[Establishment of a cell model system of herpes simplex virus type II latent infection and reactivation in SH-SY5Y cells].

OBJECTIVE

To establish an experimental culture system of herpes simplex virus type II (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 micromoL/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 degrees C, 42 degrees C, 43 degrees C, 44 degrees C and 45 degrees C, 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 micromoL/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 micromoL/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 degrees C, 1.5 h and 75 micromoL/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.