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Georgia D Tomaras,
Guido Ferrari,
Xiaoying Shen,
S Munir Alam,
Hua-Xin Liao,
Justin Pollara,
Mattia Bonsignori,
M Anthony Moody,
Youyi Fong,
Xi Chen, [......],
Robert Parks,
Jaranit Kaewkungwal,
Sorachai Nitayaphan,
Punnee Pitisuttithum,
Supachai Rerks-Ngarm,
Peter B Gilbert,
Jerome H Kim,
Nelson L Michael,
David C Montefiori,
Barton F Haynes
[show abstract]
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ABSTRACT: Analysis of correlates of risk of infection in the RV144 HIV-1 vaccine efficacy trial demonstrated that plasma IgG against the HIV-1 envelope (Env) variable region 1 and 2 inversely correlated with risk, whereas HIV-1 Env-specific plasma IgA responses directly correlated with risk. In the secondary analysis, antibody-dependent cellular cytotoxicity (ADCC) was another inverse correlate of risk, but only in the presence of low plasma IgA Env-specific antibodies. Thus, we investigated the hypothesis that IgA could attenuate the protective effect of IgG responses through competition for the same Env binding sites. We report that Env-specific plasma IgA/IgG ratios are higher in infected than in uninfected vaccine recipients in RV144. Moreover, Env-specific IgA antibodies from RV144 vaccinees blocked the binding of ADCC-mediating mAb to HIV-1 Env glycoprotein 120 (gp120). An Env-specific monomeric IgA mAb isolated from an RV144 vaccinee also inhibited the ability of natural killer cells to kill HIV-1-infected CD4(+) T cells coated with RV144-induced IgG antibodies. We show that monomeric Env-specific IgA, as part of postvaccination polyclonal antibody response, may modulate vaccine-induced immunity by diminishing ADCC effector function.
Proceedings of the National Academy of Sciences 05/2013; · 9.68 Impact Factor
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Pinghuang Liu,
Nicole L Yates,
Xiaoying Shen,
Mattia Bonsignori,
M Anthony Moody,
Hua-Xin Liao,
Youyi Fong,
S Munir Alam,
R Glenn Overman,
Thomas Denny, [......],
Punnee Pitisuttithum,
Jaranit Kaewkungwal,
Sorachai Nitayaphan,
Supachai Rerks-Ngarm,
David C Montefiori,
Peter Gilbert,
Nelson L Michael,
Jerome H Kim,
Barton F Haynes, Georgia D Tomaras
[show abstract]
[hide abstract]
ABSTRACT: The detailed examination of the antibody repertoire from RV144 provides a unique template for understanding potentially protective antibody functions. Some potential immune correlates of protection were untested in the correlates analyses due to inherent assay limitations as well as the need to keep the correlates analysis focused on a limited number of endpoints to achieve statistical power. In an RV144 pilot study, we determined that RV144 vaccination elicited antibodies that could bind infectious virions (including the vaccine strains, HIV-1 CM244 and HIV-1 MN, and an HIV-1 expressing transmitted/founder Env B.WITO.c). Among those vaccinees with the highest IgG binding antibody profile, the majority (78%) captured the infectious vaccine strain virus(CM244) while a smaller proportion of vaccinees (26%) captured HIV-1 transmitted/founder Env virus. We demonstrated that vaccine-elicited HIV-1 gp120 antibodies of multiple specificities (V3, V2, conformational C1, and gp120 conformational) mediated capture of infectious virions. Although capture of infectious HIV-1 correlated with other humoral immune responses, the extent of variation between these humoral responses and virion capture indicates that virion capture antibodies occupy unique immunological space.
