TRPV4 channel is involved in HSV-2 infection in human vaginal epithelial cells through triggering Ca2+ oscillation

Herpes simplex virus (HSV) Infection induces a rapid and transient increase in intracellular calcium concentration ([CA²⁺]_i), which plays a critical role in facilitating viral entry. T-type calcium channel blockers and EGTA, a chelate of extracellular CA²⁺, suppress HSV-2 Infection. But the cellular mechanisms mediating HSV infection-activated CA²⁺ signaling have not been completely defined. In this study we investigated whether the TRPV4 channel was involved in HSV-2 Infection in human vaginal epithelial cells. We showed that the TRPV4 channel was expressed in human vaginal epithelial cells (VK2/E6E7). Using distinct pharmacological tools, we demonstrated that activation of the TRPV4 channel induced CA²⁺ influx, and the TRPV4 channel worked as a CA²⁺-permeable channel in VK2/E6E7 cells. We detected a direct interaction between the TRPV4 channel protein and HSV-2 glycoprotein D in the plasma membrane of VK2/E6E7 cells and the vaginal tissues of HSV-2-infected mice as well as in phallic biopsies from genital herpes patients. Pretreatment with specific TRPV4 channel inhibitors, GSK2193874 (1-4 μM) and HC067047 (100 nM), or gene silence of the TRPV4 channel not only suppressed HSV-2 infectivity but also reduced HSV-2-induced cytokine and chemokine generation in VK2/E6E7 cells by blocking CA²⁺ influx through TRPV4 channel. These results reveal that the TRPV4 channel works as a CA²⁺-permeable channel to facilitate HSV-2 Infection in host epithelial cells and suggest that the design and development of novel TRPV4 channel inhibitors may help to treat HSV-2 infections.