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. 2015 Jun 15;194(12):5903-14.
doi: 10.4049/jimmunol.1500122. Epub 2015 May 11.

Diverse antibody genetic and recognition properties revealed following HIV-1 envelope glycoprotein immunization

Ganesh E Phad  1 , Néstor Vázquez Bernat  1 , Yu Feng  2 , Jidnyasa Ingale  2 , Paola Andrea Martinez Murillo  1 , Sijy O'Dell  3 , Yuxing Li  4 , John R Mascola  3 , Christopher Sundling  5 , Richard T Wyatt  2 , Gunilla B Karlsson Hedestam  6
Affiliations

Diverse antibody genetic and recognition properties revealed following HIV-1 envelope glycoprotein immunization

Ganesh E Phad et al. J Immunol. .

Abstract

Isolation of mAbs elicited by vaccination provides opportunities to define the development of effective immunity. Ab responses elicited by current HIV-1 envelope glycoprotein (Env) immunogens display narrow neutralizing activity with limited capacity to block infection by tier 2 viruses. Intense work in the field suggests that improved Env immunogens are forthcoming, and it is therefore important to concurrently develop approaches to investigate the quality of vaccine-elicited responses at a higher level of resolution. In this study, we cloned a representative set of mAbs elicited by a model Env immunogen in rhesus macaques and comprehensively characterized their genetic and functional properties. The mAbs were genetically diverse, even within groups of Abs targeting the same subregion of Env, consistent with a highly polyclonal response. mAbs directed against two subdeterminants of Env, the CD4 binding site and V region 3, could in part account for the neutralizing activity observed in the plasma of the animal from which they were cloned, demonstrating the power of mAb isolation for a detailed understanding of the elicited response. Finally, through comparative analyses of mAb binding and neutralizing capacity of HIV-1 using matched Envs, we demonstrate complex relationships between epitope recognition and accessibility, highlighting the protective quaternary packing of the HIV-1 spike relative to vaccine-induced mAbs.

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Figures

Figure 1
Figure 1. Isolation of Env-vaccine MAbs and epitope mapping
(A) Schematic of Env-specific memory B cell sorting followed by amplification, cloning, and expression of antibody HC and LC. (B) Representative ELISA binding curves showing the epitope mapping using YU2 Env ligands as follows: gp140-F and gp140-F-D368R (CD4bs-specific), gp120-F and gp120-F-ΔV3 (V3-specific), gp120-F and gp120-F-ΔV1V2 (V1V2-specific), gp140-F and gp120-F (gp41-specific), gp140-F, gp140-GCN4 and collagen foldon (foldon-specific). MAbs that could not be mapped with this set of the probes used here were defined as unknown specificity. Ova or influenza hemagglutinin-1 (HA1) were used as a controls. Titration curves are shown as Log10 dilutions (μg/ml). (C) The pie chart demonstrates the distribution of Env-specific MAbs based on their epitope specificity with the total number of MAbs indicated in the center. The color of each slice represents a different Env specificity: red - CD4bs specific, blue – V3 specific, purple – V1V2 specific, grey – gp41specific, green – foldon specific, orange – unknown specificity, and the area of the slice is proportional to the total number of MAbs against a given specificity. The numbers outside each colored slice indicates the number of unique clones for each of the specificities.
Figure 2
Figure 2. Vaccine-induced Env-specific MAbs are genetically diverse
The bars in both (A) and (B) indicate the number of cloned MAbs (Y-axis) using a given VH gene segment (A) or HCDR3 length (B). The colored dots above the bars indicate the epitope specificity of expressed MAbs for a given VH gene segment (A) or HCDR3 length (B). Larger dots indicate the number of clonally related MAbs identified. The VH gene segment expression profile by Env-specific IgG+ memory B cells isolated from the F124 and F128 together (n=383) is shown in the insert box in (A).
Figure 3
Figure 3. Summary of genetic properties of the vaccine-elicited Env-specific MAbs
V(D)J gene annotations and CDR3 regions of HC and LC of Env-specific MAbs were extracted using IgBLAST and IMGT-VQUEST respectively. Antibody HC and LC sequences with the same V and J gene, identical CDR3 length, and≥ 80% CDR3 identity at the aa level were determined as clonal variants and are denoted by special characters: GE2.JD5‡ and GE2.JG8‡; GE1.BB2† and GE1.BH6†; GE1.BH7√ and GE1.BD2√; GE1.BB4•, GE2.BA7•, and GE2.BG1•; GE2.JF4*, GE2.JB7*, GE2.JH5*, and GE2.JG5*; GE2.BB6# and GE2.BE8#.
Figure 4
Figure 4. HIV-1 neutralizing activity of Env-specific MAbs
The pre- and post-immunization plasma, and the CD4bs-directed and V3-specific MAbs isolated from animal F124 were tested for their capacities to neutralize a panel of Env-pseudoviruses. The plasma dilution and the MAb concentration inhibiting 50% of viral entry are shown as neutralization ID50 and IC50 values respectively. The ID50 values are color-coded where red indicates (>5000), orange (500–5000), yellow (50–500), and white (<50) whereas the IC50 values in red indicates (<0.1 μg/ml), dark orange (0.1–1 μg/ml), light orange (1–10 μg/ml), yellow (10–40 μg/ml) and white (>40 μg/ml).
Figure 5
Figure 5. Illustration of the epitope region targeted by the vaccine-elicited V3-directed MAbs
(A) Amino acid sequences of the full-length YU2 V3 peptide (indicated in bold letters) and twelve 15-mer peptides overlapping by 11, where each peptide is indicated by a number and a color (B) Binding curves of the isolated V3-directed MAbs to the overlapping peptides. (C) A surface rendered crystal structures of two HIV-1 Envs - CD4-complexed JRFL gp120 core (PDB ID - 2B4C) and the cleaved, soluble BG505.SOSIP gp120 in native state (PDB ID – 4NCO) (gray) generated with Chimera (66) indicate structural rearrangement of the V3 loop (blue + red) in the post-CD4 binding (JRFL gp120 core) and pre-CD4 triggered (BG505.SOSIP) conformations. The peptide region to which the V3-specific MAbs bind is shown in red while the rest of the V3 region is shown in the blue.
Figure 6
Figure 6. Comparison of MAb binding and neutralizing properties
(A) Upper panels: ELISA binding curves of the vaccine-induced CD4bs- and V3-directed MAbs to monomeric gp120 from DJ263.8, MW965.26, SS1196.01, and JRFL are shown. Lower panels: A summary of gp120 binding activity and virus neutralizing activity for DJ263.8, MW965.26, SS1196.01, and JRFL are shown. Binding activity is indicated as OD values at Ab concentration, 4 μg/ml, where red indicates High (OD >1), light orange indicates Medium (OD 1–0.5), and white indicates No/Low (OD <0.5) binding. Virus neutralizing activity is indicated as IC50 values and is color-coded as described in Figure 4. (B) Comparison of neutralizing activity for SS1196.01 and SS1196.01Δ301 and for JRFL and JRFLΔ301.
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References