Journal of Virology 05/2013; · 5.40 Impact Factor
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Josh Eckels,
Cory Nathe,
Elizabeth K Nelson,
Sara G Shoemaker,
Elizabeth Nostrand,
Nicole L Yates,
Vicki C Ashley,
Linda J Harris,
Mark Bollenbeck,
Youyi Fong, Georgia D Tomaras,
Britt Piehler
[show abstract]
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ABSTRACT: BACKGROUND: Immunoassays that employ multiplexed bead arrays produce high information content per sample. Such assays are now frequently used to evaluate humoral responses in clinical trials. Integrated software is needed for the analysis, quality control, and secure sharing of the high volume of data produced by such multiplexed assays. Software that facilitates data exchange and provides flexibility to perform customized analyses (including multiple curve fits and visualizations of assay performance over time) could increase scientists' capacity to use these immunoassays to evaluate human clinical trials. RESULTS: The HIV Vaccine Trials Network and the Statistical Center for HIV/AIDS Research and Prevention collaborated with LabKey Software to enhance the open source LabKey Server platform to facilitate workflows for multiplexed bead assays. This system now supports the management, analysis, quality control, and secure sharing of data from multiplexed immunoassays that leverage Luminex xMAP(R) technology. These assays may be custom or kit-based. Newly added features enable labs to: (i) import run data from spreadsheets output by Bio-Plex ManagerTM software; (ii) customize data processing, curve fits, and algorithms through scripts written in common languages, such as R; (iii) select script-defined calculation options through a graphical user interface; (iv) collect custom metadata for each titration, analyte, run and batch of runs; (v) calculate dose--response curves for titrations; (vi) interpolate unknown concentrations from curves for titrated standards; (vii) flag run data for exclusion from analysis; (viii) track quality control metrics across runs using Levey-Jennings plots; and (ix) automatically flag outliers based on expected values. Existing system features allow researchers to analyze, integrate, visualize, export and securely share their data, as well as to construct custom user interfaces and workflows. CONCLUSIONS: Unlike other tools tailored for Luminex immunoassays, LabKey Server allows labs to customize their Luminex analyses using scripting while still presenting users with a single, graphical interface for processing and analyzing data. The LabKey Server system also stands out among Luminex tools for enabling smooth, secure transfer of data, quality control information, and analyses between collaborators. LabKey Server and its Luminex features are freely available as open source software at http://www.labkey.com under the Apache 2.0 license.
BMC Bioinformatics 04/2013; 14(1):145. · 2.75 Impact Factor
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Genevieve G A Fouda,
Joshua D Amos,
Andrew B Wilks,
Justin Pollara,
Caroline A Ray,
Anjali Chand,
Erika L Kunz,
Brooke E Liebl,
Kaylan Whitaker,
Angela Carville, [......],
Haiyan Chen,
Celia Labranche,
Susan Barnett, Georgia D Tomaras,
Guido Ferrari,
David C Montefiori,
Hua-Xin Liao,
Norman L Letvin,
Barton F Haynes,
Sallie R Permar
[show abstract]
[hide abstract]
ABSTRACT: We previously demonstrated that vaccination of lactating rhesus monkeys with a DNA prime/vector boost strategy induces strong T cell responses, but limited Envelope (Env)-specific humoral responses in breast milk. To improve vaccine-elicited antibody responses in milk, hormone-induced lactating rhesus monkeys were vaccinated with a transmitted/founder (T/F) HIV Env immunogen in a prime-boost strategy modeled after the moderately-protective RV144 HIV vaccine. Lactating rhesus monkeys were intramuscularly primed with either recombinant DNA (n = 4) or MVA pox virus vector (n = 4) expressing the T/F HIV Env C.1086 then boosted twice intramuscularly with C.1086 gp120 and the adjuvant MF59. The vaccines induced Env-binding IgG and IgA as well as neutralizing and ADCC responses in plasma and milk of most vaccinated animals. Importantly, plasma neutralization titers against clade C HIV variants MW965 (p = 0.03) and CAP45 (p= 0.04) were significantly higher in MVA-primed than DNA-primed animals. The superior systemic prime-boost regimen was then compared to a mucosal-boost regimen, in which animals were boosted twice intranasally with C.1086 gp120 and the TLR7/8 agonist R848 following the same systemic prime. While the systemic and mucosal vaccine regimens elicited comparable levels of Env-binding IgG antibodies, mucosal immunization induced significantly stronger Env-binding IgA responses in milk (p=0.03). However, the mucosal regimen was not as potent at inducing functional IgG responses. This study shows that systemic MVA prime followed by either intranasal or systemic protein boosts can elicit strong humoral responses in breast milk, and may be a useful strategy to interrupt postnatal HIV-1 transmission.
