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. 2017 Sep 12;91(19):e00411-17.
doi: 10.1128/JVI.00411-17. Print 2017 Oct 1.

Monoclonal Antibodies, Derived From Humans Vaccinated With the RV144 HIV Vaccine Containing the HVEM Binding Domain of Herpes Simplex Virus (HSV) Glycoprotein D, Neutralize HSV Infection, Mediate Antibody-Dependent Cellular Cytotoxicity, and Protect Mice From Ocular Challenge With HSV-1

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Monoclonal Antibodies, Derived From Humans Vaccinated With the RV144 HIV Vaccine Containing the HVEM Binding Domain of Herpes Simplex Virus (HSV) Glycoprotein D, Neutralize HSV Infection, Mediate Antibody-Dependent Cellular Cytotoxicity, and Protect Mice From Ocular Challenge With HSV-1

Kening Wang et al. J Virol. .
Free PMC article

Abstract

The RV144 HIV vaccine trial included a recombinant HIV glycoprotein 120 (gp120) construct fused to a small portion of herpes simplex virus 1 (HSV-1) glycoprotein D (gD) so that the first 40 amino acids of gp120 were replaced by the signal sequence and the first 27 amino acids of the mature form of gD. This region of gD contains most of the binding site for HVEM, an HSV receptor important for virus infection of epithelial cells and lymphocytes. RV144 induced antibodies to HIV that were partially protective against infection, as well as antibodies to HSV. We derived monoclonal antibodies (MAbs) from peripheral blood B cells of recipients of the RV144 HIV vaccine and showed that these antibodies neutralized HSV-1 infection in cells expressing HVEM, but not the other major virus receptor, nectin-1. The MAbs mediated antibody-dependent cellular cytotoxicity (ADCC), and mice that received the MAbs and were then challenged by corneal inoculation with HSV-1 had reduced eye disease, shedding, and latent infection. To our knowledge, this is the first description of MAbs derived from human recipients of a vaccine that specifically target the HVEM binding site of gD. In summary, we found that monoclonal antibodies derived from humans vaccinated with the HVEM binding domain of HSV-1 gD (i) neutralized HSV-1 infection in a cell receptor-specific manner, (ii) mediated ADCC, and (iii) reduced ocular disease in virus-infected mice.IMPORTANCE Herpes simplex virus 1 (HSV-1) causes cold sores and neonatal herpes and is a leading cause of blindness. Despite many trials, no HSV vaccine has been approved. Nectin-1 and HVEM are the two major cellular receptors for HSV. These receptors are expressed at different levels in various tissues, and the role of each receptor in HSV pathogenesis is not well understood. We derived human monoclonal antibodies from persons who received the HIV RV144 vaccine that contained the HVEM binding domain of HSV-1 gD fused to HIV gp120. These antibodies were able to specifically neutralize HSV-1 infection in vitro via HVEM. Furthermore, we showed for the first time that HVEM-specific HSV-1 neutralizing antibodies protect mice from HSV-1 eye disease, indicating the critical role of HVEM in HSV-1 ocular infection.

Keywords: ADCC; HIV vaccine; HVEM; glycoprotein D; herpes simplex virus; monoclonal antibody; ocular infection.

