228 results on '"Ertl HC"'
Search Results
2. P16-09. Adenovirus 5 vector HIV vaccination does not affect mucosal homing markers on Ad5-specific CD4+ T-cells in humans
- Author
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Ertl HC, Casimiro DR, Robertson MN, Ratcliffe S, Cox K, Dubey S, Carnathan D, Hutnick N, and Betts MR
- Subjects
Immunologic diseases. Allergy ,RC581-607 - Published
- 2009
- Full Text
- View/download PDF
3. Passive immunization with polyclonal anti-SHIV IgG: partial protection or increased acquisition of heterologous tier 2 SHIV – depending on IgG dose
- Author
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Sholukh, AM, Sholukh, AM, Siddappa, NB, Shanmuganathan, V, Lakhashe, SK, Rasmussen, RA, Watkins, JD, Vyas, HK, Mukhtar, MM, Hemashettar, G, Thorat, S, Yoon, JK, Villinger, F, Novembre, FJ, Landucci, G, Forthal, DN, Ratcliffe, S, Robert-Guroff, M, Polonis, V, Montefiori, DC, Ertl, HC, Ruprecht, RM, Sholukh, AM, Sholukh, AM, Siddappa, NB, Shanmuganathan, V, Lakhashe, SK, Rasmussen, RA, Watkins, JD, Vyas, HK, Mukhtar, MM, Hemashettar, G, Thorat, S, Yoon, JK, Villinger, F, Novembre, FJ, Landucci, G, Forthal, DN, Ratcliffe, S, Robert-Guroff, M, Polonis, V, Montefiori, DC, Ertl, HC, and Ruprecht, RM
- Published
- 2012
4. P16-09. Adenovirus 5 vector HIV vaccination does not affect mucosal homing markers on Ad5-specific CD4+ T-cells in humans
- Author
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Hutnick, N, primary, Carnathan, D, additional, Dubey, S, additional, Cox, K, additional, Ratcliffe, S, additional, Robertson, MN, additional, Casimiro, DR, additional, Ertl, HC, additional, and Betts, MR, additional
- Published
- 2009
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- View/download PDF
5. Passive immunization with polyclonal anti-SHIV IgG: partial protection or increased acquisition of heterologous tier 2 SHIV – depending on IgG dose
- Author
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Siddappa, NB, Shanmuganathan, V, Hemashettar, G, Yoon, JK, Villinger, F, Novembre, FJ, Landucci, G, Forthal, DN, Ratcliffe, S, Robert-Guroff, M, Polonis, V, Montefiori, DC, Ertl, HC, Sholukh, Anton M, Lakhashe, Samir, Rasmussen, Robert Anthony, Watkins, Jennifer D, Vyas, Hemant Kumar, Mukhtar, Muhammad Mahmood, Thorat, Swati, and Ruprecht, Ruth Margrit
- Published
- 2012
- Full Text
- View/download PDF
6. A therapeutic HBV vaccine containing a checkpoint modifier enhances CD8+ T cell and antiviral responses.
- Author
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Hasanpourghadi M, Novikov M, Ambrose R, Chekaoui A, Newman D, Xiang Z, Luber AD, Currie SL, Zhou X, and Ertl HC
- Abstract
In patients who progress from acute hepatitis B virus (HBV) infection to a chronic HBV (CHB) infection, CD8+ T cells fail to eliminate the virus and become impaired. A functional cure of CHB likely requires new and highly functional CD8+ T cell responses different from those induced by the infection. Here we report preclinical immunogenicity and efficacy of an HBV therapeutic vaccine that includes herpes simplex virus (HSV) glycoprotein D (gD), a checkpoint modifier of early T cell activation, that enhances, broadens, and prolongs CD8+ T cell responses. We developed a therapeutic HBV vaccine based on a chimpanzee adenovirus serotype 6 (AdC6) vector, called AdC6-gDHBV2, that targets conserved and highly immunogenic regions of the viral polymerase (pol) and core antigens fused into HSV gD. The vaccine was tested with, and without gD, in mice for immunogenicity and in an adeno-associated virus (AAV)8-1.3HBV vector model for antiviral efficacy. The vaccine encoding the HBV antigens within gD stimulates potent and broad CD8+ T cell responses. In a surrogate model of HBV infection, a single intramuscular (i.m.) injection of AdC6-gDHBV2 achieved significant and sustained declines of circulating HBV DNA copies (cps) and HBV surface antigen (HBsAg); both inversely correlated with HBV specific CD8+ T cell frequencies in spleens and livers. AdC6-gDHBV2 is the first therapeutic vaccine to show significant reductions in levels of HBV genome copies and HBsAg when used alone, even when vaccination was delayed for months from infection.
- Published
- 2024
- Full Text
- View/download PDF
7. Heterologous chimpanzee adenovirus vector immunizations for SARS-CoV-2 spike and nucleocapsid protect hamsters against COVID-19.
- Author
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Hasanpourghadi M, Novikov M, Ambrose R, Chekaoui A, Newman D, Ding J, Giles-Davis W, Xiang Z, Zhou XY, Liu Q, Swagata K, and Ertl HC
- Subjects
- Animals, Cricetinae, Humans, SARS-CoV-2 genetics, COVID-19 Vaccines genetics, Pan troglodytes, Adenoviridae genetics, Nucleocapsid, Immunization, Antibodies, Viral, Antibodies, Neutralizing, COVID-19 prevention & control
- Abstract
Available COVID-19 vaccine only provide protection for a limited time due in part to the rapid emergence of viral variants with spike protein mutations, necessitating the generation of new vaccines to combat SARS-CoV-2. Two serologically distinct replication-defective chimpanzee-origin adenovirus (Ad) vectors (AdC) called AdC6 and AdC7 expressing early SARS-CoV-2 isolate spike (S) or nucleocapsid (N) proteins, the latter expressed as a fusion protein within herpes simplex virus glycoprotein D (gD), were tested individually or as a mixture in a hamster COVID-19 SARS-CoV-2 challenge model. The S protein expressing AdC (AdC-S) vectors induced antibodies including those with neutralizing activity that in part cross-reacted with viral variants. Hamsters vaccinated with the AdC-S vectors were protected against serious disease and showed accelerated recovery upon SARS-CoV-2 challenge. Protection was enhanced if AdC-S vectors were given together with the AdC vaccines that expressed the gD N fusion protein (AdC-gDN). In contrast hamsters that just received the AdC-gDN vaccines showed only marginal lessening of symptoms compared to control animals. These results indicate that immune response to the N protein that is less variable than the S protein may potentiate and prolong protection achieved by the currently used S protein based genetic COVID-19 vaccines., Competing Interests: Declaration of competing interest HCJE holds equity in Virion Therapeutics. She serves as a Consultant to several Gene Therapy companies., (Copyright © 2022 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)
- Published
- 2023
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8. 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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9. COVID-19 vaccination and HIV-1 acquisition.
- Author
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Logunov DY, Livermore DM, Ornelles DA, Bayer W, Marques E, Czerkinsky C, Dolzhikova IV, and Ertl HC
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- COVID-19 Vaccines, Humans, Vaccination, COVID-19 prevention & control, HIV Infections epidemiology, HIV Infections prevention & control, HIV Seropositivity, HIV-1
- Abstract
Competing Interests: DML, DAO, WB, EM, CC, and HCJE are former unpaid advisers to the Gamaleya Research Institute, Moscow, Russia. DML is on the advisory boards of and does ad hoc consultancy for Accelerate, Antabio, Centauri, Entasis, Meiji, Menarini, Mutabilis, Nordic, Paion, ParaPharm, Pfizer, QPEX, Shionogi, Summit, TAZ Corporation, VenatoRx, Wockhardt, Sumitovant and Zambon; and delivers paid lectures for bioMérieux, Beckman Coulter, Cardiome, GSK, Hikma, Merck/MSD, Menarini, Nordic, Perkin Elmer, and Shionogi. DML has relevant shareholdings or options with Dechra, GSK, Merck, and Pfizer, amounting to less than 10% of portfolio value. DML also has nominated holdings in Arecor, Avacta, Diaceutics, Evgen, Genedrive, Poolbeg, Renalytics AI, Synairgen, and Trellus (all with research and products pertinent to medicines or diagnostics) through Enterprise Investment Schemes but has no authority to trade these shares directly. HCJE has equity in Virion Therapeutics and serves as a consultant to Biogen, Takeda, Ring Therapeutics, Freeline, and Regenxbio. DYL and IVD report patents for a Sputnik V pharmaceutical agent and its method of use to prevent COVID-19. All other authors declare no competing interests.
- Published
- 2022
- Full Text
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10. Age-related changes in B cell metabolism.
- Author
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Kurupati RK, Haut LH, Schmader KE, and Ertl HC
- Subjects
- Adult, Aged, Aged, 80 and over, Energy Metabolism, Female, Humans, Male, Aging immunology, Antibody Formation, B-Lymphocytes metabolism
- Abstract
Antibody responses to vaccinations or infections decline upon aging. In this study we tested if metabolic changes in B cells may contribute to attenuation of responses to influenza vaccination in aged humans. Our data show that aging affects mitochondrial functions in B cells leading to increases in mitochondrial reactive oxygen species (MROS) and mitochondrial mass (MM) in some aged B cell subsets and decreases in expression levels of Sirtuin 1 (SIRT1), Forkhead box protein (FOX)O1 and carnitine palmitoyltransferase 1 (CPT-1). Seahorse analyses showed minor defects in glycolysis in the aged B cells after activation but a strong reduction in oxidative phosphorylation. The analyses of the transcriptome revealed further pronounced defects in one-carbon metabolism, a pathway that is essential for amino acid and nucleotide metabolism. Overall our data support the notion that the declining ability of aged B cells to increase their metabolism following activation contributes to the weakened antibody responses of the elderly.
- Published
- 2019
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11. Influenza Virus Vaccination Elicits Poorly Adapted B Cell Responses in Elderly Individuals.
- Author
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Henry C, Zheng NY, Huang M, Cabanov A, Rojas KT, Kaur K, Andrews SF, Palm AE, Chen YQ, Li Y, Hoskova K, Utset HA, Vieira MC, Wrammert J, Ahmed R, Holden-Wiltse J, Topham DJ, Treanor JJ, Ertl HC, Schmader KE, Cobey S, Krammer F, Hensley SE, Greenberg H, He XS, and Wilson PC
- Subjects
- Adult, Aged, Aged, 80 and over, Epitopes genetics, Healthy Volunteers, Humans, Influenza Vaccines administration & dosage, Middle Aged, Mutation, Orthomyxoviridae genetics, Young Adult, B-Lymphocytes immunology, Epitopes immunology, Immunity, Humoral, Influenza Vaccines immunology, Orthomyxoviridae immunology
- Abstract
Influenza is a leading cause of death in the elderly, and the vaccine protects only a fraction of this population. A key aspect of antibody-mediated anti-influenza virus immunity is adaptation to antigenically distinct epitopes on emerging strains. We examined factors contributing to reduced influenza vaccine efficacy in the elderly and uncovered a dramatic reduction in the accumulation of de novo immunoglobulin gene somatic mutations upon vaccination. This reduction is associated with a significant decrease in the capacity of antibodies to target the viral glycoprotein, hemagglutinin (HA), and critical protective epitopes surrounding the HA receptor-binding domain. Immune escape by antigenic drift, in which viruses generate mutations in key antigenic epitopes, becomes highly exaggerated. Because of this reduced adaptability, most B cells activated in the elderly cohort target highly conserved but less potent epitopes. Given these findings, vaccines driving immunoglobulin gene somatic hypermutation should be a priority to protect elderly individuals., (Copyright © 2019 Elsevier Inc. All rights reserved.)
