacteria, specifically Salmonella Typhimurium (Stm) and Mycobacterium tuberculosis (Mtb). We screened 827 ATP-competitive kinase inhibitors with known target profiles from two Published Kinase Inhibitor Sets (PKIS1 and PKIS2) using intracellular infection models for Stm and Mtb, based on human cell lines and primary macrophages. Additionally, the in vivo safety and efficacy of the compounds were assessed using zebrafish embryo infection models. Our screen identified 11 hit compounds for Stm and 17 hit compounds for Mtb that were effective against intracellular bacteria and non-toxic for host cells. Further experiments were conducted to prioritize Stm hit compounds that were able to clear the intracellular infection in primary human macrophages. From these, two structurally related Stm hit compounds, GSK1379738A and GSK1379760A, exhibited significant activity against Stm in infected zebrafish embryos. In addition, we identified compounds that were active against intracellular Mtb, including morpholino-imidazo/triazolo-pyrimidinones that target PIK3CB, as well as 2-aminobenzimidazoles targeting ABL1. Overall, this study provided insights into kinase targets acting at the host-pathogen interface and identified several kinase inhibitors as potential HDTs., (© 2024. The Author(s).)
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- 2024
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21. Cationic pH-sensitive liposome-based subunit tuberculosis vaccine induces protection in mice challenged with Mycobacterium tuberculosis.
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Szachniewicz MM, van den Eeden SJF, van Meijgaarden KE, Franken KLMC, van Veen S, Geluk A, Bouwstra JA, and Ottenhoff THM
- Subjects
- Animals, Mice, Humans, Female, Hydrogen-Ion Concentration, Mice, Inbred C57BL, Dendritic Cells immunology, Cations, Antigens, Bacterial immunology, Antigens, Bacterial administration & dosage, Cytokines metabolism, CD8-Positive T-Lymphocytes immunology, Lung immunology, Lung microbiology, CD4-Positive T-Lymphocytes immunology, Bacterial Load, Liposomes, Tuberculosis Vaccines administration & dosage, Tuberculosis Vaccines immunology, Mycobacterium tuberculosis immunology, Vaccines, Subunit immunology, Vaccines, Subunit administration & dosage, Adjuvants, Immunologic administration & dosage, Adjuvants, Immunologic pharmacology, Tuberculosis prevention & control, Tuberculosis immunology
- Abstract
Tuberculosis (TB) has been and still is a global emergency for centuries. Prevention of disease through vaccination would have a major impact on disease prevalence, but the only available current vaccine, BCG, has insufficient impact. In this article, a novel subunit vaccine against TB was developed, using the Ag85B-ESAT6-Rv2034 fusion antigen, two adjuvants - CpG and MPLA, and a cationic pH-sensitive liposome as a delivery system, representing a new TB vaccine delivery strategy not previously reported for TB. In vitro in human dendritic cells (DCs), the adjuvanted formulation induced a significant increase in the production of (innate) cytokines and chemokines compared to the liposome without additional adjuvants. In vivo, the new vaccine administrated subcutaneously significantly reduced Mycobacterium tuberculosis (Mtb) bacterial load in the lungs and spleens of mice, significantly outperforming results from mice vaccinated with the antigen mixed with adjuvants without liposomes. In-depth analysis underpinned the vaccine's effectiveness in terms of its capacity to induce polyfunctional CD4
+ and CD8+ T-cell responses, both considered essential for controlling Mtb infection. Also noteworthy was the differential abundance of various CD69+ B-cell subpopulations, which included IL17-A-producing B-cells. The vaccine stimulated robust antigen-specific antibody titers, further extending its potential as a novel protective agent against TB., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)- Published
- 2024
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22. Longitudinal soluble marker profiles reveal strong association between cytokine storms resulting from macrophage activation and disease severity in COVID-19 disease.
- Author
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van Meijgaarden KE, van Veen S, Tsonaka R, Ruibal P, Roukens AHE, Arbous SM, Manniën J, Cannegieter SC, Ottenhoff THM, and Joosten SA
- Subjects
- Humans, Male, Female, Middle Aged, SARS-CoV-2 isolation & purification, Cytokines blood, Cytokine Release Syndrome blood, Adult, Aged, Serum Amyloid P-Component metabolism, Serum Amyloid P-Component analysis, C-Reactive Protein, COVID-19 blood, COVID-19 immunology, Biomarkers blood, Severity of Illness Index, Macrophage Activation
- Abstract
SARS-CoV2 infection results in a range of disease severities, but the underlying differential pathogenesis is still not completely understood. At presentation it remains difficult to estimate and predict severity, in particular, identify individuals at greatest risk of progression towards the most severe disease-states. Here we used advanced models with circulating serum analytes as variables in combination with daily assessment of disease severity using the SCODA-score, not only at single time points but also during the course of disease, to correlate analyte levels and disease severity. We identified a remarkably strong pro-inflammatory cytokine/chemokine profile with high levels for sCD163, CCL20, HGF, CHintinase3like1 and Pentraxin3 in serum which correlated with COVID-19 disease severity and overall outcome. Although precise analyte levels differed, resulting biomarker profiles were highly similar at early and late disease stages, and even during convalescence similar biomarkers were elevated and further included CXCL3, CXCL6 and Osteopontin. Taken together, strong pro-inflammatory marker profiles were identified in patients with COVID-19 disease which correlated with overall outcome and disease severity., (© 2024. The Author(s).)
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- 2024
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23. Age and sex influence antibody profiles associated with tuberculosis progression.
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Davies LRL, Wang C, Steigler P, Bowman KA, Fischinger S, Hatherill M, Fisher M, Mbandi SK, Rodo M, Ottenhoff THM, Dockrell HM, Sutherland JS, Mayanja-Kizza H, Boom WH, Walzl G, Kaufmann SHE, Nemes E, Scriba TJ, Lauffenburger D, Alter G, and Fortune SM
- Subjects
- Humans, Male, Female, Adolescent, Sex Factors, Adult, Age Factors, South Africa epidemiology, Young Adult, Cohort Studies, Antibody Formation immunology, Mycobacterium tuberculosis immunology, Antibodies, Bacterial blood, Antibodies, Bacterial immunology, Disease Progression, Tuberculosis immunology, Tuberculosis microbiology
- Abstract
Antibody features vary with tuberculosis (TB) disease state. Whether clinical variables, such as age or sex, influence associations between Mycobacterium tuberculosis-specific antibody responses and disease state is not well explored. Here we profiled Mycobacterium tuberculosis-specific antibody responses in 140 TB-exposed South African individuals from the Adolescent Cohort Study. We identified distinct response features in individuals progressing to active TB from non-progressing, matched controls. A multivariate antibody score differentially associated with progression (SeroScore) identified progressors up to 2 years before TB diagnosis, earlier than that achieved with the RISK6 transcriptional signature of progression. We validated these antibody response features in the Grand Challenges 6-74 cohort. Both the SeroScore and RISK6 correlated better with risk of TB progression in adolescents compared with adults, and in males compared with females. This suggests that age and sex are important, underappreciated modifiers of antibody responses associated with TB progression., (© 2024. The Author(s).)
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- 2024
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24. Intrinsic immunogenicity of liposomes for tuberculosis vaccines: Effect of cationic lipid and cholesterol.