Figures

FIG 1
FIG 1
Structures of AIDSVAX B/E, HSV-1 gD, and amino acids of HSV-1 gD critical for binding of monoclonal antibodies CH42 and CH43. (A) AIDSVAX B/E has the first 27 amino acids of the mature form of HSV-1 gD fused to gp120 without the first 40 amino acids of gp120. (B) HSV-1 gD has a signal sequence and HVEM and nectin-1 binding domains. (C) HSV-1 gD amino acids recognized by MAb CH42 and CH43 were determined by binding the MAbs to peptides containing alanine substitution mutations in HSV gD (the amino acid sequence of HSV-1 gD TAG peptide is shown on the x axis), followed by ELISA. The optical density (OD) at 405 nm for binding of MAbs to gD TAG is shown on the y axes. Amino acid sequence alignment of HSV-1 gD TAG to HSV-1 and HSV-2 gD in five commonly used virus strains is shown below; the dashes represent amino acids identical to those in HSV gD TAG. The numbers indicate amino acid positions in the mature form of gD.
FIG 2
FIG 2
Neutralizing activity of MAbs CH42 and CH43 in B78H1-A10 mouse cells that express human HVEM (hHVEM) (but not nectin-1), B78H1-C10 mouse cells that express human nectin-1 (but not HVEM), Vero cells that express endogenous simian HVEM and nectin-1, and primary HFF. Stock concentrations of MAbs and human IVIG were 1 mg/ml and 10 mg/ml, respectively. Fifty percent HSV-neutralizing titers plotted as the reciprocal of the dilution are shown for HSV-1 McKrae and HSV-2 333 in the presence of MAbs CH42, CH43, and A32 (human IgG1 isotype control) or IVIG, a positive control. Similar results were obtained in two experiments, and a representative result is shown.
FIG 3
FIG 3
ADCC NK cell activation mediated by MAbs CH42 and CH43 incubated with NK-92-CD16 cells (which stably express GFP) and either HSV-1-infected SK-N-SH cells or HSV-1 gD-coated 96-well plates. (A) NK-92-CD16 cells (1 × 105) were incubated with HSV-1-infected or uninfected SK-N-SH cells in the presence or absence of MAbs at 5 μg/ml (CH42, CH43, and A32 [a human IgG1 isotype control] or no antibody [No AB]) for 5 h. Cell surface CD107a staining was then performed as a marker of NK cell activation. The percentages of NK-92-CD16 cells (GFP positive) that expressed CD107a on their surfaces are shown on the y axis. (B) Dose-response curve of ADCC using serial dilutions of MAb CH42, CH43, or isotype control A32 with HSV-1-infected or uninfected SK-N-SH cells. (C) Dose-response curve of NK-92-CD16 cell activation using serial dilutions of MAb CH42, CH43, or isotype control A32 in HSV-1 gD-coated 96-well plates. The diluted antibodies were added to the gD-coated wells and incubated for 15 min, and 5 × 105 NK-92-CD16 cells/well were added and incubated for 5 h. After washing with PBS, the cells were stained with 4 μg/ml APC-Cy7-conjugated anti-CD107a antibody and fixed with 10% paraformaldehyde. Activated degranulating NK cells (GFP+ CD107a+) were detected by flow cytometry. The percentages of CD107a+ NK-92-CD16 cells are shown. The error bars indicate standard deviations.
FIG 4
FIG 4
Survival of mice treated with MAb CH42, CH43, or isotype control (A32) or human IVIG and infected with HSV-1 by corneal scarification. (A) Eight-week-old BALB/c mice (5 animals/group) were injected i.p. with MAb CH42, CH43, or A32 or IVIG, and 2 days later, both eyes were infected with HSV-1 McKrae after corneal scarification. Eye swabs were obtained during the first 3 days after infection, disease scores were measured 4 to 9 days after infection, and trigeminal ganglia (TG) were harvested 4 weeks after infection. (B) Kaplan-Meier survival curves for days 5 to 21 after infection for mice treated with CH42 (left) or CH43 (right).
FIG 5
FIG 5
Eye scores of mice treated with MAb CH42, CH43, or isotype control (A32) and infected with HSV-1 by corneal scarification. The mice were treated and infected as described in the legend to Fig. 4. (A) Daily group mean eye disease scores in mice (5 animals/group) that received 150 μg of MAbs. The error bars indicate standard deviations. (B) Correlation of the dose of MAb (0, 6, 30, 150, and 450 μg/mouse for CH42 and 0, 6, 30, and 150 μg/mouse for CH43) with mean eye disease scores during the 6 days after infection. Each point represents the results for one mouse.
FIG 6
FIG 6
HSV-1 shedding from eyes of mice treated with MAb CH42, CH43, or isotype control (A32) and infected with HSV-1 by corneal scarification. The mice were treated and infected as described in the legend to Fig. 4. (A) Eye swabs were obtained for the first 3 days after infection, HSV-1 titers were measured in Vero cells, and daily group mean titers were determined. The error bars indicate standard deviations. (B) Correlation of the dose of MAb (0, 6, 30, 150, and 450 μg/mouse for CH42 and 0, 6, 30, and 150 μg/mouse for CH43) with the mean HSV-1 titer in eye swabs from mice during the first 3 days after infection. Each point represents the results for one mouse.
FIG 7
FIG 7
HSV-1 latent DNA loads in trigeminal ganglia of mice treated with MAb CH42, CH43, or isotype control (A32) or human IVIG and infected with HSV-1 by corneal scarification. The mice were treated and infected as described in the legend to Fig. 4; mice not infected with HSV-1 (No inf) served as a negative control. Four weeks after infection, the mice were euthanized, trigeminal ganglia were harvested, DNA was isolated, and HSV-1 DNA copies per microgram of DNA were determined by real-time PCR using primers and a probe specific for the HSV-1 gG gene. The error bars indicate standard deviations.

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