Journal of Virology 04/2013; · 5.40 Impact Factor
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Hua-Xin Liao,
Mattia Bonsignori,
S Munir Alam,
Jason S McLellan, Georgia D Tomaras,
M Anthony Moody,
Daniel M Kozink,
Kwan-Ki Hwang,
Xi Chen,
Chun-Yen Tsao, [......],
Faruk Sinangil,
Jerome H Kim,
Nelson L Michael,
Thomas B Kepler,
Peter D Kwong,
John R Mascola,
Gary J Nabel,
Abraham Pinter,
Susan Zolla-Pazner,
Barton F Haynes
[show abstract]
[hide abstract]
ABSTRACT: The RV144 HIV-1 trial of the canary pox vector (ALVAC-HIV) plus the gp120 AIDSVAX B/E vaccine demonstrated an estimated efficacy of 31%, which correlated directly with antibodies to HIV-1 envelope variable regions 1 and 2 (V1-V2). Genetic analysis of trial viruses revealed increased vaccine efficacy against viruses matching the vaccine strain at V2 residue 169. Here, we isolated four V2 monoclonal antibodies from RV144 vaccinees that recognize residue 169, neutralize laboratory-adapted HIV-1, and mediate killing of field-isolate HIV-1-infected CD4(+) T cells. Crystal structures of two of the V2 antibodies demonstrated that residue 169 can exist within divergent helical and loop conformations, which contrasted dramatically with the β strand conformation previously observed with a broadly neutralizing antibody PG9. Thus, RV144 vaccine-induced immune pressure appears to target a region that may be both sequence variable and structurally polymorphic. Variation may signal sites of HIV-1 envelope vulnerability, providing vaccine designers with new options.
Immunity 01/2013; · 21.64 Impact Factor
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Chunlai Jiang,
Nicholas F Parrish,
Craig B Wilen,
Hui Li,
Yue Chen,
Jeffrey W Pavlicek,
Anna Berg,
Xiaozhi Lu,
Hongshuo Song,
John C Tilton, [......],
Robert Schutte,
Stephanie Freel, Georgia D Tomaras,
Rebecca Nedellec,
Donald E Mosier,
Barton F Haynes,
George M Shaw,
Beatrice H Hahn,
Robert W Doms,
Feng Gao
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S Munir Alam,
Hua-Xin Liao, Georgia D Tomaras,
Mattia Bonsignori,
Chun-Yen Tsao,
Kwan-Ki Hwang,
Haiyan Chen,
Krissey E Lloyd,
Cindy Bowman,
Laura Sutherland, [......],
Sorachai Nitayaphan,
Nicos Karasavva,
Supachai Rerks-Ngarm,
Jerome H Kim,
Nelson L Michael,
Susan Zolla-Pazner,
Sampa Santra,
Norman L Letvin,
Stephen C Harrison,
Barton F Haynes
[show abstract]
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ABSTRACT: An immune correlates analysis of the RV144 HIV-1 vaccine trial revealed that antibody responses to the gp120 V1V2 region correlated inversely with infection risk. The RV144 protein immunogens (A244-rp120, MN-rgp120) were modified by an N-terminal 11 amino-acid deletion (Δ11) and addition of a HSV (Herpes simplex virus)-gD protein derived tag (gD). We investigated the effects of these modifications on gp120 expression, antigenicity and immunogenicity by comparing unmodified A244 gp120 with both Δ11 deletion and gD tag and with Δ11 only. Analysis of A244 gp120, with or without Δ11 or gD, demonstrated that the Δ11 deletion, without the addition of gD, was sufficient for enhanced antigenicity to gp120 C1 region, conformational V2 and V1/V2 gp120 conformational epitopes. RV144- vaccinee sera IgG bound more avidly to A244 gp120 Δ11 than to the unmodified gp120 and their binding was blocked by C1, V2 and V1/V2 antibodies. Rhesus macaques immunized with the three different forms of A244 gp120 proteins gave similar levels of gp120 antibody titers, although higher antibody titers developed earlier in A244 Δ11 gp120 immunized animals. Conformational V1/V2 mAbs gave significantly higher levels of blocking of plasma IgG from A244 Δ11 gp120 immunized animals than IgG from animals immunized with unmodified A244 gp120, thus indicating a qualitative difference in the V1/V2 antibodies induced by A244 Δ11 gp120. These results demonstrate that deletion of N-terminal residues in the RV144 A244 gp120 immunogen improves both envelope antigenicity and immunogenicity.