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Szachniewicz MM, Neustrup MA, van Meijgaarden KE, Jiskoot W, Bouwstra JA, Haks MC, Geluk A, and Ottenhoff THM
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- Humans, Animals, Mice, Liposomes chemistry, Adjuvants, Immunologic chemistry, Vaccines, Subunit, Lipids chemistry, Cholesterol chemistry, Mice, Inbred C57BL, Tuberculosis Vaccines chemistry, Vaccines
- Abstract
Tuberculosis (TB) is still among the deadliest infectious diseases, hence there is a pressing need for more effective TB vaccines. Cationic liposome subunit vaccines are excellent vaccine candidates offering effective protection with a better safety profile than live vaccines. In this study, we aim to explore intrinsic adjuvant properties of cationic liposomes to maximize immune activation while minimizing aspecific cytotoxicity. To achieve this, we developed a rational strategy to select liposomal formulation compositions and assessed their physicochemical and immunological properties in vitro models using human monocyte-derived dendritic cells (MDDCs). A broad selection of commercially available cationic compounds was tested to prepare liposomes containing Ag85B-ESAT6-Rv2034 (AER) fusion protein antigen. 1,2-Dioleoyl-sn‑glycero-3-ethylphosphocholine (EPC)-based liposomes exhibited the most advantageous activation profile in MDDCs as assessed by cell surface activation markers, cellular uptake, antigen-specific T-cell activation, cytokine production, and cellular viability. The addition of cholesterol to 20 mol% improved the performance of the tested formulations compared to those without it; however, when its concentration was doubled there was no further benefit, resulting in reduced cell viability. This study provides new insights into the role of cationic lipids and cholesterol in liposomal subunit vaccines., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2024. Published by Elsevier B.V.)
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- 2024
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25. Identification of kinase modulators as host-directed therapeutics against intracellular methicillin-resistant Staphylococcus aureus .
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van den Biggelaar RHGA, Walburg KV, van den Eeden SJF, van Doorn CLR, Meiler E, de Ries AS, Meijer AH, Ottenhoff THM, and Saris A
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- Humans, Animals, Staphylococcus aureus, Zebrafish, Anti-Bacterial Agents pharmacology, Anti-Bacterial Agents therapeutic use, Microbial Sensitivity Tests, Methicillin-Resistant Staphylococcus aureus, Staphylococcal Infections microbiology
- Abstract
The increasing prevalence of antimicrobial-resistant Staphylococcus aureus strains, especially methicillin-resistant S. aureus (MRSA), poses a threat to successful antibiotic treatment. Unsuccessful attempts to develop a vaccine and rising resistance to last-resort antibiotics urge the need for alternative treatments. Host-directed therapy (HDT) targeting critical intracellular stages of S. aureus emerges as a promising alternative, potentially acting synergistically with antibiotics and reducing the risk of de novo drug resistance. We assessed 201 ATP-competitive kinase inhibitors from Published Kinase Inhibitor Sets (PKIS1 and PKIS2) against intracellular MRSA. Seventeen hit compounds were identified, of which the two most effective and well-tolerated hit compounds (i.e., GW633459A and GW296115X) were selected for further analysis. The compounds did not affect planktonic bacterial cultures, while they were active in a range of human cell lines of cervical, skin, lung, breast and monocyte origin, confirming their host-directed mechanisms. GW633459A, structurally related to lapatinib, exhibited an HDT effect on intracellular MRSA independently of its known human epidermal growth factor receptor (EGFR)/(HER) kinase family targets. GW296115X activated adenosine monophosphate-activated protein kinase (AMPK), thereby enhancing bacterial degradation via autophagy. Finally, GW296115X not only reduced MRSA growth in human cells but also improved the survival rates of MRSA-infected zebrafish embryos, highlighting its potential as HDT., Competing Interests: Author EM is employed by GlasmoSmithKline. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be constructed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision, (Copyright © 2024 van den Biggelaar, Walburg, van den Eeden, van Doorn, Meiler, de Ries, Meijer, Ottenhoff and Saris.)
- Published
- 2024
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26. Whole blood RNA signatures in tuberculosis patients receiving H56:IC31 vaccine as adjunctive therapy.
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Alonso-Rodríguez N, Vianello E, van Veen S, Jenum S, Tonby K, van Riessen R, Lai X, Mortensen R, Ottenhoff THM, and Dyrhol-Riise AM
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- Adolescent, Humans, Oligodeoxyribonucleotides, Cohort Studies, RNA, Tuberculosis Vaccines therapeutic use, Tuberculosis prevention & control
- Abstract
Introduction: Therapeutic vaccination in tuberculosis (TB) represents a Host Directed Therapy strategy which enhances immune responses in order to improve clinical outcomes and shorten TB treatment. Previously, we have shown that the subunit H56:IC31 vaccine induced both humoral and cellular immune responses when administered to TB patients adjunctive to standard TB treatment (TBCOX2 study, NCT02503839). Here we present the longitudinal whole blood gene expression patterns in H56:IC31 vaccinated TB patients compared to controls receiving standard TB treatment only., Methods: The H56:IC31 group (N=11) and Control group (N=7) underwent first-line TB treatment for 182 days. The H56:IC31 group received 5 micrograms of the H56:IC31 vaccine (Statens Serum Institut; SSI, Valneva Austria GmbH) intramuscularly at day 84 and day 140. Total RNA was extracted from whole blood samples collected in PAXgene tubes on days 0, 84, 98, 140, 154, 182 and 238. The expression level of 183 immune-related genes was measured by high-throughput microfluidic qPCR (Biomark HD system, Standard BioTools)., Results: The targeted gene expression profiling unveiled the upregulation of modules such as interferon (IFN) signalling genes, pattern recognition receptors and small nucleotide guanosine triphosphate (GTP)-ases in the vaccinated group compared to controls two weeks after administration of the first H56:IC31 vaccine. Additionally, the longitudinal analysis of the Adolescent Cohort Study-Correlation of Risk (ACS-COR) signature showed a progressive downregulation in both study arms towards the end of TB treatment, in congruence with reported treatment responses and clinical improvements. Still, two months after the end of TB treatment, vaccinated patients, and especially those developing both cellular and humoral vaccine responses, showed a lower expression of the ACS-COR genes compared to controls., Discussion: Our data report gene expression patterns following H56:IC31 vaccination which might be interpreted as a lower risk of relapse in therapeutically vaccinated patients. Further studies are needed to conclude if these gene expression patterns could be used as prognostic biosignatures for therapeutic TB vaccine responses., Competing Interests: RM is employee at SSI that developed the H56:IC31 vaccine. He is not inventor of related patents and have no personal financial interests. IC31 is a trademark of Valneva Austria GmbH. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Alonso-Rodríguez, Vianello, van Veen, Jenum, Tonby, van Riessen, Lai, Mortensen, Ottenhoff and Dyrhol-Riise.)
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- 2024
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27. Mtb HLA-E-tetramer-sorted CD8 + T cells have a diverse TCR repertoire.
- Author
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Voogd L, Drittij AMHF, Dingenouts CKE, Franken KLMC, Unen VV, van Meijgaarden KE, Ruibal P, Hagedoorn RS, Leitner JA, Steinberger P, Heemskerk MHM, Davis MM, Scriba TJ, Ottenhoff THM, and Joosten SA
- Abstract
HLA-E molecules can present self- and pathogen-derived peptides to both natural killer (NK) cells and T cells. T cells that recognize HLA-E peptides via their T cell receptor (TCR) are termed donor-unrestricted T cells due to restricted allelic variation of HLA-E. The composition and repertoire of HLA-E TCRs is not known so far. We performed TCR sequencing on CD8
+ T cells from 21 individuals recognizing HLA-E tetramers (TMs) folded with two Mtb -HLA-E-restricted peptides. We sorted HLA-E Mtb TM+ and TM- CD8+ T cells directly ex vivo and performed bulk RNA-sequencing and single-cell TCR sequencing. The identified TCR repertoire was diverse and showed no conservation between and within individuals. TCRs selected from our single-cell TCR sequencing data could be activated upon HLA-E/peptide stimulation, although not robust, reflecting potentially weak interactions between HLA-E peptide complexes and TCRs. Thus, HLA-E- Mtb -specific T cells have a highly diverse TCR repertoire., Competing Interests: The authors declare that they have no competing interest., (© 2024 The Author(s).)- Published
- 2024
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28. Refined innate plasma signature after rVSVΔG-ZEBOV-GP immunization is shared among adult cohorts in Europe and North America.
