Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Aug 14;349(6249):aab1253.
doi: 10.1126/science.aab1253. Epub 2015 Jul 30.

HIV-1 VACCINES. Diversion of HIV-1 vaccine-induced immunity by gp41-microbiota cross-reactive antibodies

Wilton B Williams  1 , Hua-Xin Liao  2 , M Anthony Moody  2 , Thomas B Kepler  3 , S Munir Alam  2 , Feng Gao  2 , Kevin Wiehe  2 , Ashley M Trama  2 , Kathryn Jones  2 , Ruijun Zhang  2 , Hongshuo Song  2 , Dawn J Marshall  2 , John F Whitesides  2 , Kaitlin Sawatzki  3 , Axin Hua  3 , Pinghuang Liu  2 , Matthew Z Tay  2 , Kelly E Seaton  2 , Xiaoying Shen  2 , Andrew Foulger  2 , Krissey E Lloyd  2 , Robert Parks  2 , Justin Pollara  2 , Guido Ferrari  2 , Jae-Sung Yu  2 , Nathan Vandergrift  2 , David C Montefiori  2 , Magdalena E Sobieszczyk  4 , Scott Hammer  4 , Shelly Karuna  5 , Peter Gilbert  6 , Doug Grove  6 , Nicole Grunenberg  5 , M Juliana McElrath  5 , John R Mascola  7 , Richard A Koup  7 , Lawrence Corey  5 , Gary J Nabel  7 , Cecilia Morgan  6 , Gavin Churchyard  8 , Janine Maenza  5 , Michael Keefer  9 , Barney S Graham  7 , Lindsey R Baden  10 , Georgia D Tomaras  2 , Barton F Haynes  1
Affiliations

HIV-1 VACCINES. Diversion of HIV-1 vaccine-induced immunity by gp41-microbiota cross-reactive antibodies

Wilton B Williams et al. Science. .

Abstract

An HIV-1 DNA prime vaccine, with a recombinant adenovirus type 5 (rAd5) boost, failed to protect from HIV-1 acquisition. We studied the nature of the vaccine-induced antibody (Ab) response to HIV-1 envelope (Env). HIV-1-reactive plasma Ab titers were higher to Env gp41 than to gp120, and repertoire analysis demonstrated that 93% of HIV-1-reactive Abs from memory B cells responded to Env gp41. Vaccine-induced gp41-reactive monoclonal antibodies were non-neutralizing and frequently polyreactive with host and environmental antigens, including intestinal microbiota (IM). Next-generation sequencing of an immunoglobulin heavy chain variable region repertoire before vaccination revealed an Env-IM cross-reactive Ab that was clonally related to a subsequent vaccine-induced gp41-reactive Ab. Thus, HIV-1 Env DNA-rAd5 vaccine induced a dominant IM-polyreactive, non-neutralizing gp41-reactive Ab repertoire response that was associated with no vaccine efficacy.