- Published
- 2019
- Full Text
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12. A simian-adenovirus-vectored rabies vaccine suitable for thermostabilisation and clinical development for low-cost single-dose pre-exposure prophylaxis.
- Author
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Wang C, Dulal P, Zhou X, Xiang Z, Goharriz H, Banyard A, Green N, Brunner L, Ventura R, Collin N, Draper SJ, Hill AVS, Ashfield R, Fooks AR, Ertl HC, and Douglas AD
- Subjects
- Adjuvants, Immunologic administration & dosage, Animals, Antibodies, Neutralizing blood, Antibodies, Viral blood, Costs and Cost Analysis, Drug Stability, Female, Genetic Vectors, Immunization Schedule, Mice, Rabies Vaccines administration & dosage, Rabies Vaccines economics, Rabies Vaccines genetics, Vaccines, Synthetic administration & dosage, Vaccines, Synthetic economics, Vaccines, Synthetic genetics, Vaccines, Synthetic immunology, Adenoviruses, Simian genetics, Drug Carriers, Pre-Exposure Prophylaxis methods, Rabies prevention & control, Rabies Vaccines immunology
- Abstract
Background: Estimates of current global rabies mortality range from 26,000 to 59,000 deaths per annum. Although pre-exposure prophylaxis using inactivated rabies virus vaccines (IRVs) is effective, it requires two to three doses and is regarded as being too expensive and impractical for inclusion in routine childhood immunization programmes., Methodology/ Principal Findings: Here we report the development of a simian-adenovirus-vectored rabies vaccine intended to enable cost-effective population-wide pre-exposure prophylaxis against rabies. ChAdOx2 RabG uses the chimpanzee adenovirus serotype 68 (AdC68) backbone previously shown to achieve pre-exposure protection against rabies in non-human primates. ChAdOx2 differs from AdC68 in that it contains the human adenovirus serotype 5 (AdHu5) E4 orf6/7 region in place of the AdC68 equivalents, enhancing ease of manufacturing in cell lines which provide AdHu5 E1 proteins in trans. We show that immunogenicity of ChAdOx2 RabG in mice is comparable to that of AdC68 RabG and other adenovirus serotypes expressing rabies virus glycoprotein. High titers of rabies virus neutralizing antibody (VNA) are elicited after a single dose. The relationship between levels of VNA activity and rabies virus glycoprotein monomer-binding antibody differs after immunization with adenovirus-vectored vaccines and IRV vaccines, suggesting routes to further enhancement of the efficacy of the adenovirus-vectored candidates. We also demonstrate that ChAdOx2 RabG can be thermostabilised using a low-cost method suitable for clinical bio-manufacture and ambient-temperature distribution in tropical climates. Finally, we show that a dose-sparing effect can be achieved by formulating ChAdOx2 RabG with a simple chemical adjuvant. This approach could lower the cost of ChAdOx2 RabG and other adenovirus-vectored vaccines., Conclusions/ Significance: ChAdOx2 RabG may prove to be a useful tool to reduce the human rabies death toll. We have secured funding for Good Manufacturing Practice- compliant bio-manufacture and Phase I clinical trial of this candidate., Competing Interests: AVSH and ADD are named inventors on a patent application relating to the development of the ChAdOx2 vector. AVSH and SJD are named inventors on a patent relating to the use of the intron-containing promoter used in ChAdOx2 RabG. The University of Oxford and the Wistar Institute have entered into a partnership to share any future revenue from development of the ChAdOx2 RabG vaccine.
- Published
- 2018
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13. Coaching tumor-infiltrating CD8+ T cells to eat right.
- Author
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Kurupati R and Ertl HC
- Abstract
Competing Interests: CONFLICTS OF INTEREST The authors declare no potential conflicts of interest.
- Published
- 2018
- Full Text
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14. Poor Immunogenicity, Not Vaccine Strain Egg Adaptation, May Explain the Low H3N2 Influenza Vaccine Effectiveness in 2012-2013.
- Author
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Cobey S, Gouma S, Parkhouse K, Chambers BS, Ertl HC, Schmader KE, Halpin RA, Lin X, Stockwell TB, Das SR, Landon E, Tesic V, Youngster I, Pinsky BA, Wentworth DE, Hensley SE, and Grad YH
- Subjects
- Adaptation, Physiological, Adult, Aged, Aged, 80 and over, Animals, Antigens, Viral immunology, Cohort Studies, Cross Reactions, Eggs virology, Ferrets, Genome, Viral, High-Throughput Nucleotide Sequencing, Humans, Influenza A Virus, H3N2 Subtype isolation & purification, Influenza Vaccines therapeutic use, Influenza, Human prevention & control, Mutation, Phylogeny, Seasons, Immunogenicity, Vaccine, Influenza A Virus, H3N2 Subtype genetics, Influenza Vaccines immunology, Influenza, Human epidemiology
- Abstract
Background: Influenza vaccination aims to prevent infection by influenza virus and reduce associated morbidity and mortality; however, vaccine effectiveness (VE) can be modest, especially for subtype A(H3N2). Low VE has been attributed to mismatches between the vaccine and circulating influenza strains and to the vaccine's elicitation of protective immunity in only a subset of the population. The low H3N2 VE in the 2012-2013 season was attributed to egg-adaptive mutations that created antigenic mismatch between the actual vaccine strain (IVR-165) and both the intended vaccine strain (A/Victoria/361/2011) and the predominant circulating strains (clades 3C.2 and 3C.3)., Methods: We investigated the basis of low VE in 2012-2013 by determining whether vaccinated and unvaccinated individuals were infected by different viral strains and by assessing the serologic responses to IVR-165, A/Victoria/361/2011, and 3C.2 and 3C.3 strains in an adult cohort before and after vaccination., Results: We found no significant genetic differences between the strains that infected vaccinated and unvaccinated individuals. Vaccination increased titers to A/Victoria/361/2011 and 3C.2 and 3C.3 representative strains as much as to IVR-165. These results are consistent with the hypothesis that vaccination boosted cross-reactive immune responses instead of specific responses against unique vaccine epitopes. Only approximately one-third of the cohort achieved a ≥4-fold increase in titer., Conclusions: In contrast to analyses based on ferret studies, low H3N2 VE in 2012-2013 in adults does not appear to be due to egg adaptation of the vaccine strain. Instead, low VE might have been caused by low vaccine immunogenicity in a subset of the population.
- Published
- 2018
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15. Coaching tumor-infiltrating CD8 + T cells to eat right.
- Author
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Kurupati R and Ertl HC
- Abstract
Competing Interests: CONFLICTS OF INTEREST The authors declare no conflicts of interest.
- Published
- 2018
- Full Text
- View/download PDF
16. Spray dried human and chimpanzee adenoviral-vectored vaccines are thermally stable and immunogenic in vivo.
- Author
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Afkhami S, LeClair DA, Haddadi S, Lai R, Toniolo SP, Ertl HC, Cranston ED, Thompson MR, and Xing Z
- Subjects
- Adenoviruses, Simian, Animals, Desiccation, Drug Storage, Humans, Mannitol, Pan troglodytes, Powders, Temperature, Trehalose, Tuberculosis Vaccines administration & dosage, Tuberculosis Vaccines chemistry, Vaccines, Synthetic chemistry, Adenoviridae genetics, Immunogenicity, Vaccine, Tuberculosis Vaccines immunology, Vaccine Potency, Vaccines, Synthetic immunology
- Abstract
Cold chain-free vaccine technologies are needed to ensure effective vaccine delivery and coverage, particularly in resource-poor countries. However, the immunogenicity and thermostability of spray dried live viral vector-based vaccines such as recombinant adenoviral-vectored vaccines remain to be investigated. To address this issue, we have spray dried human adenoviral (AdHu5)- and chimpanzee adenoviral (AdCh68)-vectored tuberculosis vaccines in a mannitol and dextran matrix. Spray dried powders containing these two vaccines display the morphologic and chemical properties desired for long-term thermostability and vaccination. Upon reconstitution, they effectively transfected the cells in vitro with relatively small losses in viral infectivity related to the spray drying process. Following in vivo vaccination, AdHu5- and AdCh68-vectored vaccines were as immunogenic as the conventional fresh, cryopreserved liquid vaccine samples. Of importance, even after cold chain-free storage, at ambient temperatures and relatively low humidity for 30 and 90days, the vaccines retained their in vivo immunogenicity, while the liquid vaccine samples stored under the same conditions lost their immune-activating capability almost entirely. Our results support further development of our spray drying technologies for generating thermally stable adenoviral-vectored and other viral-vectored vaccines., (Copyright © 2017 Elsevier Ltd. All rights reserved.)
- Published
- 2017
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17. Viral vectors as vaccine carriers.
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Ertl HC
- Subjects
- Adaptive Immunity, Animals, B-Lymphocytes immunology, Clinical Trials as Topic, Humans, T-Lymphocytes immunology, Adenoviridae genetics, Dependovirus genetics, Drug Carriers, Genetic Vectors, Vaccines, Synthetic genetics, Vaccines, Synthetic immunology
- Abstract
This chapter reviews the performance of viral vectors based on adenoviruses or adeno-associated virus as vaccine carriers for infectious diseases. Replication-defective adenovirus vectors based on multiple human or non-human serotypes have consistently induced potent transgene product-specific B and T cell responses and are increasingly being explored in human clinical trials. The immunogenicity of most vectors based on adeno-associated virus vectors has been poor with the exception of a recently described hybrid vector from rhesus macaques that due to its ability to induce potent responses in mice warrant further investigation., (Copyright © 2016. Published by Elsevier B.V.)
- Published
- 2016
- Full Text
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18. Age-related changes in the gene expression profile of antigen-specific mouse CD8+ T cells can be partially reversed by blockade of the BTLA/CD160 pathways during vaccination.