- Author
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Martinez-Murillo PA, Huttner A, Lemeille S, Medaglini D, Ottenhoff THM, Harandi AM, Didierlaurent AM, and Siegrist CA
- Subjects
- Adult, Humans, Follow-Up Studies, Vaccination, Europe, North America, Democratic Republic of the Congo, Biomarkers, Antibodies, Viral, Ebola Vaccines
- Abstract
Background: During the last decade Ebola virus has caused several outbreaks in Africa. The recombinant vesicular stomatitis virus-vectored Zaire Ebola (rVSVΔG-ZEBOV-GP) vaccine has proved safe and immunogenic but is reactogenic. We previously identified the first innate plasma signature response after vaccination in Geneva as composed of five monocyte-related biomarkers peaking at day 1 post-immunization that correlates with adverse events, biological outcomes (haematological changes and viremia) and antibody titers. In this follow-up study, we sought to identify additional biomarkers in the same Geneva cohort and validate those identified markers in a US cohort., Methods: Additional biomarkers were identified using multiplexed protein biomarker platform O-link and confirmed by Luminex. Principal component analysis (PCA) evaluated if these markers could explain a higher variability of the vaccine response (and thereby refined the initial signature). Multivariable and linear regression models evaluated the correlations of the main components with adverse events, biological outcomes, and antibody titers. External validation of the refined signature was conducted in a second cohort of US vaccinees (n=142)., Results: Eleven additional biomarkers peaked at day 1 post-immunization: MCP2, MCP3, MCP4, CXCL10, OSM, CX3CL1, MCSF, CXCL11, TRAIL, RANKL and IL15. PCA analysis retained three principal components (PC) that accounted for 79% of the vaccine response variability. PC1 and PC2 were very robust and had different biomarkers that contributed to their variability. PC1 better discriminated different doses, better defined the risk of fever and myalgia, while PC2 better defined the risk of headache. We also found new biomarkers that correlated with reactogenicity, including transient arthritis (MCP-2, CXCL10, CXCL11, CX3CL1, MCSF, IL-15, OSM). Several innate biomarkers are associated with antibody levels one and six months after vaccination. Refined PC1 correlated strongly in both data sets (Geneva: r = 0.97, P < 0.001; US: r = 0.99, P< 0.001)., Conclusion: Eleven additional biomarkers refined the previously found 5-biomarker Geneva signature. The refined signature better discriminated between different doses, was strongly associated with the risk of adverse events and with antibody responses and was validated in a separate cohort., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Martinez-Murillo, Huttner, Lemeille, Medaglini, Ottenhoff, Harandi, Didierlaurent and Siegrist.)
- Published
- 2024
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29. BCG vaccination-induced acquired control of mycobacterial growth differs from growth control preexisting to BCG vaccination.
- Author
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van Meijgaarden KE, Li W, Moorlag SJCFM, Koeken VACM, Koenen HJPM, Joosten LAB, Vyakarnam A, Ahmed A, Rakshit S, Adiga V, Ottenhoff THM, Li Y, Netea MG, and Joosten SA
- Subjects
- Adult, Infant, Child, Humans, Child, Preschool, BCG Vaccine, Vaccination methods, Mycobacterium, Tuberculosis microbiology
- Abstract
Bacillus Calmette-Guèrin - vaccination induces not only protection in infants and young children against severe forms of tuberculosis, but also against non-tuberculosis related all-cause mortality. To delineate different factors influencing mycobacterial growth control, here we first investigate the effects of BCG-vaccination in healthy Dutch adults. About a quarter of individuals already control BCG-growth prior to vaccination, whereas a quarter of the vaccinees acquires the capacity to control BCG upon vaccination. This leaves half of the population incapable to control BCG-growth. Single cell RNA sequencing identifies multiple processes associated with mycobacterial growth control. These data suggest (i) that already controllers employ different mechanisms to control BCG-growth than acquired controllers, and (ii) that half of the individuals fail to develop measurable growth control irrespective of BCG-vaccination. These results shed important new light on the variable immune responses to mycobacteria in humans and may impact on improved vaccination against tuberculosis and other diseases., (© 2024. The Author(s).)
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- 2024
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30. Global blood miRNA profiling unravels early signatures of immunogenicity of Ebola vaccine rVSVΔG-ZEBOV-GP.
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Vianello E, Persson J, Andersson B, van Veen S, Dias TL, Santoro F, Östensson M, Obudulu O, Agbajogu C, Torkzadeh S, Nakaya HI, Medaglini D, Siegrist CA, Ottenhoff THM, and Harandi AM
- Abstract
The vectored Ebola vaccine rVSVΔG-ZEBOV-GP elicits protection against Ebola Virus Disease (EVD). In a study of forty-eight healthy adult volunteers who received either the rVSVΔG-ZEBOV-GP vaccine or placebo, we profiled intracellular microRNAs (miRNAs) from whole blood cells (WB) and circulating miRNAs from serum-derived extracellular vesicles (EV) at baseline and longitudinally following vaccination. Further, we identified early miRNA signatures associated with ZEBOV-specific IgG antibody responses at baseline and up to one year post-vaccination, and pinpointed target mRNA transcripts and pathways correlated to miRNAs whose expression was altered after vaccination by using systems biology approaches. Several miRNAs were differentially expressed (DE) and miRNA signatures predicted high or low IgG ZEBOV-specific antibody levels with high classification performance. The top miRNA discriminators were WB-miR-6810, EV-miR-7151-3p, and EV-miR-4426. An eight-miRNA antibody predictive signature was associated with immune-related target mRNAs and pathways. These findings provide valuable insights into early blood biomarkers associated with rVSVΔG-ZEBOV-GP vaccine-induced IgG antibody responses., Competing Interests: The authors declare no competing interests., (© 2023 The Author(s).)
- Published
- 2023
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31. Corrigendum: Defining discriminatory antibody fingerprints in active and latent tuberculosis.
- Author
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Nziza N, Cizmeci D, Davies L, Irvine EB, Jung W, Fenderson BA, de Kock M, Hanekom WA, Franken KLMC, Day CL, Ottenhoff THM, and Alter G
- Abstract
[This corrects the article DOI: 10.3389/fimmu.2022.856906.]., (Copyright © 2023 Nziza, Cizmeci, Davies, Irvine, Jung, Fenderson, de Kock, Hanekom, Franken, Day, Ottenhoff and Alter.)
- Published
- 2023
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32. Baseline gene signatures of reactogenicity to Ebola vaccination: a machine learning approach across multiple cohorts.
- Author
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Gonzalez Dias Carvalho PC, Dominguez Crespo Hirata T, Mano Alves LY, Moscardini IF, do Nascimento APB, Costa-Martins AG, Sorgi S, Harandi AM, Ferreira DM, Vianello E, Haks MC, Ottenhoff THM, Santoro F, Martinez-Murillo P, Huttner A, Siegrist CA, Medaglini D, and Nakaya HI
- Subjects
- Humans, Antibodies, Viral, Headache, Vaccination adverse effects, Vaccination methods, Clinical Trials, Phase I as Topic, Arthritis etiology, Ebola Vaccines adverse effects, Ebolavirus genetics, Hemorrhagic Fever, Ebola
- Abstract
Introduction: The rVSVDG-ZEBOV-GP (Ervebo®) vaccine is both immunogenic and protective against Ebola. However, the vaccine can cause a broad range of transient adverse reactions, from headache to arthritis. Identifying baseline reactogenicity signatures can advance personalized vaccinology and increase our understanding of the molecular factors associated with such adverse events., Methods: In this study, we developed a machine learning approach to integrate prevaccination gene expression data with adverse events that occurred within 14 days post-vaccination., Results and Discussion: We analyzed the expression of 144 genes across 343 blood samples collected from participants of 4 phase I clinical trial cohorts: Switzerland, USA, Gabon, and Kenya. Our machine learning approach revealed 22 key genes associated with adverse events such as local reactions, fatigue, headache, myalgia, fever, chills, arthralgia, nausea, and arthritis, providing insights into potential biological mechanisms linked to vaccine reactogenicity., Competing Interests: Author IM was employed by the company Microbiotec Srl. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Gonzalez Dias Carvalho, Dominguez Crespo Hirata, Mano Alves, Moscardini, do Nascimento, Costa-Martins, Sorgi, Harandi, Ferreira, Vianello, Haks, Ottenhoff, Santoro, Martinez-Murillo, Huttner, Siegrist, Medaglini and Nakaya.)
