141 results on '"Ranajit Pal"'
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2. Engagement of monocytes, NK cells, and CD4+ Th1 cells by ALVAC-SIV vaccination results in a decreased risk of SIVmac251 vaginal acquisition.
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Giacomo Gorini, Slim Fourati, Monica Vaccari, Mohammad Arif Rahman, Shari N Gordon, Dallas R Brown, Lynn Law, Jean Chang, Richard Green, Fredrik Barrenäs, Namal P M Liyanage, Melvin N Doster, Luca Schifanella, Massimiliano Bissa, Isabela Silva de Castro, Robyn Washington-Parks, Veronica Galli, Deborah H Fuller, Sampa Santra, Michael Agy, Ranajit Pal, Robert E Palermo, Georgia D Tomaras, Xiaoying Shen, Celia C LaBranche, David C Montefiori, David J Venzon, Hung V Trinh, Mangala Rao, Michael Gale, Rafick P Sekaly, and Genoveffa Franchini
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
The recombinant Canarypox ALVAC-HIV/gp120/alum vaccine regimen was the first to significantly decrease the risk of HIV acquisition in humans, with equal effectiveness in both males and females. Similarly, an equivalent SIV-based ALVAC vaccine regimen decreased the risk of virus acquisition in Indian rhesus macaques of both sexes following intrarectal exposure to low doses of SIVmac251. Here, we demonstrate that the ALVAC-SIV/gp120/alum vaccine is also efficacious in female Chinese rhesus macaques following intravaginal exposure to low doses of SIVmac251 and we confirm that CD14+ classical monocytes are a strong correlate of decreased risk of virus acquisition. Furthermore, we demonstrate that the frequency of CD14+ cells and/or their gene expression correlates with blood Type 1 CD4+ T helper cells, α4β7+ plasmablasts, and vaginal cytocidal NKG2A+ cells. To better understand the correlate of protection, we contrasted the ALVAC-SIV vaccine with a NYVAC-based SIV/gp120 regimen that used the identical immunogen. We found that NYVAC-SIV induced higher immune activation via CD4+Ki67+CD38+ and CD4+Ki67+α4β7+ T cells, higher SIV envelope-specific IFN-γ producing cells, equivalent ADCC, and did not decrease the risk of SIVmac251 acquisition. Using the systems biology approach, we demonstrate that specific expression profiles of plasmablasts, NKG2A+ cells, and monocytes elicited by the ALVAC-based regimen correlated with decreased risk of virus acquisition.
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- 2020
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3. ALVAC-HIV B/C candidate HIV vaccine efficacy dependent on neutralization profile of challenge virus and adjuvant dose and type.
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Luca Schifanella, Susan W Barnett, Massimiliano Bissa, Veronica Galli, Melvin N Doster, Monica Vaccari, Georgia D Tomaras, Xiaoying Shen, Sanjay Phogat, Ranajit Pal, David C Montefiori, Celia C LaBranche, Mangala Rao, Hung V Trinh, Robyn Washington-Parks, Namal P M Liyanage, Giacomo Gorini, Dallas R Brown, Frank Liang, Karin Loré, David J Venzon, William Magnanelli, Michelle Metrinko, Josh Kramer, Matthew Breed, Galit Alter, Ruth M Ruprecht, and Genoveffa Franchini
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
The ALVAC-HIV clade B/AE and equivalent SIV-based/gp120 + Alum vaccines successfully decreased the risk of virus acquisition in humans and macaques. Here, we tested the efficacy of HIV clade B/C ALVAC/gp120 vaccine candidates + MF59 or different doses of Aluminum hydroxide (Alum) against SHIV-Cs of varying neutralization sensitivity in macaques. Low doses of Alum induced higher mucosal V2-specific IgA that increased the risk of Tier 2 SHIV-C acquisition. High Alum dosage, in contrast, elicited serum IgG to V2 that correlated with a decreased risk of Tier 1 SHIV-C acquisition. MF59 induced negligible mucosal antibodies to V2 and an inflammatory profile with blood C-reactive Protein (CRP) levels correlating with neutralizing antibody titers. MF59 decreased the risk of Tier 1 SHIV-C acquisition. The relationship between vaccine efficacy and the neutralization profile of the challenge virus appear to be linked to the different immunological spaces created by MF59 and Alum via CXCL10 and IL-1β, respectively.
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- 2019
- Full Text
- View/download PDF
4. Antibody Fab‐Fc properties outperform titer in predictive models of SIV vaccine‐induced protection
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Srivamshi Pittala, Kenneth Bagley, Jennifer A Schwartz, Eric P Brown, Joshua A Weiner, Ilia J Prado, Wenlei Zhang, Rong Xu, Ayuko Ota‐Setlik, Ranajit Pal, Xiaoying Shen, Charles Beck, Guido Ferrari, George K Lewis, Celia C LaBranche, David C Montefiori, Georgia D Tomaras, Galit Alter, Mario Roederer, Timothy R Fouts, Margaret E Ackerman, and Chris Bailey‐Kellogg
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antibody effector function ,biomarker identification ,HIV ,protection modeling ,systems serology ,Biology (General) ,QH301-705.5 ,Medicine (General) ,R5-920 - Abstract
Abstract Characterizing the antigen‐binding and innate immune‐recruiting properties of the humoral response offers the chance to obtain deeper insights into mechanisms of protection than revealed by measuring only overall antibody titer. Here, a high‐throughput, multiplexed Fab‐Fc Array was employed to profile rhesus macaques vaccinated with a gp120‐CD4 fusion protein in combination with different genetically encoded adjuvants, and subsequently subjected to multiple heterologous simian immunodeficiency virus (SIV) challenges. Systems analyses modeling protection and adjuvant differences using Fab‐Fc Array measurements revealed a set of correlates yielding strong and robust predictive performance, while models based on measurements of response magnitude alone exhibited significantly inferior performance. At the same time, rendering Fab‐Fc measurements mathematically independent of titer had relatively little impact on predictive performance. Similar analyses for a distinct SIV vaccine study also showed that Fab‐Fc measurements performed significantly better than titer. These results suggest that predictive modeling with measurements of antibody properties can provide detailed correlates with robust predictive power, suggest directions for vaccine improvement, and potentially enable discovery of mechanistic associations.
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- 2019
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- View/download PDF
5. Mucosal B Cells Are Associated with Delayed SIV Acquisition in Vaccinated Female but Not Male Rhesus Macaques Following SIVmac251 Rectal Challenge.
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Iskra Tuero, Venkatramanan Mohanram, Thomas Musich, Leia Miller, Diego A Vargas-Inchaustegui, Thorsten Demberg, David Venzon, Irene Kalisz, V S Kalyanaraman, Ranajit Pal, Maria Grazia Ferrari, Celia LaBranche, David C Montefiori, Mangala Rao, Monica Vaccari, Genoveffa Franchini, Susan W Barnett, and Marjorie Robert-Guroff
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
Many viral infections, including HIV, exhibit sex-based pathogenic differences. However, few studies have examined vaccine-related sex differences. We compared immunogenicity and protective efficacy of monomeric SIV gp120 with oligomeric SIV gp140 in a pre-clinical rhesus macaque study and explored a subsequent sex bias in vaccine outcome. Each immunization group (16 females, 8 males) was primed twice mucosally with replication-competent Ad-recombinants encoding SIVsmH4env/rev, SIV239gag and SIV239nefΔ1-13 and boosted twice intramuscularly with SIVmac239 monomeric gp120 or oligomeric gp140 in MF59 adjuvant. Controls (7 females, 5 males) received empty Ad and MF59. Up to 9 weekly intrarectal challenges with low-dose SIVmac251 were administered until macaques became infected. We assessed vaccine-induced binding, neutralizing, and non-neutralizing antibodies, Env-specific memory B cells and plasmablasts/plasma cells (PB/PC) in bone marrow and rectal tissue, mucosal Env-specific antibodies, and Env-specific T-cells. Post-challenge, only one macaque (gp140-immunized) remained uninfected. However, SIV acquisition was significantly delayed in vaccinated females but not males, correlated with Env-specific IgA in rectal secretions, rectal Env-specific memory B cells, and PC in rectal tissue. These results extend previous correlations of mucosal antibodies and memory B cells with protective efficacy. The gp140 regimen was more immunogenic, stimulating elevated gp140 and cyclic V2 binding antibodies, ADCC and ADCP activities, bone marrow Env-specific PB/PC, and rectal gp140-specific IgG. However, immunization with gp120, the form of envelope immunogen used in RV144, the only vaccine trial to show some efficacy, provided more significant acquisition delay. Further over 40 weeks of follow-up, no gp120 immunized macaques met euthanasia criteria in contrast to 7 gp140-immunized and 2 control animals. Although males had higher binding antibodies than females, ADCC and ADCP activities were similar. The complex challenge outcomes may reflect differences in IgG subtypes, Fc glycosylation, Fc-R polymorphisms, and/or the microbiome, key areas for future studies. This first demonstration of a sex-difference in SIV vaccine-induced protection emphasizes the need for sex-balancing in vaccine trials. Our results highlight the importance of mucosal immunity and memory B cells at the SIV exposure site for protection.
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- 2015
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6. In vitro activity of cysteamine against SARS-CoV-2 variants
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Jess Thoene, Robert F. Gavin, Aaron Towne, Lauren Wattay, Maria Grazia Ferrari, Jennifer Navarrete, and Ranajit Pal
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Endocrinology ,SARS-CoV-2 ,Endocrinology, Diabetes and Metabolism ,Cysteamine ,Genetics ,Humans ,Ophthalmic Solutions ,Molecular Biology ,Biochemistry ,Pandemics ,Antiviral Agents ,COVID-19 Drug Treatment - Abstract
Global COVID-19 pandemic is caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Continuous emergence of new variants and their rapid spread are jeopardizing vaccine countermeasures to a significant extent. While currently available vaccines are effective at preventing illness associated with SARS-CoV-2 infection, these have been shown to be less effective at preventing breakthrough infection and transmission from a vaccinated individual to others. Here we demonstrate broad antiviral activity of cysteamine HCl in vitro against major emergent infectious variants of SARS-CoV-2 in a highly permissible Vero cell line. Cysteamine HCl inhibited infection of wild type, alpha, beta, gamma, delta, lambda, and omicron variants effectively. Cysteamine is a very well-tolerated US FDA-approved drug used chronically as a topical ophthalmic solution to treat ocular cystinosis in patients who receive it hourly or QID lifelong at concentrations 6 times higher than that required to inhibit SARS CoV-2 in tissue culture. Application of cysteamine as a topical nasal treatment can potentially1) mitigate existing infection 2) prevent infection in exposed individuals, and 3) limit the contagion in vulnerable populations.
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- 2022
7. Titan Atlas and the Roots of the Mediterranean cultures
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Malmi, Pasi Ilmari, Isom��ki, Risto, Iorco, Tommaso, Brohi, Muhammed Afzal, and Ranajit Pal
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- 2022
- Full Text
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8. The Effects of Ethyl Lauroyl Arginine Hydrochloride (ELAH) in Nasal Spray Formula on SAR-Cov-2
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Harshad R. Thacore, Abdul Gaffar, Seiyoung Yun, Agnes L. Chenine, Maria G. Ferrari, Ranajit Pal, Lauren Wattay, and Marnie L. Peterson
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viruses - Abstract
SARS-CoV-2 and coronaviruses, enveloped RNA viruses, are major causes of acute human respiratory diseases. The aim of the study was to investigate the broad-spectrum antiviral effects of ethyl lauroyl arginine hydrochloride (ELAH) in in vitro and in vivo assays. Cell-based assays found that the pseudovirus VSV-SARS-CoV-2 was inhibited with an EC50 of 15 micrograms/ml, with complete inhibition achieved at 110 micrograms/ml. The effects were comparable to those observed with anti-SARS-CoV-2 antibody neutralization assays against VSV-SARS-CoV-2. Intranasal administration of the Wuhan strain of SARS-CoV-2 treated in vitro with ELAH inhibited the disease symptoms caused by the virus in a Syrian hamster model compared to that caused by the same dose of virus treated in vitro with medium alone. Subgenomic RNA and total RNA viral load were concomitantly reduced in the treated animals compared with the control group. In cell-based studies, pretreatment of susceptible cells with 1-10 micrograms/ml ELAH inhibited the attachment of the virus to the cells, as measured by cytopathic and high-resolution scanning electron microscopy (SEM) effects, suggesting that the primary mode of ELAH action was due to preventing the attachment of the virus to the cells. Collectively, the data suggest that ELAH could be a promising agent for the prevention of SARS infection through nasopharyngeal surfaces.
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- 2021
9. In Vitro Activity of Cysteamine Against SARS-CoV-2 Variants
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Ranajit Pal, Robert F Gavin, Aaron Towne, Jess G. Thoene, Lauren Wattay, and Maria Grazia Ferrari
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Drug ,business.industry ,Transmission (medicine) ,media_common.quotation_subject ,Alpha (ethology) ,Breakthrough infection ,medicine.disease ,Virology ,In vitro ,chemistry.chemical_compound ,chemistry ,Cystinosis ,Vero cell ,Medicine ,Cysteamine ,business ,media_common - Abstract
Global COVID-19 pandemic is caused by infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Continuous emergence of new variants and their rapid spread are jeopardizing vaccine countermeasures to a significant extent. While currently available vaccines are effective at preventing illness associated with SARS-CoV-2 infection, these have been shown to be less effective at preventing breakthrough infection and transmission from a vaccinated individual to others. Here we demonstrate broad antiviral activity of cysteamine HCl in vitro against major emergent infectious variants of SARS-CoV-2 in a highly permissible Vero cell line. Cysteamine HCl inhibited infection of wild type, alpha, beta, gamma, delta, lambda, and omicron variants effectively. Cysteamine is a very well-tolerated US FDA-approved drug used chronically as a topical ophthalmic solution to treat ocular cystinosis in patients who receive it hourly or QID lifelong at concentrations 6 to 10 times higher than that required to completely inhibit SARS CoV-2 in tissue culture. Application of cysteamine as a topical nasal treatment can potentially1) mitigate existing infection 2) prevent infection in exposed individuals, and 3) limit the contagion in vulnerable populations.