Journal of Virology 11/2012; · 5.40 Impact Factor
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Nicos Karasavvas,
Erik Billings,
Mangala Rao,
Constance Williams,
Susan Zolla-Pazner,
Robert T Bailer,
Richard A Koup,
Sirinan Madnote,
Duangnapa Arworn,
Xiaoying Shen, [......],
Faruk Sinangil,
Bette T Korber,
David C Montefiori,
John R Mascola,
Merlin L Robb,
Barton F Haynes,
Viseth Ngauy,
Nelson L Michael,
Jerome H Kim,
Mark S de Souza For The Moph Taveg Collaboration
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ABSTRACT: Abstract The Thai Phase III clinical trial (RV144) showed modest efficacy in preventing HIV-1 acquisition. Plasma collected from HIV-1-uninfected trial participants completing all injections with ALVAC-HIV (vCP1521) prime and AIDSVAX B/E boost were tested for antibody responses against HIV-1 gp120 envelope (Env). Peptide microarray analysis from six HIV-1 subtypes and group M consensus showed that vaccination induced antibody responses to the second variable (V2) loop of gp120 of multiple subtypes. We further evaluated V2 responses by ELISA and surface plasmon resonance using cyclic (Cyc) and linear V2 loop peptides. Thirty-one of 32 vaccine recipients tested (97%) had antibody responses against Cyc V2 at 2 weeks postimmunization with a reciprocal geometric mean titer (GMT) of 1100 (range: 200-3200). The frequency of detecting plasma V2 antibodies declined to 19% at 28 weeks post-last injection (GMT: 110, range: 100-200). Antibody responses targeted the mid-region of the V2 loop that contains conserved epitopes and has the amino acid sequence KQKVHALFYKLDIVPI (HXB2 Numbering sequence 169-184). Valine at position 172 was critical for antibody binding. The frequency of V3 responses at 2 weeks postimmunization was modest (18/32, 56%) with a GMT of 185 (range: 100-800). In contrast, naturally infected HIV-1 individuals had a lower frequency of antibody responses to V2 (10/20, 50%; p=0.003) and a higher frequency of responses to V3 (19/20, 95%), with GMTs of 400 (range: 100-3200) and 3570 (range: 200-12,800), respectively. RV144 vaccination induced antibodies that targeted a region of the V2 loop that contains conserved epitopes. Early HIV-1 transmission events involve V2 loop interactions, raising the possibility that anti-V2 antibodies in RV144 may have contributed to viral inhibition.
AIDS research and human retroviruses 10/2012; · 2.18 Impact Factor
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[show abstract]
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ABSTRACT: Attempts to formulate a protective HIV-1 vaccine through classic vaccine design strategies have not been successful. Elicitation of HIV-1-specific broadly neutralizing antibodies (bnAbs) at high titers that are present before exposure might be required to achieve protection. Recently, the application of new technologies has facilitated the study of clonal lineages of HIV-1 envelope (Env) antibodies, which have provided insights into HIV-1 antibody development during infection and upon vaccination. Strategies are being developed for the analysis of infection and vaccine candidate-induced antibodies, their gene usage, and their maturation pathways such that this information can be used to attempt to guide rational vaccine design.
Trends in Microbiology 09/2012; 20(11):532-9. · 7.91 Impact Factor
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Mattia Bonsignori,
Justin Pollara,
M Anthony Moody,
Michael D Alpert,
Xi Chen,
Kwan-Ki Hwang,
Peter B Gilbert,
Ying Huang,
Thaddeus C Gurley,
Daniel M Kozink, [......],
Jerome H Kim,
Nelson L Michael, Georgia D Tomaras,
David C Montefiori,
George K Lewis,
Anthony Devico,
David T Evans,
Guido Ferrari,
Hua-Xin Liao,
Barton F Haynes
[show abstract]
[hide abstract]
ABSTRACT: The ALVAC-HIV/AIDSVAX-B/E RV144 vaccine trial showed an estimated efficacy of 31%. RV144 secondary immune correlate analysis demonstrated that the combination of low plasma anti-HIV-1 Env IgA antibodies and high levels of antibody-dependent cellular cytotoxicity (ADCC) inversely correlate with infection risk. One hypothesis is that the observed protection in RV144 is partially due to ADCC-mediating antibodies. We found that the majority (73 to 90%) of a representative group of vaccinees displayed plasma ADCC activity, usually (96.2%) blocked by competition with the C1 region-specific A32 Fab fragment. Using memory B-cell cultures and antigen-specific B-cell sorting, we isolated 23 ADCC-mediating nonclonally related antibodies from 6 vaccine recipients. These antibodies targeted A32-blockable conformational epitopes (n = 19), a non-A32-blockable conformational epitope (n = 1), and the gp120 Env variable loops (n = 3). Fourteen antibodies mediated cross-clade target cell killing. ADCC-mediating antibodies displayed modest levels of V-heavy (VH) chain somatic mutation (0.5 to 1.5%) and also displayed a disproportionate usage of VH1 family genes (74%), a phenomenon recently described for CD4-binding site broadly neutralizing antibodies (bNAbs). Maximal ADCC activity of VH1 antibodies correlated with mutation frequency. The polyclonality and low mutation frequency of these VH1 antibodies reveal fundamental differences in the regulation and maturation of these ADCC-mediating responses compared to VH1 bNAbs.