- Published
- 2023
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33. Potential business model for a European vaccine R&D infrastructure and its estimated socio-economic impact.
- Author
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Jungbluth S, Martin W, Slezak M, Depraetere H, Guzman CA, Ussi A, Morrow D, Van Heuverswyn F, Arnouts S, Carrondo MJT, Olesen O, Ottenhoff THM, Dockrell HM, Ho MM, Dobly A, Christensen D, Segalés J, Laurent F, Lantier F, Stockhofe-Zurwieden N, Morelli F, Langermans JAM, Verreck FAW, Le Grand R, Sloots A, Medaglini D, Lawrenz M, and Collin N
- Subjects
- Commerce, Socioeconomic Factors, Biomedical Research, Vaccines
- Abstract
Background: Research infrastructures are facilities or resources that have proven fundamental for supporting scientific research and innovation. However, they are also known to be very expensive in their establishment, operation and maintenance. As by far the biggest share of these costs is always borne by public funders, there is a strong interest and indeed a necessity to develop alternative business models for such infrastructures that allow them to function in a more sustainable manner that is less dependent on public financing., Methods: In this article, we describe a feasibility study we have undertaken to develop a potentially sustainable business model for a vaccine research and development (R&D) infrastructure. The model we have developed integrates two different types of business models that would provide the infrastructure with two different types of revenue streams which would facilitate its establishment and would be a measure of risk reduction. For the business model we are proposing, we have undertaken an ex ante impact assessment that estimates the expected impact for a vaccine R&D infrastructure based on the proposed models along three different dimensions: health, society and economy., Results: Our impact assessment demonstrates that such a vaccine R&D infrastructure could achieve a very significant socio-economic impact, and so its establishment is therefore considered worthwhile pursuing., Conclusions: The business model we have developed, the impact assessment and the overall process we have followed might also be of interest to other research infrastructure initiatives in the biomedical field., Competing Interests: No competing interests were disclosed., (Copyright: © 2023 Jungbluth S et al.)
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- 2023
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34. Airway epithelial cells mount an early response to mycobacterial infection.
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Barclay AM, Ninaber DK, van Veen S, Hiemstra PS, Ottenhoff THM, van der Does AM, and Joosten SA
- Subjects
- Humans, Cytokines metabolism, Epithelial Cells metabolism, Chemokines metabolism, Mycobacterium Infections, Mycobacterium tuberculosis
- Abstract
Lung epithelial cells represent the first line of host defence against foreign inhaled components, including respiratory pathogens. Their responses to these exposures may direct subsequent immune activation to these pathogens. The epithelial response to mycobacterial infections is not well characterized and may provide clues to why some mycobacterial infections are cleared, while others are persistent and pathogenic. We have utilized an air-liquid interface model of human primary bronchial epithelial cells (ALI-PBEC) to investigate the epithelial response to infection with a variety of mycobacteria: Mycobacterium tuberculosis (Mtb), M. bovis (BCG), M. avium, and M. smegmatis . Airway epithelial cells were found to be infected by all four species, albeit at low frequencies. The proportion of infected epithelial cells was lowest for Mtb and highest for M. avium . Differential gene expression analysis revealed a common epithelial host response to mycobacteria, including upregulation of BIRC3, S100A8 and DEFB4 , and downregulation of BPIFB1 at 48 h post infection. Apical secretions contained predominantly pro-inflammatory cytokines, while basal secretions contained tissue growth factors and chemokines. Finally, we show that neutrophils were attracted to both apical and basal secretions of infected ALI-PBEC. Neutrophils were attracted in high numbers to apical secretions from PBEC infected with all mycobacteria, with the exception of secretions from M. avium -infected ALI-PBEC. Taken together, our results show that airway epithelial cells are differentially infected by mycobacteria, and react rapidly by upregulation of antimicrobials, and increased secretion of inflammatory cytokines and chemokines which directly attract neutrophils. Thus, the airway epithelium may be an important immunological component in controlling and regulating mycobacterial infections., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Barclay, Ninaber, van Veen, Hiemstra, Ottenhoff, van der Does and Joosten.)
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- 2023
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35. BCG revaccination in adults enhances pro-inflammatory markers of trained immunity along with anti-inflammatory pathways.
- Author
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Ahmed A, Tripathi H, van Meijgaarden KE, Kumar NC, Adiga V, Rakshit S, Parthiban C, Eveline J S, D'Souza G, Dias M, Ottenhoff THM, Netea MG, Joosten SA, and Vyakarnam A
- Abstract
This study characterized mechanisms of Bacille Calmette-Guérin (BCG) revaccination-induced trained immunity (TI) in India. Adults, BCG vaccinated at birth, were sampled longitudinally before and after a second BCG dose. BCG revaccination significantly elevated tumor necrosis factor alpha (TNF-α), interleukin (IL)-1β, and IL-6 in HLA-DR
+ CD16- CD14hi monocytes, demonstrating induction of TI. Mycobacteria-specific CD4+ T cell interferon (IFN) γ, IL-2, and TNF-α were significantly higher in re-vaccinees and correlated positively with HLA-DR+ CD16- CD14hi TI responses. This, however, did not translate into increased mycobacterial growth control, measured by mycobacterial growth inhibition assay (MGIA). Post revaccination, elevated secreted TNF-α, IL-1β, and IL-6 to "heterologous" fungal, bacterial, and enhanced CXCL-10 and IFNα to viral stimuli were also observed concomitant with increased anti-inflammatory cytokine, IL-1RA. RNA sequencing after revaccination highlighted a BCG and LPS induced signature which included upregulated IL17 and TNF pathway genes and downregulated key inflammatory genes: CXCL11, CCL24, HLADRA, CTSS, CTSC . Our data highlight a balanced immune response comprising pro- and anti-inflammatory mediators to be a feature of BCG revaccination-induced immunity., Competing Interests: The authors declare no competing interests., (© 2023 The Authors.)- Published
- 2023
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36. Impaired resolution of blood transcriptomes through tuberculosis treatment with diabetes comorbidity.
