Human Cytomegalovirus Infection Of Mouse's Cerebral Cortex Cells

Objective: human cytomegalovirus can infect neonatal mouse's cerebral cortex cells in vitro.Virus which were latency in the mouse's cerebral cortex cells can be reactivated in vivo.Methods:In vitro:Make sure of dose of virus to infect neonatal mouse's cerebral cortex cells by TCID₅₀ .Take HCMV AD169 and three strain HCMV wilds as objects,inactive virus which were deal with high temperature be used control. Virus were put into Balb/c neonatal mouse's cerebral cortex cells and HF cells.Observing HCMV specific CPE by microscope and detecting HCMV specific early gene IE and UL83 by PCR RT-PCR.In the 1 2 3 4 5, we collected cells in the culture of neonatal mouse's cerebral cortex cells with virus. Detecting HCMV special early and later gene and UL83 by PCR RT-PCR indirect immunofluorescent; determining virus titer of cultured supernatantof neonatal mouse's cerebral cortex nerve cells;observing HCMV particles in nerve cells by Electron microscopy. In vivo:Choiced randomly five mice in the modle of human cytomegalovirus congenital latent infection There were three groups: mouse of human cytomegalovirus congenital latent infection,mouse of human cytomegalovirus congenital latent infection be reactivated ,mouse of HF cells. Put the mouse's cerebral cortex cells with HF cells by co-culture and detected HCMV specific early gene IE and UL83 by PCR RT-PCR and HCMV pp65 by indirect immunofluorescent. Observing HCMV particles in nerve cells by Electron microscopy.Result:In the vitro:In the second day,we can find the nerve cells happened shape changes in the culture of neonatal mouse's cerebral cortex nerve cells which were...