Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques
Kan Luo, … , Barton F. Haynes, M. Anthony Moody
Kan Luo, … , Barton F. Haynes, M. Anthony Moody
Published December 8, 2016
Citation Information: JCI Insight. 2016;1(20):e88522. https://doi.org/10.1172/jci.insight.88522.
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Research Article AIDS/HIV Vaccines

Tissue memory B cell repertoire analysis after ALVAC/AIDSVAX B/E gp120 immunization of rhesus macaques

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.

Authors

Kan Luo, Hua-Xin Liao, Ruijun Zhang, David Easterhoff, Kevin Wiehe, Thaddeus C. Gurley, Lawrence C. Armand, Ashley A. Allen, Tarra A. Von Holle, Dawn J. Marshall, John F. Whitesides, Jamie Pritchett, Andrew Foulger, Giovanna Hernandez, Robert Parks, Krissey E. Lloyd, Christina Stolarchuk, Sheetal Sawant, Jessica Peel, Nicole L. Yates, Erika Dunford, Sabrina Arora, Amy Wang, Cindy M. Bowman, Laura L. Sutherland, Richard M. Scearce, Shi-Mao Xia, Mattia Bonsignori, Justin Pollara, R. Whitney Edwards, Sampa Santra, Norman L. Letvin, James Tartaglia, Donald Francis, Faruk Sinangil, Carter Lee, Jaranit Kaewkungwal, Sorachai Nitayaphan, Punnee Pitisuttithum, Supachai Rerks-ngarm, Nelson L. Michael, Jerome H. Kim, S. Munir Alam, Nathan A. Vandergrift, Guido Ferrari, David C. Montefiori, Georgia D. Tomaras, Barton F. Haynes, M. Anthony Moody

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Figure 9

Epitope mapping of mAbs and characterization of mAbs with long HCDR3 loops.

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Epitope mapping of mAbs and characterization of mAbs with long HCDR3 loo...
Phylograms for clonal lineages DH614 and DH621 shown with anatomic site of origin. Epitope mapping ELISA data shown as half maximal effective concentration (EC50) and color coded as in the legend. The sequence of the 22–amino acid V2 peptide is shown across the top of the grids (A–C) and data below each amino acid are the observed EC50 for variant peptide constructs with substitutions at that position. (A) Mature lineage DH614 mAbs were most sensitive to changes at K169, H173, F176, and Y177. For this lineage, data for binding to infected cells are shown in Figure 8. (B) Mature lineage DH621 mAbs were most sensitive to changes at K168, Q170, K171, H173, K178, and D180. Lineage members bound the surface of AE.CM235-infected target cells. Lineage characteristics: VH5~117/JH4, heavy chain (HC) complementarity-determining region 3 (HCDR3) length 13, mean HC mutation frequency 5.2%; Vκ2~73/Jκ4, CDR3 length 9, mean light chain (LC) mutation frequency 4.7%. (C) V1V2-specific mAbs that did not use Vλ3~17 showed different patterns of sensitivity to amino acid substitutions. DH623 showed significantly reduced binding only for changes at H173. DH624 was not sensitive to H173 changes but was sensitive to changes at K178, L179, and D180. DH627 was sensitive to changes at H173, L175, and K178. All 3 mAbs bound the surface of AE.CM235-infected target cells. (D) Two mAbs that did use Vλ3~17 did not bind to a gp120 from RV144 breakthrough strain AE.703357, but did bind to proteins and V1V2 proteins reflective of the vaccine, and also bound to AE.CM235-infected target cells. (E) Tested long HCDR3 mAbs showed 3 patterns of binding to gp120 constructs. DH605 and DH622 showed reduced binding to gp120AE.A244 N259A/N301A/N332A. DH609 and DH611 were clonally related and showed reduced binding to gp120AE.A244 Δ371I/P363N and gp120C.YU2 D368R; DH607 did not bind clade AE proteins but was sensitive to gp120C.YU2 D368R. DH606, DH608, and DH610 potently bound all tested variants of gp120AE.A244 but differentially bound gp120C.YU2 variants. The anatomic site of origin appears after the mAb name: BL, peripheral blood; SP, spleen; IN, inguinal lymph node (LN); AP, anterior pelvic LN; PP, posterior pelvic LN; PA, periaortic LN; MT, mesenteric LN; ND, not done; MFI, mean fluorescence intensity; UCA, unmutated common ancestor.