- Author
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Eckold C, van Doorn CLR, Ruslami R, Ronacher K, Riza AL, van Veen S, Lee JS, Kumar V, Kerry-Barnard S, Malherbe ST, Kleynhans L, Stanley K, Joosten SA, Critchley JA, Hill PC, van Crevel R, Wijmenga C, Haks MC, Ioana M, Alisjahbana B, Walzl G, Ottenhoff THM, Dockrell HM, Vianello E, and Cliff JM
- Subjects
- Humans, Transcriptome genetics, Comorbidity, Gene Expression Profiling, Diabetes Mellitus, Hyperglycemia
- Abstract
Background: People with diabetes are more likely to develop tuberculosis (TB) and to have poor TB-treatment outcomes than those without. We previously showed that blood transcriptomes in people with TB-diabetes (TB-DM) co-morbidity have excessive inflammatory and reduced interferon responses at diagnosis. It is unknown whether this persists through treatment and contributes to the adverse outcomes., Methods: Pulmonary TB patients recruited in South Africa, Indonesia and Romania were classified as having TB-DM, TB with prediabetes, TB-related hyperglycaemia or TB-only, based on glycated haemoglobin concentration at TB diagnosis and after 6 months of TB treatment. Gene expression in blood at diagnosis and intervals throughout treatment was measured by unbiased RNA-Seq and targeted Multiplex Ligation-dependent Probe Amplification. Transcriptomic data were analysed by longitudinal mixed-model regression to identify whether genes were differentially expressed between clinical groups through time. Predictive models of TB-treatment response across groups were developed and cross-tested., Results: Gene expression differed between TB and TB-DM patients at diagnosis and was modulated by TB treatment in all clinical groups but to different extents, such that differences remained in TB-DM relative to TB-only throughout. Expression of some genes increased through TB treatment, whereas others decreased: some were persistently more highly expressed in TB-DM and others in TB-only patients. Genes involved in innate immune responses, anti-microbial immunity and inflammation were significantly upregulated in people with TB-DM throughout treatment. The overall pattern of change was similar across clinical groups irrespective of diabetes status, permitting models predictive of TB treatment to be developed., Conclusions: Exacerbated transcriptome changes in TB-DM take longer to resolve during TB treatment, meaning they remain different from those in uncomplicated TB after treatment completion. This may indicate a prolonged inflammatory response in TB-DM, requiring prolonged treatment or host-directed therapy for complete cure. Development of transcriptome-based biomarker signatures of TB-treatment response should include people with diabetes for use across populations., (© 2023 The Authors. Clinical and Translational Medicine published by John Wiley & Sons Australia, Ltd on behalf of Shanghai Institute of Clinical Bioinformatics.)
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- 2023
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37. Random forest and live single-cell metabolomics reveal metabolic profiles of human macrophages upon polarization.
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Tang H, Ali A, Abdelazem E, Ottenhoff THM, Heeren RMA, and Mashaghi A
- Subjects
- Humans, Metabolomics, Metabolome, Phenotype, Random Forest, Macrophages metabolism
- Abstract
Human macrophages are innate immune cells with diverse, functionally distinct phenotypes, namely, pro-inflammatory M1 and anti-inflammatory M2 macrophages. Both are involved in multiple physiological and pathological processes, including would healing, infection, and cancer. However, the metabolic differences between these phenotypes are largely unexplored at single-cell resolution. To address this knowledge gap, an untargeted live single-cell mass spectrometry-based metabolomic profiling coupled with a machine-learning data analysis approach was developed to investigate the metabolic profile of each phenotype at the single-cell level. Results show that M1 and M2 macrophages have distinct metabolic profiles, with differential levels of fatty acyls, glycerophospholipids, and sterol lipids, which are important components of plasma membrane and involved in multiple biological processes. Furthermore, we could discern several putatively annotated molecules that contribute to inflammatory response of macrophages. The combination of random forest and live single-cell metabolomics provided an in-depth profile of the metabolome of primary human M1 and M2 macrophages at the single-cell level for the first time, which will pave the way for future studies targeting the differentiation of other immune cells., (© 2023 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals LLC.)
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- 2023
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38. Identification of circulating monocytes as producers of tuberculosis disease biomarker C1q.
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Niewold P, Dijkstra DJ, Cai Y, Goletti D, Palmieri F, van Meijgaarden KE, Verreck FAW, Akkerman OW, Hofland RW, Delemarre EM, Nierkens S, Verheul MK, Pollard AJ, van Dissel JT, Ottenhoff THM, Trouw LA, and Joosten SA
- Subjects
- Animals, Humans, Complement C1q metabolism, Primates, Biomarkers metabolism, Monocytes metabolism, Tuberculosis diagnosis, Tuberculosis metabolism
- Abstract
Tuberculosis (TB) is a prevalent disease causing an estimated 1.6 million deaths and 10.6 million new cases annually. Discriminating TB disease from differential diagnoses can be complex, particularly in the field. Increased levels of complement component C1q in serum have been identified as a specific and accessible biomarker for TB disease but the source of C1q in circulation has not been identified. Here, data and samples previously collected from human cohorts, a clinical trial and a non-human primate study were used to identify cells producing C1q in circulation. Cell subset frequencies were correlated with serum C1q levels and combined with single cell RNA sequencing and flow cytometry analyses. This identified monocytes as C1q producers in circulation, with a pronounced expression of C1q in classical and intermediate monocytes and variable expression in non-classical monocytes., (© 2023. The Author(s).)
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- 2023
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39. Extensive flow cytometric immunophenotyping of human PBMC incorporating detection of chemokine receptors, cytokines and tetramers.
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van Wolfswinkel M, van Meijgaarden KE, Ottenhoff THM, Niewold P, and Joosten SA
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- Humans, Flow Cytometry methods, Immunophenotyping, Receptors, Chemokine, Chemokines, Cytokines, Leukocytes, Mononuclear
- Abstract
Characterization of immune cells is essential to advance our understanding of immunology and flow cytometry is an important tool in this context. Addressing both cellular phenotype and antigen-specific functional responses of the same cells is valuable to achieve a more integrated understanding of immune cell behavior and maximizes information obtained from precious samples. Until recently, panel size was limiting, resulting in panels generally focused on either deep immunophenotyping or functional readouts. Ongoing developments in the field of (spectral) flow cytometry have made panels of 30
+ markers more accessible, opening up possibilities for advanced integrated analyses. Here, we optimized immune phenotyping by co-detection of markers covering chemokine receptors, cytokines and specific T cell/peptide tetramer interaction using a 32-color panel. Such panels enable integrated analysis of cellular phenotypes and markers assessing the quality of immune responses and will contribute to our understanding of the immune system., (© 2023 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.)- Published
- 2023
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40. Maternal HIV infection drives altered placental Mtb -specific antibody transfer.
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Nziza N, Jung W, Mendu M, Chen T, McNamara RP, Fortune SM, Franken KLMC, Ottenhoff THM, Bryson B, Ngonzi J, Bebell LM, and Alter G
- Abstract
Introduction: Placental transfer of maternal antibodies is essential for neonatal immunity over the first months of life. In the setting of maternal HIV infection, HIV-exposed uninfected (HEU) infants are at higher risk of developing severe infections, including active tuberculosis (TB). Given our emerging appreciation for the potential role of antibodies in the control of Mycobacterium tuberculosis ( Mtb ), the bacteria that causes TB, here we aimed to determine whether maternal HIV status altered the quality of Mtb -specific placental antibody transfer., Methods: Antigen-specific antibody systems serology was performed to comprehensively characterize the Mtb -specific humoral immune response in maternal and umbilical cord blood from HIV infected and uninfected pregnant people in Uganda., Results: Significant differences were noted in overall antibody profiles in HIV positive and negative maternal plasma, resulting in heterogeneous transfer of Mtb -specific antibodies. Altered antibody transfer in HIV infected dyads was associated with impaired binding to IgG Fc-receptors, which was directly linked to HIV viral loads and CD4 counts., Conclusions: These results highlight the importance of maternal HIV status on antibody transfer, providing clues related to alterations in transferred maternal immunity that may render HEU infants more vulnerable to TB than their HIV-unexposed peers., Competing Interests: GA is a V.P. at Moderna, a founder and equity holder of SeromYx Systems, and an employee and equity holder of Leyden Labs. GA’s interests were reviewed and are managed by MH and Partners HealthCare in accordance with their conflict-of-interest policies. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Nziza, Jung, Mendu, Chen, McNamara, Fortune, Franken, Ottenhoff, Bryson, Ngonzi, Bebell and Alter.)
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- 2023
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41. Ebola virus-like particles reprogram cellular metabolism.
