57 results on '"Shi-Mao Xia"'
Search Results
2. Initiation of HIV neutralizing B cell lineages with sequential envelope immunizations
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Wilton B. Williams, Jinsong Zhang, Chuancang Jiang, Nathan I. Nicely, Daniela Fera, Kan Luo, M. Anthony Moody, Hua-Xin Liao, S. Munir Alam, Thomas B. Kepler, Akshaya Ramesh, Kevin Wiehe, James A. Holland, Todd Bradley, Nathan Vandergrift, Kevin O. Saunders, Robert Parks, Andrew Foulger, Shi-Mao Xia, Mattia Bonsignori, David C. Montefiori, Mark Louder, Amanda Eaton, Sampa Santra, Richard Scearce, Laura Sutherland, Amanda Newman, Hilary Bouton-Verville, Cindy Bowman, Howard Bomze, Feng Gao, Dawn J. Marshall, John F. Whitesides, Xiaoyan Nie, Garnett Kelsoe, Steven G. Reed, Christopher B. Fox, Kim Clary, Marguerite Koutsoukos, David Franco, John R. Mascola, Stephen C. Harrison, Barton F. Haynes, and Laurent Verkoczy
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Science - Abstract
An efficient HIV-1 vaccine will likely depend on eliciting broadly neutralizing antibodies (bnAb). Here the authors analyze the B cell repertoire in macaques and knock-in mice in response to sequential immunization with Env variants that induce a bnAb targeting the CD4-binding site of Env in a HIV-1 infected individual.
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- 2017
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3. Amino Acid Changes in the HIV-1 gp41 Membrane Proximal Region Control Virus Neutralization Sensitivity
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Todd Bradley, Ashley Trama, Nancy Tumba, Elin Gray, Xiaozhi Lu, Navid Madani, Fatemeh Jahanbakhsh, Amanda Eaton, Shi-Mao Xia, Robert Parks, Krissey E. Lloyd, Laura L. Sutherland, Richard M. Scearce, Cindy M. Bowman, Susan Barnett, Salim S. Abdool-Karim, Scott D. Boyd, Bruno Melillo, Amos B. Smith III, Joseph Sodroski, Thomas B. Kepler, S.Munir Alam, Feng Gao, Mattia Bonsignori, Hua-Xin Liao, M. Anthony Moody, David Montefiori, Sampa Santra, Lynn Morris, and Barton F. Haynes
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Medicine ,Medicine (General) ,R5-920 - Abstract
Most HIV-1 vaccines elicit neutralizing antibodies that are active against highly sensitive (tier-1) viruses or rare cases of vaccine-matched neutralization-resistant (tier-2) viruses, but no vaccine has induced antibodies that can broadly neutralize heterologous tier-2 viruses. In this study, we isolated antibodies from an HIV-1-infected individual that targeted the gp41 membrane-proximal external region (MPER) that may have selected single-residue changes in viral variants in the MPER that resulted in neutralization sensitivity to antibodies targeting distal epitopes on the HIV-1 Env. Similarly, a single change in the MPER in a second virus from another infected-individual also conferred enhanced neutralization sensitivity. These gp41 single-residue changes thus transformed tier-2 viruses into tier-1 viruses that were sensitive to vaccine-elicited tier-1 neutralizing antibodies. These data demonstrate that Env amino acid changes within the MPER bnAb epitope of naturally-selected escape viruses can increase neutralization sensitivity to multiple types of neutralizing antibodies, and underscore the critical importance of the MPER for maintaining the integrity of the tier-2 HIV-1 trimer.
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- 2016
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4. Human Non-neutralizing HIV-1 Envelope Monoclonal Antibodies Limit the Number of Founder Viruses during SHIV Mucosal Infection in Rhesus Macaques.
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Sampa Santra, Georgia D Tomaras, Ranjit Warrier, Nathan I Nicely, Hua-Xin Liao, Justin Pollara, Pinghuang Liu, S Munir Alam, Ruijun Zhang, Sarah L Cocklin, Xiaoying Shen, Ryan Duffy, Shi-Mao Xia, Robert J Schutte, Charles W Pemble Iv, S Moses Dennison, Hui Li, Andrew Chao, Kora Vidnovic, Abbey Evans, Katja Klein, Amit Kumar, James Robinson, Gary Landucci, Donald N Forthal, David C Montefiori, Jaranit Kaewkungwal, Sorachai Nitayaphan, Punnee Pitisuttithum, Supachai Rerks-Ngarm, Merlin L Robb, Nelson L Michael, Jerome H Kim, Kelly A Soderberg, Elena E Giorgi, Lily Blair, Bette T Korber, Christiane Moog, Robin J Shattock, Norman L Letvin, Joern E Schmitz, M A Moody, Feng Gao, Guido Ferrari, George M Shaw, and Barton F Haynes
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Immunologic diseases. Allergy ,RC581-607 ,Biology (General) ,QH301-705.5 - Abstract
HIV-1 mucosal transmission begins with virus or virus-infected cells moving through mucus across mucosal epithelium to infect CD4+ T cells. Although broadly neutralizing antibodies (bnAbs) are the type of HIV-1 antibodies that are most likely protective, they are not induced with current vaccine candidates. In contrast, antibodies that do not neutralize primary HIV-1 strains in the TZM-bl infection assay are readily induced by current vaccine candidates and have also been implicated as secondary correlates of decreased HIV-1 risk in the RV144 vaccine efficacy trial. Here, we have studied the capacity of anti-Env monoclonal antibodies (mAbs) against either the immunodominant region of gp41 (7B2 IgG1), the first constant region of gp120 (A32 IgG1), or the third variable loop (V3) of gp120 (CH22 IgG1) to modulate in vivo rectal mucosal transmission of a high-dose simian-human immunodeficiency virus (SHIV-BaL) in rhesus macaques. 7B2 IgG1 or A32 IgG1, each containing mutations to enhance Fc function, was administered passively to rhesus macaques but afforded no protection against productive clinical infection while the positive control antibody CH22 IgG1 prevented infection in 4 of 6 animals. Enumeration of transmitted/founder (T/F) viruses revealed that passive infusion of each of the three antibodies significantly reduced the number of T/F genomes. Thus, some antibodies that bind HIV-1 Env but fail to neutralize virus in traditional neutralization assays may limit the number of T/F viruses involved in transmission without leading to enhancement of viral infection. For one of these mAbs, gp41 mAb 7B2, we provide the first co-crystal structure in complex with a common cyclical loop motif demonstrated to be critical for infection by other retroviruses.
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- 2015
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5. IGHV1-69 B cell chronic lymphocytic leukemia antibodies cross-react with HIV-1 and hepatitis C virus antigens as well as intestinal commensal bacteria.
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Kwan-Ki Hwang, Ashley M Trama, Daniel M Kozink, Xi Chen, Kevin Wiehe, Abby J Cooper, Shi-Mao Xia, Minyue Wang, Dawn J Marshall, John Whitesides, Munir Alam, Georgia D Tomaras, Steven L Allen, Kanti R Rai, Jane McKeating, Rosa Catera, Xiao-Jie Yan, Charles C Chu, Garnett Kelsoe, Hua-Xin Liao, Nicholas Chiorazzi, and Barton F Haynes
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Medicine ,Science - Abstract
B-cell chronic lymphocytic leukemia (B-CLL) patients expressing unmutated immunoglobulin heavy variable regions (IGHVs) use the IGHV1-69 B cell receptor (BCR) in 25% of cases. Since HIV-1 envelope gp41 antibodies also frequently use IGHV1-69 gene segments, we hypothesized that IGHV1-69 B-CLL precursors may contribute to the gp41 B cell response during HIV-1 infection. To test this hypothesis, we rescued 5 IGHV1-69 unmutated antibodies as heterohybridoma IgM paraproteins and as recombinant IgG1 antibodies from B-CLL patients, determined their antigenic specificities and analyzed BCR sequences. IGHV1-69 B-CLL antibodies were enriched for reactivity with HIV-1 envelope gp41, influenza, hepatitis C virus E2 protein and intestinal commensal bacteria. These IGHV1-69 B-CLL antibodies preferentially used IGHD3 and IGHJ6 gene segments and had long heavy chain complementary determining region 3s (HCDR3s) (≥21 aa). IGHV1-69 B-CLL BCRs exhibited a phenylalanine at position 54 (F54) of the HCDR2 as do rare HIV-1 gp41 and influenza hemagglutinin stem neutralizing antibodies, while IGHV1-69 gp41 antibodies induced by HIV-1 infection predominantly used leucine (L54) allelic variants. These results demonstrate that the B-CLL cell population is an expansion of members of the innate polyreactive B cell repertoire with reactivity to a number of infectious agent antigens including intestinal commensal bacteria. The B-CLL IGHV1-69 B cell usage of F54 allelic variants strongly suggests that IGHV1-69 B-CLL gp41 antibodies derive from a restricted B cell pool that also produces rare HIV-1 gp41 and influenza hemagglutinin stem antibodies.
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- 2014
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6. Isolation of HIV-1-neutralizing mucosal monoclonal antibodies from human colostrum.
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James Friedman, S Munir Alam, Xiaoying Shen, Shi-Mao Xia, Shelley Stewart, Kara Anasti, Justin Pollara, Genevieve G Fouda, Guang Yang, Garnett Kelsoe, Guido Ferrari, Georgia D Tomaras, Barton F Haynes, Hua-Xin Liao, M Anthony Moody, and Sallie R Permar
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Medicine ,Science - Abstract
Generation of potent anti-HIV antibody responses in mucosal compartments is a potential requirement of a transmission-blocking HIV vaccine. HIV-specific, functional antibody responses are present in breast milk, and these mucosal antibody responses may play a role in protection of the majority of HIV-exposed, breastfeeding infants. Therefore, characterization of HIV-specific antibodies produced by B cells in milk could guide the development of vaccines that elicit protective mucosal antibody responses.We isolated B cells from colostrum of an HIV-infected lactating woman with a detectable neutralization response in milk and recombinantly produced and characterized the resulting HIV-1 Envelope (Env)-specific monoclonal antibodies (mAbs).The identified HIV-1 Env-specific colostrum mAbs, CH07 and CH08, represent two of the first mucosally-derived anti-HIV antibodies yet to be reported. Colostrum mAb CH07 is a highly-autoreactive, weakly-neutralizing gp140-specific mAb that binds to linear epitopes in the gp120 C5 region and gp41 fusion domain. In contrast, colostrum mAb CH08 is a nonpolyreactive CD4-inducible (CD4i) gp120-specific mAb with moderate breadth of neutralization.These novel HIV-neutralizing mAbs isolated from a mucosal compartment provide insight into the ability of mucosal B cell populations to produce functional anti-HIV antibodies that may contribute to protection against virus acquisition at mucosal surfaces.
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- 2012
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7. Induction of antibodies in rhesus macaques that recognize a fusion-intermediate conformation of HIV-1 gp41.
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S Moses Dennison, Laura L Sutherland, Frederick H Jaeger, Kara M Anasti, Robert Parks, Shelley Stewart, Cindy Bowman, Shi-Mao Xia, Ruijun Zhang, Xiaoying Shen, Richard M Scearce, Gilad Ofek, Yongping Yang, Peter D Kwong, Sampa Santra, Hua-Xin Liao, Georgia Tomaras, Norman L Letvin, Bing Chen, S Munir Alam, and Barton F Haynes
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Medicine ,Science - Abstract
A component to the problem of inducing broad neutralizing HIV-1 gp41 membrane proximal external region (MPER) antibodies is the need to focus the antibody response to the transiently exposed MPER pre-hairpin intermediate neutralization epitope. Here we describe a HIV-1 envelope (Env) gp140 oligomer prime followed by MPER peptide-liposomes boost strategy for eliciting serum antibody responses in rhesus macaques that bind to a gp41 fusion intermediate protein. This Env-liposome immunization strategy induced antibodies to the 2F5 neutralizing epitope ⁶⁶⁴DKW residues, and these antibodies preferentially bound to a gp41 fusion intermediate construct as well as to MPER scaffolds stabilized in the 2F5-bound conformation. However, no serum lipid binding activity was observed nor was serum neutralizing activity for HIV-1 pseudoviruses present. Nonetheless, the Env-liposome prime-boost immunization strategy induced antibodies that recognized a gp41 fusion intermediate protein and was successful in focusing the antibody response to the desired epitope.
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- 2011
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8. Mutation-Guided Vaccine Design: A Strategy for Developing Boosting Immunogens for HIV Broadly Neutralizing Antibody Induction
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Kevin Wiehe, Kevin O. Saunders, Victoria Stalls, Derek W. Cain, Sravani Venkatayogi, Joshua S. Martin Beem, Madison Berry, Tyler Evangelous, Rory Henderson, Bhavna Hora, Shi-Mao Xia, Chuancang Jiang, Amanda Newman, Cindy Bowman, Xiaozhi Lu, Mary E. Bryan, Joena Bal, Aja Sanzone, Haiyan Chen, Amanda Eaton, Mark A. Tomai, Christopher B. Fox, Ying Tam, Christopher Barbosa, Mattia Bonsignori, Hiromi Muramatsu, S. Munir Alam, David Montefiori, Wilton B. Williams, Norbert Pardi, Ming Tian, Drew Weissman, Frederick W. Alt, Priyamvada Acharya, and Barton F. Haynes
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A major goal of HIV-1 vaccine development is induction of broadly neutralizing antibodies (bnAbs). While success has been achieved in initiating bnAb B cell lineages, design of boosting immunogens that select for bnAb B cell receptors with improbable mutations required for bnAb affinity maturation remains difficult. Here we demonstrate a process for designing boosting immunogens for a V3-glycan bnAb B cell lineage. The immunogens induced affinity-matured antibodies by selecting for functional improbable mutations in bnAb precursor knock-in mice. Moreover, we show similar success in prime and boosting with nucleoside-modified mRNA-encoded HIV-1 envelope trimer immunogens, with improved selection by mRNA immunogens of improbable mutations required for bnAb binding to key envelope glycans. These results demonstrate the ability of both protein and mRNA prime-boost immunogens for selection of rare B cell lineage intermediates with neutralizing breadth after bnAb precursor expansion, a key proof-of concept and milestone towards development of an HIV vaccine.One-Sentence SummaryA vaccine strategy for selecting key rare antibody mutations is shown to induce HIV broadly neutralizing antibodies in mice.
