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. 2003 Mar;77(5):3307-11.
doi: 10.1128/jvi.77.5.3307-3311.2003.

Entry of herpes simplex virus type 1 into primary sensory neurons in vitro is mediated by Nectin-1/HveC

Sarah M Richart  1 , Scott A Simpson, Claude Krummenacher, J Charles Whitbeck, Lewis I Pizer, Gary H Cohen, Roselyn J Eisenberg, Christine L Wilcox
Affiliations

Entry of herpes simplex virus type 1 into primary sensory neurons in vitro is mediated by Nectin-1/HveC

Sarah M Richart et al. J Virol. 2003 Mar.

Abstract

Primary cultures of rat and mouse sensory neurons were used to study the entry of herpes simplex virus type 1 (HSV-1). Soluble, truncated nectin-1 but not HveA prevented viral entry. Antibodies against nectin-1 also blocked infection of rat neurons. These results indicate that nectin-1 is the primary receptor for HSV-1 infection of sensory neurons.

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Figures

FIG. 1.
FIG. 1.
Effects of soluble recombinant HveA or nectin-1 on viral entry into primary rat neurons. The panels shown depict representative neuronal cultures photographed using phase contrast (A, C, and E) or epifluorescence (B, D, and F) 24 h after infection. (A and B) HSVEGFP4 alone; (C and D) HSVEGFP4 with soluble HveA [HveA(200t)]; (E and F) HSVEGFP4 preincubated with soluble nectin-1 [HveC(346t)]. Images are shown at ×600 magnification.
FIG. 2.
FIG. 2.
Effect of soluble recombinant HveA or nectin-1 on viral entry into primary rat neurons, mouse neurons, and primary rat fibroblasts. (A) Virus was incubated with soluble receptor before being added to cells. An MOI of 100 PFU per cell was used for rat and mouse sensory neurons. An MOI of 10 PFU per cell was used for rat fibroblasts. Each bar represents the average percentage of GFP-positive cells per culture. Error bars indicate standard error of the mean (n = 4). (B) HSVEGFP4 virus was incubated with the indicated concentrations of recombinant HveA(200t) (open squares) or nectin-1(346t) (closed diamonds) prior to infection of rat sensory neurons. Each point represents the average percentage of GFP-positive cells per culture.
FIG. 3.
FIG. 3.
The effects of pretreatment of cells with antibodies to HveA or nectin-1 on viral entry. (A) Cells were left untreated (HSV alone) or were pretreated with rabbit antisera against HveA (R140) or nectin-1 (R166) prior to infection with HSV-1. (B) Rat or mouse neurons were left untreated (HSV alone) or were pretreated with monoclonal anti-nectin-1 antibodies (CK41) prior to infection with HSVEGFP4.
FIG. 4.
FIG. 4.
Detection of HveA and nectin-1 proteins by immunoblotting. For immunoblot analysis, 30 μg of protein from whole-cell lysates and 2 ng of soluble recombinant HveA(200t) or 10 ng of nectin-1(346t) were separated by electrophoresis and transferred to membranes for immunodetection. HveA was detected using rabbit anti-HveA serum (R140) (A), and nectin-1 was detected using rabbit anti-nectin-1 serum (R166) (B).
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