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Tang H, Abouleila Y, Saris A, Shimizu Y, Ottenhoff THM, and Mashaghi A
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- Humans, Endothelial Cells, Signal Transduction, Amino Acids, Hemorrhagic Fever, Ebola, Ebolavirus physiology
- Abstract
Ebola virus can trigger a release of pro-inflammatory cytokines with subsequent vascular leakage and impairment of clotting finally leading to multiorgan failure and shock after entering and infecting patients. Ebola virus is known to directly target endothelial cells and macrophages, even without infecting them, through direct interactions with viral proteins. These interactions affect cellular mechanics and immune processes, which are tightly linked to other key cellular functions such as metabolism. However, research regarding metabolic activity of these cells upon viral exposure remains limited, hampering our understanding of its pathophysiology and progression. Therefore, in the present study, an untargeted cellular metabolomic approach was performed to investigate the metabolic alterations of primary human endothelial cells and M1 and M2 macrophages upon exposure to Ebola virus-like particles (VLP). The results show that Ebola VLP led to metabolic changes among endothelial, M1, and M2 cells. Differential metabolite abundance and perturbed signaling pathway analysis further identified specific metabolic features, mainly in fatty acid-, steroid-, and amino acid-related metabolism pathways for all the three cell types, in a host cell specific manner. Taken together, this work characterized for the first time the metabolic alternations of endothelial cells and two primary human macrophage subtypes after Ebola VLP exposure, and identified the potential metabolites and pathways differentially affected, highlighting the important role of those host cells in disease development and progression. KEY MESSAGES: • Ebola VLP can lead to metabolic alternations in endothelial cells and M1 and M2 macrophages. • Differential abundance of metabolites, mainly including fatty acids and sterol lipids, was observed after Ebola VLP exposure. • Multiple fatty acid-, steroid-, and amino acid-related metabolism pathways were observed perturbed., (© 2023. The Author(s).)
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- 2023
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42. Differences in Inflammatory Pathways Between Dutch South Asians vs Dutch Europids With Type 2 Diabetes.
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Straat ME, Martinez-Tellez B, van Eyk HJ, Bizino MB, van Veen S, Vianello E, Stienstra R, Ottenhoff THM, Lamb HJ, Smit JWA, Jazet IM, Rensen PCN, and Boon MR
- Subjects
- Aged, Female, Humans, Male, Middle Aged, Body Mass Index, Ethnicity, South Asian People, European People, Diabetes Mellitus, Type 2 ethnology, Diabetes Mellitus, Type 2 genetics
- Abstract
Context: South Asian individuals are more prone to develop type 2 diabetes (T2D) coinciding with earlier complications than Europids. While inflammation plays a central role in the development and progression of T2D, this factor is still underexplored in South Asians., Objective: This work aimed to study whether circulating messenger RNA (mRNA) transcripts of immune genes are different between South Asian compared with Europid patients with T2D., Methods: A secondary analysis was conducted of 2 randomized controlled trials of Dutch South Asian (n = 45; age: 55 ± 10 years, body mass index [BMI]: 29 ± 4 kg/m2) and Dutch Europid (n = 44; age: 60 ± 7 years, BMI: 32 ± 4 kg/m2) patients with T2D. Main outcome measures included mRNA transcripts of 182 immune genes (microfluidic quantitative polymerase chain reaction; Fluidigm Inc) in fasted whole-blood, ingenuity pathway analyses (Qiagen)., Results: South Asians, compared to Europids, had higher mRNA levels of B-cell markers (CD19, CD79A, CD79B, CR2, CXCR5, IGHD, MS4A1, PAX5; all fold change > 1.3, false discovery rate [FDR] < 0.008) and interferon (IFN)-signaling genes (CD274, GBP1, GBP2, GBP5, FCGR1A/B/CP, IFI16, IFIT3, IFITM1, IFITM3, TAP1; all FC > 1.2, FDR < 0.05). In South Asians, the IFN signaling pathway was the top canonical pathway (z score 2.6; P < .001) and this was accompanied by higher plasma IFN-γ levels (FC = 1.5, FDR = 0.01). Notably, the ethnic difference in gene expression was larger for women (20/182 [11%]) than men (2/182 [1%])., Conclusion: South Asian patients with T2D show a more activated IFN-signaling pathway compared to Europid patients with T2D, which is more pronounced in women than men. We speculate that a more activated IFN-signaling pathway may contribute to the more rapid progression of T2D in South Asian compared with Europid individuals., (© The Author(s) 2022. Published by Oxford University Press on behalf of the Endocrine Society.)
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- 2023
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43. Thermal-exchange HLA-E multimers reveal specificity in HLA-E and NKG2A/CD94 complex interactions.
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Ruibal P, Derksen I, van Wolfswinkel M, Voogd L, Franken KLMC, El Hebieshy AF, van Hall T, Schoufour TAW, Wijdeven RH, Ottenhoff THM, Scheeren FA, and Joosten SA
- Subjects
- Protein Binding, Peptides, Receptors, Antigen, T-Cell, NK Cell Lectin-Like Receptor Subfamily D chemistry, NK Cell Lectin-Like Receptor Subfamily D metabolism, NK Cell Lectin-Like Receptor Subfamily C, HLA-E Antigens, Killer Cells, Natural, Histocompatibility Antigens Class I metabolism
- Abstract
There is growing interest in HLA-E-restricted T-cell responses as a possible novel, highly conserved, vaccination targets in the context of infectious and malignant diseases. The developing field of HLA multimers for the detection and study of peptide-specific T cells has allowed the in-depth study of TCR repertoires and molecular requirements for efficient antigen presentation and T-cell activation. In this study, we developed a method for efficient peptide thermal exchange on HLA-E monomers and multimers allowing the high-throughput production of HLA-E multimers. We optimized the thermal-mediated peptide exchange, and flow cytometry staining conditions for the detection of TCR and NKG2A/CD94 receptors, showing that this novel approach can be used for high-throughput identification and analysis of HLA-E-binding peptides which could be involved in T-cell and NK cell-mediated immune responses. Importantly, our analysis of NKG2A/CD94 interaction in the presence of modified peptides led to new molecular insights governing the interaction of HLA-E with this receptor. In particular, our results reveal that interactions of HLA-E with NKG2A/CD94 and the TCR involve different residues. Altogether, we present a novel HLA-E multimer technology based on thermal-mediated peptide exchange allowing us to investigate the molecular requirements for HLA-E/peptide interaction with its receptors., (© 2022 The Authors. Immunology published by John Wiley & Sons Ltd.)
- Published
- 2023
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44. Repurposing Tamoxifen as Potential Host-Directed Therapeutic for Tuberculosis.