- Published
- 2021
10. The effects of ethyl lauroyl arginine hydrochloride (ELAH) in nasal spray formula on SARS-Cov-2
- Author
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Maria Grazi Ferrari, Abdul Gaffar, Marnie L. Peterson, Seiyoung Yun, Harshad R. Thacore, Ranajit Pal, Lauren Wattay, and Agnès Laurence Chenine
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ARGININE HYDROCHLORIDE ,Chromatography ,Nasal spray ,Chemistry ,viruses ,medicine.medical_treatment ,Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) ,medicine - Abstract
SARS-CoV-2 and coronaviruses, enveloped RNA viruses, are major causes of acute human respiratory diseases. The aim of the study was to investigate the broad-spectrum antiviral effects of ethyl lauroyl arginine hydrochloride (ELAH) in in vitro and in vivo assays. Cell-based assays found that the pseudovirus VSV-SARS-CoV-2 was inhibited with an EC50 of 15 micrograms/ml, with complete inhibition achieved at 110 micrograms/ml. The effects were comparable to those observed with anti-SARS-CoV-2 antibody neutralization assays against VSV-SARS-CoV-2. Intranasal administration of the Wuhan strain of SARS-CoV-2 treated in vitro with ELAH inhibited the disease symptoms caused by the virus in a Syrian hamster model compared to that caused by the same dose of virus treated in vitro with medium alone. Subgenomic RNA and total RNA viral load were concomitantly reduced in the treated animals compared with the control group. In cell-based studies, pretreatment of susceptible cells with 1–10 micrograms/ml ELAH inhibited the attachment of the virus to the cells, as measured by cytopathic and high-resolution scanning electron microscopy (SEM) effects, suggesting that the primary mode of ELAH action was due to preventing the attachment of the virus to the cells. Collectively, the data suggest that ELAH could be a promising agent for the prevention of SARS infection through nasopharyngeal surfaces.
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- 2021
11. THE DATE OF THE BHARATA WAR FROM KALIBANGAN SEALS AND THE ROLE OF THE ARYANS IN THIS WAR
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Ranajit Pal
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History ,General Engineering ,Aryan race ,Ancient history - Published
- 2021
12. Titan Atlas and the Roots of the Mediterranean Cultures
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Malmi, Pasi Ilmari, Isom��ki, Risto, Iorco, Tommaso, and Ranajit Pal
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- 2021
- Full Text
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13. ALVAC-HIV B/C candidate HIV vaccine efficacy dependent on neutralization profile of challenge virus and adjuvant dose and type
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Josh Kramer, Xiaoying Shen, Karin Loré, Melvin N. Doster, Sanjay Phogat, David C. Montefiori, Monica Vaccari, David Venzon, Hung V. Trinh, Luca Schifanella, Dallas R. Brown, Frank Liang, Mangala Rao, Ranajit Pal, Michelle Metrinko, Georgia D. Tomaras, Matthew Breed, Giacomo Gorini, Massimiliano Bissa, Susan W. Barnett, Veronica Galli, Galit Alter, William Magnanelli, Genoveffa Franchini, Ruth M. Ruprecht, Namal P.M. Liyanage, Celia C. LaBranche, and Robyn Washington-Parks
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medicine.medical_treatment ,HIV Infections ,Monkeys ,Antibodies, Viral ,Graduates ,Pathology and Laboratory Medicine ,Biochemistry ,Neutralization ,0302 clinical medicine ,Immunodeficiency Viruses ,Medicine ,Public and Occupational Health ,HIV vaccine ,Biology (General) ,Mammals ,0303 health sciences ,030302 biochemistry & molecular biology ,Eukaryota ,virus diseases ,3. Good health ,Blood ,Medical Microbiology ,Viral Pathogens ,Macaque ,Primates ,QH301-705.5 ,Immunology ,Alumni ,Microbiology ,03 medical and health sciences ,Adjuvants, Immunologic ,Genetics ,Molecular Biology ,Microbial Pathogens ,Organisms ,Correction ,Proteins ,Virology ,Antibodies, Neutralizing ,chemistry ,Animal Studies ,Parasitology ,Population Groupings ,Preventive Medicine ,Immunologic diseases. Allergy ,RNA viruses ,Physiology ,viruses ,MF59 ,Simian Acquired Immunodeficiency Syndrome ,chemistry.chemical_compound ,Immunologic Adjuvants ,Immune Physiology ,Medicine and Health Sciences ,030212 general & internal medicine ,Neutralizing antibody ,AIDS Vaccines ,Vaccines ,Immune System Proteins ,biology ,SAIDS Vaccines ,Animal Models ,Vaccination and Immunization ,Body Fluids ,Infectious Diseases ,Experimental Organism Systems ,Vertebrates ,Viruses ,Alum Compounds ,Educational Status ,Female ,Simian Immunodeficiency Virus ,Anatomy ,Pathogens ,Adjuvant ,Research Article ,Infectious Disease Control ,Research and Analysis Methods ,Virus ,Antibodies ,Blood Plasma ,Old World monkeys ,Retroviruses ,Animals ,030304 developmental biology ,Biology and life sciences ,Rhesus Monkeys ,business.industry ,Alum ,Lentivirus ,HIV ,Viral Vaccines ,RC581-607 ,Vaccine efficacy ,Macaca mulatta ,13. Climate action ,Amniotes ,People and Places ,biology.protein ,business - Abstract
The ALVAC-HIV clade B/AE and equivalent SIV-based/gp120 + Alum vaccines successfully decreased the risk of virus acquisition in humans and macaques. Here, we tested the efficacy of HIV clade B/C ALVAC/gp120 vaccine candidates + MF59 or different doses of Aluminum hydroxide (Alum) against SHIV-Cs of varying neutralization sensitivity in macaques. Low doses of Alum induced higher mucosal V2-specific IgA that increased the risk of Tier 2 SHIV-C acquisition. High Alum dosage, in contrast, elicited serum IgG to V2 that correlated with a decreased risk of Tier 1 SHIV-C acquisition. MF59 induced negligible mucosal antibodies to V2 and an inflammatory profile with blood C-reactive Protein (CRP) levels correlating with neutralizing antibody titers. MF59 decreased the risk of Tier 1 SHIV-C acquisition. The relationship between vaccine efficacy and the neutralization profile of the challenge virus appear to be linked to the different immunological spaces created by MF59 and Alum via CXCL10 and IL-1β, respectively., Author summary The ALVAC-based clade C /gp120 combination is currently being tested for efficacy and licensures in South Africa in the HVTN702 HIV vaccine trial using the MF59 adjuvant rather than Alum with the intent of inducing higher neutralizing antibodies and T cell responses. Here, we present studies in 71 macaques that evaluated the relative efficacy of identical ALVAC-HIV B/C HIV vaccines with Alum or MF59 following challenge exposure to SHIV-clade C viral stocks that differed in their neutralization profiles. Our results demonstrated that use of the proprietary Novartis Alum at lower doses affects the proportion of mucosal V2-specific IgG and IgA, and that IgAs and IgGs were respectively associated with an increased or decreased risk of virus acquisition. In contrast, animals immunized with the MF59 regimen had no V2 mucosal responses, and a high level of serum neutralizing antibodies (Tier 1) to easy to neutralize viruses that were associated with a decreased risk of Tier 1 SHIV-C acquisition. In vitro studies suggest the hypothesis that the low doses of proprietary Novartis Alum used in our studies may be the underlying reason for the decreased vaccine efficacy, via lower inflammasome activation and IL-1β production.
- Published
- 2019
14. DNA prime/protein boost vaccination elicits robust humoral response in rhesus macaques using oligomeric simian immunodeficiency virus envelope and Advax delta inulin adjuvant
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Nikolai Petrovsky, Vaniambadi S. Kalyanaraman, Celia C. LaBranche, Veena Menon, Asma Ashraf, Irene Kalisz, Stephen Whitney, Victor I. Ayala, Sneha P. Rangaswamy, Lindsey Galmin, David C. Montefiori, and Ranajit Pal
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0301 basic medicine ,medicine.medical_treatment ,Immunization, Secondary ,HIV Infections ,HIV Antibodies ,HIV Envelope Protein gp120 ,Antibodies, Viral ,medicine.disease_cause ,Neutralization ,03 medical and health sciences ,Immune system ,Adjuvants, Immunologic ,Virology ,medicine ,Animals ,Humans ,AIDS Vaccines ,biology ,Immunogenicity ,Vaccination ,Inulin ,SAIDS Vaccines ,virus diseases ,Simian immunodeficiency virus ,Antibodies, Neutralizing ,Macaca mulatta ,Immunity, Humoral ,030104 developmental biology ,DNA, Viral ,Immunology ,HIV-1 ,biology.protein ,Simian Immunodeficiency Virus ,Nasal administration ,Rabbits ,Antibody ,Adjuvant ,Research Article - Abstract
The partial success of the RV144 trial underscores the importance of envelope-specific antibody responses for an effective HIV-1 vaccine. Oligomeric HIV-1 envelope proteins delivered with a potent adjuvant are expected to elicit strong antibody responses with broad neutralization specificity. To test this hypothesis, two SIV envelope proteins were formulated with delta inulin-based adjuvant (Advax) and used to immunize nonhuman primates. Oligomeric gp140-gp145 from SIVmac251 and SIVsmE660 was purified to homogeneity. Oligomers showed high-affinity interaction with CD4 and were highly immunogenic in rabbits, inducing Tier 2 SIV-neutralizing antibodies. The immunogenicity of an oligomeric Env DNA prime and protein boost together with Advax was evaluated in Chinese rhesus macaques. DNA administration elicited antibodies to both envelopes, and titres were markedly enhanced following homologous protein boosts via intranasal and intramuscular routes. Strong antibody responses were detected against the V1 and V2 domains of gp120. During peak immune responses, a low to moderate level of neutralizing activity was detected against Tier 1A/1B SIV isolates, with a moderate level noted against a Tier 2 isolate. Increased serum antibody affinity to SIVmac251 gp140 and generation of Env-specific memory B cells were observed in the immunized macaques. Animals were subjected to low-dose intravaginal challenge with SIVmac251 one week after the last protein boost. One out of three immunized animals was protected from infection. Although performed with a small number of macaques, this study demonstrates the utility of oligomeric envelopes formulated with Advax in eliciting broad antibody responses with the potential to provide protection against SIV transmission.
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- 2017
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15. Antibody Fab‐Fc properties outperform titer in predictive models of <scp>SIV</scp> vaccine‐induced protection
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Eric P. Brown, David C. Montefiori, Ranajit Pal, Margaret E. Ackerman, Chris Bailey-Kellogg, Kenneth C. Bagley, Wenlei Zhang, Ayuko Ota-Setlik, Guido Ferrari, Jennifer Schwartz, Timothy R. Fouts, Galit Alter, Mario Roederer, Celia C. LaBranche, Charles Beck, Rong Xu, Ilia Prado, Georgia D. Tomaras, Xiaoying Shen, Joshua A. Weiner, George K. Lewis, and Srivamshi Pittala
- Subjects
CD4-Positive T-Lymphocytes ,Medicine (General) ,medicine.medical_treatment ,Simian Acquired Immunodeficiency Syndrome ,Antibodies, Viral ,medicine.disease_cause ,0302 clinical medicine ,Viral Envelope Proteins ,Biology (General) ,Immunoglobulin Fragments ,0303 health sciences ,Membrane Glycoproteins ,Applied Mathematics ,SAIDS Vaccines ,Antibody titer ,Articles ,Microbiology, Virology & Host Pathogen Interaction ,antibody effector function ,Titer ,Computational Theory and Mathematics ,biomarker identification ,Simian Immunodeficiency Virus ,Antibody ,General Agricultural and Biological Sciences ,Adjuvant ,Information Systems ,QH301-705.5 ,protection modeling ,Immunology ,Heterologous ,Methods & Resources ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,03 medical and health sciences ,R5-920 ,medicine ,Animals ,Humans ,SIV Vaccine ,030304 developmental biology ,systems serology ,General Immunology and Microbiology ,HIV ,Simian immunodeficiency virus ,Macaca mulatta ,Fusion protein ,Immunity, Humoral ,Immunoglobulin G ,Multivariate Analysis ,biology.protein ,030217 neurology & neurosurgery - Abstract
Characterizing the antigen‐binding and innate immune‐recruiting properties of the humoral response offers the chance to obtain deeper insights into mechanisms of protection than revealed by measuring only overall antibody titer. Here, a high‐throughput, multiplexed Fab‐Fc Array was employed to profile rhesus macaques vaccinated with a gp120‐CD4 fusion protein in combination with different genetically encoded adjuvants, and subsequently subjected to multiple heterologous simian immunodeficiency virus (SIV) challenges. Systems analyses modeling protection and adjuvant differences using Fab‐Fc Array measurements revealed a set of correlates yielding strong and robust predictive performance, while models based on measurements of response magnitude alone exhibited significantly inferior performance. At the same time, rendering Fab‐Fc measurements mathematically independent of titer had relatively little impact on predictive performance. Similar analyses for a distinct SIV vaccine study also showed that Fab‐Fc measurements performed significantly better than titer. These results suggest that predictive modeling with measurements of antibody properties can provide detailed correlates with robust predictive power, suggest directions for vaccine improvement, and potentially enable discovery of mechanistic associations.
- Published
- 2019
16. Membrane bound Indian clade C HIV-1 envelope antigen induces antibodies to diverse and conserved epitopes upon DNA prime/protein boost in rabbits
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Veena Menon, Sneha Priya Rangasamy, Priyanka Dhopeshwarkar, Ranajit Pal, Kalyanaraman S. Vaniambadi, and Sundarasamy Mahalingam
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0301 basic medicine ,medicine.drug_class ,Immunization, Secondary ,HIV Antibodies ,HIV Envelope Protein gp120 ,Biology ,Monoclonal antibody ,Gp41 ,Epitope ,HIV Envelope Protein gp160 ,Epitopes ,03 medical and health sciences ,Antigen ,Neutralization Tests ,Vaccines, DNA ,medicine ,Animals ,Humans ,AIDS Vaccines ,General Veterinary ,General Immunology and Microbiology ,Linear epitope ,Immunogenicity ,Public Health, Environmental and Occupational Health ,Antibodies, Monoclonal ,Antibodies, Neutralizing ,Virology ,Molecular biology ,030104 developmental biology ,Infectious Diseases ,Epitope mapping ,HIV-1 ,biology.protein ,Molecular Medicine ,Binding Sites, Antibody ,Rabbits ,Antibody ,Epitope Mapping - Abstract
The partial success of RV144 human clinical trial demonstrated that ALVAC prime/envelope protein boost vaccine regimen may represent a promising strategy for the development of an effective HIV-1 vaccine. Our earlier study demonstrated that a trimeric HIV-1 envelope gp145 from an Indian clade C isolate elicited cross clade neutralizing antibodies primarily towards Tier 1 isolates. In the present study, we examined the immunogenicity of DNA prime/envelope protein boost vaccine in rabbits using gp160 DNA of the Indian clade C isolate with various cytoplasmic tail truncations and trimeric gp145 protein. Cytoplasmic tail mutants of gp160 exposed epitopes that reacted strongly with a number of broadly neutralizing human monoclonal antibodies against HIV-1. Overall, envelope specific titers were found to be similar in all rabbit groups with higher pseudovirus neutralization in protein only immunized rabbits. The complete linear epitope mapping of rabbit immune sera revealed strong binding to C1, C2, V3, C3 and C4 domains of gp145. Importantly, reactivity of gp41 ecto-domain peptides was observed in DNA prime/protein boost sera but not in the sera of rabbits immunized with protein alone. Moreover, membrane anchored but not soluble envelope encoding DNA immunization elicited antibodies against linear epitopes on the conserved gp41 ecto-domain. Together, these results suggest that priming with DNA encoding cytoplasmic domains of Env alters the quality of antibodies elicited following protein boost and hence may be utilized to generate protective immunity by HIV-1 vaccine.