Journal of Virology 08/2012; 86(21):11521-32. · 5.40 Impact Factor
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M Anthony Moody,
Nicole L Yates,
Joshua D Amos,
Mark S Drinker,
Joshua A Eudailey,
Thaddeus C Gurley,
Dawn J Marshall,
John F Whitesides,
Xi Chen,
Andrew Foulger, [......],
Joy Pickeral,
Justin Pollara,
Garnett Kelsoe,
S Munir Alam,
Guido Ferrari,
David C Montefiori,
Gerald Voss,
Hua-Xin Liao, Georgia D Tomaras,
Barton F Haynes
[show abstract]
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ABSTRACT: Most antibodies that broadly neutralize HIV-1 are highly somatically mutated in antibody clonal lineages that persist over time. Here, we describe the analysis of human antibodies induced during an HIV-1 vaccine trial (GSK PRO HIV-002) that used the clade B envelope (Env) gp120 of clone W6.1D (gp120(W6.1D)). Using dual-color antigen-specific sorting, we isolated Env-specific human monoclonal antibodies (MAbs) and studied the clonal persistence of antibodies in the setting of HIV-1 Env vaccination. We found evidence of V(H) somatic mutation induced by the vaccine but only to a modest level (3.8% ± 0.5%; range 0 to 8.2%). Analysis of 34 HIV-1-reactive MAbs recovered over four immunizations revealed evidence of both sequential recruitment of naïve B cells and restimulation of previously recruited memory B cells. These recombinant antibodies recapitulated the anti-HIV-1 activity of participant serum including pseudovirus neutralization and antibody-dependent cell-mediated cytotoxicity (ADCC). One antibody (3491) demonstrated a change in specificity following somatic mutation with binding of the inferred unmutated ancestor to a linear C2 peptide while the mutated antibody reacted only with a conformational epitope in gp120 Env. Thus, gp120(W6.1D) was strongly immunogenic but over four immunizations induced levels of affinity maturation below that of broadly neutralizing MAbs. Improved vaccination strategies will be needed to drive persistent stimulation of antibody clonal lineages to induce affinity maturation that results in highly mutated HIV-1 Env-reactive antibodies.
Journal of Virology 05/2012; 86(14):7496-507. · 5.40 Impact Factor
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Stephanie A Freel,
Ralph A Picking,
Guido Ferrari,
Haitao Ding,
Christina Ochsenbauer,
John C Kappes,
Jennifer L Kirchherr,
Kelly A Soderberg,
Kent J Weinhold,
Coleen K Cunningham,
Thomas N Denny,
John A Crump,
Myron S Cohen,
Andrew J McMichael,
Barton F Haynes, Georgia D Tomaras
[show abstract]
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ABSTRACT: CD8-mediated virus inhibition can be detected in HIV-1-positive subjects who naturally control virus replication. Characterizing the inhibitory function of CD8(+) T cells during acute HIV-1 infection (AHI) can elucidate the nature of the CD8(+) responses that can be rapidly elicited and that contribute to virus control. We examined the timing and HIV-1 antigen specificity of antiviral CD8(+) T cells during AHI. Autologous and heterologous CD8(+) T cell antiviral functions were assessed longitudinally during AHI in five donors from the CHAVI 001 cohort using a CD8(+) T cell-mediated virus inhibition assay (CD8 VIA) and transmitted/founder (T/F) viruses. Potent CD8(+) antiviral responses against heterologous T/F viruses appeared during AHI at the first time point sampled in each of the 5 donors (Fiebig stages 1/2 to 5). Inhibition of an autologous T/F virus was durable to 48 weeks; however, inhibition of heterologous responses declined concurrent with the resolution of viremia. HIV-1 viruses from 6 months postinfection were more resistant to CD8(+)-mediated virus inhibition than cognate T/F viruses, demonstrating that the virus escapes early from CD8(+) T cell-mediated inhibition of virus replication. CD8(+) T cell antigen-specific subsets mediated inhibition of T/F virus replication via soluble components, and these soluble responses were stimulated by peptide pools that include epitopes that were shown to drive HIV-1 escape during AHI. These data provide insights into the mechanisms of CD8-mediated virus inhibition and suggest that functional analyses will be important for determining whether similar antigen-specific virus inhibition can be induced by T cell-directed vaccine strategies.