- Author
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Dawany N, Parzych EM, Showe LC, and Ertl HC
- Subjects
- Animals, Antigens, CD genetics, Female, GPI-Linked Proteins genetics, GPI-Linked Proteins metabolism, Influenza A virus, Influenza Vaccines immunology, Mice, Mice, Inbred C57BL, Orthomyxoviridae Infections prevention & control, Receptors, Immunologic genetics, Vaccination, Aging physiology, Antigens, CD metabolism, CD8-Positive T-Lymphocytes physiology, Gene Expression Regulation immunology, Receptors, Immunologic metabolism, Transcriptome physiology
- Abstract
We analyzed gene expression profiles of young and aged mouse CD8
+ T cells specific for the nucleoprotein (NP) of influenza A/PR8/34 virus. CD8+ T cells were stimulated either by the NP antigen expressed in its native form or fused into the herpes virus (HSV)-1 glycoprotein D (gD) protein, which blocks signaling through the immunoinhibitory B and T lymphocyte attenuator (BTLA) and CD160 pathways. We show that NP-specific CD8+ T cells from aged mice exhibit numerous differences in gene expression compared to NP-specific CD8+ T cells from young mice, including a significant reduction of expression in genes involved in T cell receptor (TcR) and CD28 signaling. We also show that these changes can be reversed in a sub-population (~50%) of the aged mice by a BTLA/CD160 checkpoint blockade. These results suggest that BTLA/CD160 checkpoint blockade has potential value as a vaccine additive to induce better CD8+ T cell responses in the aged., Competing Interests: Conflicts of Interest: None declared.- Published
- 2016
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19. Antibody responses to prime-boost vaccination with an HIV-1 gp145 envelope protein and chimpanzee adenovirus vectors expressing HIV-1 gp140.
- Author
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Emmer KL, Wieczorek L, Tuyishime S, Molnar S, Polonis VR, and Ertl HC
- Subjects
- AIDS Vaccines administration & dosage, AIDS Vaccines genetics, Adjuvants, Immunologic administration & dosage, Alum Compounds administration & dosage, Animals, Antibodies, Neutralizing blood, Antibody Affinity, Enzyme-Linked Immunosorbent Assay, Female, Immunization Schedule, Mice, Mice, Inbred ICR, Neutralization Tests, Recombinant Proteins genetics, Treatment Outcome, Vaccines, Synthetic administration & dosage, Vaccines, Synthetic genetics, Vaccines, Synthetic immunology, env Gene Products, Human Immunodeficiency Virus genetics, AIDS Vaccines immunology, Adenoviridae genetics, Antibody Formation, Drug Carriers administration & dosage, HIV Antibodies blood, Recombinant Proteins immunology, env Gene Products, Human Immunodeficiency Virus immunology
- Abstract
Objectives: Over 2 million individuals are infected with HIV type 1 (HIV-1) each year, yet an effective vaccine remains elusive. The most successful HIV-1 vaccine to date demonstrated 31% efficacy. Immune correlate analyses associated HIV-1 envelope (Env)-specific antibodies with protection, thus providing a path toward a more effective vaccine. We sought to test the antibody response from novel prime-boost vaccination with a chimpanzee-derived adenovirus (AdC) vector expressing a subtype C Env glycoprotein (gp)140 combined with either a serologically distinct AdC vector expressing gp140 of a different subtype C isolate or an alum-adjuvanted, partially trimeric gp145 from yet another subtype C isolate., Design: Three different prime-boost regimens were tested in mice: AdC prime-protein boost, protein prime-AdC boost, and AdC prime-AdC boost. Each regimen was tested at two different doses of AdC vector in a total of six experimental groups., Methods: Sera were collected at various time points and evaluated by ELISA for Env-specific antibody binding, isotype, and avidity. Antibody functionality was assessed by pseudovirus neutralization assay., Results: Priming with AdC followed by a protein boost or sequential immunizations with two AdC vectors induced HIV-1 Env-specific binding antibodies, including those to the variable region 2, whereas priming with protein followed by an AdC boost was relatively ineffective. Antibodies that cross-neutralized tier 1 HIV-1 from different subtypes were elicited with vaccine regimens that included immunizations with protein., Conclusion: Our study warrants further investigation of AdC vector and gp145 protein prime-boost vaccines and their ability to protect against acquisition in animal challenge studies., Competing Interests: Disclaimers/Conflicts of Interest Statement: The Ertl and Polonis Labs declare no conflicts of interest. The views expressed are those of the authors and should not be construed to represent the positions of the U.S. Army or the Department of Defense.
- Published
- 2016
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20. A Partial E3 Deletion in Replication-Defective Adenoviral Vectors Allows for Stable Expression of Potentially Toxic Transgene Products.
- Author
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Haut LH, Gill AL, Kurupati RK, Bian A, Li Y, Giles-Davis W, Xiang Z, Zhou XY, and Ertl HC
- Subjects
- Animals, Genetic Vectors therapeutic use, HEK293 Cells, HIV-1 genetics, Humans, Mice, Serogroup, Transgenes genetics, Virus Replication genetics, env Gene Products, Human Immunodeficiency Virus therapeutic use, Adenoviridae genetics, Genetic Therapy, Genetic Vectors genetics, env Gene Products, Human Immunodeficiency Virus biosynthesis
- Abstract
Adenovirus (Ad) is used extensively for construction of viral vectors, most commonly with deletion in its E1 and/or E3 genomic regions. Previously, our attempts to insert envelope proteins (Env) of HIV-1 into such vectors based on chimpanzee-derived Ad (AdC) viruses were thwarted. Here, we describe that genetic instability of an E1- and E3-deleted AdC vector of serotype C6 expressing Env of HIV-1 can be overcome by reinsertion of E3 sequences with anti-apoptotic activities. This partial E3 deletion presumably delays premature death of HEK-293 packaging cell lines due to Env-induced cell apoptosis. The same partial E3 deletion also allows for the generation of stable glycoprotein 140 (gp140)- and gp160-expressing Ad vectors based on AdC7, a distinct AdC serotype. Env-expressing AdC vectors containing the partial E3 deletion are genetically stable upon serial cell culture passaging, produce yields comparable to those of other AdC vectors, and induce transgene product-specific antibody responses in mice. A partial E3 deletion thereby allows expansion of the repertoire of transgenes that can be expressed by Ad vectors., Competing Interests: Author Disclosure No competing financial interests exist.
- Published
- 2016
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21. Aging: T cell metabolism within tumors.
- Author
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Zhang Y and Ertl HC
- Subjects
- Humans, Lymphocytes, Tumor-Infiltrating immunology, Neoplasms metabolism, T-Lymphocytes immunology, Aging immunology, Immunosenescence immunology, Lymphocytes, Tumor-Infiltrating metabolism, Neoplasms immunology, T-Lymphocytes metabolism
- Published
- 2016
- Full Text
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22. Superior In vivo Transduction of Human Hepatocytes Using Engineered AAV3 Capsid.
- Author
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Vercauteren K, Hoffman BE, Zolotukhin I, Keeler GD, Xiao JW, Basner-Tschakarjan E, High KA, Ertl HC, Rice CM, Srivastava A, de Jong YP, and Herzog RW
- Subjects
- Animals, Cells, Cultured, Dependovirus metabolism, Hepatocytes metabolism, Humans, Mice, Organ Specificity, Protein Engineering, Transduction, Genetic, Capsid Proteins genetics, Dependovirus genetics, Genetic Vectors administration & dosage, Hepatocytes ultrastructure
- Abstract
Adeno-associated viral (AAV) vectors are currently being tested in multiple clinical trials for liver-directed gene transfer to treat the bleeding disorders hemophilia A and B and metabolic disorders. The optimal viral capsid for transduction of human hepatocytes has been under active investigation, but results across various models are inconsistent. We tested in vivo transduction in "humanized" mice. Methods to quantitate percent AAV transduced human and murine hepatocytes in chimeric livers were optimized using flow cytometry and confocal microscopy with image analysis. Distinct transduction efficiencies were noted following peripheral vein administration of a self-complementary vector expressing a gfp reporter gene. An engineered AAV3 capsid with two amino acid changes, S663V+T492V (AAV3-ST), showed best efficiency for human hepatocytes (~3-times, ~8-times, and ~80-times higher than for AAV9, AAV8, and AAV5, respectively). AAV5, 8, and 9 were more efficient in transducing murine than human hepatocytes. AAV8 yielded the highest transduction rate of murine hepatocytes, which was 19-times higher than that for human hepatocytes. In summary, our data show substantial differences among AAV serotypes in transduction of human and mouse hepatocytes, are the first to report on AAV5 in humanized mice, and support the use of AAV3-based vectors for human liver gene transfer.
- Published
- 2016
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23. Depletion of FAP+ cells reduces immunosuppressive cells and improves metabolism and functions CD8+T cells within tumors.
- Author
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Zhang Y and Ertl HC
- Subjects
- Animals, Dendritic Cells immunology, Endopeptidases, Female, Genetic Vectors, Humans, Melanoma, Experimental metabolism, Melanoma, Experimental therapy, Mice, Mice, Inbred C57BL, Mice, Transgenic, Stromal Cells immunology, Tumor Cells, Cultured, CD8-Positive T-Lymphocytes immunology, Cancer Vaccines administration & dosage, Gelatinases metabolism, Immunosuppression Therapy, Melanoma, Experimental immunology, Membrane Proteins metabolism, Serine Endopeptidases metabolism, Tumor Microenvironment immunology
- Abstract
The tumor stroma, which is essential to support growth and metastasis of malignant cells, provides targets for active immunotherapy of cancer. Previous studies have shown that depleting fibroblast activation protein (FAP)-expressing stromal cells reduces tumor progression and concomitantly increases tumor antigen (TA)-specific T cell responses. However the underlying pathways remain ill defined. Here we identify that immunosuppressive cells (ISCs) from tumor-bearing mice impose metabolic stress on CD8+T cells, which is associated with increased expression of the co-inhibitor PD-1. In two mouse melanoma models, depleting FAP+ stroma cells from the tumor microenvironment (TME) upon vaccination with an adenoviral-vector reduces frequencies and functions of ISCs. This is associated with changes in the cytokine/chemokine milieu in the TME and decreased activity of STAT6 signaling within ISCs. Decreases in ISCs upon FAP+stromal cell depletion is associated with reduced metabolic stress of vaccine-induced tumor infiltrating CD8+T cells and their delayed progression towards functional exhaustion, resulting in prolonged survival of tumor-bearing mice., Competing Interests: The authors declare that they have no conflict of interest.
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- 2016
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24. Age-related changes in the transcriptome of antibody-secreting cells.
- Author
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Kannan S, Dawany N, Kurupati R, Showe LC, and Ertl HC
- Subjects
- Animals, Antibody-Producing Cells cytology, B-Lymphocytes cytology, Cells, Cultured, Endoplasmic Reticulum Stress genetics, Mice, Oxidative Phosphorylation, Oxidative Stress genetics, Reactive Oxygen Species metabolism, Signal Transduction, Aging physiology, Antibody-Producing Cells metabolism, B-Lymphocytes metabolism, Transcriptome
- Abstract
We analyzed age-related defects in B cell populations from young and aged mice. Microarray analysis of bone marrow resident antibody secreting cells (ASCs) showed significant changes upon aging, affecting multiple genes, pathways and functions including those that play a role in immune regulation, humoral immune responses, chromatin structure and assembly, cell metabolism and the endoplasmic reticulum (ER) stress response. Further analysis showed upon aging defects in energy production through glucose catabolism with reduced oxidative phosphorylation. In addition aged B cells had increased levels of reactive oxygen-species (ROS), which was linked to enhanced expression of the co-inhibitor programmed cell death (PD)-1.