- Published
- 2016
17. Early SIV Dissemination After Intrarectal SIVmac251 Challenge Was Associated With Proliferating Virus-Susceptible Cells in the Colorectum
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Jay A. Berzofsky, Yongjun Sui, Ranajit Pal, Eun Mi Lee, Blake Frey, Yichuan Wang, Alison Hogg, and David Venzon
- Subjects
Male ,0301 basic medicine ,Time Factors ,Colon ,T-Lymphocytes ,viruses ,Simian Acquired Immunodeficiency Syndrome ,Rectum ,Biology ,Virus Replication ,medicine.disease_cause ,Article ,Virus ,03 medical and health sciences ,Viral entry ,medicine ,Animals ,Pharmacology (medical) ,Viral rna ,Cell Proliferation ,Cell growth ,Viral Load ,Simian immunodeficiency virus ,Macaca mulatta ,Virology ,030104 developmental biology ,Infectious Diseases ,medicine.anatomical_structure ,Viral replication ,Immunology ,RNA, Viral ,Female ,Simian Immunodeficiency Virus ,Viral load - Abstract
OBJECTIVE Few studies have examined the eclipse time of simian immunodeficiency virus/HIV infection through the anal route. We aimed to measure the eclipse time after SIVmac251 intrarectal inoculation, and to investigate the factor(s) associated with early dissemination. DESIGN Forty macaques were intrarectally challenged with SIVmac251 3 times at 2-week intervals. METHODS Plasma viral RNA was monitored at 4, 7, 11, 14, 21, and 28 days after infection. Rectal/vaginal tissues were obtained and tissue viral loads (VLs) were measured at day 14 postinfection. RESULTS Of 40 macaques 26 (65%) had first detectable viral RNAs in the plasma at day 7 after the challenge that led to productive infection. Strikingly, 6 animals (15%) had detectable viral RNA in the plasma as early as at day 4. The Ki67 viral target CD4 T cells in the colorectal tissues were significantly higher in the early or middle-transmitter groups than those in the late-transmitter group. The rectal VL did not correlate with plasma VL at 14-day postinoculation, but did positively correlate with plasma VLs at days 21 and 28 postinfection. CONCLUSIONS The median eclipse time after intrarectal challenge was 7 days, with a few early transmitters at 4 days. More rapid viral dissemination was associated with a high frequency of colorectal Ki67CCR5CD4T cells, which fuel the local viral replication. Furthermore, local viral replication in the colorectal tissue during the early stage might affect the plasma VL in a delayed manner. Therefore, to reduce/limit these target cells at the portal of viral entry is essential.
- Published
- 2016
18. Engagement of monocytes, NK cells, and CD4+ Th1 cells by ALVAC-SIV vaccination results in a decreased risk of SIVmac251 vaginal acquisition
- Author
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Genoveffa Franchini, Melvin N. Doster, Sampa Santra, Rafick Pierre Sekaly, Slim Fourati, David C. Montefiori, Robyn Washington-Parks, Namal P.M. Liyanage, Richard Green, Robert E. Palermo, Michael Gale, Monica Vaccari, Mangala Rao, Deborah H. Fuller, Celia C. LaBranche, Giacomo Gorini, Hung V. Trinh, Mohammad Arif Rahman, Xiaoying Shen, David Venzon, Dallas R. Brown, Isabela Silva de Castro, Luca Schifanella, Michael B. Agy, Lynn Law, Ranajit Pal, Fredrik Barrenäs, Jean Chang, Veronica Galli, Massimiliano Bissa, Georgia D. Tomaras, and Shari Gordon
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RNA viruses ,Physiology ,animal diseases ,viruses ,Monkeys ,CD38 ,Pathology and Laboratory Medicine ,Biochemistry ,Monocytes ,White Blood Cells ,Immunodeficiency Viruses ,Animal Cells ,Immune Physiology ,Medicine and Health Sciences ,Medicine ,Public and Occupational Health ,Biology (General) ,Mammals ,Vaccines ,0303 health sciences ,Immune System Proteins ,biology ,T Cells ,Viral Vaccine ,Vaccination ,030302 biochemistry & molecular biology ,SAIDS Vaccines ,Eukaryota ,virus diseases ,Animal Models ,Vaccination and Immunization ,ALVAC Vaccine ,3. Good health ,Killer Cells, Natural ,Infectious Diseases ,SIV ,Experimental Organism Systems ,Medical Microbiology ,Viral Pathogens ,Vertebrates ,Viruses ,Vagina ,Immunologi ,Female ,Simian Immunodeficiency Virus ,Cellular Types ,Pathogens ,Antibody ,Macaque ,Research Article ,Primates ,Canarypox ,Infectious Disease Control ,QH301-705.5 ,Immune Cells ,CD14 ,Immunology ,Research and Analysis Methods ,Microbiology ,Antibodies ,Virus ,03 medical and health sciences ,Virology ,Old World monkeys ,Retroviruses ,Genetics ,Animals ,Microbial Pathogens ,Molecular Biology ,030304 developmental biology ,Blood Cells ,Biology and life sciences ,Rhesus Monkeys ,business.industry ,Lentivirus ,Organisms ,Proteins ,Viral Vaccines ,Cell Biology ,RC581-607 ,Th1 Cells ,biology.organism_classification ,Macaca mulatta ,Amniotes ,Animal Studies ,biology.protein ,Parasitology ,Preventive Medicine ,Immunologic diseases. Allergy ,business - Abstract
The recombinant Canarypox ALVAC-HIV/gp120/alum vaccine regimen was the first to significantly decrease the risk of HIV acquisition in humans, with equal effectiveness in both males and females. Similarly, an equivalent SIV-based ALVAC vaccine regimen decreased the risk of virus acquisition in Indian rhesus macaques of both sexes following intrarectal exposure to low doses of SIVmac251. Here, we demonstrate that the ALVAC-SIV/gp120/alum vaccine is also efficacious in female Chinese rhesus macaques following intravaginal exposure to low doses of SIVmac251 and we confirm that CD14+ classical monocytes are a strong correlate of decreased risk of virus acquisition. Furthermore, we demonstrate that the frequency of CD14+ cells and/or their gene expression correlates with blood Type 1 CD4+ T helper cells, α4β7+ plasmablasts, and vaginal cytocidal NKG2A+ cells. To better understand the correlate of protection, we contrasted the ALVAC-SIV vaccine with a NYVAC-based SIV/gp120 regimen that used the identical immunogen. We found that NYVAC-SIV induced higher immune activation via CD4+Ki67+CD38+ and CD4+Ki67+α4β7+ T cells, higher SIV envelope-specific IFN-γ producing cells, equivalent ADCC, and did not decrease the risk of SIVmac251 acquisition. Using the systems biology approach, we demonstrate that specific expression profiles of plasmablasts, NKG2A+ cells, and monocytes elicited by the ALVAC-based regimen correlated with decreased risk of virus acquisition., Author summary The ALVAC-HIV/gp120/alum regimen tested in 8,197 human volunteers (61.4% males, 38.6% females) in the RV144 trial decreased the risk of HIV infection similarly in both sexes. The ALVAC-SIV/gp120/alum vaccine also reduced the risk of intrarectal SIVmac251 acquisition in both female and male vaccinated macaques at an average of 44% per exposure. In the current work, we tested whether this vaccine modality could also reduce the risk of intravaginal SIVmac251 exposure. In order to detect correlates of risk, we administered the virus by the intravaginal route and tested another vaccine regimen based on the vaccinia derivative poxvirus NYVAC in parallel. We demonstrate here that the ALVAC-SIV/gp120/alum regimen decreases the risk of vaginal SIVmac251 acquisition (50% vaccine efficacy) and, importantly, we confirmed that subsets of monocytes and CD4+ T cells are correlates of risk of acquisition. In addition, we uncovered cytotoxic vaginal NKG2A+ cells and gut-homing α4β7 positive plasmablasts as novel correlates of risk of intravaginal virus acquisition. In contrast, NYVAC-SIV vaccination induced high levels of activated T cells and did not protect against SIVmac251 acquisition. We examined the contrasting immune responses to better understand the correlate of protection and found that the unique ability of ALVAC-SIV to activate early interferon responses and the inflammasome during priming differentiates the two poxvirus vectors. This work demonstrates the reproducibility of the efficacy observed in the ALVAC-based regimen and defines novel correlates of risk in the rigorous SIVmac251 macaque model, establishing a benchmark for future improvement of this vaccine approach.
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- 2020
19. Novel Strategy To Adapt Simian-Human Immunodeficiency Virus E1 Carrying env from an RV144 Volunteer to Rhesus Macaques: Coreceptor Switch and Final Recovery of a Pathogenic Virus with Exclusive R5 Tropism
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Jennifer Delgado, Sharon Orndorff, Amanda Strickland, Celia C. LaBranche, Maria Grazia Ferrari, Deborah T. Weiss, Punnee Pitisuttithum, Sorachai Nitayaphan, Jim Treece, Hemant K. Vyas, Agnès Laurence Chenine, Anthony Griffiths, Robert McLinden, Ruth M. Ruprecht, Hanna B. Scinto, Supachai Rerks-Ngarm, Sandeep Gupta, Merlin L. Robb, Siddappa N. Byrareddy, Swati Thorat, Ranajit Pal, Jerome H. Kim, Muhammad M. Mukhtar, Elizabeth Plake, Nelson L. Michael, and David C. Montefiori
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0301 basic medicine ,viruses ,030106 microbiology ,Immunology ,virus diseases ,Viremia ,Biology ,medicine.disease ,Microbiology ,Virology ,Long terminal repeat ,Virus ,03 medical and health sciences ,Chimera (genetics) ,030104 developmental biology ,Viral replication ,Insect Science ,medicine ,biology.protein ,Antibody ,Viral load ,Tropism - Abstract
The phase III RV144 human immunodeficiency virus (HIV) vaccine trial conducted in Thailand remains the only study to show efficacy in decreasing the HIV acquisition risk. In Thailand, circulating recombinant forms of HIV clade A/E (CRF01_AE) predominate; in such viruses, env originates from clade E (HIV-E). We constructed a simian-human immunodeficiency virus (SHIV) chimera carrying env isolated from an RV144 placebo recipient in the SHIV-1157ipd3N4 backbone. The latter contains long terminal repeats (LTRs) with duplicated NF-κB sites, thus resembling HIV LTRs. We devised a novel strategy to adapt the parental infectious molecular clone (IMC), R5 SHIV-E1, to rhesus macaques: the simultaneous depletion of B and CD8+ cells followed by the intramuscular inoculation of proviral DNA and repeated administrations of cell-free virus. High-level viremia and CD4+ T-cell depletion ensued. Passage 3 virus unexpectedly caused acute, irreversible CD4+ T-cell loss; the partially adapted SHIV had become dual tropic. Virus and IMCs with exclusive R5 tropism were reisolated from earlier passages, combined, and used to complete adaptation through additional macaques. The final isolate, SHIV-E1p5, remained solely R5 tropic. It had a tier 2 neutralization phenotype, was mucosally transmissible, and was pathogenic. Deep sequencing revealed 99% Env amino acid sequence conservation; X4-only and dual-tropic strains had evolved independently from an early branch of parental SHIV-E1. To conclude, our primate model data reveal that SHIV-E1p5 recapitulates important aspects of HIV transmission and pathobiology in humans. IMPORTANCE Understanding the protective principles that lead to a safe, effective vaccine against HIV in nonhuman primate (NHP) models requires test viruses that allow the evaluation of anti-HIV envelope responses. Reduced HIV acquisition risk in RV144 has been linked to nonneutralizing IgG antibodies with a range of effector activities. Definitive experiments to decipher the mechanisms of the partial protection observed in RV144 require passive-immunization studies in NHPs with a relevant test virus. We have generated such a virus by inserting env from an RV144 placebo recipient into a SHIV backbone with HIV-like LTRs. The final SHIV-E1p5 isolate, grown in rhesus monkey peripheral blood mononuclear cells, was mucosally transmissible and pathogenic. Earlier SHIV-E passages showed a coreceptor switch, again mimicking HIV biology in humans. Thus, our series of SHIV-E strains mirrors HIV transmission and disease progression in humans. SHIV-E1p5 represents a biologically relevant tool to assess prevention strategies.
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- 2018
20. Novel Strategy To Adapt Simian-Human Immunodeficiency Virus E1 Carrying
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Hanna B, Scinto, Sandeep, Gupta, Swati, Thorat, Muhammad M, Mukhtar, Anthony, Griffiths, Jennifer, Delgado, Elizabeth, Plake, Hemant K, Vyas, Amanda, Strickland, Siddappa N, Byrareddy, David C, Montefiori, Celia, LaBranche, Ranajit, Pal, Jim, Treece, Sharon, Orndorff, Maria Grazia, Ferrari, Deborah, Weiss, Agnes-Laurence, Chenine, Robert, McLinden, Nelson, Michael, Jerome H, Kim, Merlin L, Robb, Supachai, Rerks-Ngarm, Punnee, Pitisuttithum, Sorachai, Nitayaphan, and Ruth M, Ruprecht
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Volunteers ,Receptors, CCR5 ,viruses ,Simian Acquired Immunodeficiency Syndrome ,virus diseases ,Gene Products, env ,HIV Infections ,Viral Load ,Thailand ,Virus Replication ,Macaca mulatta ,Tropism ,Proviruses ,HIV-1 ,Leukocytes, Mononuclear ,Animals ,Humans ,Pathogenesis and Immunity ,Simian Immunodeficiency Virus ,Viremia - Abstract
Understanding the protective principles that lead to a safe, effective vaccine against HIV in nonhuman primate (NHP) models requires test viruses that allow the evaluation of anti-HIV envelope responses. Reduced HIV acquisition risk in RV144 has been linked to nonneutralizing IgG antibodies with a range of effector activities. Definitive experiments to decipher the mechanisms of the partial protection observed in RV144 require passive-immunization studies in NHPs with a relevant test virus. We have generated such a virus by inserting env from an RV144 placebo recipient into a SHIV backbone with HIV-like LTRs. The final SHIV-E1p5 isolate, grown in rhesus monkey peripheral blood mononuclear cells, was mucosally transmissible and pathogenic. Earlier SHIV-E passages showed a coreceptor switch, again mimicking HIV biology in humans. Thus, our series of SHIV-E strains mirrors HIV transmission and disease progression in humans. SHIV-E1p5 represents a biologically relevant tool to assess prevention strategies.