Journal of Virology 04/2012; 86(12):6835-46. · 5.40 Impact Factor
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Barton F Haynes,
Peter B Gilbert,
M Juliana McElrath,
Susan Zolla-Pazner, Georgia D Tomaras,
S Munir Alam,
David T Evans,
David C Montefiori,
Chitraporn Karnasuta,
Ruengpueng Sutthent, [......],
Michael D Alpert,
Nicole L Yates,
Xiaoying Shen,
Richard A Koup,
Punnee Pitisuttithum,
Jaranit Kaewkungwal,
Sorachai Nitayaphan,
Supachai Rerks-Ngarm,
Nelson L Michael,
Jerome H Kim
[show abstract]
[hide abstract]
ABSTRACT: In the RV144 trial, the estimated efficacy of a vaccine regimen against human immunodeficiency virus type 1 (HIV-1) was 31.2%. We performed a case-control analysis to identify antibody and cellular immune correlates of infection risk.
In pilot studies conducted with RV144 blood samples, 17 antibody or cellular assays met prespecified criteria, of which 6 were chosen for primary analysis to determine the roles of T-cell, IgG antibody, and IgA antibody responses in the modulation of infection risk. Assays were performed on samples from 41 vaccinees who became infected and 205 uninfected vaccinees, obtained 2 weeks after final immunization, to evaluate whether immune-response variables predicted HIV-1 infection through 42 months of follow-up.
Of six primary variables, two correlated significantly with infection risk: the binding of IgG antibodies to variable regions 1 and 2 (V1V2) of HIV-1 envelope proteins (Env) correlated inversely with the rate of HIV-1 infection (estimated odds ratio, 0.57 per 1-SD increase; P=0.02; q=0.08), and the binding of plasma IgA antibodies to Env correlated directly with the rate of infection (estimated odds ratio, 1.54 per 1-SD increase; P=0.03; q=0.08). Neither low levels of V1V2 antibodies nor high levels of Env-specific IgA antibodies were associated with higher rates of infection than were found in the placebo group. Secondary analyses suggested that Env-specific IgA antibodies may mitigate the effects of potentially protective antibodies.
This immune-correlates study generated the hypotheses that V1V2 antibodies may have contributed to protection against HIV-1 infection, whereas high levels of Env-specific IgA antibodies may have mitigated the effects of protective antibodies. Vaccines that are designed to induce higher levels of V1V2 antibodies and lower levels of Env-specific IgA antibodies than are induced by the RV144 vaccine may have improved efficacy against HIV-1 infection.
New England Journal of Medicine 04/2012; 366(14):1275-86. · 53.30 Impact Factor
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[show abstract]
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ABSTRACT: Previous studies have revealed that HIV-infected individuals possess circulating CD4(+)CD8(+) double-positive (DP) T cells specific for HIV Ags. In the present study, we analyzed the proliferation and functional profile of circulating DP T cells from 30 acutely HIV-infected individuals and 10 chronically HIV-infected viral controllers. The acutely infected group had DP T cells that showed more proliferative capability and multifunctionality than did both their CD4(+) and CD8(+) T cells. DP T cells were found to exhibit greater proliferation and higher multifunctionality compared with CD4 T cells in the viral controller group. The DP T cell response represented 16% of the total anti-HIV proliferative response and >70% of the anti-HIV multifunctional response in the acutely infected subjects. Proliferating DP T cells of the acutely infected subjects responded to all HIV Ag pools with equal magnitude. Conversely, the multifunctional response was focused on the pool representing Nef, Rev, Tat, VPR, and VPU. Meanwhile, the controllers' DP T cells focused on Gag and the Nef, Rev, Tat, VPR, and VPU pool for both their proliferative and multifunctional responses. Finally, we show that the presence of proliferating DP T cells following all HIV Ag stimulations is well correlated with proliferating CD4 T cells whereas multifunctionality appears to be largely independent of multifunctionality in other T cell compartments. Therefore, DP T cells represent a highly reactive cell population during acute HIV infection, which responds independently from the traditional T cell compartments.
The Journal of Immunology 03/2012; 188(9):4289-96. · 5.79 Impact Factor
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Mattia Bonsignori,
David C Montefiori,
Xueling Wu,
Xi Chen,
Kwan-Ki Hwang,
Chun-Yen Tsao,
Daniel M Kozink,
Robert J Parks, Georgia D Tomaras,
John A Crump,
Saidi H Kapiga,
Noel E Sam,
Peter D Kwong,
Thomas B Kepler,
Hua-Xin Liao,
John R Mascola,
Barton F Haynes
[show abstract]
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ABSTRACT: Plasma from a small subset of subjects chronically infected with HIV-1 shows remarkable magnitude and breadth of neutralizing activity. From one of these individuals (CH0219), we isolated two broadly neutralizing antibodies (bnAbs), CH01 and VRC-CH31, from two clonal lineages of memory B cells with distinct specificities (variable loop 1 and 2 [V1V2] conformational specificity and CD4-binding site specificity, respectively) that recapitulate 95% of CH0219 serum neutralization breadth. These data provide proof of concept for an HIV-1 vaccine that aims to elicit bnAbs of multiple specificities.