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Boland R, Heemskerk MT, Forn-Cuní G, Korbee CJ, Walburg KV, Esselink JJ, Carvalho Dos Santos C, de Waal AM, van der Hoeven DCM, van der Sar E, de Ries AS, Xie J, Spaink HP, van der Vaart M, Haks MC, Meijer AH, and Ottenhoff THM
- Subjects
- Animals, Humans, Zebrafish, Tamoxifen pharmacology, Tamoxifen therapeutic use, Drug Repositioning, Antitubercular Agents pharmacology, Antitubercular Agents therapeutic use, Tuberculosis microbiology, Mycobacterium tuberculosis genetics
- Abstract
The global burden of tuberculosis (TB) is aggravated by the continuously increasing emergence of drug resistance, highlighting the need for innovative therapeutic options. The concept of host-directed therapy (HDT) as adjunctive to classical antibacterial therapy with antibiotics represents a novel and promising approach for treating TB. Here, we have focused on repurposing the clinically used anticancer drug tamoxifen, which was identified as a molecule with strong host-directed activity against intracellular Mycobacterium tuberculosis ( Mtb ). Using a primary human macrophage Mtb infection model, we demonstrate the potential of tamoxifen against drug-sensitive as well as drug-resistant Mtb bacteria. The therapeutic effect of tamoxifen was confirmed in an in vivo TB model based on Mycobacterium marinum infection of zebrafish larvae. Tamoxifen had no direct antimicrobial effects at the concentrations used, confirming that tamoxifen acted as an HDT drug. Furthermore, we demonstrate that the antimycobacterial effect of tamoxifen is independent of its well-known target the estrogen receptor (ER) pathway, but instead acts by modulating autophagy, in particular the lysosomal pathway. Through RNA sequencing and microscopic colocalization studies, we show that tamoxifen stimulates lysosomal activation and increases the localization of mycobacteria in lysosomes both in vitro and in vivo , while inhibition of lysosomal activity during tamoxifen treatment partly restores mycobacterial survival. Thus, our work highlights the HDT potential of tamoxifen and proposes it as a repurposed molecule for the treatment of TB. IMPORTANCE Tuberculosis (TB) is the world's most lethal infectious disease caused by a bacterial pathogen, Mycobacterium tuberculosis. This pathogen evades the immune defenses of its host and grows intracellularly in immune cells, particularly inside macrophages. There is an urgent need for novel therapeutic strategies because treatment of TB patients is increasingly complicated by rising antibiotic resistance. In this study, we explored a breast cancer drug, tamoxifen, as a potential anti-TB drug. We show that tamoxifen acts as a so-called host-directed therapeutic, which means that it does not act directly on the bacteria but helps the host macrophages combat the infection more effectively. We confirmed the antimycobacterial effect of tamoxifen in a zebrafish model for TB and showed that it functions by promoting the delivery of mycobacteria to digestive organelles, the lysosomes. These results support the high potential of tamoxifen to be repurposed to fight antibiotic-resistant TB infections by host-directed therapy.
- Published
- 2023
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45. Intranasal multivalent adenoviral-vectored vaccine protects against replicating and dormant M.tb in conventional and humanized mice.
- Author
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Afkhami S, D'Agostino MR, Vaseghi-Shanjani M, Lepard M, Yang JX, Lai R, Choi MWY, Chacon A, Zganiacz A, Franken KLMC, Ertl HC, Ottenhoff THM, Jeyanathan M, Gillgrass A, and Xing Z
- Abstract
Viral-vectored vaccines are highly amenable for respiratory mucosal delivery as a means of inducing much-needed mucosal immunity at the point of pathogen entry. Unfortunately, current monovalent viral-vectored tuberculosis (TB) vaccine candidates have failed to demonstrate satisfactory clinical protective efficacy. As such, there is a need to develop next-generation viral-vectored TB vaccine strategies which incorporate both vaccine antigen design and delivery route. In this study, we have developed a trivalent chimpanzee adenoviral-vectored vaccine to provide protective immunity against pulmonary TB through targeting antigens linked to the three different growth phases (acute/chronic/dormancy) of Mycobacterium tuberculosis (M.tb) by expressing an acute replication-associated antigen, Ag85A, a chronically expressed virulence-associated antigen, TB10.4, and a dormancy/resuscitation-associated antigen, RpfB. Single-dose respiratory mucosal immunization with our trivalent vaccine induced robust, sustained tissue-resident multifunctional CD4
+ and CD8+ T-cell responses within the lung tissues and airways, which were further quantitatively and qualitatively improved following boosting of subcutaneously BCG-primed hosts. Prophylactic and therapeutic immunization with this multivalent trivalent vaccine in conventional BALB/c mice provided significant protection against not only actively replicating M.tb bacilli but also dormant, non-replicating persisters. Importantly, when used as a booster, it also provided marked protection in the highly susceptible C3HeB/FeJ mice, and a single respiratory mucosal inoculation was capable of significant protection in a humanized mouse model. Our findings indicate the great potential of this next-generation TB vaccine strategy and support its further clinical development for both prophylactic and therapeutic applications., (© 2023. The Author(s).)- Published
- 2023
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46. Neutrophil degranulation, NETosis and platelet degranulation pathway genes are co-induced in whole blood up to six months before tuberculosis diagnosis.
- Author
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Meier S, Seddon JA, Maasdorp E, Kleynhans L, du Plessis N, Loxton AG, Malherbe ST, Zak DE, Thompson E, Duffy FJ, Kaufmann SHE, Ottenhoff THM, Scriba TJ, Suliman S, Sutherland JS, Winter J, Kuivaniemi H, Walzl G, and Tromp G
- Subjects
- Humans, Neutrophils, Positron Emission Tomography Computed Tomography, Neutrophil Activation, Mycobacterium tuberculosis genetics, Tuberculosis, Lymph Node
- Abstract
Mycobacterium tuberculosis (M.tb) causes tuberculosis (TB) and remains one of the leading causes of mortality due to an infectious pathogen. Host immune responses have been implicated in driving the progression from infection to severe lung disease. We analyzed longitudinal RNA sequencing (RNAseq) data from the whole blood of 74 TB progressors whose samples were grouped into four six-month intervals preceding diagnosis (the GC6-74 study). We additionally analyzed RNAseq data from an independent cohort of 90 TB patients with positron emission tomography-computed tomography (PET-CT) scan results which were used to categorize them into groups with high and low levels of lung damage (the Catalysis TB Biomarker study). These groups were compared to non-TB controls to obtain a complete whole blood transcriptional profile for individuals spanning from early stages of M.tb infection to TB diagnosis. The results revealed a steady increase in the number of genes that were differentially expressed in progressors at time points closer to diagnosis with 278 genes at 13-18 months, 742 at 7-12 months and 5,131 detected 1-6 months before diagnosis and 9,205 detected in TB patients. A total of 2,144 differentially expressed genes were detected when comparing TB patients with high and low levels of lung damage. There was a large overlap in the genes upregulated in progressors 1-6 months before diagnosis (86%) with those in TB patients. A comprehensive pathway analysis revealed a potent activation of neutrophil and platelet mediated defenses including neutrophil and platelet degranulation, and NET formation at both time points. These pathways were also enriched in TB patients with high levels of lung damage compared to those with low. These findings suggest that neutrophils and platelets play a critical role in TB pathogenesis, and provide details of the timing of specific effector mechanisms that may contribute to TB lung pathology., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Meier et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2022
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47. Inflammatory responses in SARS-CoV-2 associated Multisystem Inflammatory Syndrome and Kawasaki Disease in children: An observational study.
- Author
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Biesbroek G, Kapitein B, Kuipers IM, Gruppen MP, van Stijn D, Peros TE, van Veenendaal M, Jansen MHA, van der Zee CW, van der Kuip M, von Asmuth EGJ, Mooij MG, den Boer MEJ, Landman GW, van Houten MA, Schonenberg-Meinema D, Tutu van Furth AM, Boele van Hensbroek M, Scherpbier H, van Meijgaarden KE, Ottenhoff THM, Joosten SA, Ketharanathan N, Blink M, Brackel CLH, Zaaijer HL, Hombrink P, van den Berg JM, Buddingh EP, and Kuijpers TW
- Subjects
- Child, Humans, Antibodies, Viral, Cytokines, Inflammation, Interleukin-6, SARS-CoV-2, Connective Tissue Diseases, COVID-19 complications, Mucocutaneous Lymph Node Syndrome complications
- Abstract
Multisystem Inflammatory Syndrome in Children (MIS-C) is a severe inflammatory disease in children related to SARS-CoV-2 with multisystem involvement including marked cardiac dysfunction and clinical symptoms that can resemble Kawasaki Disease (KD). We hypothesized that MIS-C and KD might have commonalities as well as unique inflammatory responses and studied these responses in both diseases. In total, fourteen children with MIS-C (n=8) and KD (n=6) were included in the period of March-June 2020. Clinical and routine blood parameters, cardiac follow-up, SARS-CoV-2-specific antibodies and CD4+ T-cell responses, and cytokine-profiles were determined in both groups. In contrast to KD patients, all MIS-C patients had positive Spike protein-specific CD3+CD4+ T-cell responses. MIS-C and KD patients displayed marked hyper-inflammation with high expression of serum cytokines, including the drug-targetable interleukin (IL)-6 and IFN-γ associated chemokines CXCL9, 10 and 11, which decreased at follow-up. No statistical differences were observed between groups. Clinical outcomes were all favourable without cardiac sequelae at 6 months follow-up. In conclusion, MIS-C and KD-patients both displayed cytokine-associated hyper-inflammation with several high levels of drug-targetable cytokines., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2022 Biesbroek et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)
- Published
- 2022
- Full Text
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48. Identification of HLA-E Binding Mycobacterium tuberculosis -Derived Epitopes through Improved Prediction Models.