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- 2018
21. Two-Year Follow-Up of Macaques Developing Intermittent Control of the Human Immunodeficiency Virus Homolog Simian Immunodeficiency Virus SIVmac251 in the Chronic Phase of Infection
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Marjorie Robert-Guroff, Iart Luca Shytaj, Hye Kyung Chung, Celia C. LaBranche, Eric J. Arts, Anna Teresa Palamara, David C. Montefiori, Ranajit Pal, Mauro Biffoni, Mark G. Lewis, Diego A. Vargas-Inchaustegui, Gabrielle Nickel, Andrea Savarino, Nicholas Farrell, and Jonah B. Sacha
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CD4-Positive T-Lymphocytes ,gag ,Auranofin ,viruses ,animals ,antiviral agents ,cd4-positive t-lymphocytes ,disease models ,animal ,follow-up studies ,gene products ,hiv infections ,humans ,macaca mulatta ,simian acquired immunodeficiency syndrome ,simian immunodeficiency virus ,viral load ,immunology ,virology ,Immunology ,Simian Acquired Immunodeficiency Syndrome ,Gene Products, gag ,HIV Infections ,Viremia ,Biology ,medicine.disease_cause ,Antiviral Agents ,Microbiology ,Virus ,chemistry.chemical_compound ,Immune system ,Acquired immunodeficiency syndrome (AIDS) ,Virology ,medicine ,Animals ,Humans ,Buthionine sulfoximine ,Viral Load ,Simian immunodeficiency virus ,medicine.disease ,Macaca mulatta ,Disease Models, Animal ,chemistry ,Insect Science ,Pathogenesis and Immunity ,Simian Immunodeficiency Virus ,Viral load ,Follow-Up Studies ,medicine.drug - Abstract
Off-therapy control of viremia by HIV-infected individuals has been associated with two likely players: a restricted viral reservoir and an efficient cell-mediated immune response. We previously showed that a combination of highly suppressive antiretroviral therapy and two experimental drugs, i.e., auranofin and buthionine sulfoximine, was able to reduce the viral reservoir, elicit efficient cell-mediated antiviral responses, and induce intermittent posttherapy viral load control in chronically SIVmac251-infected macaques. We here show that the macaques that had received this drug combination and then stopped antiretroviral therapy were also able to maintain low numbers of activated CD4 + T cells at viral rebound. Moreover, these macaques consistently displayed low-level simian immunodeficiency virus (SIV) diversity, which was in line with the strong and broadly reactive cell-mediated immune responses against conserved Gag antigens. Extended follow-up showed that the two macaques that had received the complete drug combination remained healthy and did not develop AIDS in 2 years of follow-up after therapy suspension. This disease-free survival is longer than twice the average time of progression to AIDS in SIVmac251-infected rhesus macaques. These results suggest that limited numbers of activated T cells at viral rebound and subsequent development of broadly reactive cell-mediated responses may be interrelated in reducing the viral reservoir. IMPORTANCE The HIV reservoir in CD4 + T cells represents one main obstacle to HIV eradication. Recent studies, however, show that a drastic reduction of this reservoir is insufficient for inducing a functional cure of AIDS. In the present work, we thoroughly studied and subjected to long-term follow-up two macaques showing intermittent control of the virus following suspension of antiretroviral therapy plus an experimental antireservoir treatment, i.e., the gold salt auranofin and the investigational chemotherapeutic agent buthionione sulfoximine (BSO). We found that these drugs were able to decrease the number of activated CD4 + T cells, which are preferential targets for HIV infection. Then, efficient immune responses against the virus were developed in the macaques, which remained healthy during 2 years of follow-up. This result may furnish another building block for future attempts to cure HIV/AIDS.
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- 2015
22. Characterization of protective immune response elicited by a trimeric envelope protein from an Indian clade C HIV-1 isolate in rhesus macaques
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Veena Menon, Ranajit Pal, Rangasamy Sneha Priya, Celia C. LaBranche, Vaniambadi S. Kalyanaraman, David C. Montefiori, and Sundarasamy Mahalingam
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Antigenicity ,Immunogen ,medicine.drug_class ,medicine.medical_treatment ,animal experiment ,Human immunodeficiency virus type 1 subtype C ,Molecular Sequence Data ,Human immunodeficiency virus (HIV) ,rhesus monkey ,Monoclonal antibody ,medicine.disease_cause ,antibody titer ,immune response ,Neutralization ,Immune system ,Viral Envelope Proteins ,protein purification ,medicine ,Animals ,antigen binding ,bio-layer interferometry ,controlled study ,virus envelope protein ,virus isolation ,nonhuman ,General Veterinary ,biology ,animal model ,gp145 protein ,Monkey Diseases ,antibody response ,Sequence Analysis, DNA ,interferometry ,Macaca mulatta ,Virology ,unclassified drug ,priority journal ,monoclonal antibody ,antigenicity ,virus neutralization ,HIV-1 ,biology.protein ,Animal Science and Zoology ,Antibody ,CD4 antigen ,Adjuvant - Abstract
Background: Recent preclinical studies have demonstrated the use of properly folded trimeric HIV-1 envelope proteins as immunogen for eliciting protecting immune response in macaques. Methods: Trimeric gp145 protein of Indian clade C HIV-1 (93IN101) was characterized for antigenicity by evaluating its binding to sCD4, and several monoclonal antibodies to HIV-1 by bio-layer interferometry. Ten macaques were immunized four times with purified gp145 in adjuplex adjuvant, and serum antibodies were characterized for binding to gp145 and neutralization. Immunized macaques were subjected to weekly low-dose vaginal challenge with SHIV1157-ipEL-p for 8 weeks. Results: Env protein elicited strong antibody response in macaques. Following challenge, seven of ten immunized macaques resisted challenge, while six of eight control animals were infected. Conclusions: Env proteins from a clade C Indian isolate can elicit protective immune response and therefore may be a candidate for inclusion in a multiclade-based HIV-1 vaccine. � 2015 John Wiley & Sons A/S.
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- 2015
23. Therapeutic envelope vaccination in combination with antiretroviral therapy temporarily rescues SIV-specific CD4+T-cell-dependent natural killer cell effector responses in chronically infected rhesus macaques
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Peng Xiao, Marjorie Robert-Guroff, Diego A. Vargas-Inchaustegui, Thorsten Demberg, and Ranajit Pal
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CD4-Positive T-Lymphocytes ,Male ,Time Factors ,Necrosis ,animal diseases ,viruses ,Immunology ,Cell ,Simian Acquired Immunodeficiency Syndrome ,Biology ,medicine.disease_cause ,Natural killer cell ,Immune system ,medicine ,Animals ,Immunology and Allergy ,Immunity, Cellular ,Tumor Necrosis Factor-alpha ,Vaccination ,Gene Products, env ,Original Articles ,Simian immunodeficiency virus ,Macaca mulatta ,Virology ,CD4 Lymphocyte Count ,Killer Cells, Natural ,medicine.anatomical_structure ,Immunization ,Female ,Simian Immunodeficiency Virus ,medicine.symptom ,Viral load - Abstract
Summary Natural killer (NK) cells are essential components of the immune system, and due to their rapid response potential, can have a great impact during early anti-viral immune responses. We have previously shown that interleukin-2-dependent NK and CD4+ T-cell co-operative immune responses exist in long-term simian immunodeficiency virus (SIV) -infected controlling macaques and can be rescued in SIV-infected non-controlling macaques by a short course of antiretroviral therapy (ART). Given that co-operative responses may play an important role in disease prevention and therapeutic treatment, in the present study we sought to determine if these responses can be enhanced in chronically SIV-infected macaques by vaccination with a single-dose of envelope protein given during ART. To this end, we treated 14 chronically SIV-infected macaques with ART for 11 weeks and gave 10 of these macaques a single intramuscular dose of SIV gp120 at week 9 of treatment. ART significantly decreased plasma and mucosal viral loads, increased the numbers of circulating CD4+ T cells in all macaques, and increased T-cell-dependent envelope- and gag-specific interferon-γ and tumour necrosis factor-α production by circulatory CD56+ NK cells. The therapeutic envelope immunization resulted in higher envelope-specific responses compared with those in macaques that received ART only. Functional T-cell responses restored by ART and therapeutic Env immunization were correlated with transiently reduced plasma viraemia levels following ART release. Collectively our results indicate that SIV-specific T-cell-dependent NK cell responses can be efficiently rescued by ART in chronically SIV-infected macaques and that therapeutic immunization may be beneficial in previously vaccinated individuals.
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- 2015
24. Corrigendum to: Infection of monkeys by simian-human immunodeficiency viruses with transmitted/founder clade C HIV-1 envelopes [Virology 475 (2015) 37–45]
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Hanne Anderson, Matthias Pauthner, Khoa Le, Beatrice H. Hahn, Ranajit Pal, Christie Brinkley, Mark G. Lewis, Joseph Sodroski, Devin Sok, Sampa Santra, Barton F. Haynes, Christy L. Lavine, Burton, Seaman, Mohammed Asmal, Linh Mach, Corinne Luedemann, George M. Shaw, Harikrishnan Balachandran, Thomas N. Denny, and Norman L. Letvin
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viruses ,Virology ,Human immunodeficiency virus (HIV) ,medicine ,virus diseases ,Biology ,Simian ,medicine.disease_cause ,biology.organism_classification - Published
- 2015
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25. Infection of monkeys by simian-human immunodeficiency viruses with transmitted/founder clade C HIV-1 envelopes
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Christy L. Lavine, Sampa Santra, Hanne Anderson, George M. Shaw, Matthias Pauthner, Christie Brinkley, Barton F. Haynes, Khoa Le, Michael S. Seaman, Dennis R. Burton, Thomas N. Denny, Linh Mach, Norman L. Letvin, Joseph Sodroski, Mark G. Lewis, Beatrice H. Hahn, Ranajit Pal, Mohammed Asmal, Harikrishnan Balachandran, Corinne Luedemann, and Devin Sok
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Gene Expression Regulation, Viral ,animal diseases ,viruses ,Simian Acquired Immunodeficiency Syndrome ,Heterologous ,Viremia ,Biology ,Simian ,medicine.disease_cause ,Neutralization ,Article ,Viral Envelope Proteins ,stomatognathic system ,Virology ,medicine ,Animals ,Humans ,Transmitted/founder Env ,Gene ,Immunodeficiency ,Phylogeny ,HIV-1 Clade C ,virus diseases ,Simian immunodeficiency virus ,biology.organism_classification ,medicine.disease ,Mucosal transmission ,3. Good health ,HEK293 Cells ,SHIV ,Mutation ,biology.protein ,HIV-1 ,Simian Immunodeficiency Virus ,Antibody - Abstract
Simian-human immunodeficiency viruses (SHIVs) that mirror natural transmitted/founder (T/F) viruses in man are needed for evaluation of HIV-1 vaccine candidates in nonhuman primates. Currently available SHIVs contain HIV-1 env genes from chronically-infected individuals and do not reflect the characteristics of biologically relevant HIV-1 strains that mediate human transmission. We chose to develop clade C SHIVs, as clade C is the major infecting subtype of HIV-1 in the world. We constructed 10 clade C SHIVs expressing Env proteins from T/F viruses. Three of these ten clade C SHIVs (SHIV KB9 C3, SHIV KB9 C4 and SHIV KB9 C5) replicated in naïve rhesus monkeys. These three SHIVs are mucosally transmissible and are neutralized by sCD4 and several HIV-1 broadly neutralizing antibodies. However, like natural T/F viruses, they exhibit low Env reactivity and a Tier 2 neutralization sensitivity. Of note, none of the clade C T/F SHIVs elicited detectable autologous neutralizing antibodies in the infected monkeys, even though antibodies that neutralized a heterologous Tier 1 HIV-1 were generated. Challenge with these three new clade C SHIVs will provide biologically relevant tests for vaccine protection in rhesus macaques.
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- 2015
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26. Fabrication and evaluation of structural film adhesive using oxazolidinone modified novolac epoxy resin
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Suraj Sudhi, Ranajit Pal, and Rajeev Raghavan
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Fabrication ,Materials science ,Polymers and Plastics ,visual_art ,Materials Chemistry ,visual_art.visual_art_medium ,General Chemistry ,Adhesive ,Epoxy ,Composite material ,Surfaces, Coatings and Films - Published
- 2019
27. Immune targeting of PD-1hi expressing cells during and after antiretroviral therapy in SIV-infected rhesus macaques
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Solomon Langermann, Eun Mi Lee, Katherine McKinnon, Thorsten Demberg, Alison Hogg, Linda Liu, Lauren Hudacik, Yongjun Sui, Peng Xiao, Janet DiPasquale, Ranajit Pal, David Venzon, Jay A. Berzofsky, Marjorie Robert-Guroff, Diego A. Vargas-Inchaustegui, and Egidio Brocca-Cofano
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medicine.medical_treatment ,T cell ,T-Lymphocytes ,Programmed Cell Death 1 Receptor ,Simian Acquired Immunodeficiency Syndrome ,Viremia ,Biology ,Peripheral blood mononuclear cell ,Article ,Immunomodulation ,Antigen ,Antiretroviral Therapy, Highly Active ,Virology ,PD-1 ,medicine ,Animals ,Immunologic Factors ,Immunotherapy ,Viral Load ,medicine.disease ,Macaca mulatta ,Treg ,medicine.anatomical_structure ,Anti-Retroviral Agents ,Immunology ,Lymph ,Viral load ,CD8 ,ART - Abstract
High-level T cell expression of PD-1 during SIV infection is correlated with impaired proliferation and function. We evaluated the phenotype and distribution of T cells and Tregs during antiretroviral therapy plus PD-1 modulation (using a B7-DC-Ig fusion protein) and post-ART. Chronically SIV-infected rhesus macaques received: 11 weeks of ART (Group A); 11 weeks of ART plus B7-DC-Ig (Group B); 11 weeks of ART plus B7-DC-Ig, then 12 weeks of B7-DC-Ig alone (Group C). Continuous B7-DC-Ig treatment (Group C) decreased rebound viremia post-ART compared to pre-ART levels, associated with decreased PD-1(hi) expressing T cells and Tregs in PBMCs, and PD-1(hi) Tregs in lymph nodes. It transiently decreased expression of Ki67 and α4β7 in PBMC CD4(+) and CD8(+) Tregs for up to 8 weeks post-ART and maintained Ag-specific T-cell responses at low levels. Continued immune modulation targeting PD-1(hi) cells during and post-ART helps maintain lower viremia, keeps a favorable T cell/Treg repertoire and modulates antigen-specific responses.
