Establishment of a cell model system of herpes simplex virus typeⅡ latent infection and reactivation in SH-SY5Y cells

Abstract:

Abstract: [Objective] To establish an experimental culture system of herpes simplex virus typeⅡ(HSV-2) latent infection and reactivation in SH-SY5Y cells. [Methods] Changes of biological character were observed after 20, 40, 60, 80,100, 120 and 140 μmoL/L ACV were added into cell cultures, and also the morphological observation was detected with phase-contrast microscopy after HSV-2 was inoculated into SH-SY5Y cells using MOI of 0.1, 1, 10 and 100. The optimum condition of time and temperature was approached using temperature of 41℃, 42℃, 43℃, 44℃ and 45℃, and time of 0.5 h, 1.0 h, 1.5 h, 2.0 h and 2.5 h to induce HSV-2 reactivation. The optimum concentration of Forskolin was also decided using 25, 50, 75, 100 and 125 μmoL/L to activate the virus from latency. PCR was used to authenticate HSV-2 latent infection and reactivation. The morphological changes were observed when the virus was reactivated from latency. [Results] The optimum concentration of ACV was 60 μmoL/L to establish latency in SH-SY5Y cells. Suitable infective dose of HSV-2 was 1-10 MOI to construct latency and reactivation in SH-SY5Y cells. The time of virus latency in SH-SY5Y cells could reach up to 14 d. Heat stress of 43℃, 1.5 h and 75 μmoL/L Forskolin were the optimum condition to induce virus reactivation. The morphologic changes in SH-SY5Y cells recurred by HSV-2: Cytopathic effects were more and more obvious with time lasting from 24 h to 72 h after reactivation from latency. PCR and results of electrophoresis proved the cell model of latent infection and reactivation was set up successfully. [Conclusions]The cell model system of HSV-2 latent infection and reactivation in SH-SY5Y cells was established. The research provided usefulness for study on HSV-2 of latency, reactivation and pathogenic mechanism.