- Author
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Ruibal P, Franken KLMC, van Meijgaarden KE, van Wolfswinkel M, Derksen I, Scheeren FA, Janssen GMC, van Veelen PA, Sarfas C, White AD, Sharpe SA, Palmieri F, Petrone L, Goletti D, Abeel T, Ottenhoff THM, and Joosten SA
- Subjects
- Antigens, Bacterial, CD8-Positive T-Lymphocytes, Epitopes, T-Lymphocyte, Histocompatibility Antigens Class I, Humans, Peptides, HLA-E Antigens, Mycobacterium tuberculosis, Tuberculosis
- Abstract
Tuberculosis (TB) remains one of the deadliest infectious diseases worldwide, posing great social and economic burden to affected countries. Novel vaccine approaches are needed to increase protective immunity against the causative agent Mycobacterium tuberculosis (Mtb) and to reduce the development of active TB disease in latently infected individuals. Donor-unrestricted T cell responses represent such novel potential vaccine targets. HLA-E-restricted T cell responses have been shown to play an important role in protection against TB and other infections, and recent studies have demonstrated that these cells can be primed in vitro. However, the identification of novel pathogen-derived HLA-E binding peptides presented by infected target cells has been limited by the lack of accurate prediction algorithms for HLA-E binding. In this study, we developed an improved HLA-E binding peptide prediction algorithm and implemented it to identify (to our knowledge) novel Mtb-derived peptides with capacity to induce CD8
+ T cell activation and that were recognized by specific HLA-E-restricted T cells in Mycobacterium -exposed humans. Altogether, we present a novel algorithm for the identification of pathogen- or self-derived HLA-E-presented peptides., (Copyright © 2022 by The American Association of Immunologists, Inc.)- Published
- 2022
- Full Text
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49. Evidence for the heterologous benefits of prior BCG vaccination on COVISHIELD™ vaccine-induced immune responses in SARS-CoV-2 seronegative young Indian adults.
- Author
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Rakshit S, Adiga V, Ahmed A, Parthiban C, Chetan Kumar N, Dwarkanath P, Shivalingaiah S, Rao S, D'Souza G, Dias M, Maguire TJA, Doores KJ, Zoodsma M, Geckin B, Dasgupta P, Babji S, van Meijgaarden KE, Joosten SA, Ottenhoff THM, Li Y, Netea MG, Stuart KD, De Rosa SC, McElrath MJ, and Vyakarnam A
- Subjects
- Humans, Young Adult, Adjuvants, Immunologic, Chromatin, Immunity, Interleukin-2, SARS-CoV-2, Tumor Necrosis Factor-alpha, Vaccination, BCG Vaccine, COVID-19 prevention & control, COVID-19 Vaccines immunology
- Abstract
This proof-of-concept study tested if prior BCG revaccination can qualitatively and quantitively enhance antibody and T-cell responses induced by Oxford/AstraZeneca ChAdOx1nCoV-19 or COVISHIELD™, an efficacious and the most widely distributed vaccine in India. We compared COVISHIELD™ induced longitudinal immune responses in 21 BCG re-vaccinees (BCG-RV) and 13 BCG-non-revaccinees (BCG-NRV), all of whom were BCG vaccinated at birth; latent tuberculosis negative and SARS-CoV-2 seronegative prior to COVISHIELD™ vaccination. Compared to BCG-NRV, BCG-RV displayed significantly higher and persistent spike-specific neutralizing (n) Ab titers and polyfunctional CD4+ and CD8+ T-cells for eight months post COVISHIELD™ booster, including distinct CD4+IFN-γ+ and CD4+IFN-γ- effector memory (EM) subsets co-expressing IL-2, TNF-α and activation induced markers (AIM) CD154/CD137 as well as CD8+IFN-γ+ EM,TEMRA (T cell EM expressing RA) subset combinations co-expressing TNF-α and AIM CD137/CD69. Additionally, elevated nAb and T-cell responses to the Delta mutant in BCG-RV highlighted greater immune response breadth. Mechanistically, these BCG adjuvant effects were associated with elevated markers of trained immunity, including higher IL-1β and TNF-α expression in CD14+HLA-DR+monocytes and changes in chromatin accessibility highlighting BCG-induced epigenetic changes. This study provides first in-depth analysis of both antibody and memory T-cell responses induced by COVISHIELD™ in SARS-CoV-2 seronegative young adults in India with strong evidence of a BCG-induced booster effect and therefore a rational basis to validate BCG, a low-cost and globally available vaccine, as an adjuvant to enhance heterologous adaptive immune responses to current and emerging COVID-19 vaccines., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Rakshit, Adiga, Ahmed, Parthiban, Chetan Kumar, Dwarkanath, Shivalingaiah, Rao, D’Souza, Dias, Maguire, Doores, Zoodsma, Geckin, Dasgupta, Babji, van Meijgaarden, Joosten, Ottenhoff, Li, Netea, Stuart, De Rosa, McElrath and Vyakarnam.)
- Published
- 2022
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50. Immune Determinants of Viral Clearance in Hospitalised COVID-19 Patients: Reduced Circulating Naïve CD4+ T Cell Counts Correspond with Delayed Viral Clearance.
- Author
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Zlei M, Sidorov IA, Joosten SA, Heemskerk MHM, Myeni SK, Pothast CR, de Brouwer CS, Boomaars-van der Zanden AL, van Meijgaarden KE, Morales ST, Wessels E, Janse JJ, Goeman JJ, Cobbaert CM, Kroes ACM, Cannegieter SC, Roestenberg M, Visser LG, Kikkert M, Feltkamp MCW, Arbous SM, Staal FJT, Ottenhoff THM, van Dongen JJM, Roukens AHE, de Vries JJC, In Collaboration With Beat-Covid, and In Collaboration With Lumc Covid
- Subjects
- Antibodies, Viral, CD4-Positive T-Lymphocytes, Critical Illness, Humans, SARS-CoV-2, COVID-19
- Abstract
Virus-specific cellular and humoral responses are major determinants for protection from critical illness after SARS-CoV-2 infection. However, the magnitude of the contribution of each of the components to viral clearance remains unclear. Here, we studied the timing of viral clearance in relation to 122 immune parameters in 102 hospitalised patients with moderate and severe COVID-19 in a longitudinal design. Delayed viral clearance was associated with more severe disease and was associated with higher levels of SARS-CoV-2-specific (neutralising) antibodies over time, increased numbers of neutrophils, monocytes, basophils, and a range of pro-inflammatory cyto-/chemokines illustrating ongoing, partially Th2 dominating, immune activation. In contrast, early viral clearance and less critical illness correlated with the peak of neutralising antibodies, higher levels of CD4 T cells, and in particular naïve CD4+ T cells, suggesting their role in early control of SARS-CoV-2 possibly by proving appropriate B cell help. Higher counts of naïve CD4+ T cells also correlated with lower levels of MIF, IL-9, and TNF-beta, suggesting an indirect role in averting prolonged virus-induced tissue damage. Collectively, our data show that naïve CD4+ T cell play a critical role in rapid viral T cell control, obviating aberrant antibody and cytokine profiles and disease deterioration. These data may help in guiding risk stratification for severe COVID-19.
- Published
- 2022
- Full Text
- View/download PDF
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