- Published
- 2016
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25. Starved and Asphyxiated: How Can CD8(+) T Cells within a Tumor Microenvironment Prevent Tumor Progression.
- Author
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Zhang Y and Ertl HC
- Abstract
Although cancer immunotherapy has achieved significant breakthroughs in recent years, its overall efficacy remains limited in the majority of patients. One major barrier is exhaustion of tumor antigen-specific CD8(+) tumor-infiltrating lymphocytes (TILs), which conventionally has been attributed to persistent stimulation with antigen within the tumor microenvironment (TME). A series of recent studies have highlighted that the TME poses significant metabolic challenges to TILs, which may contribute to their functional exhaustion. Hypoxia increases the expression of coinhibitors on activated CD8(+) T cells, which in general reduces the T cells' effector functions. It also impairs the cells' ability to gain energy through oxidative phosphorylation. Glucose limitation increases the expression of programed cell death protein-1 and reduces functions of activated CD8(+) T cells. A combination of hypoxia and hypoglycemia, as is common in solid tumors, places CD8(+) TILs at dual metabolic jeopardy by affecting both major pathways of energy production. Recently, a number of studies addressed the effects of metabolic stress on modulating CD8(+) T cell metabolism, differentiation, and functions. Here, we discuss recent findings on how different types of metabolic stress within the TME shape the tumor-killing capacity of CD8(+) T cells. We propose that manipulating the metabolism of TILs to more efficiently utilize nutrients, especially during intermittent periods of hypoxia could maximize their performance, prolong their survival and improve the efficacy of active cancer immunotherapy.
- Published
- 2016
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26. A shortened interval between vaccinations with the trivalent inactivated influenza vaccine increases responsiveness in the aged.
- Author
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Kannan S, Kossenkov A, Kurupati RK, Xiang JZ, Doyle SA, Schmader KE, Schowe L, and Ertl HC
- Subjects
- Aged, Aged, 80 and over, Female, Gene Expression Regulation immunology, Humans, Immunization Schedule, Influenza A Virus, H1N1 Subtype immunology, Influenza A Virus, H3N2 Subtype immunology, Influenza Vaccines administration & dosage, Influenza, Human blood, Male, Antibodies, Viral blood, Influenza Vaccines immunology, Influenza, Human prevention & control
- Abstract
We tested antibody responses to the trivalent inactivated influenza vaccine (TIV) in 34 aged individuals (>65 yrs) during the 2012/13 vaccination seasons. Nearly all had been vaccinated the previous year although the time interval between the two vaccine doses differed. One subgroup was re-vaccinated in 2012/13 within 6-9 months of their 2011/12 vaccination, the other received the two doses of vaccine in the typical ~12 month interval. Unexpectedly the sub-cohort with early revaccination exhibited significantly increased response rates and antibody titers to TIV compared to their normally re-vaccinated aged counter parts. Microarray analyses of gene expression in whole blood RNA taken at the day of the 2012/13 re-vaccination revealed statistically significant differences in expression of 754 genes between the individuals with early re-vaccination compared to subjects vaccinated in a normal 12 month interval. These observations suggest that TIV has long-lasting effects on the immune system affecting B cell responses as well as the transcriptome of peripheral blood mononuclear cells and this residual effect may augment vaccination response in patients where the effect of the previous vaccination has not yet diminished.
- Published
- 2015
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27. Novel chimpanzee adenovirus-vectored respiratory mucosal tuberculosis vaccine: overcoming local anti-human adenovirus immunity for potent TB protection.
- Author
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Jeyanathan M, Thanthrige-Don N, Afkhami S, Lai R, Damjanovic D, Zganiacz A, Feng X, Yao XD, Rosenthal KL, Medina MF, Gauldie J, Ertl HC, and Xing Z
- Subjects
- Adenoviridae, Animals, Antibodies, Bacterial blood, Antibodies, Bacterial immunology, Antibodies, Neutralizing blood, Antibodies, Neutralizing immunology, Disease Models, Animal, Enzyme-Linked Immunosorbent Assay, Flow Cytometry, Genetic Vectors, Humans, Mice, Mice, Inbred BALB C, Pan troglodytes, Tuberculosis Vaccines immunology, Tuberculosis, Pulmonary immunology
- Abstract
Pulmonary tuberculosis (TB) remains to be a major global health problem despite many decades of parenteral use of Bacillus Calmette-Guérin (BCG) vaccine. Developing safe and effective respiratory mucosal TB vaccines represents a unique challenge. Over the past decade or so, the human serotype 5 adenovirus (AdHu5)-based TB vaccine has emerged as one of the most promising candidates based on a plethora of preclinical and early clinical studies. However, anti-AdHu5 immunity widely present in the lung of humans poses a serious gap and limitation to its real-world applications. In this study we have developed a novel chimpanzee adenovirus 68 (AdCh68)-vectored TB vaccine amenable to the respiratory route of vaccination. We have evaluated AdCh68-based TB vaccine for its safety, T-cell immunogenicity, and protective efficacy in relevant animal models of human pulmonary TB with or without parenteral BCG priming. We have also compared AdCh68-based TB vaccine with its AdHu5 counterpart in both naive animals and those with preexisting anti-AdHu5 immunity in the lung. We provide compelling evidence that AdCh68-based TB vaccine is not only safe when delivered to the respiratory tract but, importantly, is also superior to its AdHu5 counterpart in induction of T-cell responses and immune protection, and limiting lung immunopathology in the presence of preexisting anti-AdHu5 immunity in the lung. Our findings thus suggest AdCh68-based TB vaccine to be an ideal candidate for respiratory mucosal immunization, endorsing its further clinical development in humans.
- Published
- 2015
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28. BTLA expression declines on B cells of the aged and is associated with low responsiveness to the trivalent influenza vaccine.
- Author
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Kannan S, Kurupati RK, Doyle SA, Freeman GJ, Schmader KE, and Ertl HC
- Subjects
- Adult, Age Factors, Aged, Aged, 80 and over, Antibodies, Neutralizing blood, Antibodies, Viral blood, B-Lymphocytes immunology, B-Lymphocytes metabolism, Biomarkers blood, Down-Regulation, Female, Flow Cytometry, Humans, Immunoglobulin G blood, Immunologic Memory, Immunophenotyping methods, Male, Receptors, Immunologic immunology, Time Factors, Vaccination, Aging immunology, Aging metabolism, B-Lymphocytes drug effects, Influenza A Virus, H1N1 Subtype immunology, Influenza Vaccines administration & dosage, Receptors, Immunologic metabolism
- Abstract
Virus-neutralizing antibody and B cell responses to influenza A viruses were measured in 35 aged and 28 middle-aged individuals following vaccination with the 2012 and 2013 trivalent inactivated influenza vaccines. Antibody responses to the vaccine strains were lower in the aged. An analysis of B cell subsets by flow cytometry with stains for immunoregulators showed that B cells of multiple subsets from the aged as compared to younger human subjects showed differences in the expression of the co-inhibitor B and T lymphocyte attenuator (BTLA). Expression of BTLA inversely correlated with age and appears to be linked to shifting the nature of the response from IgM to IgG. High BTLA expression on mature B cells was linked to higher IgG responses to the H1N1 virus. Finally, high BTLA expression on isotype switched memory B cells was linked to better preservation of virus neutralizing antibody titers and improved recall responses to vaccination given the following year.
- Published
- 2015
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29. Repeated Low-Dose Influenza Virus Infection Causes Severe Disease in Mice: a Model for Vaccine Evaluation.
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Song Y, Wang X, Zhang H, Tang X, Li M, Yao J, Jin X, Ertl HC, and Zhou D
- Subjects
- Animals, Antibodies, Viral immunology, Chick Embryo, Female, Humans, Influenza A Virus, H1N1 Subtype genetics, Influenza A Virus, H1N1 Subtype immunology, Influenza Vaccines administration & dosage, Influenza, Human immunology, Influenza, Human pathology, Influenza, Human virology, Lung immunology, Lung pathology, Lung virology, Mice, Inbred C57BL, Disease Models, Animal, Influenza A Virus, H1N1 Subtype physiology, Influenza Vaccines immunology, Influenza, Human prevention & control, Mice
- Abstract
Unlabelled: Influenza infection causes severe disease and death in humans. In traditional vaccine research and development, a single high-dose virus challenge of animals is used to evaluate vaccine efficacy. This type of challenge model may have limitations. In the present study, we developed a novel challenge model by infecting mice repeatedly in short intervals with low doses of influenza A virus. Our results show that compared to a single high-dose infection, mice that received repeated low-dose challenges showed earlier morbidity and mortality and more severe disease. They developed higher vial loads, more severe lung pathology, and greater inflammatory responses and generated only limited influenza A virus-specific B and T cell responses. A commercial trivalent influenza vaccine protected mice against a single high and lethal dose of influenza A virus but was ineffective against repeated low-dose virus challenges. Overall, our data show that the repeated low-dose influenza A virus infection mouse model is more stringent and may thus be more suitable to select for highly efficacious influenza vaccines., Importance: Influenza epidemics and pandemics pose serious threats to public health. Animal models are crucial for evaluating the efficacy of influenza vaccines. Traditional models based on a single high-dose virus challenge may have limitations. Here, we describe a new mouse model based on repeated low-dose influenza A virus challenges given within a short period. Repeated low-dose challenges caused more severe disease in mice, associated with higher viral loads and increased lung inflammation and reduced influenza A virus-specific B and T cell responses. A commercial influenza vaccine that was shown to protect mice from high-dose challenge was ineffective against repeated low-dose challenges. Overall, our results show that the low-dose repeated-challenge model is more stringent and may therefore be better suited for preclinical vaccine efficacy studies., (Copyright © 2015, American Society for Microbiology. All Rights Reserved.)
- Published
- 2015
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30. Adenovirus-mediated artificial MicroRNAs targeting matrix or nucleoprotein genes protect mice against lethal influenza virus challenge.