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- 2013
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28. Impact of antibody quality and anamnestic response on viremia control post-challenge in a combined Tat/Env vaccine regimen in rhesus macaques
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Diego A. Vargas-Inchaustegui, Indresh K. Srivastava, David Venzon, Eun Mi Lee, Vaniambadi S. Kalyanaraman, David C. Montefiori, Janet DiPasquale, Thorsten Demberg, Irene Kalisz, Ruth M. Ruprecht, Stanley Aladi, Susan W. Barnett, Seraphin Kuate, Ranajit Pal, Marjorie Robert-Guroff, and Egidio Brocca-Cofano
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HIV vaccine ,HIV Tat ,viruses ,Simian Acquired Immunodeficiency Syndrome ,chemical and pharmacologic phenomena ,Viremia ,Antibodies, Viral ,Article ,03 medical and health sciences ,0302 clinical medicine ,Adjuvants, Immunologic ,Virology ,Rhesus macaque ,SHIV challenge ,medicine ,Animals ,Humans ,Avidity ,Alum adjuvant ,Neutralizing antibody ,030304 developmental biology ,0303 health sciences ,biology ,Immunogenicity ,SAIDS Vaccines ,Gene Products, env ,virus diseases ,medicine.disease ,Antibodies, Neutralizing ,Macaca mulatta ,3. Good health ,HIV Envelope ,Gene Products, tat ,Immunology ,Humoral immunity ,biology.protein ,Alum Compounds ,Simian Immunodeficiency Virus ,Antibody ,ADCC ,Immunologic Memory ,030215 immunology - Abstract
Previously, priming rhesus macaques with Adenovirus type 5 host range mutant-recombinants encoding Tat and Env and boosting with Tat and Env protein in MPL-SE controlled chronic viremia by 4 logs following homologous intravenous SHIV89.6P challenge. Here we evaluated Tat, Env, and Tat/Env regimens for immunogenicity and protective efficacy using clade C Env, alum adjuvant, and a heterologous intrarectal SHIV1157ipd3N4 challenge. Despite induction of strong cellular and humoral immunity, Tat/Env group T and B-cell memory responses were not significantly enhanced over Tat- or Env-only groups. Lack of viremia control post-challenge was attributed to lower avidity Env antibodies and no anamnestic ADCC response or SHIV1157ipd3N4 neutralizing antibody development post-challenge. Poor biologic activity of the Tat immunogen may have impaired Tat immunity. In the absence of sterilizing immunity, strong anamnestic responses to heterologous virus can help control viremia. Both antibody breadth and optimal adjuvanticity are needed to elicit high-quality antibody for protective efficacy.
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- 2013
29. Comparative Characterization of Transfection- and Infection-Derived Simian Immunodeficiency Virus Challenge Stocks for In Vivo Nonhuman Primate Studies
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Ronald C. Desrosiers, Brandon F. Keele, Elena Chertova, Gregory Q. Del Prete, Carolyn Reid, Christopher J. Miller, Matthew Scarlotta, Luis D. Giavedoni, Laura Newman, Laura M. Parodi, Dan H. Barouch, James D. Roser, Ranajit Pal, Michael Piatak, Jeffrey D. Lifson, Kelli Oswald, David H. O’Connor, and Preston A. Marx
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Virus Cultivation ,viruses ,Immunology ,Simian Acquired Immunodeficiency Syndrome ,Biology ,Transfection ,medicine.disease_cause ,Microbiology ,Virus ,In vivo ,Virology ,medicine ,Animals ,Infectivity ,Primate Diseases ,Genetic Variation ,Sequence Analysis, DNA ,Viral Load ,Simian immunodeficiency virus ,Titer ,Genetic Diversity and Evolution ,Insect Science ,Simian Immunodeficiency Virus ,Viral load - Abstract
Simian immunodeficiency virus (SIV) stocks for in vivo nonhuman primate models of AIDS are typically generated by transfection of 293T cells with molecularly cloned viral genomes or by expansion in productively infected T cells. Although titers of stocks are determined for infectivity in vitro prior to in vivo inoculation, virus production methods may differentially affect stock features that are not routinely analyzed but may impact in vivo infectivity, mucosal transmissibility, and early infection events. We performed a detailed analysis of nine SIV stocks, comprising five infection-derived SIVmac251 viral swarm stocks and paired infection- and transfected-293T-cell-derived stocks of both SIVmac239 and SIVmac766. Representative stocks were evaluated for (i) virus content, (ii) infectious titer, (iii) sequence diversity and polymorphism frequency by single-genome amplification and 454 pyrosequencing, (iv) virion-associated Env content, and (v) cytokine and chemokine content by 36-plex Luminex analysis. Regardless of production method, all stocks had comparable particle/infectivity ratios, with the transfected-293T stocks possessing the highest overall virus content and infectivity titers despite containing markedly lower levels of virion-associated Env than infection-derived viruses. Transfected-293T stocks also contained fewer and lower levels of cytokines and chemokines than infection-derived stocks, which had elevated levels of multiple analytes, with substantial variability among stocks. Sequencing of the infection-derived SIVmac251 stocks revealed variable levels of viral diversity between stocks, with evidence of stock-specific selection and expansion of unique viral lineages. These analyses suggest that there may be underappreciated features of SIV in vivo challenge stocks with the potential to impact early infection events, which may merit consideration when selecting virus stocks for in vivo studies.
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- 2013
30. Antibodies with High Avidity to the gp120 Envelope Protein in Protection from Simian Immunodeficiency Virus SIV mac251 Acquisition in an Immunization Regimen That Mimics the RV-144 Thai Trial
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Ranajit Pal, Poonam Pegu, Georgia D. Tomaras, Guido Ferrari, Nelson L. Michael, Yongjun Guan, Mangala Rao, Monica Vaccari, Jeffrey D. Lifson, Jim Tartaglia, David Venzon, Jerome H. Kim, Maria Grazia Ferrari, S. Munir Alam, Melvin N. Doster, Brandon F. Keele, Lauren Hudacik, Shari N. Gordon, David C. Montefiori, Claudio Fenizia, Erik Billings, Genoveffa Franchini, Stephen Whitney, and Donald Stablein
- Subjects
CD4-Positive T-Lymphocytes ,viruses ,Immunology ,Antibody Affinity ,Simian Acquired Immunodeficiency Syndrome ,CD8-Positive T-Lymphocytes ,Biology ,Antibodies, Viral ,medicine.disease_cause ,Microbiology ,Virus ,Viral Envelope Proteins ,Virology ,Vaccines and Antiviral Agents ,medicine ,Animals ,Avidity ,RV 144 ,Membrane Glycoproteins ,SAIDS Vaccines ,virus diseases ,Simian immunodeficiency virus ,Vaccination ,Immunization ,Insect Science ,biology.protein ,Macaca ,Antibody - Abstract
The recombinant canarypox vector, ALVAC-HIV, together with human immunodeficiency virus (HIV) gp120 envelope glycoprotein, has protected 31.2% of Thai individuals from HIV acquisition in the RV144 HIV vaccine trial. This outcome was unexpected, given the limited ability of the vaccine components to induce CD8 + T-cell responses or broadly neutralizing antibodies. We vaccinated macaques with an immunization regimen intended to mimic the RV144 trial and exposed them intrarectally to a dose of the simian immunodeficiency virus SIV mac251 that transmits few virus variants, similar to HIV transmission to humans. Vaccination induced anti-envelope antibodies in all vaccinees and CD4 + and CD8 + T-cell responses. Three of the 11 macaques vaccinated with ALVAC-SIV/gp120 were protected from SIV mac251 acquisition, but the result was not significant. The remaining vaccinees were infected and progressed to disease. The magnitudes of vaccine-induced SIV mac251 -specific T-cell responses and binding antibodies were not significantly different between protected and infected animals. However, sera from protected animals had higher avidity antibodies to gp120, recognized the variable envelope regions V1/V2, and reduced SIV mac251 infectivity in cells that express high levels of α 4 β 7 integrins, suggesting a functional role of antibodies to V2. The current results emphasize the utility of determining the titer of repeated mucosal challenge in the preclinical evaluation of HIV vaccines.
- Published
- 2013
31. Derivation and Characterization of Pathogenic Transmitted/Founder Molecular Clones from Simian Immunodeficiency Virus SIVsmE660 and SIVmac251 following Mucosal Infection
- Author
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Carolyn Reid, Brandon F. Keele, Michael J. Lopker, Hui Li, Laura Newman, Gregory Q. Del Prete, Leslie Lipkey, Julie M. Decker, Beatrice H. Hahn, Jeffrey D. Lifson, George M. Shaw, Deborah E. Weiss, Celine Camus, Katharine J. Bar, Shuyi Wang, Ranajit Pal, Gerald H. Learn, Juliet L. Easlick, and Jacob D. Estes
- Subjects
0301 basic medicine ,Genotype ,T cell ,viruses ,Immunology ,Simian Acquired Immunodeficiency Syndrome ,Biology ,medicine.disease_cause ,Virus Replication ,Microbiology ,Virus ,Pathogenesis ,03 medical and health sciences ,Virology ,medicine ,Animals ,Immunodeficiency ,Transmission (medicine) ,Simian immunodeficiency virus ,medicine.disease ,Phenotype ,Macaca mulatta ,030104 developmental biology ,medicine.anatomical_structure ,Lymphatic system ,Genetic Diversity and Evolution ,Insect Science ,Simian Immunodeficiency Virus - Abstract
Currently available simian immunodeficiency virus (SIV) infectious molecular clones (IMCs) and isolates used in nonhuman primate (NHP) models of AIDS were originally derived from infected macaques during chronic infection or end stage disease and may not authentically recapitulate features of transmitted/founder (T/F) genomes that are of particular interest in transmission, pathogenesis, prevention, and treatment studies. We therefore generated and characterized T/F IMCs from genetically and biologically heterogeneous challenge stocks of SIVmac251 and SIVsmE660. Single-genome amplification (SGA) was used to identify full-length T/F genomes present in plasma during acute infection resulting from atraumatic rectal inoculation of Indian rhesus macaques with low doses of SIVmac251 or SIVsmE660. All 8 T/F clones yielded viruses that were infectious and replication competent in vitro , with replication kinetics similar to those of the widely used chronic-infection-derived IMCs SIVmac239 and SIVsmE543. Phenotypically, the new T/F virus strains exhibited a range of neutralization sensitivity profiles. Four T/F virus strains were inoculated into rhesus macaques, and each exhibited typical SIV replication kinetics. The SIVsm T/F viruses were sensitive to TRIM5α restriction. All T/F viruses were pathogenic in rhesus macaques, resulting in progressive CD4 + T cell loss in gastrointestinal tissues, peripheral blood, and lymphatic tissues. The animals developed pathological immune activation; lymphoid tissue damage, including fibrosis; and clinically significant immunodeficiency leading to AIDS-defining clinical endpoints. These T/F clones represent a new molecular platform for the analysis of virus transmission and immunopathogenesis and for the generation of novel “bar-coded” challenge viruses and next-generation simian-human immunodeficiency viruses that may advance the HIV/AIDS vaccine agenda. IMPORTANCE Nonhuman primate research has relied on only a few infectious molecular clones for a myriad of diverse research projects, including pathogenesis, preclinical vaccine evaluations, transmission, and host-versus-pathogen interactions. With new data suggesting a selected phenotype of the virus that causes infection (i.e., the transmitted/founder virus), we sought to generate and characterize infectious molecular clones from two widely used simian immunodeficiency virus lineages (SIVmac251 and SIVsmE660). Although the exact requirements necessary to be a T/F virus are not yet fully understood, we generated cloned viruses with all the necessary characteristic of a successful T/F virus. The cloned viruses revealed typical acute and set point viral-load dynamics with pathological immune activation, lymphoid tissue damage progressing to significant immunodeficiency, and AIDS-defining clinical endpoints in some animals. These T/F clones represent a new molecular platform for studies requiring authentic T/F viruses.
- Published
- 2016
32. Development of real-time PCR for quantitation of simian immunodeficiency virus 2-LTR circles
- Author
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Ranajit Pal, Hye Kyung Chung, Michael Lee, Marcelo J. Kuroda, and Woong-Ki Kim
- Subjects
0301 basic medicine ,Male ,viruses ,030106 microbiology ,Simian Acquired Immunodeficiency Syndrome ,Proviral dna ,Biology ,medicine.disease_cause ,Real-Time Polymerase Chain Reaction ,Peripheral blood mononuclear cell ,Macaque ,Pathogenesis ,03 medical and health sciences ,Proviruses ,biology.animal ,medicine ,Animals ,General Veterinary ,Base Sequence ,Brain ,Reproducibility of Results ,Simian immunodeficiency virus ,medicine.disease ,Virology ,Orders of magnitude (mass) ,030104 developmental biology ,Real-time polymerase chain reaction ,Immunology ,DNA, Viral ,Leukocytes, Mononuclear ,Animal Science and Zoology ,Female ,Simian Immunodeficiency Virus ,DNA, Circular ,Sequence Alignment ,Encephalitis - Abstract
Background Non-human primates infected with simian immunodeficiency virus (SIV) represent a robust model to evaluate pre-clinical efficacy of HIV-1 preventive strategies and to determine the size of reservoir. Methods We developed a real-time qPCR assay to specifically quantify episomal 2-LTR circular DNA in peripheral blood mononuclear cells and brain tissues from SIV-infected macaques. Results This assay has sensitivity, accuracy and reproducibility over seven orders of magnitude. High copy numbers of SIV 2-LTR circles were correlated to high proviral DNA levels in brains of two SIV encephalitic animals. In contrast, no 2-LTR circles were detectable in two SIV-infected animals with no sign of encephalitis or two animals that had mild encephalitis with low levels of proviral DNA. Conclusions These results suggest that simultaneous application of total proviral DNA and 2-LTR circle assays provides quantitative evaluation of pathogenesis and outcome of SIV infection in macaques.