Journal of Virology 02/2012; 86(8):4688-92. · 5.40 Impact Factor
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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,
Sallie R Permar
[show abstract]
[hide abstract]
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.
PLoS ONE 01/2012; 7(5):e37648. · 4.09 Impact Factor
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Hua-Xin Liao,
Xi Chen,
Supriya Munshaw,
Ruijun Zhang,
Dawn J Marshall,
Nathan Vandergrift,
John F Whitesides,
Xiaozhi Lu,
Jae-Sung Yu,
Kwan-Ki Hwang, [......],
Michael Egholm,
Birgitte B Simen,
Bozena Hanczaruk,
Andrew Z Fire,
Gerald Voss,
Garnett Kelsoe, Georgia D Tomaras,
M Anthony Moody,
Thomas B Kepler,
Barton F Haynes
[show abstract]
[hide abstract]
ABSTRACT: The initial antibody response to HIV-1 is targeted to envelope (Env) gp41, and is nonneutralizing and ineffective in controlling viremia. To understand the origins and characteristics of gp41-binding antibodies produced shortly after HIV-1 transmission, we isolated and studied gp41-reactive plasma cells from subjects acutely infected with HIV-1. The frequencies of somatic mutations were relatively high in these gp41-reactive antibodies. Reverted unmutated ancestors of gp41-reactive antibodies derived from subjects acutely infected with HIV-1 frequently did not react with autologous HIV-1 Env; however, these antibodies were polyreactive and frequently bound to host or bacterial antigens. In one large clonal lineage of gp41-reactive antibodies, reactivity to HIV-1 Env was acquired only after somatic mutations. Polyreactive gp41-binding antibodies were also isolated from uninfected individuals. These data suggest that the majority of gp41-binding antibodies produced after acute HIV-1 infection are cross-reactive responses generated by stimulating memory B cells that have previously been activated by non-HIV-1 antigens.
Journal of Experimental Medicine 10/2011; 208(11):2237-49. · 13.85 Impact Factor
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Lindsey R Baden,
William A Blattner,
Cecilia Morgan,
Yunda Huang,
Olivier D Defawe,
Magdalena E Sobieszczyk,
Nidhi Kochar, Georgia D Tomaras,
M Juliana McElrath,
Nina Russell,
Kara Brandariz,
Massimo Cardinali,
Barney S Graham,
Dan H Barouch,
Raphael Dolin
[show abstract]
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ABSTRACT: To investigate the potential immunostimulatory effect of interleukin (IL) 2 as a human immunodeficiency virus type 1 (HIV-1) vaccine adjuvant, we conducted a study of a plasmid coding for a fusion protein of IL-2 and immunoglobulin (IL-2/Ig).
This phase I trial evaluated an HIV-1 DNA vaccine with the plasmid cytokine adjuvant (IL-2/Ig) in 70 HIV-negative adults. Subjects received placebo (group C), adjuvant alone (group A), vaccine alone (group D), increasing doses of adjuvant concurrent with vaccine (groups T1-T4), or adjuvant given 2 days after vaccine (group T5).
No significant differences in adverse events were observed between treatment groups. Cellular immune responses to envelope protein EnvA peptides were detected by interferon (IFN) γ and IL-2 enzyme-linked immunospot (ELISPOT) assays in 50% and 40% of subjects, respectively, in T4, and in 100% and 80% in T5. The median responses for groups T4 and T5, respectively, were 90 and 193 spot-forming cells (SFCs)/10⁶ peripheral blood mononuclear cells (P = .004; T4 vs T5) for the IL-2 ELISPOT assay and 103 and 380 SFCs/10⁶ PBMCs (P = .003; T4 vs T5) for the IFN-γ ELISPOT assay. A trend to more durable cellular immune responses in T5 was observed at 1 year (T5 vs T4/D; P = .07). Higher anti-Env antibody responses were detected with T5 than with T4.