    1. Wibmer CK, Bhiman JN, Gray ES, Tumba N, Abdool Karim SS, Williamson C, Morris L, Moore PL. Viral escape from HIV-1 neutralizing antibodies drives increased plasma neutralization breadth through sequential recognition of multiple epitopes and immunotypes. PLoS Pathog. 2013;9:e1003738. - PMC - PubMed
    1. Moore PL, Gray ES, Wibmer CK, Bhiman JN, Nonyane M, Sheward DJ, Hermanus T, Bajimaya S, Tumba NL, Abrahams MR, Lambson BE, Ranchobe N, Ping L, Ngandu N, Abdool Karim Q, Abdool Karim SS, Swanstrom RI, Seaman MS, Williamson C, Morris L. Evolution of an HIV glycan-dependent broadly neutralizing antibody epitope through immune escape. Nat Med. 2012;18:1688–1692. - PMC - PubMed
    1. Liao HX, Lynch R, Zhou T, Gao F, Alam SM, Boyd SD, Fire AZ, Roskin KM, Schramm CA, Zhang Z, Zhu J, Shapiro L, Mullikin JC, Gnanakaran S, Hraber P, Wiehe K, Kelsoe G, Yang G, Xia SM, Montefiori DC, Parks R, Lloyd KE, Scearce RM, Soderberg KA, Cohen M, Kamanga G, Louder MK, Tran LM, Chen Y, Cai F, Chen S, Moquin S, Du X, Joyce MG, Srivatsan S, Zhang B, Zheng A, Shaw GM, Hahn BH, Kepler TB, Korber BT, Kwong PD, Mascola JR, Haynes BF. Co-evolution of a broadly neutralizing HIV-1 antibody and founder virus. Nature. 2013;496:469–476. - PMC - PubMed
    1. Doria-Rose NA, Schramm CA, Gorman J, Moore PL, Bhiman JN, DeKosky BJ, Ernandes MJ, Georgiev IS, Kim HJ, Pancera M, Staupe RP, Altae-Tran HR, Bailer RT, Crooks ET, Cupo A, Druz A, Garrett NJ, Hoi KH, Kong R, Louder MK, Longo NS, McKee K, Nonyane M, O’Dell S, Roark RS, Rudicell RS, Schmidt SD, Sheward DJ, Soto C, Wibmer CK, Yang Y, Zhang Z, Mullikin JC, Binley JM, Sanders RW, Wilson IA, Moore JP, Ward AB, Georgiou G, Williamson C, Abdool Karim SS, Morris L, Kwong PD, Shapiro L, Mascola JR. Developmental pathway for potent V1V2-directed HIV-neutralizing antibodies. Nature. 2014;509:55–62. - PMC - PubMed
    1. Gao F, Bonsignori M, Liao HX, Kumar A, Xia SM, Lu X, Cai F, Hwang KK, Song H, Zhou T, Lynch RM, Alam SM, Moody MA, Ferrari G, Berrong M, Kelsoe G, Shaw GM, Hahn BH, Montefiori DC, Kamanga G, Cohen MS, Hraber P, Kwong PD, Korber BT, Mascola JR, Kepler TB, Haynes BF. Cooperation of B cell lineages in induction of HIV-1-broadly neutralizing antibodies. Cell. 2014;158:481–491. - PMC - PubMed

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