- Author
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Zhang H, Tang X, Zhu C, Song Y, Yin J, Xu J, Ertl HC, and Zhou D
- Subjects
- Adenoviridae genetics, Adenoviridae immunology, Adenoviridae metabolism, Animals, Cell Line, Tumor, Female, Genetic Vectors, HEK293 Cells, Humans, Influenza A Virus, H5N1 Subtype metabolism, Influenza A Virus, H9N2 Subtype metabolism, Mice, Inbred C57BL, MicroRNAs immunology, Nuclear Matrix genetics, Nucleoproteins genetics, Influenza A Virus, H1N1 Subtype metabolism, MicroRNAs metabolism, Nuclear Matrix metabolism, Nucleoproteins metabolism
- Abstract
Influenza virus (IV) infection is a major public health problem, causing millions of cases of severe illness and as many as 500 000 deaths each year worldwide. Given the limitations of current prevention or treatment of acute influenza, novel therapies are needed. RNA interference (RNAi) through microRNAs (miRNA) is an emerging technology that can suppress virus replication in vitro and in vivo. Here, we describe a novel strategy for the treatment of infuenza based on RNAi delivered by a replication-defective adenovirus (Ad) vector, derived from chimpanzee serotype 68 (AdC68). Our results showed that artificial miRNAs (amiRNAs) specifically targeting conserved regions of the IV genome could effectively inhibit virus replication in human embryonic kidney 293 cells. Moreover, our results demonstrated that prophylactic treatment with AdC68 expressing amiRNAs directed against M1, M2 or nucleoprotein genes of IV completely protected mice from homologous A/PR8 virus challenge and partially protected the mice from heterologous influenza A virus strains such as H9N2 and H5N1. Collectively, our data demonstrate that amiRNAs targeting the conserved regions of influenza A virus delivered by Ad vectors should be pursued as a novel strategy for prophylaxis of IV infection in humans and animals.
- Published
- 2015
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31. Activated CD4+CCR5+ T cells in the rectum predict increased SIV acquisition in SIVGag/Tat-vaccinated rhesus macaques.
- Author
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Carnathan DG, Wetzel KS, Yu J, Lee ST, Johnson BA, Paiardini M, Yan J, Morrow MP, Sardesai NY, Weiner DB, Ertl HC, and Silvestri G
- Subjects
- Animals, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes virology, Gene Products, gag immunology, Gene Products, tat immunology, Humans, Immunity, Cellular, Immunity, Mucosal, Lymphocyte Activation, Macaca mulatta immunology, Macaca mulatta virology, Receptors, CCR5 metabolism, Rectum immunology, Rectum virology, Simian Acquired Immunodeficiency Syndrome immunology, Simian Acquired Immunodeficiency Syndrome prevention & control, Simian Immunodeficiency Virus pathogenicity, Vaccination methods, Viremia immunology, Viremia prevention & control, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes virology, SAIDS Vaccines immunology, Simian Immunodeficiency Virus immunology, T-Lymphocyte Subsets immunology, T-Lymphocyte Subsets virology
- Abstract
An effective T-cell-based AIDS vaccine should induce strong HIV-specific CD8(+) T cells in mucosal tissues without increasing the availability of target cells for the virus. Here, we evaluated five immunization strategies that include Human adenovirus-5 (AdHu5), Chimpanzee adenovirus-6 (AdC6) or -7 (AdC7), Vaccinia virus (VV), and DNA given by electroporation (DNA/EP), all expressing Simian immunodeficiency virus group specific antigen/transactivator of transcription (SIV(mac239Gag/Tat)). Five groups of six rhesus macaques (RMs) each were vaccinated with DNA/EP-AdC6-AdC7, VV-AdC6-AdC7, DNA/-EP-VV-AdC6, DNA/EP-VV-AdC7, or AdHu5-AdHu5-AdHu5 and were challenged repeatedly with low-dose intrarectal SIVmac239. Upon challenge, there were no significant differences among study groups in terms of virus acquisition or viral load after infection. When taken together, the immunization regimens did not protect against SIV acquisition compared with controls but did result in an ∼ 1.6-log decline in set-point viremia. Although all immunized RMs had detectable SIV-specific CD8(+) T cells in blood and rectal mucosa, we found no correlation between the number or function of these SIV-specific CD8(+) T cells and protection against SIV acquisition. Interestingly, RMs experiencing breakthrough infection showed significantly higher prechallenge levels of CD4(+)C-C chemokine receptor type 5 (CCR5)(+)HLA-DR(+) T cells in the rectal biopsies (RB) than animals that remained uninfected. In addition, among the infected RMs, the percentage of CD4(+)CCR5(+)Ki-67(+) T cells in RBs prechallenge correlated with higher early viremia. Overall, these data suggest that the levels of activated CD4(+)CCR5(+) target T cells in the rectal mucosa may predict the risk of SIV acquisition in RMs vaccinated with vectors that express SIVGag/Tat.
- Published
- 2015
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32. Gene therapy and gene transfer approaches to prevent or treat chronic virus infections.
- Author
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Berkout B, Ertl HC, and Weinberg MS
- Subjects
- Animals, Chronic Disease, Genetic Therapy trends, HIV Infections therapy, HIV-1 genetics, HIV-1 immunology, Hepatitis B therapy, Hepatitis C therapy, Humans, Vaccines, DNA therapeutic use, Viral Vaccines genetics, Virus Diseases genetics, Virus Diseases prevention & control, Gene Transfer Techniques trends, Genetic Therapy methods, Viral Vaccines therapeutic use, Virus Diseases therapy
- Published
- 2015
33. Unique Roles of TLR9- and MyD88-Dependent and -Independent Pathways in Adaptive Immune Responses to AAV-Mediated Gene Transfer.
- Author
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Rogers GL, Suzuki M, Zolotukhin I, Markusic DM, Morel LM, Lee B, Ertl HC, and Herzog RW
- Subjects
- Animals, B-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Dependovirus genetics, Immunoglobulin G genetics, Immunoglobulin G immunology, Mice, Mice, Knockout, Myeloid Differentiation Factor 88 genetics, Toll-Like Receptor 9 genetics, Adaptive Immunity, Antibody Formation, Dependovirus immunology, Myeloid Differentiation Factor 88 immunology, Toll-Like Receptor 9 immunology, Transduction, Genetic
- Abstract
The immune system represents a significant barrier to successful gene therapy with adeno-associated viral (AAV) vectors. In particular, adaptive immune responses to the viral capsid or the transgene product are of concern. The sensing of AAV by toll-like receptors (TLRs) TLR2 and TLR9 has been suggested to play a role in innate immunity to the virus and may also shape subsequent adaptive immune responses. Here, we investigated the functions of TLR2, TLR9 and the downstream signaling adaptor MyD88 in antibody and CD8+ T-cell responses. Antibody formation against the transgene product occurred largely independently of TLR signaling following gene transfer with AAV1 or AAV2 vectors, whereas loss of signaling through the TLR9-MyD88 pathway substantially reduced CD8+ T-cell responses. In contrast, MyD88 (but neither of the TLRs) regulated antibody responses to capsid. B cell-intrinsic MyD88 was required for the formation of anti-capsid IgG2c independently of vector serotype or route of administration. However, MyD88(-/-) mice instead produced anti-capsid IgG1 that emerged with delayed kinetics but nonetheless completely prevented in vivo readministration. We conclude that there are distinct roles for TLR9 and MyD88 in promoting adaptive immune responses to AAV-mediated gene transfer and that there are redundant MyD88-dependent and MyD88-independent mechanisms that stimulate neutralizing antibody formation against AAV., (© 2015 S. Karger AG, Basel.)
- Published
- 2015
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34. Survivin-targeting Artificial MicroRNAs Mediated by Adenovirus Suppress Tumor Activity in Cancer Cells and Xenograft Models.
- Author
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Chi Y, Wang X, Yang Y, Zhang C, Ertl HC, and Zhou D
- Abstract
Survivin is highly expressed in most human tumors and fetal tissue, and absent in terminally differentiated cells. It promotes tumor cell proliferation by negatively regulating cell apoptosis and facilitating cell division. Survivin's selective expression pattern suggests that it might be a suitable target for cancer therapy, which would promote death of transformed but not normal cells. This was tested using artificial microRNAs (amiRNAs) targeting survivin. After screening, two effective amiRNAs, which knocked down survivin expression, were identified and cloned into a replication-defective adenoviral vector. Tumor cells infected with the recombinant vector downregulated expression of survivin and underwent apoptotic cell death. Further studies showed that apoptosis was associated with increases in caspase 3 and cleaved Poly (ADP-ribose) polymerase, and activation of the p53 signaling pathway. Furthermore, amiRNA treatment caused blockade of mitosis and cell cycle arrest at the G2/M phase. In vivo, survivin-targeting amiRNAs expressed by adenoviral vectors effectively delayed growth of hepatocellular and cervical carcinomas in mouse xenograft models. These results indicate that silencing of survivin by amiRNA has potential for treatment of cancer.
- Published
- 2014
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35. The effect of adenovirus-specific antibodies on adenoviral vector-induced, transgene product-specific T cell responses.
- Author
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Small JC, Haut LH, Bian A, and Ertl HC
- Subjects
- Adenoviridae classification, Adenoviridae Infections immunology, Adenoviridae Infections prevention & control, Animals, Antibody Specificity immunology, Cross Reactions immunology, Epitopes immunology, Female, Humans, Immune Sera immunology, Immunization, Passive, Immunologic Memory, Immunophenotyping, Mice, Phenotype, T-Cell Antigen Receptor Specificity immunology, T-Lymphocyte Subsets metabolism, Transduction, Genetic, gag Gene Products, Human Immunodeficiency Virus genetics, gag Gene Products, Human Immunodeficiency Virus immunology, Adenoviridae genetics, Adenoviridae immunology, Antibodies, Viral immunology, Genetic Vectors genetics, Genetic Vectors immunology, T-Lymphocyte Subsets immunology, Transgenes immunology
- Abstract
In this study, we tested the effect of neutralizing Abs to different serotypes of E1-deleted Ad vectors on the immunogenicity of the homologous Ad vector or a vector derived from a heterologous serotype. Our results showed that, as expected, even low titers of passively transferred neutralizing Abs significantly reduced the homologous vectors' ability to elicit transgene-specific CD8(+) T cell responses. In addition, Abs changed the fate of transgene product-specific CD8(+) T cells by promoting their transition into the central memory cell pool, which resulted in markedly enhanced expansion of transgene product-specific CD8(+) T cells after a boost with a heterologous Ad vector. Non-neutralizing Abs specific to a distinct Ad serotype had no effect on the magnitude of transgene product-specific CD8(+) T cells induced by a heterologous Ad vector, nor did such Abs promote induction of more resting memory CD8(+) T cells. These results show that Abs to an Ad vaccine carrier affect not only the magnitude but also the profile of a vector-induced CD8(+) T cell response., (© 2014 Society for Leukocyte Biology.)
- Published
- 2014
- Full Text
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36. The effect of adjuvanting cancer vaccines with herpes simplex virus glycoprotein D on melanoma-driven CD8+ T cell exhaustion.