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- 2022
9. RAB11FIP5-Deficient Mice Exhibit Cytokine-Related Transcriptomic Signatures
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Maggie Barr, Robert Parks, Yunfei Wang, Gregory D. Sempowski, Yue Chen, Barton F. Haynes, Maria Aggelakopoulou, Dapeng Li, Todd Bradley, Shi-Mao Xia, Bhavna Hora, Kristina J. Riebe, Isabela Pedroza-Pacheco, Amanda Newman, Kevin O. Saunders, Persephone Borrow, Richard M. Scearce, Grace Stevens, Cindy M. Bowman, and Derek W. Cain
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medicine.medical_treatment ,Immunology ,Cell ,General Medicine ,Biology ,Article ,medicine.anatomical_structure ,Immune system ,Cytokine ,Transcytosis ,medicine ,Immunology and Allergy ,Tumor necrosis factor alpha ,Interleukin 4 ,CD8 ,Gene knockout - Abstract
Rab11 recycling endosomes are involved in immunological synaptic functions, but the roles of Rab11 family–interacting protein 5 (Rab11Fip5), one of the Rab11 effectors, in the immune system remain obscure. Our previous study demonstrated that RAB11FIP5 transcripts are significantly elevated in PBMCs from HIV-1–infected individuals, making broadly HIV-1–neutralizing Abs compared with those without broadly neutralizing Abs; however, the role of Rab11FiP5 in immune functions remains unclear. In this study, a RAB11FIP5 gene knockout (RAB11FIP5−/−) mouse model was employed to study the role of Rab11Fip5 in immune responses. RAB11FIP5−/− mice exhibited no perturbation in lymphoid tissue cell subsets, and Rab11Fip5 was not required for serum Ab induction following HIV-1 envelope immunization, Ab transcytosis to mucosal sites, or survival after influenza challenge. However, differences were observed in multiple transcripts, including cytokine genes, in lymphocyte subsets from envelope-immunized RAB11FIP5−/− versus control mice. These included alterations in several genes in NK cells that mirrored observations in NKs from HIV-infected humans expressing less RAB11FIP5, although Rab11Fip5 was dispensable for NK cell cytolytic activity. Notably, immunized RAB11FIP5−/− mice had lower IL4 expression in CD4+ T follicular helper cells and showed lower TNF expression in CD8+ T cells. Likewise, TNF-α production by human CD8+ T cells correlated with PBMC RAB11FIP5 expression. These observations in RAB11FIP5−/− mice suggest a role for Rab11Fip5 in regulating cytokine responses.
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- 2020
10. Ability of nucleoside-modified mRNA to encode HIV-1 envelope trimer nanoparticles
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Christopher Barbosa, Amanda Newman, Sravani Venkatayogi, Shi-Mao Xia, Rory Henderson, Wilton B. Williams, Qifeng Han, Barton F. Haynes, Robert J. Edwards, Robert Parks, Kevin O. Saunders, Kevin Wiehe, Mattia Bonsignori, Katayoun Mansouri, Norbert Pardi, Xiaozhi Lu, David C. Montefiori, Tyler Evangelous, Zekun Mu, Wes Rountree, Diana Martik, Yunfei Wang, Drew Weissman, Derek W. Cain, Ying Tam, and S. Munir Alam
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Messenger RNA ,biology ,Chemistry ,medicine.drug_class ,mRNA ,broadly neutralizing antibodies ,knock-in mice ,Priming (immunology) ,Heterologous ,lipid nanoparticles ,Monoclonal antibody ,Virology ,Article ,Ferritin ,mRNA-LNP ,medicine.anatomical_structure ,Antigen ,vaccine ,HIV-1 ,medicine ,biology.protein ,Nucleoside ,B cell - Abstract
SUMMARYThe success of nucleoside-modified mRNAs in lipid nanoparticles (mRNA-LNP) as COVID-19 vaccines heralded a new era of vaccine development. For HIV-1, multivalent envelope (Env) trimer protein nanoparticles are superior immunogens compared to trimers alone for priming of broadly neutralizing antibody (bnAb) B cell lineages. The successful expression of complex multivalent nanoparticle immunogens with mRNAs has not been demonstrated. Here we show that mRNAs can encode antigenic Env trimers on ferritin nanoparticles that initiate bnAb precursor B cell expansion and induce serum autologous tier 2 neutralizing activity in bnAb precursor VH + VL knock-in mice. Next generation sequencing demonstrated acquisition of critical mutations, and monoclonal antibodies that neutralized heterologous HIV-1 isolates were isolated. Thus, mRNA- LNP can encode complex immunogens and are of use in design of germline-targeting and sequential boosting immunogens for HIV-1 vaccine development.
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- 2021
11. mRNA-encoded HIV-1 Env trimer ferritin nanoparticles induce monoclonal antibodies that neutralize heterologous HIV-1 isolates in mice
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Zekun Mu, Kevin Wiehe, Kevin O. Saunders, Rory Henderson, Derek W. Cain, Robert Parks, Diana Martik, Katayoun Mansouri, Robert J. Edwards, Amanda Newman, Xiaozhi Lu, Shi-Mao Xia, Amanda Eaton, Mattia Bonsignori, David Montefiori, Qifeng Han, Sravani Venkatayogi, Tyler Evangelous, Yunfei Wang, Wes Rountree, Bette Korber, Kshitij Wagh, Ying Tam, Christopher Barbosa, S. Munir Alam, Wilton B. Williams, Ming Tian, Frederick W. Alt, Norbert Pardi, Drew Weissman, and Barton F. Haynes
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AIDS Vaccines ,COVID-19 Vaccines ,env Gene Products, Human Immunodeficiency Virus ,Antibodies, Monoclonal ,COVID-19 ,HIV Antibodies ,Antibodies, Neutralizing ,General Biochemistry, Genetics and Molecular Biology ,Epitopes ,Mice ,Ferritins ,Liposomes ,HIV-1 ,Animals ,Humans ,Nanoparticles ,RNA, Messenger - Abstract
The success of nucleoside-modified mRNAs in lipid nanoparticles (mRNA-LNP) as COVID-19 vaccines heralded a new era of vaccine development. For HIV-1, multivalent envelope (Env) trimer protein nanoparticles are superior immunogens compared with trimers alone for priming of broadly neutralizing antibody (bnAb) B cell lineages. The successful expression of complex multivalent nanoparticle immunogens with mRNAs has not been demonstrated. Here, we show that mRNAs can encode antigenic Env trimers on ferritin nanoparticles that initiate bnAb precursor B cell expansion and induce serum autologous tier 2 neutralizing activity in bnAb precursor V
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- 2021
12. Rapid selection of HIV envelopes that bind to neutralizing antibody B cell lineage members with functional improbable mutations
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Kevin Wiehe, Shi-Mao Xia, Brianna Rhodes, Barton F. Haynes, Robert Parks, Olivia Swanson, Aja Sanzone, Mattia Bonsignori, Bob C. Lin, Avivah Wang, Kevin O. Saunders, Mihai L. Azoitei, Melissa Cooper, Mark K. Louder, and Nicole A. Doria-Rose
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medicine.anatomical_structure ,Immune system ,Lineage (genetic) ,biology ,Somatic cell ,medicine ,biology.protein ,Priming (immunology) ,HIV vaccine ,Antibody ,Neutralizing antibody ,Virology ,B cell - Abstract
SummaryElicitation of broadly neutralizing antibodies (bnAbs) by an HIV vaccine will involve priming the immune system to activate antibody precursors, followed by boosting immunizations to select for antibodies with functional features required for neutralization breadth. The higher the number of acquired mutations necessary for function, the more convoluted are the antibody developmental pathways. HIV bnAbs acquire a large number of somatic mutations, but not all mutations are functionally important. Here we identified a minimal subset of mutations sufficient for the function of the naturally occurring V3-glycan bnAb DH270.6. Using antibody library screening, candidate envelope immunogens that interacted with DH270.6-like antibodies containing this set of key mutations were identified and selectedin vitro. Our results demonstrate that less complex B cell evolutionary pathways than those naturally observed exist for the induction of HIV bnAbs by vaccination, and establish rational approaches to identify boosting sequential envelope candidate immunogens.
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- 2021
13. Development of a recombinant yellow fever vector expressing a HIV clade C founder envelope gp120
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Robert Parks, Jae-Sung Yu, Wilton B. Williams, Mattia Bonsignori, Callie Vivian, Melissa Cooper, Steven G. Reed, Cindy M. Bowman, Hua-Xin Liao, Charles M. Rice, Nathan Vandergrift, Barton F. Haynes, Meng Chen, Shi-Mao Xia, and Jamie Pritchett
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0301 basic medicine ,viruses ,Genetic Vectors ,Immunization, Secondary ,HIV Infections ,HIV Antibodies ,HIV Envelope Protein gp120 ,Vaccines, Attenuated ,Article ,Neutralization ,Virus ,law.invention ,Mice ,03 medical and health sciences ,0302 clinical medicine ,Neutralization Tests ,law ,Virology ,Chlorocebus aethiops ,Vaccines, DNA ,medicine ,Animals ,030212 general & internal medicine ,Vero Cells ,AIDS Vaccines ,Attenuated vaccine ,biology ,Yellow fever ,virus diseases ,medicine.disease ,Antibodies, Neutralizing ,Transmembrane protein ,030104 developmental biology ,HIV-1 ,biology.protein ,Recombinant DNA ,Vero cell ,Yellow fever virus ,Antibody - Abstract
Development of a HIV-1 vaccine is a major global priority. The yellow fever virus (YFV) attenuated vaccine 17D is among the most effective of currently used vaccines. However, the stability of the YFV17D vector when carrying non-flavivirus genes has been problematic. We have constructed and expressed HIV-1 Env in YFV17D with either single transmembrane (STM) or double transmembrane (DTM) YFV E protein domains for the development of anti-HIV antibodies. Here we describe modifications of the YFV17D vector such that HIV-1 Env gp120 is expressed in up to 5 passages in Vero cells. Immunization with recombinant YFV17D vector prime followed by HIV-1 CH505 TF gp120 protein boosts were able to induce neutralizing antibodies for a HIV-1 tier 1 isolate in mice. This modified YFV vector may be a starting point for constructing HIV-1 vaccine candidate priming vectors.
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- 2017
14. Immunodominance of Antibody Recognition of the HIV Envelope V2 Region in Ig-Humanized Mice
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Macdonald Lynn, Kara Anasti, Barton F. Haynes, Ryan Duffy, S. Munir Alam, M. Anthony Moody, Christos A. Kyratsous, Andrew J. Murphy, Krissey E. Lloyd, Cindy M. Bowman, Shi-Mao Xia, Nathan I. Nicely, Richard M. Scearce, Robert Parks, Masayuki Kuraoka, Christina Stolarchuk, Laurent Verkoczy, Garnett Kelsoe, Kevin Wiehe, Xiaoying Shen, Bradley Lockwood, Georgia D. Tomaras, and Sabrina Arora
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0301 basic medicine ,Subdominant ,Amino Acid Motifs ,Immunology ,Immunodominance ,HIV Antibodies ,HIV Envelope Protein gp120 ,Biology ,Article ,Epitope ,Epitopes ,Mice ,03 medical and health sciences ,Immunoglobulin lambda-Chains ,Aspartic acid ,Animals ,Humans ,Immunology and Allergy ,AIDS Vaccines ,030102 biochemistry & molecular biology ,Linear epitope ,Glutamic acid ,Antibodies, Neutralizing ,Virology ,030104 developmental biology ,HIV-1 ,biology.protein ,Immunoglobulin heavy chain ,Antibody ,Immunoglobulin Heavy Chains - Abstract
In the RV144 gp120 HIV vaccine trial, decreased transmission risk was correlated with Abs that reacted with a linear epitope at a lysine residue at position 169 (K169) in the HIV-1 envelope (Env) V2 region. The K169 V2 response was restricted to Abs bearing Vλ rearrangements that expressed aspartic acid/glutamic acid in CDR L2. The AE.A244 gp120 in AIDSVAX B/E also bound to the unmutated ancestor of a V2-glycan broadly neutralizing Ab, but this Ab type was not induced in the RV144 trial. In this study, we sought to determine whether immunodominance of the V2 linear epitope could be overcome in the absence of human Vλ rearrangements. We immunized IgH- and Igκ-humanized mice with the AE.A244 gp120 Env. In these mice, the V2 Ab response was focused on a linear epitope that did not include K169. V2 Abs were isolated that used the same human VH gene segment as an RV144 V2 Ab but paired with a mouse λ L chain. Structural characterization of one of these V2 Abs revealed how the linear V2 epitope could be engaged, despite the lack of aspartic acid/glutamic acid encoded in the mouse repertoire. Thus, despite the absence of the human Vλ locus in these humanized mice, the dominance of Vλ pairing with human VH for HIV-1 Env V2 recognition resulted in human VH pairing with mouse λ L chains instead of allowing otherwise subdominant V2-glycan broadly neutralizing Abs to develop.
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- 2017
15. Neutralization Takes Precedence Over IgG or IgA Isotype-related Functions in Mucosal HIV-1 Antibody-mediated Protection
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Hua-Xin Liao, Guido Ferrari, Punnee Pitisuttithum, Thomas N. Denny, Justin Pollara, Nicole L. Yates, Georgia D. Tomaras, Florian Hladik, George M. Shaw, Barton F. Haynes, M. Juliana McElrath, Georgeanna Morton, Katharine Westerberg, Tiffany Hensley-McBain, Shi-Mao Xia, Linh Mach, Sampa Santra, Benjamin Mildenberg, Gregory J. Mize, Ranjit Warrier, Rena D. Astronomo, Jaranit Kaewkungwal, Christina Ochsenbauer, Sorachai Nitayapan, Supachai Rerks-Ngarm, Lamar Ballweber-Fleming, Laura L. Sutherland, and David C. Montefiori
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0301 basic medicine ,Immunoglobulin A ,Non-human primate rectal challenge model ,Vaginal explants ,Simian Acquired Immunodeficiency Syndrome ,pIgR, Polymeric IgA receptor ,GI, Gastrointestinal ,HIV Infections ,ADCC, Antibody-dependent cell-mediated cytotoxicity ,HIV Antibodies ,Immunoglobulin G ,sIgA, Secretory IgA ,Leukocytes ,T/F, Transmitted/founder ,CD4bs, CD4 binding site ,Antibody-dependent cell-mediated cytotoxicity ,biology ,General Medicine ,Isotype ,3. Good health ,mIgA, Monomeric IgA ,dIgA, Dimeric IgA ,Vagina ,Mucosal immunology ,Female ,Simian Immunodeficiency Virus ,Antibody ,MPER, Membrane-proximal external region ,IDV, Indinivir ,IgA ,Research Paper ,AZT, Zidovudine ,ARV, Anti-retroviral ,IgG ,nnAb, Non-neutralizing antibody ,030106 microbiology ,LED, Lowest effective dose ,Neutralizing antibodies ,General Biochemistry, Genetics and Molecular Biology ,snLuc, Secreted nanoluciferase ,Antibodies ,Immunophenotyping ,GalCer, Galactosyl ceramide ,03 medical and health sciences ,Neutralization Tests ,SC, Secretory component ,Animals ,Humans ,FcRn, Neonatal Fc receptor ,Immunity, Mucosal ,IMC, Infectious molecular clone ,bnAb, Broadly neutralizing antibody ,Mucous Membrane ,Virology ,Antibodies, Neutralizing ,Macaca mulatta ,Disease Models, Animal ,030104 developmental biology ,Immunoglobulin class switching ,Immunology ,biology.protein ,HIV-1 ,mAb, Monoclonal antibody ,GU, Genitourinary ,Ex vivo ,Biomarkers - Abstract
HIV-1 infection occurs primarily through mucosal transmission. Application of biologically relevant mucosal models can advance understanding of the functional properties of antibodies that mediate HIV protection, thereby guiding antibody-based vaccine development. Here, we employed a human ex vivo vaginal HIV-1 infection model and a rhesus macaque in vivo intrarectal SHIV challenge model to probe the protective capacity of monoclonal broadly-neutralizing (bnAb) and non-neutralizing Abs (nnAbs) that were functionally modified by isotype switching. For human vaginal explants, we developed a replication-competent, secreted NanoLuc reporter virus system and showed that CD4 binding site bnAbs b12 IgG1 and CH31 IgG1 and IgA2 isoforms potently blocked HIV-1JR-CSF and HIV-1Bal26 infection. However, IgG1 and IgA nnAbs, either alone or together, did not inhibit infection despite the presence of FcR-expressing effector cells in the tissue. In macaques, the CH31 IgG1 and IgA2 isoforms infused before high-dose SHIV challenge were completely to partially protective, respectively, while nnAbs (CH54 IgG1 and CH38 mIgA2) were non-protective. Importantly, in both mucosal models IgG1 isotype bnAbs were more protective than the IgA2 isotypes, attributable in part to greater neutralization activity of the IgG1 variants. These findings underscore the importance of potent bnAb induction as a primary goal of HIV-1 vaccine development., Highlights • Only broadly-neutralizing IgG and IgA Abs were protective in two relevant mucosal models of HIV-1 infection. • IgG isotype variants were typically more protective than the IgA isotype variants against vaginal or rectal challenge. • Neutralization rather than IgA-specific functions may afford better protection against mucosal HIV-1 infection. Antibodies are likely to play a key role in preventing HIV-1 infection following mucosal exposure. We used two mucosal models, in conjunction with IgG and IgA versions of the same antibodies, to identify protective antibody properties relevant to vaginal and rectal transmission. Antibodies that can potently neutralize most HIV-1 strains were protective against viral challenge, but those without this capability were non-protective. The IgA antibodies were no more protective than their IgG counterparts even though IgA antibody forms are more abundant in the mucosa. These findings can help prioritize vaccine designs to those that induce potent broadly neutralizing IgG antibodies.