PubMed Disclaimer

Figures

Figure 1
Figure 1. Characteristics of HIV-1-reactive antibodies (Abs) induced by DNA prime, rAd5 boost vaccine
A) Plasma Ab titers (AUC, area under the curve) to gp120 and gp41 proteins in a subset of 40 HVTN 505 vaccine recipients; red circles represent vaccine responders (percents) to the antigens tested, and blue represent non-responders. (B) Plasma binding Abs from 8 HVTN 082 and 204 trial participants (vaccine responders) were screened for gp120 and gp41 reactivity via binding Ab multiplex assay (BAMA). (C) HIV-1-reactive Abs were isolated from single memory B cells of the 8 HVTN 082 and 204 trial participants and screened for binding gp120 and gp41 proteins via ELISA. Statistics: (A) *** p < 0.0001, Wilcoxon signed rank test, B.MN gp41 vs. gp120 (maximum; which is the highest value of all gp120 Envs for each participant), *p < 0.05 McNemar’s test, B.MN gp41 vs. each gp120 (not shown); (B) *p < 0.05**p < 0.01, Wilcoxon signed rank test [Graphpad Prism].
Figure 2
Figure 2. Polyreactivity of vaccine-induced gp41-reactive monoclonal antibodies (mAbs)
Five representative polyreactive and gp41-reactive mAbs demonstrated binding with auto-antigens (ANA) (A), Hep-2 cells (immunofluorescence staining) (B), and anaerobe and aerobe whole cell lysates of intestinal microbiota (IM), bacterial (E. coli) RNA polymerase, MN gp41, and vaccine-strain VRC-A gp140 antigens (BAMA) (C). (B) All images were taken on an Olympus AX70 fluorescence microscope using a 40x objective with a SPOT Flex camera. All images were prepared using 50 µg/mL of mAb. Images were acquired for 5 sec (2F5, 17b) or 10 sec (DH432, DH438, DH440, DH443, DH444). Size bars are 25 µm. (A, B) Control antibodies: 4E10 (gp41), 2F5 (gp41), 17b (gp41), and palivizumab (RSV). (C) MAb binding was reported as MFI-background-blank, where background is plate specific background binding and blank represents non-specific sample binding to a negative control bead. Positivity cutoff for binding was 100 MFI as previously reported (2); AbCLL1324 as an IM-reactive control was used to validate IM binding and gp41 mAb 7B2 that does not react with IM was used as a negative control. Data shown were generated from the same commercially available kits (A, B), and a single BAMA experiment (C), which was in agreement with Ab-IM reactivity in western blots (Figure S10 or not shown).
Figure 3
Figure 3. Characteristics of clonally-related antibodies (Abs)
Pre- (DH477) and post- (DH476) vaccine clonally-related Abs found in vaccinee 082-003 had the same VHDJH recombination. IGHV, IGHD and IGHJ Ab segments were statistically inferred (A, B); nucleotides in the 3’ end of the V-gene and 5’ end of J-gene shown with rearrangement junctions (gray-shaded nucleotides - NN1, NN2), and HCDR3 (underlined nucleotides) indicated. The lower case letters show nucleotides that have been removed from the germline gene during VHDJH rearrangement. Pre- (DH477) and post- (DH476) vaccine heavy chain sequences were paired with the natural light chain (IGκV3–20*01, IGκJ1*01, 1.7% mutated nt, 10 aa CDR3) of the post-vaccine Ab DH476. (C) Ab binding to recombinant HIV-1 Env gp140, 5-Helix gp41 and MN gp41 proteins was determined by surface plasmon resonance (SPR) analysis, and (D) the antibody binding Kd determined by rate constants or steady-state analysis measurements. (E) Binding of DH477 to VRC-A gp140 expressed on the surface of 293i cells. Anti-V3 mAb (19B) and a representative gp41-reactive Ab (DH440) from this study, both of which bind recombinant VRC-A gp140, were used as positive controls, while palivizumab (RSV Ab) was used as a negative IgG1 control. (F) The cross-reactivity of pre-(DH477) and post-(DH476) vaccine Abs with intestinal microbiota (IM) whole cell lysates (WCLs) was determined via western blot and SPR as previously described (2). DH438 (gp41-reactive Ab) and palivizumab were used as positive and negative control Abs, respectively, for western blots, and the arrows indicate candidate IM antigens bound by DH476 and/or DH477. In SPR analysis, IM WCLs were diluted in PBS buffer as indicated and injected over mAbs captured on an anti-IgFc immobilized sensor surface. Specific binding of the lysate proteins to the mAbs in SPR were measured after subtraction of non-specific binding to anti-HA mAb CH65. Abbreviations - nt, nucleotides; aa, amino acid. Data shown (C–F) were representative of duplicate experiments, except SPR screen of DH477 and DH476 for binding rgp140 proteins that was performed once (C).
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Tomaras GD, et al. Initial B-cell responses to transmitted human immunodeficiency virus type 1: virion-binding immunoglobulin M (IgM) and IgG antibodies followed by plasma anti-gp41 antibodies with ineffective control of initial viremia. J Virol. 2008 Dec;82:12449. - PMC - PubMed
    1. Trama AM, et al. HIV-1 Envelope gp41 Antibodies Can Originate from Terminal Ileum B Cells that Share Cross-Reactivity with Commensal Bacteria. Cell Host Microbe. 2014 Aug;16:215. - PMC - PubMed
    1. Liao HX, et al. Initial antibodies binding to HIV-1 gp41 in acutely infected subjects are polyreactive and highly mutated. J Exp Med. 2011 Oct;208:2237. - PMC - PubMed
    1. Catanzaro AT, et al. Phase I clinical evaluation of a six-plasmid multiclade HIV-1 DNA candidate vaccine. Vaccine. 2007 May;25:4085. - PubMed
    1. Churchyard GJ, et al. A phase IIA randomized clinical trial of a multiclade HIV-1 DNA prime followed by a multiclade rAd5 HIV-1 vaccine boost in healthy adults (HVTN204) PLoS One. 2011;6:e21225. - PMC - PubMed

Publication types

MeSH terms

-