- Author
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Zhang Y and Ertl HC
- Subjects
- Adjuvants, Immunologic, Animals, Antigens, CD biosynthesis, CD4-Positive T-Lymphocytes immunology, Cell Line, Chemotherapy, Adjuvant, Epitopes, T-Lymphocyte immunology, Female, Hemagglutinins, Viral genetics, Hemagglutinins, Viral immunology, Herpesvirus 1, Human immunology, Lymphocyte Activation immunology, Melanoma immunology, Melanoma prevention & control, Mice, Mice, Inbred C57BL, Programmed Cell Death 1 Receptor biosynthesis, Receptors, Immunologic biosynthesis, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins immunology, Signaling Lymphocytic Activation Molecule Family, Viral Envelope Proteins genetics, Lymphocyte Activation Gene 3 Protein, Antigens, Neoplasm immunology, CD8-Positive T-Lymphocytes immunology, Cancer Vaccines immunology, Melanoma therapy, Viral Envelope Proteins immunology
- Abstract
Two vaccines expressing CD4(+) and CD8(+) T cell epitopes of melanoma-associated Ags (MAAs) by a chimpanzee-derived replication-defective AdC68 vector were compared in a mouse model of melanoma. In one vaccine, termed AdC68-gDMelapoly, the epitopes were expressed as a fusion protein within HSV-1 glycoprotein D (gD), which blocks immunoinhibitory signaling through the herpes virus entry mediator pathway. The other vaccine, termed AdC68-Melapoly, expressed only the MAA epitopes. AdC68-gDMelapoly induced more potent MAA-specific CD8(+) T cell responses especially to the subdominant MAA epitopes. Upon prophylactic vaccination, mice that developed CD8(+) T cell responses to the two vaccines that were comparable in magnitude showed equal protection against tumor challenge. When mice were first challenged with tumor cells and then vaccinated results differed. In animals with comparable CD8(+) T cell responses, the AdC68-gDMelapoly vaccine was more efficacious compared with the AdC68-Melapoly vaccine in delaying tumor growth. This effect was linked to reduced expression of 2B4, LAG-3, and programmed death-1 on tumor-infiltrating MAA-specific CD8(+) T cells elicited by the gD-adjuvanted vaccine, suggesting that CD8(+) T cells induced in presence of gD are less susceptible to tumor-driven exhaustion., (Copyright © 2014 by The American Association of Immunologists, Inc.)
- Published
- 2014
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37. Enhancement of recombinant adenovirus vaccine-induced primary but not secondary systemic and mucosal immune responses by all-trans retinoic acid.
- Author
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Tuyishime S, Haut LH, Zhu C, and Ertl HC
- Subjects
- Adenoviridae immunology, Adjuvants, Immunologic administration & dosage, Animals, Female, Immunization, Secondary, Mice, Inbred BALB C, Receptors, CCR metabolism, Spleen immunology, Vaccines, Synthetic immunology, gag Gene Products, Human Immunodeficiency Virus immunology, AIDS Vaccines immunology, CD8-Positive T-Lymphocytes immunology, Genetic Vectors immunology, Immunity, Mucosal, Tretinoin immunology
- Abstract
Vaccination is an important tool for enhancing immune responses against mucosal pathogens. Intramuscularly administered adenovirus (Ad) vectors have been demonstrated to be strong inducers of both systemic and mucosal immune responses. Further enhancement of immune responses following Ad vaccination is highly desirable. All-trans retinoic acid (ATRA), a biologically active vitamin A metabolite, has been explored as an adjuvant for primary immune responses following vaccination. In this study, we investigated the effect of ATRA on a heterologous Ad prime boost regimen. ATRA co-administration during priming increased mucosal and systemic antibody responses as well as mucosal but not systemic CD8(+) T cell responses. However, this effect was no longer apparent after boosting regardless of whether ATRA was administered at the time of priming, at the time of boosting, or at both immunizations. Our findings confirm ATRA as an adjuvant for primary immune responses and suggest that the adjuvant effect does not extend to secondary immune responses., (Copyright © 2014. Published by Elsevier Ltd.)
- Published
- 2014
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38. Adenovirus-based vaccines against rhesus lymphocryptovirus EBNA-1 induce expansion of specific CD8+ and CD4+ T cells in persistently infected rhesus macaques.
- Author
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Leskowitz R, Fogg MH, Zhou XY, Kaur A, Silveira EL, Villinger F, Lieberman PM, Wang F, and Ertl HC
- Subjects
- Adenoviruses, Simian genetics, Animals, Drug Carriers, Female, Genetic Vectors, Herpesviridae Infections immunology, Herpesvirus Vaccines administration & dosage, Herpesvirus Vaccines genetics, Lymphocryptovirus genetics, Macaca mulatta, Vaccination methods, Viral Proteins genetics, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Herpesviridae Infections veterinary, Herpesvirus Vaccines immunology, Lymphocryptovirus immunology, Viral Proteins immunology
- Abstract
Unlabelled: The impact of Epstein-Barr virus (EBV) on human health is substantial, but vaccines that prevent primary EBV infections or treat EBV-associated diseases are not yet available. The Epstein-Barr nuclear antigen 1 (EBNA-1) is an important target for vaccination because it is the only protein expressed in all EBV-associated malignancies. We have designed and tested two therapeutic EBV vaccines that target the rhesus (rh) lymphocryptovirus (LCV) EBNA-1 to determine if ongoing T cell responses during persistent rhLCV infection in rhesus macaques can be expanded upon vaccination. Vaccines were based on two serotypes of E1-deleted simian adenovirus and were administered in a prime-boost regimen. To further modulate the response, rhEBNA-1 was fused to herpes simplex virus glycoprotein D (HSV-gD), which acts to block an inhibitory signaling pathway during T cell activation. We found that vaccines expressing rhEBNA-1 with or without functional HSV-gD led to expansion of rhEBNA-1-specific CD8(+) and CD4(+) T cells in 33% and 83% of the vaccinated animals, respectively. Additional animals developed significant changes within T cell subsets without changes in total numbers. Vaccination did not increase T cell responses to rhBZLF-1, an immediate early lytic phase antigen of rhLCV, thus indicating that increases of rhEBNA-1-specific responses were a direct result of vaccination. Vaccine-induced rhEBNA-1-specific T cells were highly functional and produced various combinations of cytokines as well as the cytolytic molecule granzyme B. These results serve as an important proof of principle that functional EBNA-1-specific T cells can be expanded by vaccination., Importance: EBV is a common human pathogen that establishes a persistent infection through latency in B cells, where it occasionally reactivates. EBV infection is typically benign and is well controlled by the host adaptive immune system; however, it is considered carcinogenic due to its strong association with lymphoid and epithelial cell malignancies. Latent EBNA-1 is a promising target for a therapeutic vaccine, as it is the only antigen expressed in all EBV-associated malignancies. The goal was to determine if rhEBNA-1-specific T cells could be expanded upon vaccination of infected animals. Results were obtained with vaccines that target EBNA-1 of rhLCV, a virus closely related to EBV. We found that vaccination led to expansion of rhEBNA-1 immune cells that exhibited functions fit for controlling viral infection. This confirms that rhEBNA-1 is a suitable target for therapeutic vaccines. Future work should aim to generate more-robust T cell responses through modified vaccines.
- Published
- 2014
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39. Construction and characterization of E1- and E3-deleted adenovirus vectors expressing two antigens from two separate expression cassettes.
- Author
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Small JC, Kurupati RK, Zhou X, Bian A, Chi E, Li Y, Xiang Z, and Ertl HC
- Subjects
- Adenoviridae immunology, Animals, Antigens immunology, CD8-Positive T-Lymphocytes immunology, Cell Line, Female, Gene Expression, Gene Order, Genetic Vectors immunology, Genome, Viral, Genomic Instability, Humans, Mice, Transgenes immunology, Adenoviridae genetics, Adenovirus E1 Proteins genetics, Adenovirus E3 Proteins genetics, Antigens genetics, Gene Deletion, Genetic Vectors genetics, Transgenes genetics
- Abstract
Here we describe a series of replication-defective adenovirus vectors designed to express transgene products from two expression cassettes placed into the deleted E1 and E3 domains. Vectors that contained an E1 cassette with a cytomegalovirus promoter in the forward orientation and an E3 cassette with the chicken β-actin promoter in the reverse orientation grew to acceptable yields and expressed both transgenes. Additionally, they elicited immune responses to both transgene products. Levels of expression and the vectors' immunogenicity were influenced by the presence of regulatory elements shared between the two expression cassettes. Specifically, vectors that carried the same intron and enhancer in both expression cassettes could be rescued and expanded, but they were poorly immunogenic. Deletion of the enhancer or both the enhancer and the intron from the E3 cassette increased T- and B-cell responses to both transgene products.
- Published
- 2014
- Full Text
- View/download PDF
40. Protection of non-human primates against rabies with an adenovirus recombinant vaccine.
- Author
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Xiang ZQ, Greenberg L, Ertl HC, and Rupprecht CE
- Subjects
- Adenoviridae genetics, Adenoviridae metabolism, Animals, Antibodies, Viral immunology, Female, Gene Expression, Genetic Vectors genetics, Genetic Vectors metabolism, Humans, Macaca fascicularis, Macaca mulatta, Male, Rabies immunology, Rabies virology, Rabies Vaccines genetics, Rabies Vaccines immunology, Rabies virus genetics, Vaccination, Vaccines, Synthetic administration & dosage, Vaccines, Synthetic genetics, Vaccines, Synthetic immunology, Viral Proteins administration & dosage, Viral Proteins genetics, Viral Proteins immunology, Rabies prevention & control, Rabies Vaccines administration & dosage, Rabies virus immunology
- Abstract
Rabies remains a major neglected global zoonosis. New vaccine strategies are needed for human rabies prophylaxis. A single intramuscular immunization with a moderate dose of an experimental chimpanzee adenovirus (Ad) vector serotype SAd-V24, also termed AdC68, expressing the rabies virus glycoprotein, resulted in sustained titers of rabies virus neutralizing antibodies and protection against a lethal rabies virus challenge infection in a non-human primate model. Taken together, these data demonstrate the safety, immunogenicity, and efficacy of the recombinant Ad-rabies vector for further consideration in human clinical trials., (Copyright © 2013 Elsevier Inc. All rights reserved.)
- Published
- 2014
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41. Immunogenicity of a prime-boost vaccine containing the circumsporozoite proteins of Plasmodium vivax in rodents.