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- 2016
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16. Amino Acid Changes in the HIV-1 gp41 Membrane Proximal Region Control Virus Neutralization Sensitivity
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Nancy Tumba, Krissey E. Lloyd, Fatemeh Jahanbakhsh, Salim Abdool-Karim, Amanda Eaton, Shi-Mao Xia, Navid Madani, M. Anthony Moody, Feng Gao, S. Munir Alam, Bruno Melillo, Robert Parks, Richard M. Scearce, Amos B. Smith, David C. Montefiori, Mattia Bonsignori, Barton F. Haynes, Scott D. Boyd, Hua-Xin Liao, Laura L. Sutherland, Susan W. Barnett, Todd Bradley, Sampa Santra, Cindy M. Bowman, Joseph Sodroski, Lynn Morris, Xiaozhi Lu, Elin S. Gray, Thomas B. Kepler, and Ashley M. Trama
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0301 basic medicine ,viruses ,education ,lcsh:Medicine ,Gene Expression ,Heterologous ,HIV Infections ,HIV Antibodies ,HIV Envelope Protein gp120 ,Gp41 ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Neutralization ,Epitope ,03 medical and health sciences ,Neutralization Tests ,Antigenic variation ,Animals ,Humans ,Protein Interaction Domains and Motifs ,Amino Acid Sequence ,Antibodies, Blocking ,Peptide sequence ,health care economics and organizations ,AIDS Vaccines ,lcsh:R5-920 ,biology ,lcsh:R ,General Medicine ,Antibodies, Neutralizing ,Antigenic Variation ,Macaca mulatta ,Virology ,HIV Envelope Protein gp41 ,3. Good health ,Disease Models, Animal ,030104 developmental biology ,Amino Acid Substitution ,Mutation ,HIV-1 ,biology.protein ,Immunization ,Antibody ,lcsh:Medicine (General) ,Research Paper - Abstract
Most HIV-1 vaccines elicit neutralizing antibodies that are active against highly sensitive (tier-1) viruses or rare cases of vaccine-matched neutralization-resistant (tier-2) viruses, but no vaccine has induced antibodies that can broadly neutralize heterologous tier-2 viruses. In this study, we isolated antibodies from an HIV-1-infected individual that targeted the gp41 membrane-proximal external region (MPER) that may have selected single-residue changes in viral variants in the MPER that resulted in neutralization sensitivity to antibodies targeting distal epitopes on the HIV-1 Env. Similarly, a single change in the MPER in a second virus from another infected-individual also conferred enhanced neutralization sensitivity. These gp41 single-residue changes thus transformed tier-2 viruses into tier-1 viruses that were sensitive to vaccine-elicited tier-1 neutralizing antibodies. These data demonstrate that Env amino acid changes within the MPER bnAb epitope of naturally-selected escape viruses can increase neutralization sensitivity to multiple types of neutralizing antibodies, and underscore the critical importance of the MPER for maintaining the integrity of the tier-2 HIV-1 trimer., Highlights • Amino acid changes in the HIV gp41 MPER can regulate neutralization sensitivity of distal epitopes. • MPER antibodies isolated early are resistant to MPER changes that enhance neutralization sensitivity. • HIV gp41 MPER is critical for determining overall HIV envelope conformations. The HIV-1 envelope protein (Env) is the primary target for neutralizing antibodies. Most HIV-1 vaccines elicit neutralizing antibodies that are active against highly neutralization-sensitive (tier-1) or rare vaccine-matched more neutralization-resistant (tier-2) viruses, but no vaccine has induced antibodies that can broadly neutralize heterologous tier-2 viruses. In this study, we identified changes that occurred in two HIV-1-infected individuals in the membrane proximal region of the HIV-1 Env that resulted in neutralization sensitivity to antibodies targeting distal epitopes on the HIV Env. These single-residue changes thus transformed tier-2 viruses into tier-1 viruses, highlighting the importance of MPER residues in maintaining neutralization-resistant virus., Graphical Abstract Image 2
- Published
- 2016
17. Selection of immunoglobulin elbow region mutations impacts interdomain conformational flexibility in HIV-1 broadly neutralizing antibodies
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Robert Parks, S. Munir Alam, Mattia Bonsignori, Shi-Mao Xia, Ashley M. Trama, Kevin Wiehe, Brian E. Watts, Hua-Xin Liao, Kevin O. Saunders, Barton F. Haynes, Rory Henderson, Hieu N. Ergin, and Kara Anasti
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0301 basic medicine ,Flexibility (anatomy) ,Lineage (genetic) ,Science ,Reversion ,General Physics and Astronomy ,Heterologous ,02 engineering and technology ,HIV Antibodies ,Molecular Dynamics Simulation ,Article ,General Biochemistry, Genetics and Molecular Biology ,Virus ,Neutralization ,03 medical and health sciences ,Neutralization Tests ,medicine ,Humans ,lcsh:Science ,Genetics ,Multidisciplinary ,Calorimetry, Differential Scanning ,biology ,Circular Dichroism ,General Chemistry ,Surface Plasmon Resonance ,021001 nanoscience & nanotechnology ,Antibodies, Neutralizing ,3. Good health ,030104 developmental biology ,medicine.anatomical_structure ,Mutation ,HIV-1 ,biology.protein ,lcsh:Q ,Paratope ,Antibody ,0210 nano-technology - Abstract
Somatic mutations within antibody variable and framework regions (FWR) can alter thermostability and structural flexibility, but their impact on functional potency is unclear. Here we study thermostability and use molecular dynamics (MD) simulations to assess the role of FWR mutations during maturation of HIV-1 broadly neutralizing antibodies (bnAbs). The tested bnAbs show lower thermostability than their unmutated ancestor antibodies. FWR mutations in the Fab elbow region are frequently observed in HIV-1 bnAbs and MD simulations show that such FWR mutations alter interdomain flexibility in two HIV-1 bnAbs. In a CD4-binding site lineage, reversion mutations result in a loss of neutralization potency in an early intermediate and affinity-matured bnAb against autologous and heterologous Tier-2 viruses, respectively. Elbow region reversion mutations in a glycan-V3 bnAb modestly reduces potency against an autologous virus isolate. Thus, selection of mutations in the Fab elbow region impacts interdomain conformational flexibility and paratope plasticity during bnAb development., Somatic mutations within antibody framework regions (FWR) can alter structural flexibility, but their role in maturation of broadly neutralizing antibodies (bnAbs) is unclear. Here the authors show how FWR mutations impact interdomain conformational flexibility and paratope plasticity during bnAb development.
- Published
- 2019
18. Inference of the HIV-1 VRC01 Antibody Lineage Unmutated Common Ancestor Reveals Alternative Pathways to Overcome a Key Glycan Barrier
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Mattia, Bonsignori, Eric, Scott, Kevin, Wiehe, David, Easterhoff, S Munir, Alam, Kwan-Ki, Hwang, Melissa, Cooper, Shi-Mao, Xia, Ruijun, Zhang, David C, Montefiori, Rory, Henderson, Xiaoyan, Nie, Garnett, Kelsoe, M Anthony, Moody, Xuejun, Chen, M Gordon, Joyce, Peter D, Kwong, Mark, Connors, John R, Mascola, Andrew T, McGuire, Leonidas, Stamatatos, Max, Medina-Ramírez, Rogier W, Sanders, Kevin O, Saunders, Thomas B, Kepler, Barton F, Haynes, Medical Microbiology and Infection Prevention, and AII - Infectious diseases
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monoclonal antibody VRC01 ,immunogen design ,clonal evolution ,HIV Infections ,HIV Antibodies ,HIV Envelope Protein gp120 ,Article ,Polysaccharides ,Humans ,neutralizing antibodies ,Amino Acid Sequence ,indel mutation ,cell lineage ,Phylogeny ,AIDS Vaccines ,B-Lymphocytes ,Binding Sites ,Sequence Homology, Amino Acid ,Antibodies, Monoclonal ,vaccines ,Antibodies, Neutralizing ,CD4 Antigens ,HIV-1 ,glycans ,Broadly Neutralizing Antibodies ,B lymphocytes - Abstract
Summary Elicitation of VRC01-class broadly neutralizing antibodies (bnAbs) is an appealing approach for a preventative HIV-1 vaccine. Despite extensive investigations, strategies to induce VRC01-class bnAbs and overcome the barrier posed by the envelope N276 glycan have not been successful. Here, we inferred a high-probability unmutated common ancestor (UCA) of the VRC01 lineage and reconstructed the stages of lineage maturation. Env immunogens designed on reverted VRC01-class bnAbs bound to VRC01 UCA with affinity sufficient to activate naive B cells. Early mutations defined maturation pathways toward limited or broad neutralization, suggesting that focusing the immune response is likely required to steer B cell maturation toward the development of neutralization breadth. Finally, VRC01 lineage bnAbs with long CDR H3s overcame the HIV-1 N276 glycan barrier without shortening their CDR L1, revealing a solution for broad neutralization in which the heavy chain, not CDR L1, is the determinant to accommodate the N276 glycan., Graphical Abstract, Highlights • A high-probability VRC01 lineage UCA was inferred and CDRH3 evolution defined • Env immunogens bind to VRC01 UCA with affinity sufficient to activate naive B cells • Early mutations defined maturation pathways toward limited or broad neutralization • Antibodies with long CDRH3s achieved neutralization breadth without shortening CDRL1s, Understanding how HIV-1 VRC01-class broadly neutralizing antibodies overcome maturation barriers is key for vaccine development. Bonsignori et al. inferred the unmutated common ancestor of the VRC01 lineage, reconstructed the stages of lineage maturation, and identified multiple solutions adopted by evolving B cells to overcome the N276 glycan barrier and achieve broad neutralization.
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- 2018
19. Initiation of HIV neutralizing B cell lineages with sequential envelope immunizations
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Barton F. Haynes, Hua-Xin Liao, Cindy M. Bowman, Kevin O. Saunders, Nathan Vandergrift, Akshaya Ramesh, Christopher B. Fox, Nathan I. Nicely, Todd Bradley, John R. Mascola, Laura L. Sutherland, Sampa Santra, Dawn J. Marshall, Garnett Kelsoe, Robert Parks, David Franco, Amanda Newman, S. Munir Alam, Hilary Bouton-Verville, Laurent Verkoczy, Kan Luo, James A. Holland, Richard M. Scearce, Marguerite Koutsoukos, Jinsong Zhang, Feng Gao, M. Anthony Moody, Xiaoyan Nie, Steven G. Reed, Thomas B. Kepler, Kevin Wiehe, Amanda Eaton, Mark K. Louder, David C. Montefiori, Wilton B. Williams, Chuancang Jiang, Andrew Foulger, Stephen C. Harrison, John F. Whitesides, Kim Clary, Mattia Bonsignori, Shi-Mao Xia, Howard M. Bomze, and Daniela Fera
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Male ,Models, Molecular ,0301 basic medicine ,Immunogen ,Science ,viruses ,General Physics and Astronomy ,Mice, Transgenic ,HIV Antibodies ,Biology ,Article ,General Biochemistry, Genetics and Molecular Biology ,Immune tolerance ,Affinity maturation ,Mice ,03 medical and health sciences ,Immune Tolerance ,medicine ,Animals ,Humans ,Cell Lineage ,Gene Knock-In Techniques ,B cell ,AIDS Vaccines ,Clonal Anergy ,B-Lymphocytes ,Multidisciplinary ,env Gene Products, Human Immunodeficiency Virus ,virus diseases ,Germinal center ,Receptor editing ,General Chemistry ,Antibodies, Neutralizing ,Macaca mulatta ,Virology ,3. Good health ,Vaccination ,030104 developmental biology ,medicine.anatomical_structure ,Immunization ,CD4 Antigens ,HIV-1 ,Female ,Binding Sites, Antibody - Abstract
A strategy for HIV-1 vaccine development is to define envelope (Env) evolution of broadly neutralizing antibodies (bnAbs) in infection and to recreate those events by vaccination. Here, we report host tolerance mechanisms that limit the development of CD4-binding site (CD4bs), HCDR3-binder bnAbs via sequential HIV-1 Env vaccination. Vaccine-induced macaque CD4bs antibodies neutralize 7% of HIV-1 strains, recognize open Env trimers, and accumulate relatively modest somatic mutations. In naive CD4bs, unmutated common ancestor knock-in mice Env+B cell clones develop anergy and partial deletion at the transitional to mature B cell stage, but become Env− upon receptor editing. In comparison with repetitive Env immunizations, sequential Env administration rescue anergic Env+ (non-edited) precursor B cells. Thus, stepwise immunization initiates CD4bs-bnAb responses, but immune tolerance mechanisms restrict their development, suggesting that sequential immunogen-based vaccine regimens will likely need to incorporate strategies to expand bnAb precursor pools., An efficient HIV-1 vaccine will likely depend on eliciting broadly neutralizing antibodies (bnAb). Here the authors analyze the B cell repertoire in macaques and knock-in mice in response to sequential immunization with Env variants that induce a bnAb targeting the CD4-binding site of Env in a HIV-1 infected individual.