- Published
- 2016
33. An HIV gp120-CD4 Immunogen Does Not Elicit Autoimmune Antibody Responses in Cynomolgus Macaques
- Author
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Anthony L. DeVico, Jennifer Schwartz, Johnathan Misamore, Ilia Prado, Deborah T. Weiss, Timothy R. Fouts, George K. Lewis, Anthony D. Cristillo, Robert C. Gallo, Maria Cecilia Huaman, Jesse Francis, and Ranajit Pal
- Subjects
0301 basic medicine ,Microbiology (medical) ,Male ,Immunogen ,T cell ,Clinical Biochemistry ,Immunology ,Enzyme-Linked Immunosorbent Assay ,Biology ,HIV Antibodies ,HIV Envelope Protein gp120 ,medicine.disease_cause ,Epitope ,03 medical and health sciences ,0302 clinical medicine ,Antigen ,T-Lymphocyte Subsets ,medicine ,Immunology and Allergy ,Animals ,030212 general & internal medicine ,Autoantibodies ,Vaccines ,Autoantibody ,Antibody titer ,Simian immunodeficiency virus ,Virology ,Recombinant Proteins ,CD4 Lymphocyte Count ,Macaca fascicularis ,030104 developmental biology ,medicine.anatomical_structure ,CD4 Antigens ,biology.protein ,Female ,Antibody - Abstract
A promising concept for human immunodeficiency virus (HIV) vaccines focuses immunity on the highly conserved transition state structures and epitopes that appear when the HIV glycoprotein gp120 binds to its receptor, CD4. We are developing chimeric antigens (full-length single chain, or FLSC) in which gp120 and CD4 sequences are flexibly linked to allow stable intrachain complex formation between the two moieties (A. DeVico et al., Proc Natl Acad Sci U S A 104:17477–17482, 2007, doi: 10.1073/pnas.0707399104 ; T. R. Fouts et al., J Virol 74:11427–11436, 2000, doi:10.1128/JVI.74.24.11427-11436.2000 ). Proof of concept studies with nonhuman primates show that FLSC elicited heterologous protection against simian-human immunodeficiency virus (SHIV)/simian immunodeficiency virus (SIV) (T. R. Fouts et al., Proc Natl Acad Sci U S A 112:E992–E999, 2016, doi:10.1073/pnas.1423669112 ), which correlated with antibodies against transition state gp120 epitopes. Nevertheless, advancement of any vaccine that comprises gp120-CD4 complexes must consider whether the CD4 component breaks tolerance and becomes immunogenic in the autologous host. To address this, we performed an immunotoxicology study with cynomolgus macaques vaccinated with either FLSC or a rhesus variant of FLSC containing macaque CD4 sequences (rhFLSC). Enzyme-linked immunosorbent assay (ELISA) binding titers, primary CD3 + T cell staining, and temporal trends in T cell subset frequencies served to assess whether anti-CD4 autoantibody responses were elicited by vaccination. We find that immunization with multiple high doses of rhFLSC did not elicit detectable antibody titers despite robust responses to rhFLSC. In accordance with these findings, immunized animals had no changes in circulating CD4 + T cell counts or evidence of autoantibody reactivity with cell surface CD4 on primary naive macaque T cells. Collectively, these studies show that antigens using CD4 sequences to stabilize transition state gp120 structures are unlikely to elicit autoimmune antibody responses, supporting the advancement of gp120-CD4 complex-based antigens, such as FLSC, into clinical testing.
- Published
- 2016
34. Replicating Adenovirus-Simian Immunodeficiency Virus (SIV) Recombinant Priming and Envelope Protein Boosting Elicits Localized, Mucosal IgA Immunity in Rhesus Macaques Correlated with Delayed Acquisition following a Repeated Low-Dose Rectal SIV mac251 Challenge
- Author
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Marjorie Robert-Guroff, Jun Zhao, Brandon F. Keele, Seraphin Kuate, Peng Xiao, David C. Montefiori, Eun Mi Lee, Vaniambadi S. Kalyanaraman, Irene Kalisz, Claudio Fenizia, Michael A. Thomas, Janet DiPasquale, David Venzon, L. Jean Patterson, Egidio Brocca-Cofano, and Ranajit Pal
- Subjects
Antibody-dependent cell-mediated cytotoxicity ,Cellular immunity ,Immunology ,chemical and pharmacologic phenomena ,Viremia ,Simian immunodeficiency virus ,Biology ,medicine.disease_cause ,medicine.disease ,Microbiology ,Virology ,Macaque ,Immunity ,Insect Science ,biology.animal ,Humoral immunity ,medicine ,Avidity - Abstract
We have shown that sequential replicating adenovirus type 5 host range mutant human immunodeficiency virus/simian immunodeficiency virus (HIV/SIV) recombinant priming delivered first intranasally (i.n.) plus orally and then intratracheally (i.t.), followed by envelope protein boosting, elicits broad cellular immunity and functional, envelope-specific serum and mucosal antibodies that correlate with protection from high-dose SIV and simian/human immunodeficiency virus (SHIV) challenges in rhesus macaques. Here we extended these studies to compare the standard i.n./i.t. regimen with additional mucosal administration routes, including sublingual, rectal, and vaginal routes. Similar systemic cellular and humoral immunity was elicited by all immunization routes. Central and effector memory T cell responses were also elicited by the four immunization routes in bronchoalveolar lavage fluid and jejunal, rectal, and vaginal tissue samples. Cellular responses in vaginal tissue were more compartmentalized, being induced primarily by intravaginal administration. In contrast, all immunization routes elicited secretory IgA (sIgA) responses at multiple mucosal sites. Following a repeated low-dose intrarectal (i.r.) challenge with SIV mac251 at a dose transmitting one or two variants, protection against acquisition was not achieved except in one macaque in the i.r. immunized group. All immunized macaques exhibited reduced peak viremia compared to that of controls, correlated inversely with prechallenge serum antienvelope avidity, antibody-dependent cellular cytotoxicity (ADCC) titers, and percent antibody-dependent cell-mediated viral inhibition. Both antibody avidity and ADCC titers were correlated with the number of exposures required for infection. Notably, we show for the first time a significant correlation of vaccine-induced sIgA titers in rectal secretions with delayed acquisition. Further investigation of the characteristics and properties of the sIgA should elucidate the mechanism leading to this protective effect.
- Published
- 2012
35. Prevention of vaginal SHIV transmission in macaques by a live recombinant Lactobacillus
- Author
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Brigitte E Sanders-Beer, Rosa R. Yu, Laurel A. Lagenaur, Beda Brichacek, Dean H. Hamer, Peter P. Lee, Xiaowen Liu, Ranajit Pal, David Venzon, and Yang Liu
- Subjects
Recombinant Fusion Proteins ,Immunology ,Antiviral protein ,HIV Infections ,Biology ,medicine.disease_cause ,Article ,Microbiology ,03 medical and health sciences ,Bacterial Proteins ,Lactobacillus ,medicine ,Animals ,Humans ,Immunology and Allergy ,Immunity, Mucosal ,030304 developmental biology ,0303 health sciences ,030306 microbiology ,Lactobacillus jensenii ,HIV ,Breakthrough infection ,Viral Load ,Virus Internalization ,Simian immunodeficiency virus ,biology.organism_classification ,Macaca mulatta ,Virology ,3. Good health ,Entry inhibitor ,Administration, Intravaginal ,Disease Models, Animal ,medicine.anatomical_structure ,Vagina ,Cytokines ,Female ,Simian Immunodeficiency Virus ,Carrier Proteins ,Genetic Engineering ,Viral load ,medicine.drug - Abstract
Most human immunodeficiency virus (HIV) transmissions in women occur through the cervicovaginal mucosa, which is coated by a bacterial biofilm including Lactobacillus. This commensal bacterium has a role in maintaining a healthy mucosa and can be genetically engineered to produce antiviral peptides. Here, we report a 63% reduction in transmission of a chimeric simian/HIV (SHIV(SF162P3)) after repeated vaginal challenges of macaques treated with Lactobacillus jensenii expressing the HIV-1 entry inhibitor cyanovirin-N. Furthermore, peak viral loads in colonized macaques with breakthrough infection were reduced sixfold. Colonization and prolonged antiviral protein secretion by the genetically engineered lactobacilli did not cause any increase in proinflammatory markers. These findings lay the foundation for an accessible and durable approach to reduce heterosexual transmission of HIV in women, which is coitally independent, inexpensive, and enhances the natural protective effects of the vaginal microflora.
- Published
- 2011
36. Synergistic inhibition of R5 HIV-1 by maraviroc and CCR5 antibody HGS004 in primary cells: implications for treatment and prevention
- Author
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Nhut Le, Alonso Heredia, Anthony L. DeVico, Robert R. Redfield, Ranajit Pal, Olga Latinovic, James S. Foulke, and Marvin S. Reitz
- Subjects
Receptors, CCR5 ,viruses ,Immunology ,Integrase inhibitor ,HIV Infections ,CCR5 receptor antagonist ,Biology ,Pharmacology ,Article ,Maraviroc ,chemistry.chemical_compound ,Cyclohexanes ,Drug Resistance, Viral ,medicine ,Humans ,Immunology and Allergy ,Potency ,EC50 ,virus diseases ,Triazoles ,Raltegravir ,Virology ,body regions ,Infectious Diseases ,chemistry ,PRO 140 ,CCR5 Receptor Antagonists ,HIV-1 ,Drug Therapy, Combination ,Antagonism ,medicine.drug - Abstract
CCR5 serves as the main coreceptor for the predominantly transmitted and most persistent HIV-1, the R5 strains. Maraviroc (MVC) is the only licensed CCR5 inhibitor [1, 2]. CCR5 antibodies PRO 140 and HGS004 have significantly reduced HIV-1 RNA in clinical trials of patients infected with R5 HIV-1 only [3-6]. However, HGS004 reduced HIV-1 RNA in only 54% of patients. Retrospectively, it was found that sensitivity of the patients’ viruses to HGS004 predicted antiviral responses [6]. We tested the hypothesis that the combination of HGS004 and MVC has antiviral synergy, thereby increasing the potency of both drugs and reducing their effective doses. This hypothesis is based on the observation that HGS004 and MVC recognize separate regions of CCR5, with HGS004 binding to the second extracellular loop [6], and MVC to the transmembrane region [7-9]. We tested the antiviral activity of HGS004 alone and in the presence of varying concentrations of MVC. For comparison, we evaluated HGS004 in combination with the integrase inhibitor Raltegravir (RAL). Both MVC and RAL were used at concentrations spanning their EC50s. We performed these assays using PHA-activated peripheral blood mononuclear cells (PBMCs) infected with the R5 HIV-1 strains BaL or CC1/85. Data for HIV-1 BaL are shown in Fig 1a (upper panels). In the absence of MVC or RAL, HGS004 inhibited HIV-1 BaL with an EC50 value of 31.3 µg/ml. However, in the presence of increasing concentrations of MVC, HGS004 EC50s were successively lowered to 4.4, 0.7, 0.1, 0.05 and 0.03 μg/ml. In contrast, RAL had little effect on HGS004 potency, with HGS004 EC50s of 26, 24, 22, 15 and 9.8 μg/ml in the presence of increasing RAL. Neither the drug alone nor the drug combinations treatments were toxic to cells as demonstrated by MTT assays (not shown). Together, the marked leftwards shift of the HGS004 viral inhibition curves in the presence of MVC compared to RAL suggested that HGS004 and MVC are synergistic against HIV-1 BaL. Figure 1 Antiviral interactions between HGS004 and MVC versus HGS004 and RAL against R5 HIV-1 in primary cells To determine whether inhibition by HGS004 and MVC or RAL might indeed be synergistic, we performed a three-dimensional analysis using the method of Prichard and Shipman [10]. For the HGS004/MVC and HGS004/RAL combinations, the 96% confidence synergy plots, after Bonferroni adjustment, are shown in Fig 1a (lower panels). The combination of HGS004 and MVC had antiviral synergy across the entire concentration grid (synergy volume of 522). In contrast, the combination of HGS004 and RAL was mainly additive (synergy volume of only 19). Similar results to those with HIV-1 BaL were obtained with HIV-1 CC1/85, a R5 HIV-1 primary patient isolate [11] (Fig 1b, upper panels). HGS004 EC50s were 40 μg/ml in antibody alone treatment, but only 3.75, 0.22, 0.04 and 0.03 μg/ml in combinations containing the indicated MVC concentrations. HGS004 EC50s were 30, 26, 59, 9.2 and 1.5 nM in combinations containing RAL, suggesting some increased antibody potency at high concentrations of RAL (Fig 1b, upper panels). Inhibition of CC1/85 by the HGS004/MVC and HGS004/RAL combinations gave synergy volumes of 502 and 54, respectively (Fig 1b, lower panels). To confirm the synergy data, we analyzed the drug combinations by the Median Effect Principle [12], which evaluates drug combinations at fixed ratios and allows calculation of Combination Indices (CI). CI=1 indicates additivity, CI>1 indicates antagonism, and CI
- Published
- 2011
37. Induction of mucosal and systemic antibody and T-cell responses following prime-boost immunization with novel adjuvanted human immunodeficiency virus-1-vaccine formulations
- Author
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Brad Lewis, Lauren Hudacik, Maria Grazia Ferrari, Nikolai Petrovsky, Phillip D. Markham, DeVon Thompson, Ranajit Pal, Anthony D. Cristillo, Britany Bowen, and Lindsey Galmin
- Subjects
Immunoglobulin A ,Cellular immunity ,Sexual transmission ,T-Lymphocytes ,medicine.medical_treatment ,Immunization, Secondary ,HIV Antibodies ,HIV Envelope Protein gp120 ,Injections, Intramuscular ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Adjuvants, Immunologic ,Neutralization Tests ,Immunity ,Virology ,Vaccines, DNA ,medicine ,Animals ,Immunity, Mucosal ,Administration, Intranasal ,030304 developmental biology ,AIDS Vaccines ,Mice, Inbred BALB C ,0303 health sciences ,biology ,Animal ,Vaccination ,3. Good health ,Immunoglobulin G ,Vaccines, Subunit ,Immunology ,HIV-1 ,biology.protein ,Female ,Nasal administration ,Adjuvant ,030215 immunology - Abstract
As sexual transmission of human immunodeficiency virus-1 (HIV-1) occurs via the mucosa, an ideal HIV-1 vaccine should induce both mucosal and systemic immunity. We therefore sought to evaluate the induction of mucosal responses using a DNA env prime–gp120 protein boost approach in which sequential nasal and parenteral protein administration was performed with two novel carbohydrate-based adjuvants. These adjuvants, Advax-M and Advax-P, were specifically designed for mucosal and systemic immune enhancement, respectively. Murine intranasal immunization with gp120/Advax-M adjuvant elicited gp120-specific IgA in serum and mucosal secretions that was markedly enhanced by DNA priming. Boosting of DNA-primed mice with gp120/Advax-M and gp120/Advax-P by sequential intranasal and intramuscular immunization, or vice versa, elicited persistent mucosal gp120-specific IgA, systemic IgG and memory T- and B-cell responses. Induction of homologous, but not heterologous, neutralizing activity was noted in the sera of all immunized groups. While confirmation of efficacy is required in challenge studies using non-human primates, these results suggest that the combination of DNA priming with sequential nasal and parenteral protein boosting, with appropriate mucosal and systemic adjuvants, could generate strong mucosal and systemic immunity and may block HIV-1 mucosal transmission and infection.