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Teixeira LH, Tararam CA, Lasaro MO, Camacho AG, Ersching J, Leal MT, Herrera S, Bruna-Romero O, Soares IS, Nussenzweig RS, Ertl HC, Nussenzweig V, and Rodrigues MM
- Subjects
- Adjuvants, Immunologic administration & dosage, Animals, Antibodies, Protozoan blood, Female, Immunoglobulin G blood, Malaria Vaccines administration & dosage, Malaria Vaccines genetics, Malaria, Vivax immunology, Mice, Mice, Inbred C57BL, Plasmodium vivax genetics, Poly I-C administration & dosage, Protozoan Proteins genetics, Vaccines, Synthetic administration & dosage, Vaccines, Synthetic genetics, Vaccines, Synthetic immunology, Malaria Vaccines immunology, Malaria, Vivax prevention & control, Plasmodium vivax immunology, Protozoan Proteins immunology, Vaccination methods
- Abstract
Plasmodium vivax is the most widespread and the second most prevalent malaria-causing species in the world. Current measures used to control the transmission of this disease would benefit from the development of an efficacious vaccine. In the case of the deadly parasite P. falciparum, the recombinant RTS,S vaccine containing the circumsporozoite antigen (CSP) consistently protects 30 to 50% of human volunteers against infection and is undergoing phase III clinical trials in Africa with similar efficacy. These findings encouraged us to develop a P. vivax vaccine containing the three circulating allelic forms of P. vivax CSP. Toward this goal, we generated three recombinant bacterial proteins representing the CSP alleles, as well as a hybrid polypeptide called PvCSP-All-CSP-epitopes. This hybrid contains the conserved N and C termini of P. vivax CSP and the three variant repeat domains in tandem. We also generated simian and human recombinant replication-defective adenovirus vectors expressing PvCSP-All-CSP-epitopes. Mice immunized with the mixture of recombinant proteins in a formulation containing the adjuvant poly(I·C) developed high and long-lasting serum IgG titers comparable to those elicited by proteins emulsified in complete Freund's adjuvant. Antibody titers were similar in mice immunized with homologous (protein-protein) and heterologous (adenovirus-protein) vaccine regimens. The antibodies recognized the three allelic forms of CSP, reacted to the repeated and nonrepeated regions of CSP, and recognized sporozoites expressing the alleles VK210 and VK247. The vaccine formulations described in this work should be useful for the further development of an anti-P. vivax vaccine.
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- 2014
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42. Role of the vector genome and underlying factor IX mutation in immune responses to AAV gene therapy for hemophilia B.
- Author
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Rogers GL, Martino AT, Zolotukhin I, Ertl HC, and Herzog RW
- Subjects
- Animals, Antibodies, Viral immunology, CD8-Positive T-Lymphocytes immunology, Capsid immunology, Codon, Nonsense genetics, Gene Transfer Techniques, Genome genetics, Hemophilia B genetics, Hemophilia B immunology, Humans, Mice, Dependovirus genetics, Factor IX genetics, Factor IX therapeutic use, Genetic Therapy, Genetic Vectors genetics, Hemophilia B therapy, Immunity genetics
- Abstract
Background: Self-complementary adeno-associated virus (scAAV) vectors have become a desirable vector for therapeutic gene transfer due to their ability to produce greater levels of transgene than single-stranded AAV (ssAAV). However, recent reports have suggested that scAAV vectors are more immunogenic than ssAAV. In this study, we investigated the effects of a self-complementary genome during gene therapy with a therapeutic protein, human factor IX (hF.IX)., Methods: Hemophilia B mice were injected intramuscularly with ss or scAAV1 vectors expressing hF.IX. The outcome of gene transfer was assessed, including transgene expression as well as antibody and CD8⁺ T cell responses to hF.IX., Results: Self-complementary AAV1 vectors induced similar antibody responses (which eliminated systemic hF.IX expression) but stronger CD8⁺ T cell responses to hF.IX relative to ssAAV1 in mice with F9 gene deletion. As a result, hF.IX-expressing muscle fibers were effectively eliminated in scAAV-treated mice. In contrast, mice with F9 nonsense mutation (late stop codon) lacked antibody or T cell responses, thus showing long-term expression regardless of the vector genome., Conclusions: The nature of the AAV genome can impact the CD8⁺ T cell response to the therapeutic transgene product. In mice with endogenous hF.IX expression, however, this enhanced immunogenicity did not break tolerance to hF.IX, suggesting that the underlying mutation is a more important risk factor for transgene-specific immunity than the molecular form of the AAV genome.
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- 2014
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43. Passive immunization of macaques with polyclonal anti-SHIV IgG against a heterologous tier 2 SHIV: outcome depends on IgG dose.
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Sholukh AM, Byrareddy SN, Shanmuganathan V, Hemashettar G, Lakhashe SK, Rasmussen RA, Watkins JD, Vyas HK, Thorat S, Brandstoetter T, Mukhtar MM, Yoon JK, Novembre FJ, Villinger F, Landucci G, Forthal DN, Ratcliffe S, Tuero I, Robert-Guroff M, Polonis VR, Bilska M, Montefiori DC, Johnson WE, Ertl HC, and Ruprecht RM
- Subjects
- Acquired Immunodeficiency Syndrome virology, Animals, Disease Models, Animal, Macaca mulatta, Simian Immunodeficiency Virus immunology, Treatment Outcome, Acquired Immunodeficiency Syndrome prevention & control, Antibodies, Neutralizing administration & dosage, Cross Protection, HIV Antibodies administration & dosage, HIV-1 immunology, Immunization, Passive methods, Immunoglobulin G administration & dosage
- Abstract
Background: A key goal for HIV-1 envelope immunogen design is the induction of cross-reactive neutralizing antibodies (nAbs). As AIDS vaccine recipients will not be exposed to strains exactly matching any immunogens due to multiple HIV-1 quasispecies circulating in the human population worldwide, heterologous SHIV challenges are essential for realistic vaccine efficacy testing in primates. We assessed whether polyclonal IgG, isolated from rhesus monkeys (RMs) with high-titer nAbs (termed SHIVIG), could protect RMs against the R5-tropic tier-2 SHIV-2873Nip, which was heterologous to the viruses or HIV-1 envelopes that had elicited SHIVIG., Results: SHIVIG demonstrated binding to HIV Gag, Tat, and Env of different clades and competed with the broadly neutralizing antibodies b12, VRC01, 4E10, and 17b. SHIVIG neutralized tier 1 and tier 2 viruses, including SHIV-2873Nip. NK-cell depletion decreased the neutralizing activity of SHIVIG 20-fold in PBMC assays. Although SHIVIG neutralized SHIV-2873Nip in vitro, this polyclonal IgG preparation failed to prevent acquisition after repeated intrarectal low-dose virus challenges, but at a dose of 400 mg/kg, it significantly lowered peak viremia (P = 0.001). Unexpectedly, single-genome analysis revealed a higher number of transmitted variants at the low dose of 25 mg/kg, implying increased acquisition at low SHIVIG levels. In vitro, SHIVIG demonstrated complement-mediated Ab-dependent enhancement of infection (C'-ADE) at concentrations similar to those observed in plasmas of RMs treated with 25 mg/kg of SHIVIG., Conclusion: Our primate model data suggest a dual role for polyclonal anti-HIV-1 Abs depending on plasma levels upon virus encounter.
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- 2014
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44. CD8+ T cell recognition of epitopes within the capsid of adeno-associated virus 8-based gene transfer vectors depends on vectors' genome.
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Wu TL, Li H, Faust SM, Chi E, Zhou S, Wright F, High KA, and Ertl HC
- Subjects
- Animals, Capsid Proteins immunology, Dependovirus genetics, Epitopes, T-Lymphocyte, Genetic Vectors standards, Humans, Immunologic Memory, Lymphocyte Activation immunology, Male, Mice, Quality Control, T-Cell Antigen Receptor Specificity, Transduction, Genetic, CD8-Positive T-Lymphocytes immunology, Capsid immunology, Dependovirus immunology, Genetic Vectors genetics, Genetic Vectors immunology, Genome
- Abstract
Self-complementary adeno-associated viral (AAV) vectors expressing human factor IX (hF.IX) have achieved transient or sustained correction of hemophilia B in human volunteers. High doses of AAV2 or AAV8 vectors delivered to the liver caused in several patients an increase in transaminases accompanied by a rise in AAV capsid-specific T cells and a decrease in circulating hF.IX levels suggesting immune-mediated destruction of vector-transduced cells. Kinetics of these adverse events differed in patients receiving AAV2 or AAV8 vectors causing rise in transaminases at 3 versus 8 weeks after vector injection, respectively. To test if CD8+ T cells to AAV8 vectors, which are similar to AAV2 vectors are fully-gutted vectors and thereby fail to encode structural viral proteins, could cause damage at this late time point, we tested in a series of mouse studies how long major histocompatibility (MHC) class I epitopes within AAV8 capsid can be presented to CD8+ T cells. Our results clearly show that depending on the vectors' genome, CD8+ T cells can detect such epitopes on AAV8's capsid for up to 6 months indicating that the capsid of AAV8 degrades slowly in mice.
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- 2014
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45. Therapeutic vaccination against the rhesus lymphocryptovirus EBNA-1 homologue, rhEBNA-1, elicits T cell responses to novel epitopes in rhesus macaques.
- Author
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Silveira EL, Fogg MH, Leskowitz RM, Ertl HC, Wiseman RW, O'Connor DH, Lieberman P, Wang F, and Villinger F
- Subjects
- Animals, Epitope Mapping, Epitopes, T-Lymphocyte chemistry, Epitopes, T-Lymphocyte genetics, Epitopes, T-Lymphocyte immunology, Epstein-Barr Virus Nuclear Antigens administration & dosage, Epstein-Barr Virus Nuclear Antigens chemistry, Epstein-Barr Virus Nuclear Antigens genetics, Female, Herpesviridae Infections drug therapy, Herpesviridae Infections immunology, Herpesviridae Infections virology, Lymphocryptovirus genetics, Male, Primate Diseases drug therapy, Primate Diseases virology, T-Lymphocytes virology, Vaccination, Viral Vaccines administration & dosage, Viral Vaccines chemistry, Viral Vaccines genetics, Viral Vaccines immunology, Epstein-Barr Virus Nuclear Antigens immunology, Herpesviridae Infections veterinary, Lymphocryptovirus immunology, Macaca mulatta genetics, Macaca mulatta immunology, Macaca mulatta virology, Primate Diseases immunology, T-Lymphocytes immunology
- Abstract
Epstein-Barr virus (EBV) is a vaccine/immunotherapy target due to its association with several human malignancies. EBNA-1 is an EBV protein consistently expressed in all EBV-associated cancers. Herein, EBNA-1-specific T cell epitopes were evaluated after AdC-rhEBNA-1 immunizations in chronically lymphocryptovirus-infected rhesus macaques, an EBV infection model. Preexisting rhEBNA-1-specific responses were augmented in 4/12 animals, and new epitopes were recognized in 5/12 animals after vaccinations. This study demonstrated that EBNA-1-specific T cells can be expanded by vaccination.
- Published
- 2013
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46. Vaccine-induced boosting of influenza virus-specific CD4 T cells in younger and aged humans.