- Published
- 2017
20. Maternal HIV-1 envelope–specific antibody responses and reduced risk of perinatal transmission
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Fangping Cai, Marcella Sarzotti-Kelsoe, M. Anthony Moody, Shi Mao Xia, David C. Montefiori, Peter B. Gilbert, Robert Parks, Amit Kumar, Emma Friberg, Raul Louzao, Guido Ferrari, Krissey E. Lloyd, Thaddeus C. Gurley, Hua-Xin Liao, Thomas N. Denny, Tarra Von Holle, Youyi Fong, Samantha Wilkes, Dawn J. Marshall, Erika Solomon, Xiaoying Shen, Kaylan Whitaker, Genevieve G. Fouda, David R. Martinez, Frederick H. Jaeger, S. Munir Alam, Saheli Datta, Jamie Pritchett, Brooke E. Liebl, Nathan Vandergrift, Feng Gao, Xiaozhi Lu, Sallie R. Permar, Georgia D. Tomaras, John F. Whitesides, Barton F. Haynes, Amanda Martelli, Nicole L. Yates, Justin Pollara, R. Glenn Overman, and Haiyan Chen
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Multivariate analysis ,HIV Infections ,HIV Antibodies ,HIV Envelope Protein gp120 ,Logistic regression ,Immunoglobulin G ,Neutralization ,law.invention ,Cohort Studies ,Antibody Specificity ,Pregnancy ,Risk Factors ,law ,medicine ,Humans ,Pregnancy Complications, Infectious ,Antigens, Viral ,AIDS Vaccines ,biology ,business.industry ,Brief Report ,Infant, Newborn ,Vaccine trial ,Infant ,virus diseases ,General Medicine ,medicine.disease ,Antibodies, Neutralizing ,Virology ,Infectious Disease Transmission, Vertical ,Peptide Fragments ,Logistic Models ,Transmission (mechanics) ,Multivariate Analysis ,Immunology ,HIV-1 ,biology.protein ,Female ,business ,Cohort study - Abstract
Despite the wide availability of antiretroviral drugs, more than 250,000 infants are vertically infected with HIV-1 annually, emphasizing the need for additional interventions to eliminate pediatric HIV-1 infections. Here, we aimed to define humoral immune correlates of risk of mother-to-child transmission (MTCT) of HIV-1, including responses associated with protection in the RV144 vaccine trial. Eighty-three untreated, HIV-1–transmitting mothers and 165 propensity score–matched nontransmitting mothers were selected from the Women and Infants Transmission Study (WITS) of US nonbreastfeeding, HIV-1–infected mothers. In a multivariable logistic regression model, the magnitude of the maternal IgG responses specific for the third variable loop (V3) of the HIV-1 envelope was predictive of a reduced risk of MTCT. Neutralizing Ab responses against easy-to-neutralize (tier 1) HIV-1 strains also predicted a reduced risk of peripartum transmission in secondary analyses. Moreover, recombinant maternal V3–specific IgG mAbs mediated neutralization of autologous HIV-1 isolates. Thus, common V3-specific Ab responses in maternal plasma predicted a reduced risk of MTCT and mediated autologous virus neutralization, suggesting that boosting these maternal Ab responses may further reduce HIV-1 MTCT.
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- 2015
21. Potent and broad HIV-neutralizing antibodies in memory B cells and plasma
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Barton F. Haynes, Kshitij Wagh, Krisha McKee, Michael S. Seaman, Bette T. Korber, Xiaozhi Lu, Nathan I. Nicely, M. Anthony Moody, Robert Parks, Georgia D. Tomaras, John R. Mascola, Feng Gao, Gilad Ofek, Jonathan R. McDaniel, Kevin Wiehe, Dawn J. Marshall, Robert T. Bailer, Kara Anasti, Caleb S. Lougheed, Shi Mao Xia, Mattia Bonsignori, Ivelin S. Georgiev, Lynn Morris, Guang Yang, Kevin O. Saunders, Gift Kamanga, Sebastian Schätzle, Todd Bradley, Thomas B. Kepler, George Georgiou, Hua-Xin Liao, Xiaoyan Nie, Myron S. Cohen, Mark K. Louder, La Tonya D. Williams, S. Munir Alam, Sijy O'Dell, Nancy L. Tumba, Liming Wu, Garnett Kelsoe, and David C. Montefiori
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0301 basic medicine ,Lineage (genetic) ,biology ,Immunology ,General Medicine ,Gp41 ,Virology ,Epitope ,Article ,Immune tolerance ,03 medical and health sciences ,030104 developmental biology ,0302 clinical medicine ,Immune system ,medicine.anatomical_structure ,biology.protein ,medicine ,Antibody ,Memory B cell ,B cell ,030215 immunology - Abstract
Induction of broadly neutralizing antibodies (bnAbs) is a goal of HIV-1 vaccine development. Antibody 10E8, reactive with the distal portion of the membrane-proximal external region (MPER) of HIV-1 gp41, is broadly neutralizing. However, the ontogeny of distal MPER antibodies and the relationship of memory B cell to plasma bnAbs are poorly understood. HIV-1–specific memory B cell flow sorting and proteomic identification of anti-MPER plasma antibodies from an HIV-1–infected individual were used to isolate broadly neutralizing distal MPER bnAbs of the same B cell clonal lineage. Structural analysis demonstrated that antibodies from memory B cells and plasma recognized the envelope gp41 bnAb epitope in a distinct orientation compared with other distal MPER bnAbs. The unmutated common ancestor of this distal MPER bnAb was autoreactive, suggesting lineage immune tolerance control. Construction of chimeric antibodies of memory B cell and plasma antibodies yielded a bnAb that potently neutralized most HIV-1 strains.
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- 2017
22. Staged induction of HIV-1 glycan–dependent broadly neutralizing antibodies
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S. Munir Alam, Mark K. Louder, Kevin Wiehe, Brendan W. Pier, Feng Gao, Barton F. Haynes, Kshitij Wagh, John R. Mascola, Morgan A. Gladden, Kwan-Ki Hwang, Peter T. Hraber, Ruijun Zhang, William E. Walkowicz, George M. Shaw, Stephen C. Harrison, Anthony Monroe, Beatrice H. Hahn, M. Anthony Moody, Hua-Xin Liao, Amit Kumar, Lynn Morris, Gift Kamanga, R. Ryan Meyerhoff, Kevin O. Saunders, Myron S. Cohen, Edward F. Kreider, Todd Bradley, Bette T. Korber, Robert T. Bailer, Mattia Bonsignori, John C. Kappes, Ashley M. Trama, Wilton B. Williams, Garnett Kelsoe, Daniela Fera, Shi-Mao Xia, Thomas B. Kepler, Claudia A. Jette, Samuel J. Danishefsky, Krisha McKee, Baptiste Aussedat, Melissa Cooper, and David C. Montefiori
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Male ,0301 basic medicine ,Cytidine deaminase activity ,Lineage (genetic) ,Somatic hypermutation ,Crystallography, X-Ray ,Article ,Epitope ,Virus ,Affinity maturation ,03 medical and health sciences ,Neutralization Tests ,Polysaccharides ,Humans ,Neutralizing antibody ,Phylogeny ,biology ,env Gene Products, Human Immunodeficiency Virus ,General Medicine ,Antibodies, Neutralizing ,Virology ,030104 developmental biology ,Mutation ,HIV-1 ,biology.protein ,Antibody ,Protein Binding - Abstract
A preventive HIV-1 vaccine should induce HIV-1-specific broadly neutralizing antibodies (bnAbs). However, bnAbs generally require high levels of somatic hypermutation (SHM) to acquire breadth, and current vaccine strategies have not been successful in inducing bnAbs. Because bnAbs directed against a glycosylated site adjacent to the third variable loop (V3) of the HIV-1 envelope protein require limited SHM, the V3-glycan epitope is an attractive vaccine target. By studying the cooperation among multiple V3-glycan B cell lineages and their coevolution with autologous virus throughout 5 years of infection, we identify key events in the ontogeny of a V3-glycan bnAb. Two autologous neutralizing antibody lineages selected for virus escape mutations and consequently allowed initiation and affinity maturation of a V3-glycan bnAb lineage. The nucleotide substitution required to initiate the bnAb lineage occurred at a low-probability site for activation-induced cytidine deaminase activity. Cooperation of B cell lineages and an improbable mutation critical for bnAb activity defined the necessary events leading to breadth in this V3-glycan bnAb lineage. These findings may, in part, explain why initiation of V3-glycan bnAbs is rare, and suggest an immunization strategy for inducing similar V3-glycan bnAbs.
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- 2017
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23. Cooperation of B Cell Lineages in Induction of HIV-1-Broadly Neutralizing Antibodies
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Hua-Xin Liao, Shi Mao Xia, Hongshuo Song, M. Anthony Moody, John R. Mascola, Rebecca M. Lynch, Barton F. Haynes, Kwan Ki Hwang, S. Munir Alam, Fangping Cai, Gift Kamanga, Peter D. Kwong, David C. Montefiori, Xiaozhi Lu, Myron S. Cohen, Feng Gao, Mattia Bonsignori, Tongqing Zhou, Amit Kumar, Bette T. Korber, Guido Ferrari, George M. Shaw, Peter T. Hraber, Mark Berrong, Thomas B. Kepler, Beatrice H. Hahn, and Garnett Kelsoe
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Models, Molecular ,Immunogen ,Lineage (evolution) ,Mutant ,Molecular Sequence Data ,Biology ,HIV Antibodies ,medicine.disease_cause ,General Biochemistry, Genetics and Molecular Biology ,Virus ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Amino Acid Sequence ,B cell ,030304 developmental biology ,Immune Evasion ,AIDS Vaccines ,0303 health sciences ,Mutation ,B-Lymphocytes ,Biochemistry, Genetics and Molecular Biology(all) ,env Gene Products, Human Immunodeficiency Virus ,Virology ,Antibodies, Neutralizing ,3. Good health ,medicine.anatomical_structure ,biology.protein ,HIV-1 ,Antibody ,Sequence Alignment ,030217 neurology & neurosurgery - Abstract
SummaryDevelopment of strategies for induction of HIV-1 broadly neutralizing antibodies (bnAbs) by vaccines is a priority. Determining the steps of bnAb induction in HIV-1-infected individuals who make bnAbs is a key strategy for immunogen design. Here, we study the B cell response in a bnAb-producing individual and report cooperation between two B cell lineages to drive bnAb development. We isolated a virus-neutralizing antibody lineage that targeted an envelope region (loop D) and selected virus escape mutants that resulted in both enhanced bnAb lineage envelope binding and escape mutant neutralization—traits associated with increased B cell antigen drive. Thus, in this individual, two B cell lineages cooperated to induce the development of bnAbs. Design of vaccine immunogens that simultaneously drive both helper and broadly neutralizing B cell lineages may be important for vaccine-induced recapitulation of events that transpire during the maturation of neutralizing antibodies in HIV-1-infected individuals.
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- 2014
- Full Text
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24. Toll-Like Receptor 7/8 (TLR7/8) and TLR9 Agonists Cooperate To Enhance HIV-1 Envelope Antibody Responses in Rhesus Macaques
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Mark S. Drinker, Barton F. Haynes, Brandy M. Ward, Laura L. Sutherland, Ashley A. Allen, Thaddeus C. Gurley, Robert Parks, Hua-Xin Liao, Robert A. Seder, David Easterhoff, Krissey E. Lloyd, David C. Montefiori, R. Ryan Meyerhoff, M. Anthony Moody, Norman L. Letvin, Sampa Santra, Georgia D. Tomaras, Guido Ferrari, S. Munir Alam, Shi-Mao Xia, and Nathan Vandergrift
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Male ,Immunogen ,medicine.medical_treatment ,Immunology ,HIV Infections ,HIV Antibodies ,Ligands ,Microbiology ,Adjuvants, Immunologic ,Virology ,Vaccines and Antiviral Agents ,medicine ,Animals ,Humans ,Toll-like receptor ,biology ,Immunogenicity ,env Gene Products, Human Immunodeficiency Virus ,TLR9 ,TLR7 ,Acquired immune system ,Macaca mulatta ,Toll-Like Receptor 7 ,Toll-Like Receptor 8 ,Toll-Like Receptor 9 ,Insect Science ,HIV-1 ,biology.protein ,Female ,Immunization ,Antibody ,Adjuvant - Abstract
The development of a vaccine that can induce high titers of functional antibodies against HIV-1 remains a high priority. We have developed an adjuvant based on an oil-in-water emulsion that incorporates Toll-like receptor (TLR) ligands to test whether triggering multiple pathogen-associated molecular pattern receptors could enhance immunogenicity. Compared to single TLR agonists or other pairwise combinations, TLR7/8 and TLR9 agonists combined were able to elicit the highest titers of binding, neutralizing, and antibody-dependent cellular cytotoxicity-mediating antibodies against the protein immunogen, transmitted/founder HIV-1 envelope gp140 (B.63521). We further found that the combination of TLR7/8 and TLR9 agonists was associated with the release of CXCL10 (IP-10), suggesting that this adjuvant formulation may have optimally stimulated innate and adaptive immunity to elicit high titers of antibodies. IMPORTANCE Combining TLR agonists in an adjuvant formulation resulted in higher antibody levels compared to an adjuvant without TLR agonists. Adjuvants that combine TLR agonists may be useful for enhancing antibody responses to HIV-1 vaccines.
- Published
- 2014
25. Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques
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Laura L. Sutherland, Guido Ferrari, Giovanna Hernandez, Jamie Pritchett, Norman L. Letvin, Supachai Rerks-Ngarm, Faruk Sinangil, Ashley A. Allen, Nelson L. Michael, Robert Parks, Erika Dunford, Donald P. Francis, Mattia Bonsignori, Georgia D. Tomaras, John F. Whitesides, Punnee Pitisuttithum, David Easterhoff, Christina Stolarchuk, R. Whitney Edwards, M. Anthony Moody, Krissey E. Lloyd, Thaddeus C. Gurley, Hua-Xin Liao, Barton F. Haynes, Carter Lee, Andrew Foulger, Tarra Von Holle, Sorachai Nitayaphan, Dawn J. Marshall, Jaranit Kaewkungwal, Sampa Santra, Ruijun Zhang, Kan Luo, Shi-Mao Xia, Kevin Wiehe, Richard M. Scearce, Sabrina Arora, Jerome H. Kim, Sheetal Sawant, David C. Montefiori, Cindy M. Bowman, Lawrence C. Armand, S. Munir Alam, James Tartaglia, Nathan Vandergrift, Amy Wang, Nicole L. Yates, Jessica N. Peel, and Justin Pollara
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0301 basic medicine ,Immunization, Secondary ,HIV Infections ,HIV Antibodies ,HIV Envelope Protein gp120 ,03 medical and health sciences ,0302 clinical medicine ,Immunity ,medicine ,Animals ,Memory B cell ,Immunity, Mucosal ,B cell ,AIDS Vaccines ,Antibody-dependent cell-mediated cytotoxicity ,B-Lymphocytes ,biology ,Vaccine trial ,General Medicine ,Macaca mulatta ,Virology ,3. Good health ,030104 developmental biology ,medicine.anatomical_structure ,Immunization ,Immunoglobulin G ,030220 oncology & carcinogenesis ,AIDSVAX ,Immunology ,biology.protein ,Antibody ,Research Article - Abstract
The ALVAC prime/ALVAC + AIDSVAX B/E boost RV144 vaccine trial induced an estimated 31% efficacy in a low-risk cohort where HIV‑1 exposures were likely at mucosal surfaces. An immune correlates study demonstrated that antibodies targeting the V2 region and in a secondary analysis antibody-dependent cellular cytotoxicity (ADCC), in the presence of low envelope-specific (Env-specific) IgA, correlated with decreased risk of infection. Thus, understanding the B cell repertoires induced by this vaccine in systemic and mucosal compartments are key to understanding the potential protective mechanisms of this vaccine regimen. We immunized rhesus macaques with the ALVAC/AIDSVAX B/E gp120 vaccine regimen given in RV144, and then gave a boost 6 months later, after which the animals were necropsied. We isolated systemic and intestinal vaccine Env-specific memory B cells. Whereas Env-specific B cell clonal lineages were shared between spleen, draining inguinal, anterior pelvic, posterior pelvic, and periaortic lymph nodes, members of Env‑specific B cell clonal lineages were absent in the terminal ileum. Env‑specific antibodies were detectable in rectal fluids, suggesting that IgG antibodies present at mucosal sites were likely systemically produced and transported to intestinal mucosal sites.