- Published
- 2010
38. Molecular methods for evaluation of virological status of nonhuman primates challenged with simian immunodeficiency or simian-human immunodeficiency viruses
- Author
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Ranajit Pal, Lindsay Finke, Britany Bowen, Phillip D. Markham, Hye-Kyung Chung, Lauren Hudacik, Anthony D. Cristillo, Jessica Livesay, John J. Suschak, Eun Mi Lee, and Lindsey Galmin
- Subjects
Lymphoid Tissue ,viruses ,Biology ,medicine.disease_cause ,Recombinant virus ,Sensitivity and Specificity ,Virus ,Plasma ,Proviruses ,Virology ,medicine ,Animals ,Self-Sustained Sequence Replication ,Immunodeficiency ,Temperature ,Reproducibility of Results ,RNA ,Viral Load ,Provirus ,Simian immunodeficiency virus ,medicine.disease ,biology.organism_classification ,Macaca mulatta ,NASBA ,DNA, Viral ,Immunology ,Lentivirus ,HIV-1 ,Leukocytes, Mononuclear ,RNA, Viral ,Simian Immunodeficiency Virus - Abstract
Nonhuman primates represent a robust model to evaluate preclinical efficacy of HIV-1 vaccine and therapeutic strategies. Plasma and tissue viral RNA as well as tissue proviral DNA load are key parameters in assessing efficacy of vaccines and therapeutics against simian immunodeficiency virus (SIV) or simian-human immunodeficiency virus (SHIV) challenge. To quantitate SIV RNA in plasma and tissues, an isothermal nucleic acid sequence-based amplification (NASBA) method using real-time detection of amplified RNA with molecular beacons was developed. This assay has accuracy and reproducibility over seven orders of magnitude and has advantages over the electrochemiluminescence-based NASBA assay described previously, both in terms of higher throughput and sensitivity. Reproducibility and accuracy were also demonstrated for a TaqMan real-time PCR assay for quantitating proviral DNA load in PBMCs and lymphoid tissues. In infected macaques, the level of plasma viremia correlated with the tissue viral RNA but not always with proviral DNA loads. Further, animals with undetectable levels of viral RNA in plasma and proviral DNA in tissues, showed no sign of seroconversion and activation of Gag-specific CD8+ or CD4+ T cells in peripheral blood. These results suggest that simultaneous application of real-time NASBA and PCR assays provides quantitative evaluation of challenge outcome in macaques.
- Published
- 2010
39. Novolac epoxy resin from 4,4′-dihydroxybenzophenone: Thermal, thermomechanical, interfacial, and cure kinetics with DGEBA/DICY blend
- Author
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Suraj Sudhi, Ranajit Pal, Monisha Baby, and Nimmy K. Francis
- Subjects
Materials science ,Polymers and Plastics ,Kinetics ,02 engineering and technology ,General Chemistry ,Epoxy ,010402 general chemistry ,021001 nanoscience & nanotechnology ,01 natural sciences ,0104 chemical sciences ,Surfaces, Coatings and Films ,4,4'-Dihydroxybenzophenone ,chemistry.chemical_compound ,chemistry ,visual_art ,Thermal ,Materials Chemistry ,visual_art.visual_art_medium ,Composite material ,0210 nano-technology ,Glass transition - Published
- 2017
40. Characterization of vaginal transmission of a simian human immunodeficiency virus (SHIV) encoding the reverse transcriptase gene from HIV-1 in Chinese rhesus macaques
- Author
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Ranajit Pal, Hye-Kyung Chung, Joseph Romano, Deborah T. Weiss, Lindsey Galmin, and Jeremy Nuttall
- Subjects
Nevirapine ,Efavirenz ,Anti-HIV Agents ,viruses ,Dapivirine ,Simian Acquired Immunodeficiency Syndrome ,HIV Infections ,Spleen ,Viremia ,Microbial Sensitivity Tests ,Biology ,Virus ,law.invention ,chemistry.chemical_compound ,law ,Virology ,medicine ,Animals ,Humans ,HIV ,virus diseases ,medicine.disease ,Macaca mulatta ,HIV Reverse Transcriptase ,medicine.anatomical_structure ,chemistry ,Vagina ,Immunology ,Recombinant DNA ,RNA, Viral ,Female ,Simian Immunodeficiency Virus ,Lymph Nodes ,Lymph ,medicine.drug - Abstract
Replication competent recombinant simian-human immunodeficiency virus encoding the reverse transcriptase gene (RT SHIV) from HIV-1 was characterized for vaginal transmission in rhesus macaques. RT SHIV was shown to transmit efficiently via the vaginal route in macaques with detectable plasma viremia persisting for a year in some animals. Analyses of virus load in tissues of infected animals revealed accumulation of viral RNA in lymph nodes and spleen with levels correlating with plasma viremia. RT-SHIV was inhibited by dapivirine, nevirapine, efavirenz and tenofovir in vitro, although the effect was less pronounced with tenofovir. Virus isolated from infected animals at early and later time points had limited changes in RT sequences and exhibited similar sensitivity to RT inhibitors as the challenge virus. The vaginal transmission of RT SHIV demonstrated here suggests this virus may possibly be used in the nonhuman primate model for limited evaluation of RT inhibitors applied vaginally.
- Published
- 2009
41. Differential pathogenicity of SHIVSF162 P4infection in pig-tailed and rhesus macaques
- Author
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Susan W. Barnett, Patricia Polacino, Kay Larsen, Ranajit Pal, Kristen Bost, Zane Kraft, Atmaram H. Bandivdekar, Leonidas Stamatatos, Shiu Lok Hu, John J. Suschak, Christopher J. Miller, Lindsey Galmin, and David Anderson
- Subjects
Infectivity ,General Veterinary ,biology ,T cell ,Viremia ,Simian immunodeficiency virus ,Vaccine efficacy ,medicine.disease_cause ,biology.organism_classification ,medicine.disease ,Macaque ,Virology ,medicine.anatomical_structure ,In vivo ,biology.animal ,Lentivirus ,Immunology ,medicine ,Animal Science and Zoology - Abstract
Background Differential pathogenicity has been observed in cynomolgus and rhesus macaques following primate lentivirus infection. However, little is known about the comparative susceptibility of pig-tailed macaques to lentivirus infection and diseases. Methods We compared the in vivo infectivity and pathogenicity of a CCR5-tropic SHIVSF162 P4 after intravenous, intravaginal or intrarectal inoculation in rhesus and pig-tailed macaques. Plasma viral load, peripheral blood CD4+ T cell counts and clinical signs were monitored. Results Both rhesus and pig-tailed macaques are similarly susceptible to SHIVSF162 P4 infection by intravenous and mucosal routes. However, infection was significantly more robust in pig-tailed macaques than in rhesus, resulting in persistent viremia in 9/21 pig-tails vs. 2/24 rhesus (P
- Published
- 2008
42. The safety and tolerability of an HIV-1 DNA prime–protein boost vaccine (DP6-001) in healthy adult volunteers
- Author
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Ranajit Pal, Melissa A. O'Neill, Janice P. Adams, Shan Lu, Renita Johnson-Leva, Jaclyn K. Longtine, Mary Dawn T. Co, Phillip D. Markham, Sharone Green, Qiao Yu, Shixia Wang, Jeffrey S. Kennedy, Alan L. Rothman, and Karen J. Longtine
- Subjects
Adult ,Male ,Vasculitis ,medicine.medical_treatment ,Immunization, Secondary ,Biology ,Article ,Immune system ,Adjuvants, Immunologic ,Vaccines, DNA ,medicine ,Humans ,Hypersensitivity, Delayed ,Adverse effect ,Skin ,AIDS Vaccines ,General Veterinary ,General Immunology and Microbiology ,Public Health, Environmental and Occupational Health ,Saponins ,medicine.disease ,biology.organism_classification ,Virology ,QS21 ,Vaccination ,Human Experimentation ,Infectious Diseases ,Tolerability ,Vaccines, Subunit ,Immunology ,Lentivirus ,Molecular Medicine ,Female ,Adjuvant - Abstract
This report describes the safety observations following administration of a polyvalent DNA prime–protein boost HIV-1 vaccine formulated with adjuvant QS21. Local injection site reactions were the most common (65% of subjects), and included type IV delayed-type hypersensitivity (DTH) reactions at prior DNA inoculation sites in 12 of 28 (43%) subjects following protein vaccination. Systemic reactions revealed two cases of vasculitis temporally related to inoculation with recombinant Env protein + QS21 adjuvant. Questions remain regarding the cause of the vasculitis, but the unique DTH observation may have contributed to the high level of immune responses previously reported for this vaccine.
- Published
- 2008
43. Comparative study of Tat vaccine regimens in Mauritian cynomolgus and Indian rhesus macaques: Influence of Mauritian MHC haplotypes on susceptibility/resistance to SHIV89.6P infection
- Author
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David Venzon, Marjorie Robert-Guroff, David H. O’Connor, Ruth H. Florese, Fausto Titti, Ranajit Pal, L. Jean Patterson, Megan J. Heath, Vaniambadi S. Kalyanaraman, Thorsten Demberg, Aurelio Cafaro, Roger W. Wiseman, Leon Flanary, Julie A. Karl, Kay Larsen, and Barbara Ensoli
- Subjects
Simian Acquired Immunodeficiency Syndrome ,Viremia ,Major histocompatibility complex ,Article ,Virus ,MHC class I ,medicine ,Animals ,AIDS Vaccines ,General Veterinary ,General Immunology and Microbiology ,biology ,Histocompatibility Antigens Class I ,Histocompatibility Antigens Class II ,Public Health, Environmental and Occupational Health ,Viral Load ,biology.organism_classification ,medicine.disease ,Macaca mulatta ,Virology ,Macaca fascicularis ,Regimen ,Infectious Diseases ,Haplotypes ,Lentivirus ,Immunology ,biology.protein ,Molecular Medicine ,tat Gene Products, Human Immunodeficiency Virus ,Disease Susceptibility ,Viral load - Abstract
Protection afforded by HIV Tat-based vaccines has differed in Indian rhesus and Mauritian cynomolgus macaques. We evaluated native Tat and Ad-HIVtat priming/Tat-boosting regimens in both species. Both vaccines were immunogenic. Only the Ad-tat regimen modestly reduced acute viremia in rhesus macaques after SHIV(89.6P) challenge. Confounding variables uncovered in Mauritian macaques included significant associations of susceptibility to infection with MHC class IB and class II H2 and H5 haplotypes, and resistance to infection with class IB haplotypes H3 and H6. Although protection here was limited, Tat-based vaccines incorporating other HIV components have shown greater efficacy. Combination strategies should be further explored.
- Published
- 2008
44. HIV-1 Env vaccine comprised of electroporated DNA and protein co-administered with Talabostat
- Author
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Lindsey Galmin, Phillip D. Markham, Ruxandra Draghia-Akli, Susana Restrepo, John J. Suschak, Deborah T. Weiss, Ranajit Pal, Brad Lewis, Lauren Hudacik, Nazneen Aziz, and Anthony D. Cristillo
- Subjects
Cellular immunity ,Biophysics ,HIV Envelope Protein gp120 ,Biochemistry ,law.invention ,Mice ,Th2 Cells ,law ,Vaccines, DNA ,Animals ,Molecular Biology ,AIDS Vaccines ,Mice, Inbred BALB C ,biology ,Immunogenicity ,Antibody titer ,virus diseases ,Dipeptides ,Cell Biology ,Th1 Cells ,Boronic Acids ,Virology ,Recombinant Proteins ,CTL ,Electroporation ,Antibody Formation ,DNA, Viral ,Humoral immunity ,Immunology ,HIV-1 ,biology.protein ,Recombinant DNA ,Cytokines ,Female ,Antibody ,CD8 - Abstract
Selection of potent yet low reactogenic adjuvants for protein immunization is important for HIV-1 vaccine development. Immunogenicity of electroporated DNA (HIV env) and recombinant gp120, administered with either QS-21 or the orally administered immunomodulator, Talabostat, was evaluated in BALB/c mice. Electroporation of low dose DNA elicited Th1 cytokines and anti-envelope antibodies. Immunization with gp120 protein alone with or without Talabostat elicited lower Th1 and Th2 cytokine levels but comparable anti-gp120 antibodies to QS-21-formulated protein. Boosting of DNA-primed mice with gp120/Talabostat induced similar anti-gp120 antibody titers and slightly higher levels of Th1 and Th2 cytokines relative to QS-21-formulated protein. Induction of CD8(+) and CD4(+) T cells and functional CTL activity was noted. These results highlight the potential use of orally administered Talabostat for efficient protein boosting of antibody and T-cell responses primed by DNA.
- Published
- 2008
45. Persistent antibody and T cell responses induced by HIV-1 DNA vaccine delivered by electroporation
- Author
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Susana Restrepo, Ruxandra Draghia-Akli, Ranajit Pal, John J. Suschak, Lindsey Galmin, Phillip D. Markham, Deborah T. Weiss, Anthony D. Cristillo, and Lauren Hudacik
- Subjects
Cellular immunity ,T-Lymphocytes ,Biophysics ,chemical and pharmacologic phenomena ,Biology ,Transfection ,Biochemistry ,Antibodies ,DNA vaccination ,Immune system ,Immunity ,Vaccines, DNA ,Animals ,Molecular Biology ,AIDS Vaccines ,Electroporation ,Immunogenicity ,Cell Biology ,Virology ,Macaca fascicularis ,Immunology ,Humoral immunity ,HIV-1 ,biology.protein ,Rabbits ,Antibody - Abstract
Intramuscular needle injection of HIV-1 DNA vaccines typically elicits weak immune responses in immunized individuals. To improve such responses, the immunogenicity of a vaccine consisting of electroporated DNA followed by intramuscular protein boost was evaluated in rabbits and macaques. In macaques, electroporation of low dose DNA encoding HIV-1 env followed by gp120 protein elicited Th1 cytokines and functional CTL that persisted for over 1 year. In both macaques and rabbits, robust anti-envelope antibodies, elicited by electroporated DNA, were augmented by gp120 protein and such responses neutralized sensitive SHIV isolates. These findings highlight efficient priming of immune responses by electroporated DNA that in conjunction with protein boost may give rise to long-term immunity in immunized hosts.