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Dolfi DV, Mansfield KD, Kurupati RK, Kannan S, Doyle SA, Ertl HC, Schmader KE, and Wherry EJ
- Subjects
- Adult, Aged, Antibodies, Neutralizing, Cell Differentiation immunology, Cell Proliferation, Demography, Female, Humans, Kinetics, Male, Species Specificity, Vaccines, Inactivated immunology, Viral Proteins immunology, Aging immunology, CD4-Positive T-Lymphocytes immunology, Influenza Vaccines immunology, Orthomyxoviridae immunology
- Abstract
Current yearly influenza virus vaccines induce strain-specific neutralizing antibody (NAb) responses providing protective immunity to closely matched viruses. However, these vaccines are often poorly effective in high-risk groups such as the elderly and challenges exist in predicting yearly or emerging pandemic influenza virus strains to include in the vaccines. Thus, there has been considerable emphasis on understanding broadly protective immunological mechanisms for influenza virus. Recent studies have implicated memory CD4 T cells in heterotypic immunity in animal models and in human challenge studies. Here we examined how influenza virus vaccination boosted CD4 T cell responses in younger versus aged humans. Our results demonstrate that while the magnitude of the vaccine-induced CD4 T cell response and number of subjects responding on day 7 did not differ between younger and aged subjects, fewer aged subjects had peak responses on day 14. While CD4 T cell responses were inefficiently boosted against NA, both HA and especially nucleocaspid protein- and matrix-(NP+M) specific responses were robustly boosted. Pre-existing CD4 T cell responses were associated with more robust responses to influenza virus NP+M, but not H1 or H3. Finally pre-existing strain-specific NAb decreased the boosting of CD4 T cell responses. Thus, accumulation of pre-existing influenza virus-specific immunity in the form of NAb and cross-reactive T cells to conserved virus proteins (e.g. NP and M) over a lifetime of exposure to infection and vaccination may influence vaccine-induced CD4 T cell responses in the aged.
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- 2013
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47. Immunological and virological analyses of rhesus macaques immunized with chimpanzee adenoviruses expressing the simian immunodeficiency virus Gag/Tat fusion protein and challenged intrarectally with repeated low doses of SIVmac.
- Author
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Cervasi B, Carnathan DG, Sheehan KM, Micci L, Paiardini M, Kurupati R, Tuyishime S, Zhou XY, Else JG, Ratcliffe SJ, Ertl HC, and Silvestri G
- Subjects
- AIDS Vaccines genetics, AIDS Vaccines immunology, Adenoviruses, Simian genetics, Animals, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Gene Products, gag genetics, Gene Products, tat genetics, Genetic Vectors, Humans, Immunization, Secondary, Pan troglodytes virology, Recombinant Fusion Proteins genetics, Recombinant Fusion Proteins immunology, Rectal Diseases, SAIDS Vaccines administration & dosage, SAIDS Vaccines genetics, SAIDS Vaccines immunology, Simian Acquired Immunodeficiency Syndrome immunology, Simian Acquired Immunodeficiency Syndrome prevention & control, Simian Acquired Immunodeficiency Syndrome virology, Simian Immunodeficiency Virus genetics, Simian Immunodeficiency Virus physiology, Virus Replication, Adenoviruses, Simian immunology, Gene Products, gag immunology, Gene Products, tat immunology, Macaca mulatta immunology, Macaca mulatta virology, Simian Immunodeficiency Virus immunology
- Abstract
Human adenovirus (AdHu)-based candidate AIDS vaccine can provide protection from simian immunodeficiency virus (SIV) transmission and disease progression. However, their potential use may be limited by widespread preexisting immunity to the vector. In contrast, preexisting immunity to chimpanzee adenoviruses (AdC) is relatively rare. In this study, we utilized two regimens of prime-boost immunizations with AdC serotype SAd-V23 (also called AdC6) and SAd-V24 (also called AdC7) expressing SIV Gag/Tat to test their immunogenicity and ability to protect rhesus macaques (RMs) from a repeated low-dose SIVmac239 challenge. Both AdC6 followed by AdC7 (AdC6/7) and AdC7 followed by AdC6 (AdC7/6) induced robust SIV Gag/Tat-specific T cell responses as measured by tetramer staining and functional assays. However, no significant protection from SIV transmission was observed in either AdC7/6- or AdC7/6-vaccinated RMs. Interestingly, in the RMs showing breakthrough infections, AdC7/6-SIV immunization was associated with a transient but significant (P = 0.035 at day 90 and P = 0.033 at day 120 postinfection) reduction in the setpoint viral load compared to unvaccinated controls. None of the measured immunological markers (i.e., number or functionality of SIV-specific CD8(+) and CD4(+) T cell responses and level of activated and/or CCR5(+) CD4(+) target cells) at the time of challenge correlated with protection from SIV transmission in the AdC-SIV-vaccinated RMs. The robust immunogenicity observed in all AdC-immunized RMs and the transient signal of protection from SIV replication exhibited by AdC7/6-vaccinated RMs even in the absence of any envelope immunogen suggest that AdC-based vectors may represent a promising platform for candidate AIDS vaccines.
- Published
- 2013
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48. CD4+ and CD8+ T-cell responses to latent antigen EBNA-1 and lytic antigen BZLF-1 during persistent lymphocryptovirus infection of rhesus macaques.
- Author
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Leskowitz RM, Zhou XY, Villinger F, Fogg MH, Kaur A, Lieberman PM, Wang F, and Ertl HC
- Subjects
- Animals, Cytokines metabolism, Disease Models, Animal, Female, Herpesviridae Infections virology, Macaca mulatta, Tumor Virus Infections virology, CD4-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes immunology, Epstein-Barr Virus Nuclear Antigens immunology, Herpesviridae Infections immunology, Lymphocryptovirus immunology, Trans-Activators immunology, Tumor Virus Infections immunology
- Abstract
Epstein-Barr virus (EBV) infection leads to lifelong viral persistence through its latency in B cells. EBV-specific T cells control reactivations and prevent the development of EBV-associated malignancies in most healthy carriers, but infection can sometimes cause chronic disease and malignant transformation. Epstein-Barr nuclear antigen 1 (EBNA-1) is the only viral protein consistently expressed during all forms of latency and in all EBV-associated malignancies and is a promising target for a therapeutic vaccine. Here, we studied the EBNA-1-specific immune response using the EBV-homologous rhesus lymphocryptovirus (rhLCV) infection in rhesus macaques. We assessed the frequency, phenotype, and cytokine production profiles of rhLCV EBNA-1 (rhEBNA-1)-specific T cells in 15 rhesus macaques and compared them to the lytic antigen of rhLCV BZLF-1 (rhBZLF-1). We were able to detect rhEBNA-1-specific CD4(+) and/or CD8(+) T cells in 14 of the 15 animals screened. In comparison, all 15 animals had detectable rhBZLF-1 responses. Most peptide-specific CD4(+) T cells exhibited a resting phenotype of central memory (TCM), while peptide-specific CD8(+) T cells showed a more activated phenotype, belonging mainly to the effector cell subset. By comparing our results to the human EBV immune response, we demonstrate that the rhLCV model is a valid system for studying chronic EBV infection and for the preclinical development of therapeutic vaccines.
- Published
- 2013
- Full Text
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49. Effects of immunosuppression on circulating adeno-associated virus capsid-specific T cells in humans.
- Author
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Parzych EM, Li H, Yin X, Liu Q, Wu TL, Podsakoff GM, High KA, Levine MH, and Ertl HC
- Subjects
- Adult, Aged, Antigens, Viral immunology, CD4-Positive T-Lymphocytes immunology, CD4-Positive T-Lymphocytes metabolism, CD4-Positive T-Lymphocytes pathology, CD8-Positive T-Lymphocytes immunology, CD8-Positive T-Lymphocytes metabolism, CD8-Positive T-Lymphocytes pathology, Capsid immunology, Case-Control Studies, Dependovirus immunology, Female, Genetic Vectors, Humans, Immune Tolerance, Kidney Transplantation, Liver immunology, Liver metabolism, Liver Transplantation, Male, Middle Aged, Transplantation Immunology, Dependovirus genetics, Immunosuppression Therapy
- Abstract
In humans adeno-associated virus (AAV)-mediated gene transfer is followed by expansion of AAV capsid-specific T cells, evidence of cell damage, and loss of transgene product expression, implicating immunological rejection of vector-transduced cells, which may be prevented by immunosuppressive drugs. We undertook this study to assess the effect of immunosuppression (IS) used for organ transplantation on immune responses to AAV capsid antigens. Recipients of liver or kidney transplants were tested before and 4 weeks after induction of IS in comparison with matched samples from healthy human adults and an additional cohort with comorbid conditions similar to those of the transplant patients. Our data show that transplant patients and comorbid control subjects have markedly higher frequencies of circulating AAV capsid-specific T cells compared with healthy adults. On average, IS resulted in a reduction of AAV-specific CD4⁺ T cells, whereas numbers of circulating CD8⁺ effector and central memory T cells tended to increase. Independent of the type of transplant or the IS regimens, the trend of AAV capsid-specific T cell responses after drug treatment varied; in some patients responses were unaffected whereas others showed decreases or even pronounced increases, casting doubt on the usefulness of prophylactic IS for AAV vector recipients.
- Published
- 2013
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50. Engineered AAV vector minimizes in vivo targeting of transduced hepatocytes by capsid-specific CD8+ T cells.
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Martino AT, Basner-Tschakarjan E, Markusic DM, Finn JD, Hinderer C, Zhou S, Ostrov DA, Srivastava A, Ertl HC, Terhorst C, High KA, Mingozzi F, and Herzog RW
- Subjects
- Adoptive Transfer methods, Animals, CD8-Positive T-Lymphocytes metabolism, Capsid Proteins genetics, Capsid Proteins metabolism, Cells, Cultured, Dependovirus genetics, Dependovirus immunology, Dependovirus metabolism, Genetic Engineering, Genetic Vectors genetics, Hepatocytes metabolism, Humans, Male, Mice, Mice, Inbred BALB C, T-Cell Antigen Receptor Specificity genetics, Transduction, Genetic, CD8-Positive T-Lymphocytes immunology, Capsid Proteins immunology, Dependovirus physiology, Genetic Therapy methods, Genetic Vectors physiology, Hepatocytes immunology
- Abstract
Recent clinical trials have shown that evasion of CD8(+) T-cell responses against viral capsid is critical for successful liver-directed gene therapy with adeno-associated viral (AAV) vectors for hemophilia. Preclinical models to test whether use of alternate serotypes or capsid variants could avoid this deleterious response have been lacking. Here, the ability of CD8(+) T cells ("cap-CD8," specific for a capsid epitope presented by human B*0702 or murine H2-L(d) molecules) to target AAV-infected hepatocytes was investigated. In a murine model based on adoptive transfer of ex vivo expanded cap-CD8, AAV2-transduced livers showed CD8(+) T-cell infiltrates, transaminitis, significant reduction in factor IX transgene expression, and loss of transduced hepatocytes. AAV8 gene transfer resulted in prolonged susceptibility to cap-CD8, consistent with recent clinical findings. In contrast, using an AAV2(Y-F) mutant capsid, which is known to be less degraded by proteasomes, preserved transgene expression and largely avoided hepatotoxicity. In vitro assays confirmed reduced major histocompatibility complex class I presentation of this capsid and killing of human or murine hepatocytes compared with AAV2. In conclusion, AAV capsids can be engineered to substantially reduce the risk of destruction by cytotoxic T lymphocytes, whereas use of alternative serotypes per se does not circumvent this obstacle.
- Published
- 2013
- Full Text
- View/download PDF
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