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- 2016
26. Induction of HIV-1 Broad Neutralizing Antibodies in 2F5 Knock-in Mice: Selection against Membrane Proximal External Region–Associated Autoreactivity Limits T-Dependent Responses
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S. Moses Dennison, Hua-Xin Liao, Richard M. Scearce, Barton F. Haynes, Yao Chen, Laurent Verkoczy, Shi-Mao Xia, Amanda Newman, Bradley Lockwood, Hilary Bouton-Verville, Kwan-Ki Hwang, David C. Montefiori, S. Munir Alam, and Jinsong Zhang
- Subjects
Immunogen ,T-Lymphocytes ,Immunology ,Enzyme-Linked Immunosorbent Assay ,HIV Antibodies ,Lymphocyte Activation ,Gp41 ,Article ,Epitope ,Clonal deletion ,Mice ,Negative selection ,Neutralization Tests ,Gene knockin ,Animals ,Humans ,Immunology and Allergy ,Gene Knock-In Techniques ,Antigens, Viral ,AIDS Vaccines ,Mice, Knockout ,B-Lymphocytes ,biology ,breakpoint cluster region ,Antibodies, Monoclonal ,Antibodies, Neutralizing ,Molecular biology ,HIV Envelope Protein gp41 ,HIV-1 ,biology.protein ,Antibody ,Broadly Neutralizing Antibodies - Abstract
A goal of HIV-1 vaccine development is to elicit broadly neutralizing Abs (BnAbs). Using a knock-in (KI) model of 2F5, a human HIV-1 gp41 membrane proximal external region (MPER)–specific BnAb, we previously demonstrated that a key obstacle to BnAb induction is clonal deletion of BnAb-expressing B cells. In this study of this model, we provide a proof-of-principle that robust serum neutralizing IgG responses can be induced from pre-existing, residual, self-reactive BnAb-expressing B cells in vivo using a structurally compatible gp41 MPER immunogen. Furthermore, in CD40L-deficient 2F5 KI mice, we demonstrate that these BnAb responses are elicited via a type II T-independent pathway, coinciding with expansion and activation of transitional splenic B cells specific for 2F5's nominal gp41 MPER-binding epitope (containing the 2F5 neutralization domain ELDKWA). In contrast, constitutive production of nonneutralizing serum IgGs in 2F5 KI mice is T dependent and originates from a subset of splenic mature B2 cells that have lost their ability to bind 2F5's gp41 MPER epitope. These results suggest that residual, mature B cells expressing autoreactive BnAbs, like 2F5 as BCR, may be limited in their ability to participate in T-dependent responses by purifying selection that selectively eliminates reactivity for neutralization epitope-containing/mimicked host Ags.
- Published
- 2013
27. Common Tolerance Mechanisms, but Distinct Cross-Reactivities Associated with gp41 and Lipids, Limit Production of HIV-1 Broad Neutralizing Antibodies 2F5 and 4E10
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Yao Chen, Kwan-Ki Hwang, Barton F. Haynes, Hilary Bouton-Verville, Laurent Verkoczy, Ying-Bin Ouyang, Jinsong Zhang, Shi-Mao Xia, and Amanda Newman
- Subjects
medicine.drug_class ,Cellular differentiation ,Central B cell deletion ,Immunology ,Receptors, Antigen, B-Cell ,Cross Reactions ,HIV Antibodies ,Biology ,Monoclonal antibody ,Article ,Immunoglobulin G ,Immune tolerance ,Mice ,Antigen ,Immune Tolerance ,medicine ,Animals ,Immunology and Allergy ,Gene Knock-In Techniques ,AIDS Vaccines ,Mice, Knockout ,B-Lymphocytes ,breakpoint cluster region ,Antibodies, Monoclonal ,Cell Differentiation ,Antibodies, Neutralizing ,Lipids ,Molecular biology ,HIV Envelope Protein gp41 ,Mice, Inbred C57BL ,HIV-1 ,biology.protein ,Antibody ,Broadly Neutralizing Antibodies - Abstract
Developing an HIV-1 vaccine has been hampered by the inability of immunogens to induce broadly neutralizing Abs (BnAbs) that protect against infection. Previously, we used knockin (KI) mice expressing a prototypical gp41-specific BnAb, 2F5, to demonstrate that immunological tolerance triggered by self-reactivity of the 2F5 H chain impedes BnAb induction. In this study, we generate KI models expressing H chains from two other HIV-1 Abs, 4E10 (another self-/polyreactive, anti-gp41 BnAb) and 48d (an anti-CD4 inducible, nonpolyreactive Ab), and find a similar developmental blockade consistent with central B cell deletion in 4E10, but not in 48d VH KI mice. Furthermore, in KI strains expressing the complete 2F5 and 4E10 Abs as BCRs, we find that residual splenic B cells arrest at distinct developmental stages, yet exhibit uniformly low BCR densities, elevated basal activation, and profoundly muted responses to BCR ligation and, when captured as hybridoma mAb lines, maintain their dual (gp41/lipid) affinities and capacities to neutralize HIV-1, establishing a key role for anergy in suppressing residual 2F5- or 4E10-expressing B cells. Importantly, serum IgGs from naive 2F5 and 4E10 KI strains selectively eliminate gp41 and lipid binding, respectively, suggesting B cells expressing 2F5 or 4E10 as BCRs exhibit specificity for a distinct spectrum of host Ags, including selective interactions by 2F5 BCR+ B cells (i.e., and not 4E10 BCR+ B cells) with those mimicked by its gp41 neutralization epitope.
- Published
- 2013
28. Antigenicity and Immunogenicity of Transmitted/Founder, Consensus, and Chronic Envelope Glycoproteins of Human Immunodeficiency Virus Type 1
- Author
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Alan Lapedes, R. Swanstrom, Julie Blinn, Andrew Foulger, Chun-Yen Tsao, Sydeaka Watson, Mark Muldoon, Hua-Xin Liao, Beatrice H. Hahn, Richard M. Scearce, Bette T. Korber, Xiaozhi Lu, Robert Parks, Shi-Mao Xia, Ben-Jiang Ma, George M. Shaw, Laura L. Sutherland, Haiyan Chen, Barton F. Haynes, Feng Gao, Cindy M. Bowman, Nathan Vandergrift, S. Munir Alam, and David C. Montefiori
- Subjects
Antigenicity ,medicine.drug_class ,viruses ,Guinea Pigs ,Immunology ,Antibody Affinity ,Human immunodeficiency virus (HIV) ,Cross Reactions ,HIV Antibodies ,Monoclonal antibody ,medicine.disease_cause ,Microbiology ,Virology ,Vaccines and Antiviral Agents ,medicine ,Animals ,Humans ,Potency ,Binding site ,Antigens, Viral ,Glycoproteins ,chemistry.chemical_classification ,biology ,Immunogenicity ,env Gene Products, Human Immunodeficiency Virus ,virus diseases ,Antibodies, Neutralizing ,chemistry ,Insect Science ,HIV-1 ,biology.protein ,Antibody ,Glycoprotein - Abstract
Human immunodeficiency virus type 1 (HIV-1) vaccine development requires selection of appropriate envelope (Env) immunogens. Twenty HIV-1 Env glycoproteins were examined for their ability to bind human anti-HIV-1 monoclonal antibodies (MAbs) and then used as immunogens in guinea pigs to identify promising immunogens. These included five Envs derived from chronically infected individuals, each representing one of five common clades and eight consensus Envs based on these five clades, as well as the consensus of the entire HIV-1 M group, and seven transmitted/founder (T/F) Envs from clades B and C. Sera from immunized guinea pigs were tested for neutralizing activity using 36 HIV-1 Env-pseudotyped viruses. All Envs bound to CD4 binding site, membrane-proximal, and V1/V2 MAbs with similar apparent affinities, although the T/F Envs bound with higher affinity to the MAb 17b, a CCR5 coreceptor binding site antibody. However, the various Envs differed in their ability to induce neutralizing antibodies. Consensus Envs elicited the most potent responses, but neutralized only a subset of viruses, including mostly easy-to-neutralize tier 1 and some more-difficult-to-neutralize tier 2 viruses. T/F Envs elicited fewer potent neutralizing antibodies but exhibited greater breadth than chronic or consensus Envs. Finally, chronic Envs elicited the lowest level and most limited breadth of neutralizing antibodies overall. Thus, each group of Env immunogens elicited a different antibody response profile. The complementary benefits of consensus and T/F Env immunogens raise the possibility that vaccines utilizing a combination of consensus and T/F Envs may be able to induce neutralizing responses with greater breadth and potency than single Env immunogens.
- Published
- 2013
29. Co-evolution of a broadly neutralizing HIV-1 antibody and founder virus
- Author
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Barton F. Haynes, Nisc Comparative Sequencing Program, John R. Mascola, Chaim A. Schramm, Krissey E. Lloyd, Robert Parks, Xiulian Du, Jiang Zhu, Lillian Tran, James C. Mullikin, Peter T. Hraber, Richard M. Scearce, Lawrence Shapiro, Shi-Mao Xia, Hua-Xin Liao, Peter D. Kwong, Fangping Cai, Zhenhai Zhang, Garnett Kelsoe, Sandrasegaram Gnanakaran, Feng Gao, Myron S. Cohen, Beatrice H. Hahn, M. Gordon Joyce, Anqi Zheng, Kevin Wiehe, Baoshan Zhang, Scott D. Boyd, Sanjay Srivatsan, Andrew Fire, Stephanie Moquin, Mark K. Louder, Guang Yang, S. Munir Alam, Gift Kamanga, Tongqing Zhou, Thomas B. Kepler, David C. Montefiori, Yue Chen, Bette T. Korber, Krishna M. Roskin, George M. Shaw, Rebecca M. Lynch, Kelly A. Soderberg, and Sheri Chen
- Subjects
Models, Molecular ,Molecular Sequence Data ,Cross Reactions ,HIV Antibodies ,HIV Envelope Protein gp120 ,Crystallography, X-Ray ,medicine.disease_cause ,Epitope ,Virus ,Neutralization ,Article ,Evolution, Molecular ,Epitopes ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Neutralization Tests ,medicine ,Humans ,Cell Lineage ,Amino Acid Sequence ,Cells, Cultured ,Phylogeny ,030304 developmental biology ,AIDS Vaccines ,0303 health sciences ,Mutation ,Multidisciplinary ,biology ,Antibodies, Monoclonal ,Antibodies, Neutralizing ,Virology ,Clone Cells ,Protein Structure, Tertiary ,3. Good health ,Vaccination ,Viral evolution ,Africa ,CD4 Antigens ,HIV-1 ,biology.protein ,Antibody ,030217 neurology & neurosurgery - Abstract
Current human immunodeficiency virus-1 (HIV-1) vaccines elicit strain-specific neutralizing antibodies. However, cross-reactive neutralizing antibodies arise in approximately 20% of HIV-1-infected individuals, and details of their generation could provide a blueprint for effective vaccination. Here we report the isolation, evolution and structure of a broadly neutralizing antibody from an African donor followed from the time of infection. The mature antibody, CH103, neutralized approximately 55% of HIV-1 isolates, and its co-crystal structure with the HIV-1 envelope protein gp120 revealed a new loop-based mechanism of CD4-binding-site recognition. Virus and antibody gene sequencing revealed concomitant virus evolution and antibody maturation. Notably, the unmutated common ancestor of the CH103 lineage avidly bound the transmitted/founder HIV-1 envelope glycoprotein, and evolution of antibody neutralization breadth was preceded by extensive viral diversification in and near the CH103 epitope. These data determine the viral and antibody evolution leading to induction of a lineage of HIV-1 broadly neutralizing antibodies, and provide insights into strategies to elicit similar antibodies by vaccination.
- Published
- 2013
30. Nonneutralizing HIV-1 gp41 Envelope Cluster II Human Monoclonal Antibodies Show Polyreactivity for Binding to Phospholipids and Protein Autoantigens
- Author
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S. Munir Alam, Hua-Xin Liao, Robert Parks, Laura L. Sutherland, Richard M. Scearce, S. Moses Dennison, Susan Zolla-Pazner, Miroslaw K. Gorny, Barton F. Haynes, Kara Anasti, and Shi-Mao Xia
- Subjects
medicine.drug_class ,viruses ,Immunology ,Plasma protein binding ,HIV Antibodies ,Gp41 ,Monoclonal antibody ,Autoantigens ,Microbiology ,Neutralization ,Epitope ,Virology ,medicine ,Humans ,Phospholipids ,biology ,Binding protein ,Antibodies, Monoclonal ,virus diseases ,Primary and secondary antibodies ,Molecular biology ,HIV Envelope Protein gp41 ,Kinetics ,Insect Science ,biology.protein ,Pathogenesis and Immunity ,Antibody ,Protein Binding - Abstract
HIV-1 gp41 envelope antibodies, which are frequently induced in HIV-1-infected individuals, are predominantly nonneutralizing. The rare and difficult-to-induce neutralizing antibodies (2F5 and 4E10) that target gp41 membrane-proximal epitopes (MPER) are polyspecific and require lipid binding for HIV-1 neutralization. These results raise the questions of how prevalent polyreactivity is among gp41 antibodies and how the binding properties of gp41-nonneutralizing antibodies differ from those of antibodies that are broadly neutralizing. In this study, we have characterized a panel of human gp41 antibodies with binding specificities within the immunodominant cluster I (gp41 amino acids [aa] 579 to 613) or cluster II (gp41 aa 644 to 667) for reactivity to autoantigens, to the gp140 protein, and with MPER peptide-lipid conjugates. We report that while none of the gp41 cluster I antibodies studied were polyspecific, all three gp41 cluster II antibodies bound either to lipids or autoantigens, thus showing the propensity of cluster II antibodies to manifest polyreactivity. All cluster II gp41 monoclonal antibodies (MAbs), including those that were lipid reactive, failed to bind to gp41 MPER peptide-lipid complexes. Cluster II antibodies bound strongly with nanomolar binding affinity (dissociation constant [ K d ]) to oligomeric gp140 proteins, and thus, they recognize conformational epitopes on gp41 that are distinct from those of neutralizing gp41 antibodies. These results demonstrate that lipid-reactive gp41 cluster II antibodies are nonneutralizing due to their inability to bind to the relevant neutralizing epitopes on gp41.