Plasmid IL-2/Ig significantly increased immune responses when administered 2 days after the DNA vaccine, compared with simultaneous administration. These observations have important implications for the development of cytokine augmentation strategies. Clinical Trials Registration: NCT00069030.
The Journal of Infectious Diseases 09/2011; 204(10):1541-9. · 6.41 Impact Factor
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S Munir Alam,
Hua-Xin Liao,
S Moses Dennison,
Frederick Jaeger,
Robert Parks,
Kara Anasti,
Andrew Foulger,
Michele Donathan,
Judith Lucas,
Laurent Verkoczy,
Nathan Nicely, Georgia D Tomaras,
Garnett Kelsoe,
Bing Chen,
Thomas B Kepler,
Barton F Haynes
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ABSTRACT: Genetic factors, as well as antigenic stimuli, can influence antibody repertoire formation. Moreover, the affinity of antigen for unmutated naïve B cell receptors determines the threshold for activation of germinal center antibody responses. The gp41 2F5 broadly neutralizing antibody (bNAb) uses the V(H)2-5 gene, which has 10 distinct alleles that use either a heavy-chain complementarity-determining region 2 (HCDR2) aspartic acid (D(H54)) or an HCDR2 asparagine (N(H54)) residue. The 2F5 HCDR2 D(H54) residue has been shown to form a salt bridge with gp41 (665)K; the V(H)2-5 germ line allele variant containing N(H54) cannot do so and thus should bind less avidly to gp41. Thus, the induction of 2F5 bNAb is dependent on both genetic and structural factors that could affect antigen affinity of unmutated naïve B cell receptors. Here, we studied allelic variants of the V(H)2-5 inferred germ line forms of the HIV-1 gp41 bNAb 2F5 for their antigen binding affinities to gp41 linear peptide and conformational protein antigens. Both V(H)2-5 2F5 inferred germ line variants bound to gp41 peptides and protein, including the fusion intermediate protein mimic, although more weakly than the mature 2F5 antibody. As predicted, the affinity of the N(H54) variant for fusion-intermediate conformation was an order of magnitude lower than that of the D(H54) V(H)2-5 germ line antibody, demonstrating that allelic variants of 2F5 germ line antibodies differentially bind to gp41. Thus, these data demonstrate a genetically determined trait that may affect host responses to HIV-1 envelope epitopes recognized by broadly neutralizing antibodies and has implications for unmutated ancestor-based immunogen design.
Journal of Virology 09/2011; 85(22):11725-31. · 5.40 Impact Factor
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Ben-Jiang Ma,
S Munir Alam,
Eden P Go,
Xiaozhi Lu,
Heather Desaire, Georgia D Tomaras,
Cindy Bowman,
Laura L Sutherland,
Richard M Scearce,
Sampa Santra,
Norman L Letvin,
Thomas B Kepler,
Hua-Xin Liao,
Barton F Haynes
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ABSTRACT: The HIV-1 gp41 envelope (Env) membrane proximal external region (MPER) is an important vaccine target that in rare subjects can elicit neutralizing antibodies. One mechanism proposed for rarity of MPER neutralizing antibody generation is lack of reverted unmutated ancestor (putative naive B cell receptor) antibody reactivity with HIV-1 envelope. We have studied the effect of partial deglycosylation under non-denaturing (native) conditions on gp140 Env antigenicity for MPER neutralizing antibodies and their reverted unmutated ancestor antibodies. We found that native deglycosylation of clade B JRFL gp140 as well as group M consensus gp140 Env CON-S selectively increased the reactivity of Env with the broad neutralizing human mAbs, 2F5 and 4E10. Whereas fully glycosylated gp140 Env either did not bind (JRFL), or weakly bound (CON-S), 2F5 and 4E10 reverted unmutated ancestors, natively deglycosylated JRFL and CON-S gp140 Envs did bind well to these putative mimics of naive B cell receptors. These data predict that partially deglycoslated Env would bind better than fully glycosylated Env to gp41-specific naïve B cells with improved immunogenicity. In this regard, immunization of rhesus macaques demonstrated enhanced immunogenicity of the 2F5 MPER epitope on deglyosylated JRFL gp140 compared to glycosylated JRFL gp140. Thus, the lack of 2F5 and 4E10 reverted unmutated ancestor binding to gp140 Env may not always be due to lack of unmutated ancestor antibody reactivity with gp41 peptide epitopes, but rather, may be due to glycan interference of binding of unmutated ancestor antibodies of broad neutralizing mAb to Env gp41.
PLoS Pathogens 09/2011; 7(9):e1002200. · 9.13 Impact Factor