- Published
- 2008
46. Protection of macaques against vaginal SHIV challenge by systemic or mucosal and systemic vaccinations with HIV-envelope
- Author
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Indresh K. Srivastava, Amanda Goodsell, Anthony D. Cristillo, Elaine Kan, Ranajit Pal, Michael Vajdy, Susan W. Barnett, Maria Grazia Ferrai, Deborah E. Weiss, Norman L. Letvin, Fengmin Zhou, and David C. Montefiori
- Subjects
CD4-Positive T-Lymphocytes ,Immunology ,Simian Acquired Immunodeficiency Syndrome ,HIV Infections ,Antibodies, Viral ,medicine.disease_cause ,Injections, Intramuscular ,Virus ,Acquired immunodeficiency syndrome (AIDS) ,medicine ,Animals ,Humans ,Immunology and Allergy ,Seroconversion ,Administration, Intranasal ,AIDS Vaccines ,biology ,env Gene Products, Human Immunodeficiency Virus ,Simian immunodeficiency virus ,biology.organism_classification ,medicine.disease ,Macaca mulatta ,Virology ,Vaccination ,Rhesus macaque ,Infectious Diseases ,Vagina ,Lentivirus ,HIV-1 ,Female ,Immunization ,Simian Immunodeficiency Virus ,Intravaginal administration - Abstract
Background: Worldwide, the majority of human immunodeficiency virus (HIV) infections occur by heterosexual transmission. Thus, the development of a vaccine that can prevent intravaginal HIV infection is an important goal of AIDS vaccine research. Objectives: To determine which single or combination of systemic and mucosal routes of immunizations of female rhesus macaques with an HIV-1SF162 envelope protein vaccine induced protection against intravaginal challenge with SHIV. Design: Female rhesus macaques were immunized with an HIV-1SF162 envelope protein vaccine administered systemically (intramuscularly), or mucosally (intranasally), or as a sequential combination of both routes. The macaques were then challenged intravaginally with SHIVSF162P4, expressing an envelope that is closely matched (homologous) to the vaccine. Results: Macaques receiving intramuscular immunizations, alone or in combination with intranasal immunizations, were protected from infection, with no detectable plasma viral RNA, provirus, or seroconversion to nonvaccine viral proteins, and better preservation of intestinal CD4þ T cells. Serum neutralizing antibodies against the challenge virus appeared to correlate with protection. Conclusions: The results of this study demonstrate that, in the nonhuman primate model, it is possible for vaccine-elicited immune responses to prevent infection after intravaginal administration of virus. 2008 Wolters Kluwer Health | Lippincott Williams & Wilkins AIDS 2008, 22:339‐348
- Published
- 2008
47. Antibodies to CD4-induced sites in HIV gp120 correlate with the control of SHIV challenge in macaques vaccinated with subunit immunogens
- Author
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Ranajit Pal, Ilia Harris, George K. Lewis, Anthony L. DeVico, Robert C. Gallo, Lindsey Galmin, Karla Godfrey, Timothy R. Fouts, and Manhattan Charurat
- Subjects
Immunogen ,Heterologous ,Viremia ,HIV Antibodies ,HIV Envelope Protein gp120 ,Biology ,medicine.disease_cause ,Epitope ,Cell Line ,Viral envelope ,medicine ,Animals ,Humans ,HIV vaccine ,AIDS Vaccines ,Multidisciplinary ,Antibodies, Monoclonal ,virus diseases ,Biological Sciences ,Simian immunodeficiency virus ,medicine.disease ,Virology ,CD4 Antigens ,Vaccines, Subunit ,Immunology ,Lentivirus Infections ,biology.protein ,Macaca ,RNA, Viral ,Simian Immunodeficiency Virus ,Antibody - Abstract
Epitopes located in and around the coreceptor binding site of HIV-1 envelope glycoprotein (gp120) exhibit enhanced exposure after attachment to the CD4 receptor and comprise some of the most conserved and functionally important residues on the viral envelope. Therefore, antibody responses to these epitopes [designated as CD4-induced (CD4i)] should be highly cross-reactive and potentially useful for HIV vaccine development. To address this question, rhesus macaques were vaccinated with subunit immunogens designed to raise humoral responses against CD4i epitopes and challenged rectally with SHIV 162P3 , which encodes a heterologous envelope versus the immunogen. We found that animals vaccinated with a rhesus full-length single-chain (rhFLSC) complex exhibited significantly accelerated clearance of plasma viremia and an absence of long-term tissue viremia compared with unvaccinated control animals. Such control of infection correlated with stronger responses to CD4i epitopes in the rhFLSC-vaccinated animals, compared with macaques immunized with gp120, cross-linked gp120–CD4 complexes, or soluble CD4 alone. These responses were strongly boosted in the rhFLSC-vaccinated animals by SHIV 162P3 infection. The control of infection was not associated with anti-CD4 responses, overall anti-gp120-binding titers, or neutralizing activity measured in conventional assays. Vaccine-naive animals also developed anti-CD4i epitope responses after simian/ human immunodeficiency virus (SHIV) challenge, which appeared later than the overall anti-gp120 responses and in concert with the decline of viremia to a low set point. Collectively, these data suggest that antibodies to CD4i epitopes may play a role in controlling SHIV infection and provide insights for HIV vaccine development.
- Published
- 2007
48. A Replication-Competent Adenovirus-Human Immunodeficiency Virus (Ad-HIV)tatand Ad-HIVenvPriming/Tat and Envelope Protein Boosting Regimen Elicits Enhanced Protective Efficacy against Simian/Human Immunodeficiency Virus SHIV89.6PChallenge in Rhesus Macaques
- Author
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L. Jean Patterson, Vaniambadi S. Kalyanaraman, Thorsten Demberg, Megan J. Heath, Kay Larsen, Norman L. Letvin, Birgit Korioth-Schmitz, Irene Kalisz, Adam P. Buzby, Eun Mi Lee, David Venzon, Barbara Ensoli, Richard Grant, Ruth H. Florese, Aurelio Cafaro, Dilani Dombagoda, Ranajit Pal, David C. Montefiori, and Marjorie Robert-Guroff
- Subjects
Cellular immunity ,viruses ,Immunology ,Viremia ,Antibodies, Viral ,Virus Replication ,medicine.disease_cause ,Microbiology ,Virus ,Adenoviridae ,Virology ,Vaccines and Antiviral Agents ,medicine ,Animals ,Lymphocytes ,biology ,Immunogenicity ,Gene Products, env ,HIV ,virus diseases ,Simian immunodeficiency virus ,biology.organism_classification ,medicine.disease ,Macaca mulatta ,Viral replication ,Insect Science ,Gene Products, tat ,Lentivirus ,Simian Immunodeficiency Virus ,tat Gene Products, Human Immunodeficiency Virus ,Immunologic Memory - Abstract
We previously demonstrated that replication-competent adenovirus (Ad)-simian immunodeficiency virus (SIV) recombinant prime/protein boost regimens elicit potent immunogenicity and strong, durable protection of rhesus macaques against SIVmac251. Additionally, native Tat vaccines have conferred strong protection against simian/human immunodeficiency virus SHIV89.6Pchallenge of cynomolgus monkeys, while native, inactivated, or vectored Tat vaccines have failed to elicit similar protective efficacy in rhesus macaques. Here we asked if priming rhesus macaques with replicating Ad-human immunodeficiency virus (HIV)tatand boosting with the Tat protein would elicit protection against SHIV89.6P. We also evaluated a Tat/Env regimen, adding an Ad-HIVenvrecombinant and envelope protein boost to test whether envelope antibodies would augment acute-phase protection. Further, expecting cellular immunity to enhance chronic viremia control, we tested a multigenic group: Ad-HIVtat, -HIVenv, -SIVgag, and -SIVnefrecombinants and Tat, Env, and Nef proteins. All regimens were immunogenic. A hierarchy was observed in enzyme-linked immunospot responses (with the strongest response for Env, followed by Gag, followed by Nef, followed by Tat) and antibody titers (with the highest titer for Env, followed by Tat, followed by Nef, followed by Gag). Following intravenous SHIV89.6Pchallenge, all macaques became infected. Compared to controls, no protection was seen in the Tat-only group, confirming previous reports for rhesus macaques. However, the multigenic group blunted acute viremia by approximately 1 log (P= 0.017), and both the multigenic and Tat/Env groups reduced chronic viremia by 3 and 4 logs, respectively, compared to controls (multigenic,P= 0.0003; Tat/Env,P< 0.0001). The strikingly greater reduction in the Tat/Env group than in the multigenic group (P= 0.014) was correlated with Tat and Env binding antibodies. Since prechallenge anti-Env antibodies lacked SHIV89.6P-neutralizing activity, other functional anti-Env and anti-Tat activities are under investigation, as is a possible synergy between the Tat and Env immunogens.
- Published
- 2007
49. Mucosal B Cells Are Associated with Delayed SIV Acquisition in Vaccinated Female but Not Male Rhesus Macaques Following SIVmac251 Rectal Challenge
- Author
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Monica Vaccari, Maria Grazia Ferrari, Venkatramanan Mohanram, Iskra Tuero, Diego A. Vargas-Inchaustegui, Marjorie Robert-Guroff, David C. Montefiori, Celia C. LaBranche, Ranajit Pal, Leia Miller, David Venzon, Genoveffa Franchini, Thomas Musich, Susan W. Barnett, Vaniambadi S. Kalyanaraman, Thorsten Demberg, Mangala Rao, and Irene Kalisz
- Subjects
Male ,lcsh:Immunologic diseases. Allergy ,medicine.medical_treatment ,viruses ,Immunology ,Simian Acquired Immunodeficiency Syndrome ,Antibodies, Viral ,medicine.disease_cause ,Microbiology ,Macaque ,Sex Factors ,Immune system ,Immunity ,Virology ,biology.animal ,Genetics ,medicine ,Animals ,Intestinal Mucosa ,Immunity, Mucosal ,Molecular Biology ,lcsh:QH301-705.5 ,B-Lymphocytes ,Immunity, Cellular ,biology ,Rectum ,SAIDS Vaccines ,Vaccine trial ,virus diseases ,Simian immunodeficiency virus ,Macaca mulatta ,Immunization ,lcsh:Biology (General) ,biology.protein ,Female ,Simian Immunodeficiency Virus ,Parasitology ,Antibody ,lcsh:RC581-607 ,Adjuvant ,Research Article - Abstract
Many viral infections, including HIV, exhibit sex-based pathogenic differences. However, few studies have examined vaccine-related sex differences. We compared immunogenicity and protective efficacy of monomeric SIV gp120 with oligomeric SIV gp140 in a pre-clinical rhesus macaque study and explored a subsequent sex bias in vaccine outcome. Each immunization group (16 females, 8 males) was primed twice mucosally with replication-competent Ad-recombinants encoding SIVsmH4 env/rev, SIV239 gag and SIV239 nefΔ1–13 and boosted twice intramuscularly with SIVmac239 monomeric gp120 or oligomeric gp140 in MF59 adjuvant. Controls (7 females, 5 males) received empty Ad and MF59. Up to 9 weekly intrarectal challenges with low-dose SIVmac251 were administered until macaques became infected. We assessed vaccine-induced binding, neutralizing, and non-neutralizing antibodies, Env-specific memory B cells and plasmablasts/plasma cells (PB/PC) in bone marrow and rectal tissue, mucosal Env-specific antibodies, and Env-specific T-cells. Post-challenge, only one macaque (gp140-immunized) remained uninfected. However, SIV acquisition was significantly delayed in vaccinated females but not males, correlated with Env-specific IgA in rectal secretions, rectal Env-specific memory B cells, and PC in rectal tissue. These results extend previous correlations of mucosal antibodies and memory B cells with protective efficacy. The gp140 regimen was more immunogenic, stimulating elevated gp140 and cyclic V2 binding antibodies, ADCC and ADCP activities, bone marrow Env-specific PB/PC, and rectal gp140-specific IgG. However, immunization with gp120, the form of envelope immunogen used in RV144, the only vaccine trial to show some efficacy, provided more significant acquisition delay. Further over 40 weeks of follow-up, no gp120 immunized macaques met euthanasia criteria in contrast to 7 gp140-immunized and 2 control animals. Although males had higher binding antibodies than females, ADCC and ADCP activities were similar. The complex challenge outcomes may reflect differences in IgG subtypes, Fc glycosylation, Fc-R polymorphisms, and/or the microbiome, key areas for future studies. This first demonstration of a sex-difference in SIV vaccine-induced protection emphasizes the need for sex-balancing in vaccine trials. Our results highlight the importance of mucosal immunity and memory B cells at the SIV exposure site for protection., Author Summary Viral infections can have different disease courses in men and women. Following HIV infection, women generally exhibit lower viral loads and higher CD4 counts than men, but paradoxically progress faster to AIDS. Sex differences result from effects of X-linked genes and hormonal influences, and are believed to be largely based on immune response differences. Nevertheless, little is known about potential sex differences following vaccination. Here we report for the first time a sex bias in response to a SIV vaccine in rhesus macaques, showing that female animals were better protected against acquisition of SIV compared to males. The vaccine-induced immune responses that contributed to this better protection were viral-specific antibodies and immune antibody-secreting B cells, both at the local rectal site of SIV exposure. These results suggest that HIV/SIV vaccines should be better designed to target mucosal exposure sites. Additionally, they indicate that more vaccine studies should include animals of both sexes to address potential differences. Our study also illustrates that inclusion of both sexes can lead to greater complexity in vaccine trial outcomes, necessitating more in depth analyses. However, we believe sex balancing to be particularly important, as approximately 50% of HIV infections worldwide occur in women.
- Published
- 2015
50. Transcription profiling of CD4⁺ T cells in rhesus macaques that infected with simian-human immunodeficiency virus and re-challenged with SIVmac251
- Author
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Cynthia A. Pise-Masison, Jae Ho Lee, Hye Kyung Chung, Annamalai Muthiah, Eun Mi Lee, Ranajit Pal, and Michael F. Radonovich
- Subjects
Male ,Simian Acquired Immunodeficiency Syndrome ,Viremia ,Biology ,medicine.disease_cause ,Antibodies, Viral ,Virus ,Gene expression ,medicine ,Animals ,Gene ,Immunodeficiency ,General Veterinary ,Gene Expression Profiling ,Simian immunodeficiency virus ,medicine.disease ,Virology ,Macaca mulatta ,CD4 Lymphocyte Count ,Gene expression profiling ,Viral replication ,Immunology ,Leukocytes, Mononuclear ,Animal Science and Zoology ,Simian Immunodeficiency Virus - Abstract
Background Insights into the host factors that contribute to an effective antiviral immune response may be obtained by examining global gene expression in simian-human immunodeficiency virus (SHIV)-infected non-human primates that exhibit different virological outcomes. Methods Six chronically SHIV-infected macaques were rectally challenged with SIVmac251. Viral RNA and proviral DNA load in blood were measured. Gene expression profiles in CD4+ T cells were examined and compared between animals with different levels of infection following challenge. Results and Conclusions Viral RNA was markedly controlled in four challenged animals, whereas two animals had persistent high viremia. Analysis of the gene expression profiles at early infection revealed gene expression signatures between protectors and non-protectors and identified potential protective biomarkers. Pathway analyses revealed that IFN pathway genes are down-regulated in protectors compared to unprotectors. This study suggests that high levels of expression of type 1 IFN-related genes may paradoxically promote virus replication.
- Published
- 2015
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