- Published
- 2011
31. Anti-phospholipid human monoclonal antibodies inhibit CCR5-tropic HIV-1 and induce β-chemokines
- Author
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Barton F. Haynes, Christina Ochsenbauer-Jambor, Kwan Ki Hwang, Ruijun Zhang, Anita Kavlie, Gary Landucci, John C. Kappes, M. Anthony Moody, Tara G. Edmonds, Steven W. King, Ian Giles, Connie Chang, Pojen P. Chen, Mark S. Drinker, Melina Soares, Thomas N. Denny, Georgia D. Tomaras, Donald N. Forthal, Richard M. Scearce, Philip E. Thorpe, Daniel M. Kozink, Gustavo Barbero, M. Kelly Plonk, S. Munir Alam, Shi Mao Xia, Hua-Xin Liao, Laura L. Sutherland, and David C. Montefiori
- Subjects
CD4-Positive T-Lymphocytes ,Chemokine ,Giant Cells ,Neutralization ,Monocytes ,Cell Fusion ,0302 clinical medicine ,Immunology and Allergy ,Chemokine CCL4 ,Chemokine CCL3 ,0303 health sciences ,biology ,env Gene Products, Human Immunodeficiency Virus ,virus diseases ,Antibodies, Monoclonal ,3. Good health ,beta 2-Glycoprotein I ,Chemokines, CC ,Antibodies, Antiphospholipid ,Antibody ,Chemokines ,Receptors, CCR5 ,medicine.drug_class ,Cardiolipins ,Immunology ,Phosphatidylserines ,Monoclonal antibody ,Peripheral blood mononuclear cell ,Article ,03 medical and health sciences ,Immunoglobulin Fab Fragments ,Viral entry ,medicine ,Beta 2-Glycoprotein I ,Humans ,Primary isolate ,030304 developmental biology ,Phosphatidylethanolamines ,Epithelial Cells ,Virus Internalization ,Virology ,Complementarity Determining Regions ,Immunity, Innate ,Immunoglobulin Fc Fragments ,Endotoxins ,Kinetics ,Viral Tropism ,Culture Media, Conditioned ,Mutation ,biology.protein ,HIV-1 ,Leukocytes, Mononuclear ,030215 immunology - Abstract
Traditional antibody-mediated neutralization of HIV-1 infection is thought to result from the binding of antibodies to virions, thus preventing virus entry. However, antibodies that broadly neutralize HIV-1 are rare and are not induced by current vaccines. We report that four human anti-phospholipid monoclonal antibodies (mAbs) (PGN632, P1, IS4, and CL1) inhibit HIV-1 CCR5-tropic (R5) primary isolate infection of peripheral blood mononuclear cells (PBMCs) with 80% inhibitory concentrations of
- Published
- 2010
32. Role of HIV membrane in neutralization by two broadly neutralizing antibodies
- Author
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Bing Chen, Sophia Rits-Volloch, Hua-Xin Liao, Shi-Mao Xia, Ruijun Zhang, S. Moses Dennison, Stephen C. Harrison, Li Sun, S. Munir Alam, Marco Morelli, and Barton F. Haynes
- Subjects
Models, Molecular ,medicine.drug_class ,HIV Infections ,Monoclonal antibody ,Gp41 ,Neutralization ,Epitope ,Neutralization Tests ,Viral entry ,medicine ,Humans ,Multidisciplinary ,biology ,Chemistry ,Viral Vaccine ,Antibodies, Monoclonal ,Membranes, Artificial ,Viral Vaccines ,Biological Sciences ,Viral membrane ,Antibodies, Neutralizing ,Virology ,HIV Envelope Protein gp41 ,Mutation ,HIV-1 ,biology.protein ,Antibody ,Protein Binding - Abstract
Induction of effective antibody responses against HIV-1 infection remains an elusive goal for vaccine development. Progress may require in-depth understanding of the molecular mechanisms of neutralization by monoclonal antibodies. We have analyzed the molecular actions of two rare, broadly neutralizing, human monoclonal antibodies, 4E10 and 2F5, which target the transiently exposed epitopes in the membrane proximal external region (MPER) of HIV-1 gp41 envelope during viral entry. Both have long CDR H3 loops with a hydrophobic surface facing away from the peptide epitope. We find that the hydrophobic residues of 4E10 mediate a reversible attachment to the viral membrane and that they are essential for neutralization, but not for interaction with gp41. We propose that these antibodies associate with the viral membrane in a required first step and are thereby poised to capture the transient gp41 fusion intermediate. These results bear directly on strategies for rational design of HIV-1 envelope immunogens.
- Published
- 2009
33. High throughput functional analysis of HIV-1 env genes without cloning
- Author
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Hua-Xin Liao, Barton F. Haynes, Jennifer L. Kirchherr, Lawrence Mwananyanda, Richard M. Scearce, Xiaozhi Lu, Webster Kasongo, Victor Chalwe, Feng Gao, David C. Montefiori, Rosemary Musonda, and Shi-Mao Xia
- Subjects
medicine.drug_class ,Cytomegalovirus ,Zambia ,Biology ,Monoclonal antibody ,Genes, env ,Polymerase Chain Reaction ,Article ,Neutralization ,Virus ,Cell Line ,law.invention ,Pseudovirion ,law ,Virology ,medicine ,Humans ,Promoter Regions, Genetic ,Gene ,Polymerase chain reaction ,Cloning ,Acquired Immunodeficiency Syndrome ,Gene Products, env ,virus diseases ,Molecular biology ,Transformation (genetics) ,HIV-1 - Abstract
Functional human immunodeficiency virus type 1 (HIV-1) env genes have been widely used for vaccine design, neutralization assays, and pathogenesis studies. However, obtaining bona fide functional env clones is a time consuming and labor intensive process. A new high throughput method has been developed to characterize HIV-1 env genes. Multiple rev/env gene cassettes were obtained from each of seven HIV-1 strains using single genome amplification (SGA) PCR. The cytomegalovirus (CMV) promoter was amplified separately by PCR. A promoter PCR (pPCR) method was developed to link both PCR products using an overlapping PCR method. Pseudovirions were generated by cotransfection of pPCR products and pSG3 Delta env backbone into 293T cells. After infecting TZM-bl cells, 75 out of 87 (86%) of the rev/env gene cassettes were functional. Pseudoviruses generated with pPCR products or corresponding plasmid DNA showed similar sensitivity to six HIV-1 positive sera and three monoclonal antibodies, suggesting neutralization properties are not altered in pPCR pseudovirions. Furthermore, sufficient amounts of pseudovirions can be obtained for a large number of neutralization assays. The new pPCR method eliminates cloning, transformation, and plasmid DNA preparation steps in the generation of HIV-1 pseudovirions. This allows for quick analysis of multiple env genes from HIV-1 infected individuals.
- Published
- 2007
34. Human Non-neutralizing HIV-1 Envelope Monoclonal Antibodies Limit the Number of Founder Viruses during SHIV Mucosal Infection in Rhesus Macaques
- Author
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Robin J. Shattock, Christiane Moog, Norman L. Letvin, Gary Landucci, Merlin L. Robb, Ryan Duffy, Georgia D. Tomaras, Elena E. Giorgi, Pinghuang Liu, Lily M. Blair, Guido Ferrari, Robert J. Schutte, Shi-Mao Xia, Supachai Rerks-Ngarm, M. A. Moody, Kelly A. Soderberg, S. Munir Alam, Nathan I. Nicely, Punnee Pitisuttithum, David C. Montefiori, Donald N. Forthal, Hui Li, Ruijun Zhang, Jerome H. Kim, Andrew Chao, Ranjit Warrier, Bette T. Korber, George M. Shaw, Amit Kumar, Nelson L. Michael, Sarah L. Cocklin, Kora Vidnovic, Hua-Xin Liao, Justin Pollara, James E. Robinson, Katja Klein, Jaranit Kaewkungwal, Charles W. Pemble, S. Moses Dennison, Joern E. Schmitz, Xiaoying Shen, Abbey Evans, Sorachai Nitayaphan, Feng Gao, Sampa Santra, and Barton F. Haynes
- Subjects
lcsh:Immunologic diseases. Allergy ,CD4-Positive T-Lymphocytes ,medicine.drug_class ,Protein Conformation ,viruses ,Immunology ,Simian Acquired Immunodeficiency Syndrome ,Fluorescent Antibody Technique ,medicine.disease_cause ,Gp41 ,Monoclonal antibody ,Antibodies, Viral ,Microbiology ,Virus ,Immune system ,Viral Envelope Proteins ,Virology ,Genetics ,medicine ,Animals ,Humans ,Intestinal Mucosa ,Molecular Biology ,lcsh:QH301-705.5 ,biology ,Reverse Transcriptase Polymerase Chain Reaction ,Rectum ,Antibodies, Monoclonal ,Simian immunodeficiency virus ,Surface Plasmon Resonance ,Vaccine efficacy ,Macaca mulatta ,3. Good health ,Mucosal Infection ,lcsh:Biology (General) ,biology.protein ,HIV-1 ,Parasitology ,Simian Immunodeficiency Virus ,Antibody ,lcsh:RC581-607 ,Research Article - Abstract
HIV-1 mucosal transmission begins with virus or virus-infected cells moving through mucus across mucosal epithelium to infect CD4+ T cells. Although broadly neutralizing antibodies (bnAbs) are the type of HIV-1 antibodies that are most likely protective, they are not induced with current vaccine candidates. In contrast, antibodies that do not neutralize primary HIV-1 strains in the TZM-bl infection assay are readily induced by current vaccine candidates and have also been implicated as secondary correlates of decreased HIV-1 risk in the RV144 vaccine efficacy trial. Here, we have studied the capacity of anti-Env monoclonal antibodies (mAbs) against either the immunodominant region of gp41 (7B2 IgG1), the first constant region of gp120 (A32 IgG1), or the third variable loop (V3) of gp120 (CH22 IgG1) to modulate in vivo rectal mucosal transmission of a high-dose simian-human immunodeficiency virus (SHIV-BaL) in rhesus macaques. 7B2 IgG1 or A32 IgG1, each containing mutations to enhance Fc function, was administered passively to rhesus macaques but afforded no protection against productive clinical infection while the positive control antibody CH22 IgG1 prevented infection in 4 of 6 animals. Enumeration of transmitted/founder (T/F) viruses revealed that passive infusion of each of the three antibodies significantly reduced the number of T/F genomes. Thus, some antibodies that bind HIV-1 Env but fail to neutralize virus in traditional neutralization assays may limit the number of T/F viruses involved in transmission without leading to enhancement of viral infection. For one of these mAbs, gp41 mAb 7B2, we provide the first co-crystal structure in complex with a common cyclical loop motif demonstrated to be critical for infection by other retroviruses., Author Summary Antibodies specifically recognize antigenic sites on pathogens and can mediate multiple antiviral functions through engagement of effector cells via their Fc region. Current HIV-1 vaccine candidates induce polyclonal antibody responses with multiple antiviral functions, but do not induce broadly neutralizing antibodies. An improved understanding of whether certain types of non-neutralizing HIV-1 specific antibodies can individually protect against HIV-1 infection may facilitate vaccine development. Here, we test whether non-neutralizing antibodies with multiple antiviral functions mediated through FcR engagement and recognition of virus particles or virus-infected cells can limit infection, despite lacking classical virus neutralization activity. In a passive antibody infusion-rhesus macaque challenge model, we tested the ability of non-neutralizing monoclonal antibodies to limit virus acquisition. We demonstrate that two different types of non-neutralizing antibodies, one that recognizes both virus particles and infected cells (7B2) and another that recognizes only infected cells (A32) were capable of decreasing the number of transmitted founder viruses. Further, we provide the structure of 7B2 in complex with the gp41 cyclical loop motif, a motif critical for entry. These findings provide insights into the role that antibodies with antiviral properties, including virion capture and FcR mediated effector function, may play in protecting against HIV-1 acquisition.
- Published
- 2015
35. Analysis of HIV-1 subtype B third variable region peptide motifs for induction of neutralizing antibodies against HIV-1 primary isolates
- Author
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Terri Wrin, Hua-Xin Liao, Richard M. Scearce, Barton F. Haynes, Shi Mao Xia, Christos J. Petropoulos, Laura L. Sutherland, Ben-Jiang Ma, David C. Montefiori, Bette T. Korber, and Cathrine Denton
- Subjects
Immunogen ,T cell ,Amino Acid Motifs ,Blotting, Western ,Guinea Pigs ,HIV-1 vaccine ,Peptide ,Cross Reactions ,HIV Envelope Protein gp120 ,V3 loop ,Antibodies, Viral ,Neutralization ,Guinea pig ,03 medical and health sciences ,Neutralization Tests ,Virology ,Consensus Sequence ,Consensus sequence ,medicine ,Animals ,Humans ,Amino Acid Sequence ,Antibody ,030304 developmental biology ,chemistry.chemical_classification ,0303 health sciences ,Sequence Homology, Amino Acid ,biology ,030302 biochemistry & molecular biology ,Peptide Fragments ,HIV-1 envelope ,3. Good health ,medicine.anatomical_structure ,chemistry ,HIV-1 ,biology.protein - Abstract
The HIV-1 gp120 V3 loop is a potent inducer of neutralizing antibodies for T cell line adapted-HIV-1, but less so for primary isolates. We hypothesized that peptides representative of the diversity of natural HIV-1 V3 loop variants might capture elements of conserved higher order structures and so stimulate broadly reactive neutralizing antibodies. We designed a panel of 29 subtype B V3 sequences postulated to reflect the range of V3 diversity. These peptides were used to immunize guinea pigs. The most effective peptide (62.19) clustered around the subtype B consensus sequence and induced antibodies that reproducibly neutralized 31% of the subtype B HIV-1 primary isolates evaluated, but exhibited limited cross-neutralization of non-subtype B HIV-1 strains. Taken together, these data demonstrated that the limited neutralization profile of antibodies induced by optimal subtype B V3 motifs likely represents the maximum breadth of neutralization of subtype B HIV-1 primary isolates attainable by anti-V3 peptide antibodies.
- Published
- 2006
36. A polymorphism in the HCP5 gene associated with HLA-B*5701 does not restrict HIV-1 in vitro
- Author
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Jacques Fellay, Wei Huang, David Goldstein, Ben-Jiang Ma, Shi-Mao Xia, Barton F. Haynes, Hua-Xin Liao, Ruijun Zhang, Woohyun Yoon, and Kevin V. Shianna
- Subjects
Infectivity ,Genetics ,RNA, Untranslated ,Immunology ,Haplotype ,HIV Infections ,Single-nucleotide polymorphism ,Viral Load ,Biology ,Major histocompatibility complex ,Polymorphism, Single Nucleotide ,Virology ,Article ,Major Histocompatibility Complex ,Infectious Diseases ,HLA-B Antigens ,HIV-1 ,biology.protein ,Humans ,Immunology and Allergy ,Coding region ,RNA, Long Noncoding ,Gene ,Viral load - Abstract
A single-nucleotide polymorphism (rs2395029) in the HCP5 gene associated with HLA-B*5701 is correlated with lower HIV-1 viral set point. The two allelic forms of coding region were ectopically expressed in TZM-bl cells for an effect on HIV-1 replication. No significant HIV-1 restriction was observed in the cells with infectivity assays throughout HIV-1 life cycle, suggesting that the association of HCP5 variant with viral control is likely due to HLA-B*5701-related effect or other functional variants in the haplotype or both.
