HVEM-LIGHT signaling promotes antibody-dependent neutrophil FcγR-mediated trogocytosis against herpes simplex virus infection
Matthew S. Gromisch, Masayuki Kuraoka, Carl F. Ware, Steven C. Almo, Betsy C. Herold
Matthew S. Gromisch, Masayuki Kuraoka, Carl F. Ware, Steven C. Almo, Betsy C. Herold
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Research Article Immunology Infectious disease Virology

HVEM-LIGHT signaling promotes antibody-dependent neutrophil FcγR-mediated trogocytosis against herpes simplex virus infection

Abstract

Studies with a candidate vaccine deleted in glycoprotein D (ΔgD-2) for herpes simplex virus (HSV) prevention uncovered a role for herpes virus entry mediator (HVEM) in mediating antibody-dependent cell-mediated killing (ADCK) of virally infected cells. Antibodies elicited by ΔgD-2 passively protect WT but not Fc γ receptor (FcγR) or HVEM knockout (KO) mice. The goals of this study were to identify which cells mediate ADCK and the role of HVEM signaling. Using HVEM ligand and conditional cell-type–specific HVEM-KO mice combined with in vitro mouse and human cytolytic assays, we demonstrate that ADCK of HSV-infected cells is mediated primarily by neutrophils and requires their expression of HVEM and its ligand, LIGHT. Cytolysis is not associated with granzyme and perforin production but occurs by a trogocytosis-like pathway. Pharmacological inhibition of myosin light-chain kinase (MLCK), which mediates trogocytosis, inhibits cytolysis. Similar results were obtained when human neutrophils were cocultured with HSV-infected cells opsonized with ADCK-containing human immune serum or with breast cancer cells treated with an anti-HER2 trogocytosis mediating antibody. Killing was significantly reduced when an MLCK inhibitor or blocking antibodies to CD16a, HVEM, or LIGHT were added. Together, these results define a mechanism of HVEM-enhanced FcγR-mediated neutrophil-dependent ADCK of targets cells.

Authors

Matthew S. Gromisch, Masayuki Kuraoka, Carl F. Ware, Steven C. Almo, Betsy C. Herold

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Figure 3

Neutrophils generate RNOS but not granzyme or perforin when cocultured with antibody-coated HSV-infected target cells.

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Neutrophils generate RNOS but not granzyme or perforin when cocultured w...
Immune cells isolated from bone marrow of WT mice were incubated for 4 hours with HSV-2–infected Vero cells (targets) that had been pretreated with control or ΔgD-2 immune serum. The percentage of immune cells staining positive for intracellular granzyme B (A) or perforin (B) or cell surface CD107a (C) after coculture with ΔgD-2 serum–opsonized targets minus control-opsonized targets (n = 6). (D) Representative flow plots of rhodamine 123 expression of indicated immune cell populations either with (red/orange) or without (black) DHR123 staining following incubation with ΔgD-2 serum–opsonized targets (n = 6). (E) Geometric mean fluorescent intensity (GeoMFI) of Rhodamine123 staining in represented immune cell populations isolated from total bone marrow cocultured with ΔgD-2 serum–treated HSV-2–infected Vero cells. Results were compared with B cells (FcγR negative) by 1-way ANOVA. (F) Representative flow plots of Rhodamine123 expression in neutrophils or macrophages/monocytes cocultured with HSV-infected Vero cells treated with either control (black) or ΔgD-2 (red) serum. (G) Change in fluorescent intensity of DHR123 staining in isolated neutrophils or macrophages/monocytes in the ΔgD-2 immune serum–treated HSV-2 infected Vero cells compared with control serum–treated targets (n = 5). The difference in RNOS production was compared by 1-way ANOVA with Holm-Šidák’s multiple comparisons test (***P < 0.001, ****P < 0.0001).