- Published
- 2010
37. Initiation of HIV neutralizing B cell lineages with sequential envelope immunizations.
- Author
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Williams, Wilton B., Jinsong Zhang, Chuancang Jiang, Nicely, Nathan I., Fera, Daniela, Kan Luo, Moody, M. Anthony, Hua-Xin Liao, Alam, S. Munir, Kepler, Thomas B., Ramesh, Akshaya, Wiehe, Kevin, Holland, James A., Bradley, Todd, Vandergrift, Nathan, Saunders, Kevin O., Parks, Robert, Foulger, Andrew, Shi-Mao Xia, and Bonsignori, Mattia
- Abstract
A strategy for HIV-1 vaccine development is to define envelope (Env) evolution of broadly neutralizing antibodies (bnAbs) in infection and to recreate those events by vaccination. Here, we report host tolerance mechanisms that limit the development of CD4-binding site (CD4bs), HCDR3-binder bnAbs via sequential HIV-1 Env vaccination. Vaccine-induced macaque CD4bs antibodies neutralize 7% of HIV-1 strains, recognize open Env trimers, and accumulate relatively modest somatic mutations. In naive CD4bs, unmutated common ancestor knock-in mice Env
+ B cell clones develop anergy and partial deletion at the transitional to mature B cell stage, but become Env− upon receptor editing. In comparison with repetitive Env immunizations, sequential Env administration rescue anergic Env+ (non-edited) precursor B cells. Thus, stepwise immunization initiates CD4bs-bnAb responses, but immune tolerance mechanisms restrict their development, suggesting that sequential immunogenbased vaccine regimens will likely need to incorporate strategies to expand bnAb precursor pools. [ABSTRACT FROM AUTHOR]- Published
- 2017
- Full Text
- View/download PDF
38. Isolation of HIV-1-Neutralizing Mucosal Monoclonal Antibodies from Human Colostrum
- Author
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Kara Anasti, Garnett Kelsoe, Barton F. Haynes, Guido Ferrari, Shi-Mao Xia, Guang Yang, Hua-Xin Liao, M. Anthony Moody, Justin Pollara, Xiaoying Shen, James Friedman, Georgia D. Tomaras, Genevieve G. Fouda, Shelley Stewart, S. Munir Alam, and Sallie R. Permar
- Subjects
B Cells ,lcsh:Medicine ,HIV Antibodies ,HIV Envelope Protein gp120 ,Neutralization ,Epitope ,0302 clinical medicine ,Immunodeficiency Viruses ,Pregnancy ,HIV vaccine ,lcsh:Science ,Fluorescent Antibody Technique, Indirect ,0303 health sciences ,B-Lymphocytes ,Multidisciplinary ,biology ,virus diseases ,Antibodies, Monoclonal ,3. Good health ,medicine.anatomical_structure ,Medicine ,Infectious diseases ,Epitopes, B-Lymphocyte ,Female ,Antibody ,Research Article ,medicine.drug_class ,Immune Cells ,Immunology ,HIV prevention ,Immunoglobulins ,Viral diseases ,Gp41 ,Monoclonal antibody ,Microbiology ,03 medical and health sciences ,Neutralization Tests ,Virology ,Retroviruses ,medicine ,Humans ,Antibody-Producing Cells ,Biology ,B cell ,030304 developmental biology ,Colostrum ,lcsh:R ,HIV ,Viral Vaccines ,Antibodies, Neutralizing ,Viral Classification ,biology.protein ,lcsh:Q ,030215 immunology - Abstract
Background: Generation of potent anti-HIV antibody responses in mucosal compartments is a potential requirement of a transmission-blocking HIV vaccine. HIV-specific, functional antibody responses are present in breast milk, and these mucosal antibody responses may play a role in protection of the majority of HIV-exposed, breastfeeding infants. Therefore, characterization of HIV-specific antibodies produced by B cells in milk could guide the development of vaccines that elicit protective mucosal antibody responses. Methods: We isolated B cells from colostrum of an HIV-infected lactating woman with a detectable neutralization response in milk and recombinantly produced and characterized the resulting HIV-1 Envelope (Env)-specific monoclonal antibodies (mAbs). Results: The identified HIV-1 Env-specific colostrum mAbs, CH07 and CH08, represent two of the first mucosally-derived anti-HIV antibodies yet to be reported. Colostrum mAb CH07 is a highly-autoreactive, weakly-neutralizing gp140-specific mAb that binds to linear epitopes in the gp120 C5 region and gp41 fusion domain. In contrast, colostrum mAb CH08 is a nonpolyreactive CD4-inducible (CD4i) gp120-specific mAb with moderate breadth of neutralization. Conclusions: These novel HIV-neutralizing mAbs isolated from a mucosal compartment provide insight into the ability of mucosal B cell populations to produce functional anti-HIV antibodies that may contribute to protection against virus acquisition at mucosal surfaces.
- Published
- 2012
39. Human Immunodeficiency Virus Type 1 gp41 Antibodies That Mask Membrane Proximal Region Epitopes: Antibody Binding Kinetics, Induction, and Potential for Regulation in Acute Infection▿
- Author
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Miroslaw K. Gorny, M. Anthony Moody, Hua-Xin Liao, Shi-Mao Xia, Laura L. Sutherland, Steven G. Plonk, Kent J. Weinhold, Susan Zolla-Pazner, George M. Shaw, Charles B. Hicks, David C. Montefiori, James E. Robinson, Robert Parks, S. Munir Alam, Stacie Vanleeuwen, Salim S. Abdool Karim, Richard M. Scearce, Kelly Plonk, Georgia D. Tomaras, and Barton F. Haynes
- Subjects
medicine.drug_class ,Immunology ,HIV Infections ,Biology ,Cross Reactions ,HIV Antibodies ,Gp41 ,Monoclonal antibody ,Microbiology ,Epitope ,chemistry.chemical_compound ,Epitopes ,Mice ,Non-competitive inhibition ,Neutralization Tests ,Virology ,medicine ,Cardiolipin ,Animals ,Humans ,virus diseases ,Antibodies, Monoclonal ,Phosphatidylserine ,Molecular biology ,Lipids ,HIV Envelope Protein gp41 ,Kinetics ,chemistry ,Immunization ,Insect Science ,biology.protein ,HIV-1 ,Pathogenesis and Immunity ,Antibody ,Protein Binding - Abstract
Two human monoclonal antibodies (MAbs) (2F5 and 4E10) against the human immunodeficiency virus type 1 (HIV-1) envelope g41 cluster II membrane proximal external region (MPER) broadly neutralize HIV-1 primary isolates. However, these antibody specificities are rare, are not induced by Env immunization or HIV-1 infection, and are polyspecific and also react with lipids such as cardiolipin or phosphatidylserine. To probe MPER anti-gp41 antibodies that are produced in HIV-1 infection, we have made two novel murine MAbs, 5A9 and 13H11, against HIV-1 gp41 envelope that partially cross-blocked 2F5 MAb binding to Env but did not neutralize HIV-1 primary isolates or bind host lipids. Competitive inhibition assays using labeled 13H11 MAb and HIV-1-positive patient plasma samples demonstrated that cluster II 13H11-blocking plasma antibodies were made in 83% of chronically HIV-1 infected patients and were acquired between 5 to 10 weeks after acute HIV-1 infection. Both the mouse 13H11 MAb and the three prototypic cluster II human MAbs (98-6, 126-6, and 167-D) blocked 2F5 binding to gp41 epitopes to variable degrees; the combination of 98-6 and 13H11 completely blocked 2F5 binding. These data provide support for the hypothesis that in some patients, B cells make nonneutralizing cluster II antibodies that may mask or otherwise down-modulate B-cell responses to immunogenic regions of gp41 that could be recognized by B cells capable of producing antibodies like 2F5.
- Published
- 2007
40. Staged induction of HIV-1 glycan-dependent broadly neutralizing antibodies.
- Author
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Bonsignori, Mattia, Kreider, Edward F., Fera, Daniela, Meyerhoff, R. Ryan, Bradley, Todd, Wiehe, Kevin, Alam, S. Munir, Aussedat, Baptiste, Walkowicz, William E., Kwan-Ki Hwang, Saunders, Kevin O., Ruijun Zhang, Gladden, Morgan A., Monroe, Anthony, Kumar, Amit, Shi-Mao Xia, Cooper, Melissa, Louder, Mark K., McKee, Krisha, and Bailer, Robert T.
- Subjects
DIAGNOSIS of HIV infections ,AIDS vaccines ,IMMUNOGLOBULINS ,IMMUNOGENETICS ,GLYCANS - Abstract
The article reports on the induction of HIV-1 glycan–dependent in neutralizing antibodies (bnAbs ). It cites on a vaccine to prevent HIV-1 infection that include immunogens, citing the five major targets for bnAbs including glycan-rich apex of the HIV-1 envelope (Env) trimer and the base of the third variable loop (V3).
- Published
- 2017
- Full Text
- View/download PDF
41. IGHV1-69 B Cell Chronic Lymphocytic Leukemia Antibodies Cross-React with HIV-1 and Hepatitis C Virus Antigens as Well as Intestinal Commensal Bacteria
- Author
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Munir Alam, Barton F. Haynes, Xiao-Jie Yan, Kevin Wiehe, Georgia D. Tomaras, Jane A. McKeating, Nicholas Chiorazzi, Kanti R. Rai, Ashley M. Trama, Kwan-Ki Hwang, John Whitesides, Xi Chen, Shi-Mao Xia, Daniel M. Kozink, Steven L. Allen, Abby J. Cooper, Hua-Xin Liao, Charles C. Chu, Garnett Kelsoe, Minyue Wang, Rosa Catera, and Dawn J. Marshall
- Subjects
Viral Diseases ,Anatomy and Physiology ,B Cells ,Anti-nuclear antibody ,Antibodies, Neoplasm ,HIV Antigens ,Chronic lymphocytic leukemia ,Immunoglobulin Variable Region ,lcsh:Medicine ,HIV Infections ,Hepacivirus ,Hematologic Cancers and Related Disorders ,Immune Physiology ,hemic and lymphatic diseases ,lcsh:Science ,Chronic Lymphoblastic Leukemia ,Multidisciplinary ,biology ,Antibodies, Monoclonal ,virus diseases ,Recombinant Proteins ,3. Good health ,Intestines ,Treatment Outcome ,Infectious Diseases ,medicine.anatomical_structure ,Oncology ,Medicine ,Antibody ,Immunoglobulin Heavy Chains ,Protein Binding ,Research Article ,Immune Cells ,Molecular Sequence Data ,B-cell receptor ,Receptors, Antigen, B-Cell ,Hemagglutinin (influenza) ,Cross Reactions ,Antibodies ,Antigen ,Cell Line, Tumor ,Leukemias ,medicine ,Humans ,Amino Acid Sequence ,Symbiosis ,Immunity to Infections ,Alleles ,B cell ,Hybridomas ,Bacteria ,lcsh:R ,Immunity ,HIV ,Cancers and Neoplasms ,medicine.disease ,Leukemia, Lymphocytic, Chronic, B-Cell ,Virology ,HIV-1 ,biology.protein ,Immunoglobulin heavy chain ,Clinical Immunology ,lcsh:Q ,Hepatitis C Antigens ,Sequence Alignment ,Paraproteins - Abstract
B-cell chronic lymphocytic leukemia (B-CLL) patients expressing unmutated immunoglobulin heavy variable regions (IGHVs) use the IGHV1-69 B cell receptor (BCR) in 25% of cases. Since HIV-1 envelope gp41 antibodies also frequently use IGHV1-69 gene segments, we hypothesized that IGHV1-69 B-CLL precursors may contribute to the gp41 B cell response during HIV-1 infection. To test this hypothesis, we rescued 5 IGHV1-69 unmutated antibodies as heterohybridoma IgM paraproteins and as recombinant IgG1 antibodies from B-CLL patients, determined their antigenic specificities and analyzed BCR sequences. IGHV1-69 B-CLL antibodies were enriched for reactivity with HIV-1 envelope gp41, influenza, hepatitis C virus E2 protein and intestinal commensal bacteria. These IGHV1-69 B-CLL antibodies preferentially used IGHD3 and IGHJ6 gene segments and had long heavy chain complementary determining region 3s (HCDR3s) (≥21 aa). IGHV1-69 B-CLL BCRs exhibited a phenylalanine at position 54 (F54) of the HCDR2 as do rare HIV-1 gp41 and influenza hemagglutinin stem neutralizing antibodies, while IGHV1-69 gp41 antibodies induced by HIV-1 infection predominantly used leucine (L54) allelic variants. These results demonstrate that the B-CLL cell population is an expansion of members of the innate polyreactive B cell repertoire with reactivity to a number of infectious agent antigens including intestinal commensal bacteria. The B-CLL IGHV1-69 B cell usage of F54 allelic variants strongly suggests that IGHV1-69 B-CLL gp41 antibodies derive from a restricted B cell pool that also produces rare HIV-1 gp41 and influenza hemagglutinin stem antibodies.
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- 2014
42. Induction of Antibodies in Rhesus Macaques That Recognize a Fusion-Intermediate Conformation of HIV-1 gp41
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Shi-Mao Xia, S. Moses Dennison, Robert Parks, Gilad Ofek, Ruijun Zhang, Peter D. Kwong, S. Munir Alam, Frederick H. Jaeger, Laura L. Sutherland, Richard M. Scearce, Xiaoying Shen, Hua-Xin Liao, Cindy M. Bowman, Yongping Yang, Shelley Stewart, Bing Chen, Norman L. Letvin, Georgia D. Tomaras, Sampa Santra, Barton F. Haynes, and Kara Anasti
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B Cells ,viruses ,lcsh:Medicine ,HIV Antibodies ,Macaque ,Epitope ,Epitopes ,Protein structure ,Antibody Specificity ,lcsh:Science ,Immune Response ,Vaccines ,0303 health sciences ,Liposome ,Multidisciplinary ,biology ,Vaccination ,030302 biochemistry & molecular biology ,virus diseases ,Animal Models ,HIV Envelope Protein gp41 ,3. Good health ,Medicine ,Infectious diseases ,Antibody ,Research Article ,Recombinant Fusion Proteins ,Immune Cells ,Guinea Pigs ,Immunology ,HIV prevention ,Immunoglobulins ,Viral diseases ,Gp41 ,03 medical and health sciences ,Cross-Priming ,Model Organisms ,Immune system ,Neutralization Tests ,biology.animal ,Animals ,Protein Structure, Quaternary ,Biology ,030304 developmental biology ,lcsh:R ,Immunity ,HIV ,Macaca mulatta ,Virology ,Molecular biology ,Immunization ,Antibody Formation ,Liposomes ,HIV-1 ,biology.protein ,lcsh:Q ,Clinical Immunology ,Peptides - Abstract
A component to the problem of inducing broad neutralizing HIV-1 gp41 membrane proximal external region (MPER) antibodies is the need to focus the antibody response to the transiently exposed MPER pre-hairpin intermediate neutralization epitope. Here we describe a HIV-1 envelope (Env) gp140 oligomer prime followed by MPER peptide-liposomes boost strategy for eliciting serum antibody responses in rhesus macaques that bind to a gp41 fusion intermediate protein. This Env-liposome immunization strategy induced antibodies to the 2F5 neutralizing epitope ⁶⁶⁴DKW residues, and these antibodies preferentially bound to a gp41 fusion intermediate construct as well as to MPER scaffolds stabilized in the 2F5-bound conformation. However, no serum lipid binding activity was observed nor was serum neutralizing activity for HIV-1 pseudoviruses present. Nonetheless, the Env-liposome prime-boost immunization strategy induced antibodies that recognized a gp41 fusion intermediate protein and was successful in focusing the antibody response to the desired epitope.
- Published
- 2011
43. Isolation of HIV-1-Neutralizing Mucosal Monoclonal Antibodies from Human Colostrum.
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Friedman, James, Alam, S. Munir, Xiaoying Shen, Shi-Mao Xia, Stewart, Shelley, Anasti, Kara, Pollara, Justin, Fouda, Genevieve G., Guang Yang, Kelsoe, Garnett, Ferrari, Guido, Tomaras, Georgia D., Haynes, Barton F., Hua-Xin Liao, Moody, M. Anthony, and Permar, Sallie R.
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COLOSTRUM ,ANTI-HIV agents ,HIV antibodies ,MONOCLONAL antibodies ,B cells ,BREASTFEEDING ,CD4 antigen - Abstract
Background: Generation of potent anti-HIV antibody responses in mucosal compartments is a potential requirement of a transmission-blocking HIV vaccine. HIV-specific, functional antibody responses are present in breast milk, and these mucosal antibody responses may play a role in protection of the majority of HIV-exposed, breastfeeding infants. Therefore, characterization of HIV-specific antibodies produced by B cells in milk could guide the development of vaccines that elicit protective mucosal antibody responses. Methods: We isolated B cells from colostrum of an HIV-infected lactating woman with a detectable neutralization response in milk and recombinantly produced and characterized the resulting HIV-1 Envelope (Env)-specific monoclonal antibodies (mAbs). Results: The identified HIV-1 Env-specific colostrum mAbs, CH07 and CH08, represent two of the first mucosally-derived anti-HIV antibodies yet to be reported. Colostrum mAb CH07 is a highly-autoreactive, weakly-neutralizing gp140-specific mAb that binds to linear epitopes in the gp120 C5 region and gp41 fusion domain. In contrast, colostrum mAb CH08 is a nonpolyreactive CD4-inducible (CD4i) gp120-specific mAb with moderate breadth of neutralization. Conclusions: These novel HIV-neutralizing mAbs isolated from a mucosal compartment provide insight into the ability of mucosal B cell populations to produce functional anti-HIV antibodies that may contribute to protection against virus acquisition at mucosal surfaces. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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44. Role of HIV membrane in neutralization by two broadly neutralizing antibodies.
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AIam, S. Munir, MoreIIi, Marco, Dennison, S. Moses, Hua-Xin Liao, Ruijun Zhang, Shi-Mao Xia, Rits-VoIIoch, Sophia, Li Sun, Harrison, Stephen C., Haynes, Barton F., and Chen, Bing
- Subjects
HIV infections ,VACCINE biotechnology ,NEUTRALIZATION (Chemistry) ,MONOCLONAL antibodies ,IMMUNOGENETICS - Abstract
Induction of effective antibody responses against HIV-1 infection remains an elusive goal for vaccine development. Progress may require in-depth understanding of the molecular mechanisms of neutralization by monoclonal antibodies. We have analyzed the molecular actions of two rare, broadly neutralizing, human monoclonal antibodies, 4E10 and 2F5, which target the transiently exposed epitopes in the membrane proximal external region (MPER) of HlV-1 gp41 envelope during viral entry. Both have long CDR H3 loops with a hydrophobic surface facing away from the peptide epitope. We find that the hydrophobic residues of 4E10 mediate a reversible attachment to the viral membrane and that they are essential for neutralization, but not for interaction with gp41. We propose that these antibodies associate with the viral membrane in a required first step and are thereby poised to capture the transient gp41 fusion intermediate. These results bear directly on strategies for rational design of HIV-1 envelope immunogens. [ABSTRACT FROM AUTHOR]
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- 2009
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45. A group M consensus envelope glycoprotein induces antibodies that neutralize subsets of subtype B and C HIV-1 primary viruses
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Feng Gao, Shi Mao Xia, Bette T. Korber, Ben Jiang Ma, David C. Montefiori, Barton F. Haynes, Hua-Xin Liao, Richard M. Scearce, Mildred W. Mcadams, Laura L. Sutherland, Yingying Li, Eric A. Weaver, Beatrice H. Hahn, Mary E. Brock, Julie M. Decker, S. Munir Alam, Stacie Vanleeuwen, Zenaido T. Camacho, and Gary J. Nabel
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Guinea Pigs ,Molecular Sequence Data ,HIV Infections ,Cross Reactions ,HIV Antibodies ,Biology ,Consensus immunogens ,Article ,Immunophenotyping ,03 medical and health sciences ,Neutralization Tests ,Virology ,Consensus Sequence ,Animals, Outbred Strains ,Consensus sequence ,Neutralization antibody ,Animals ,Amino Acid Sequence ,Gene ,Peptide sequence ,030304 developmental biology ,AIDS Vaccines ,chemistry.chemical_classification ,0303 health sciences ,030306 microbiology ,Immunogenicity ,env Gene Products, Human Immunodeficiency Virus ,Gene Products, env ,virus diseases ,3. Good health ,Titer ,chemistry ,biology.protein ,HIV-1 ,Antibody ,Glycoprotein ,Vaccine - Abstract
HIV-1 subtype C is the most common HIV-1 group M subtype in Africa and many parts of Asia. However, to date HIV-1 vaccine candidate immunogens have not induced potent and broadly neutralizing antibodies against subtype C primary isolates. We have used a centralized gene strategy to address HIV-1 diversity, and generated a group M consensus envelope gene with shortened consensus variable loops (CON-S) for comparative studies with wildtype (WT) Env immunogens. Our results indicate that the consensus HIV-1 group M CON-S Env elicited cross-subtype neutralizing antibodies of similar or greater breadth and titer than the WT Envs tested, indicating the utility of a centralized gene strategy. Our study also shows the feasibility of iterative improvements in Env immunogenicity by rational design of centralized genes.
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46. Cross-reactive monoclonal antibodies to multiple HIV-1 subtype and SIVcpz envelope glycoproteins
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Robert Parks, Barton F. Haynes, Julie E. Hohm, Myron S. Cohen, Richard M. Scearce, Vernon C. Maino, M. Anthony Moody, Georgia D. Tomaras, Damon F. Ogburn, Hua-Xin Liao, Feng Gao, Woodrow E. Lomas, S. Munir Alam, Beatrice H. Hahn, Susan A. Fiscus, Bhavna Hora, Shi-Mao Xia, Emily Park, and Bette T. Korber
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Monoclonal antibody ,medicine.drug_class ,viruses ,Antibody Affinity ,Subtype ,Envelope glycoprotein ,Plasma protein binding ,Cross Reactions ,HIV Antibodies ,Biology ,medicine.disease_cause ,Cross-reactivity ,Article ,Epitope ,Mice ,03 medical and health sciences ,Western blot ,Virology ,medicine ,Animals ,030304 developmental biology ,chemistry.chemical_classification ,Mice, Inbred BALB C ,0303 health sciences ,medicine.diagnostic_test ,030306 microbiology ,env Gene Products, Human Immunodeficiency Virus ,Antibodies, Monoclonal ,virus diseases ,Surface Plasmon Resonance ,Simian immunodeficiency virus ,Molecular biology ,3. Good health ,chemistry ,biology.protein ,HIV-1 ,Simian Immunodeficiency Virus ,Antibody ,Glycoprotein ,Protein Binding - Abstract
The extraordinarily high level of genetic variation of HIV-1 env genes poses a challenge to obtain antibodies that cross-react with multiple subtype Env glycoproteins. To determine if cross-reactive monoclonal antibodies (mAbs) to highly conserved epitopes in HIV-1 envelope glycoproteins can be induced, we immunized mice with wild-type or consensus HIV-1 Env proteins and characterized a panel of ten mAbs that reacted with varying breadth to subtypes A, B, C, D, F, G, CRF01_AE, and a highly divergent SIVcpzUS Env proteins by ELISA and Western blot analysis. Two mAbs (3B3 and 16H3) cross-reacted with all tested Env proteins, including SIVcpzUS Env. Surface plasmon resonance analyses showed both 3B3 and 16H3 bound Env proteins with high affinity. However, neither neutralized primary HIV-1 pseudoviruses. These data indicate that broadly reactive non-neutralizing monoclonal antibodies can be elicited, but that the conserved epitopes that they recognize are not present on functional virion trimers. Nonetheless, such mAbs represent valuable reagents to study the biochemistry and structural biology of Env protein oligomers.
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47. Immunodominance of Antibody Recognition of the HIV Envelope V2 Region in Ig-Humanized Mice.
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Wiehe, Kevin, Nicely, Nathan I., Lockwood, Bradley, Kuraoka, Masayuki, Anasti, Kara, Arora, Sabrina, Bowman, Cindy M., Stolarchuk, Christina, Parks, Robert, Lloyd, Krissey E., Shi-Mao Xia, Duffy, Ryan, Xiaoying Shen, Kyratsous, Christos A., Macdonald, Lynn E., Murphy, Andrew J., Scearce, Richard M., Moody, M. Anthony, Alam, S. Munir, and Verkoczy, Laurent
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HIV infections , *THERAPEUTICS , *LABORATORY mice , *LYSINE , *GLUTAMIC acid , *EPITOPES - Abstract
In the RV144 gp120 HIV vaccine trial, decreased transmission risk was correlated with Abs that reacted with a linear epitope at a lysine residue at position 169 (K169) in the HIV-1 envelope (Env) V2 region. The K169 V2 response was restricted to Abs bearing Vl rearrangements that expressed aspartic acid/glutamic acid in CDR L2. The AE.A244 gp120 in AIDSVAX B/E also bound to the unmutated ancestor of a V2-glycan broadly neutralizing Ab, but this Ab type was not induced in the RV144 trial. In this study, we sought to determine whether immunodominance of the V2 linear epitope could be overcome in the absence of human Vλ rearrangements. We immunized IgH- and Igk-humanized mice with the AE.A244 gp120 Env. In these mice, the V2 Ab response was focused on a linear epitope that did not include K169. V2 Abs were isolated that used the same human VH gene segment as an RV144 V2 Ab but paired with a mouse λ L chain. Structural characterization of one of these V2 Abs revealed how the linear V2 epitope could be engaged, despite the lack of aspartic acid/glutamic acid encoded in the mouse repertoire. Thus, despite the absence of the human Vλ locus in these humanized mice, the dominance of Vλ pairing with human VH for HIV-1 Env V2 recognition resulted in human VH pairing with mouse λ L chains instead of allowing otherwise subdominant V2-glycan broadly neutralizing Abs to develop. [ABSTRACT FROM AUTHOR]
- Published
- 2017
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48. Cooperation of B Cell Lineages in Induction of HIV-1-Broadly Neutralizing Antibodies.
- Author
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Feng Gao, Bonsignori, Mattia, Hua-Xin Liao, Kumar, Amit, Shi-Mao Xia, Xiaozhi Lu, Fangping Cai, Kwan-Ki Hwang, Hongshuo Song, Tongqing Zhou, Lynch, Rebecca M., Alam, S. Munir, Moody, M. Anthony, Ferrari, Guido, Berrong, Mark, Kelsoe, Garnett, Shaw, George M., Hahn, Beatrice H., Montefiori, David C., and Kamanga, Gift
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B cells , *ANTIGEN-antibody reactions , *NEUTRALIZATION (Chemistry) , *HIV prevention , *HIV antibodies , *AIDS vaccines - Abstract
Development of strategies for induction of HIV-1 broadly neutralizing antibodies (bnAbs) by vaccines is a priority. Determining the steps of bnAb induction in HIV-1-infected individuals who make bnAbs is a key strategy for immunogen design. Here, we study the B cell response in a bnAb-producing individual and report cooperation between two B cell lineages to drive bnAb development. We isolated a virus-neutralizing antibody lineage that targeted an envelope region (loop D) and selected virus escape mutants that resulted in both enhanced bnAb lineage envelope binding and escape mutant neutralization--traits associated with increased B cell antigen drive. Thus, in this individual, two B cell lineages cooperated to induce the development of bnAbs. Design of vaccine immunogens that simultaneously drive both helper and broadly neutralizing B cell lineages may be important for vaccine-induced recapitulation of events that transpire during the maturation of neutralizing antibodies in HIV-1-infected individuals. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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49. Toll-Like Receptor 7/8 (TLR7/8) and TLR9 Agonists Cooperate To Enhance HIV-1 Envelope Antibody Responses in Rhesus Macaques.
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Moody, M. Anthony, Santra, Sampa, Vandergrift, Nathan A., Sutherland, Laura L., Gurley, Thaddeus C., Drinker, Mark S., Allen, Ashley A., Shi-Mao Xia, Meyerhoff, R. Ryan, Parks, Robert, Lloyd, Krissey E., Easterhoff, David, Alam, S. Munir, Hua-Xin Liao, Ward, Brandy M., Ferrari, Guido, Montefiori, David C., Tomaras, Georgia D., Seder, Robert A., and Letvin, Norman L.
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- *
TOLL-like receptors , *IMMUNOLOGICAL adjuvants , *CHEMICAL agonists , *CELL-mediated cytotoxicity , *IMMUNITY - Abstract
The development of a vaccine that can induce high titers of functional antibodies against HIV-1 remains a high priority. We have developed an adjuvant based on an oil-in-water emulsion that incorporates Toll-like receptor (TLR) ligands to test whether triggering multiple pathogen-associated molecular pattern receptors could enhance immunogenicity. Compared to single TLR agonists or other pairwise combinations, TLR7/8 and TLR9 agonists combined were able to elicit the highest titers of binding, neutralizing, and antibody-dependent cellular cytotoxicity-mediating antibodies against the protein immunogen, transmitted/ founder HIV-1 envelope gp140 (B.63521). We further found that the combination of TLR7/8 and TLR9 agonists was associated with the release of CXCL10 (IP-10), suggesting that this adjuvant formulation may have optimally stimulated innate and adaptive immunity to elicit high titers of antibodies. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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50. Induction of HIV-1 Broad Neutralizing Antibodies in 2F5 Knock-in Mice: Selection against Membrane Proximal External Region-Associated Autoreactivity Limits T-Dependent Responses.
- Author
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Verkoczy, Laurent, Yao Chen, Jinsong Zhang, Bouton-Verville, Hilary, Newman, Amanda, Lockwood, Bradley, Scearce, Richard M., Montefiori, David C., Dennison, S. Moses, Shi-Mao Xia, Kwan-Ki Hwang, Hua-Xin Liao, Alam, S. Munir, and Haynes, Barton F.
- Subjects
- *
HIV antibodies , *AIDS vaccines , *IMMUNOREGULATION , *B cells , *IMMUNIZATION , *EPITOPES , *T cells - Abstract
A goal of HIV-1 vaccine development is to elicit broadly neutralizing Abs (BnAbs). Using a knock-in (KI) model of 2F5, a human HIV-1 gp41 membrane proximal external region (MPER)-specific BnAb, we previously demonstrated that a key obstacle to BnAb induction is clonal deletion of BnAb-expressing B cells. In this study of this model, we provide a proof-of-principle that robust serum neutralizing IgG responses can be induced from pre-existing, residual, self-reactive BnAb-expressing B cells in vivo using a structurally compatible gp41 MPER immunogen. Furthermore, in CD40L-deficient 2F5 KI mice, we demonstrate that these BnAb responses are elicited via a type II T-independent pathway, coinciding with expansion and activation of transitional splenic B cells specific for 2F5's nominal gp41 MPER-binding epitope (containing the 2F5 neutralization domain ELDKWA). In contrast, constitutive production of nonneutralizing serum IgGs in 2F5 KI mice is T dependent and originates from a subset of splenic mature B2 cells that have lost their ability to bind 2F5's gp41 MPER epitope. These results suggest that residual, mature B cells expressing autoreactive BnAbs, like 2F5 as BCR, may be limited in their ability to participate in T-dependent responses by purifying selection that selectively eliminates reactivity for neutralization epitope-containing/mimicked host Ags. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
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