Co-culture For Reactivation Of Cerebral Cortex And Lung From Aged Mouse With Human Cytomegalovirus Latency In Vitro

Objective we selected the cerebral cortex and lung of aged mouse with HCMV latency. Then, we established a co-cultivation techique of cell from cerebral cortex and lung together with HF in vitro to observe HCMV reactivation from latency. We aim at providing a new idea for the research on HCMV latency, reactivation and its mechanism in the study.Methods On the basis of mice brain and lung with HCMV latency, we study the HCMV reactivation from latency by co-cultivation technique. The cerebral cortex and lung tissues from 6 young mice with congenital HCMV latency or from DMEM control were minced into small pieces with sharp scalpels and grind until cell concentration is 1 ×10~6/ ml, allowed to adhere to flask after which 1 × 10~6 human embro fibroblasts were added. At the same time, we established HCMV infection in HF as positive control and HF cell as negative control. The cells were collected in the 1,3,7,14,28 and 35days after co-culture to detection as follows: (1) Observation of CPE in HF . (2) HCMV IE , UL83 gene and transcripts were analyzed by polymerase chain reaction (PCR) and RT-PCR. (3) The fixed cell slices with acetone in co-cultivation experiment were to detect the expression of HCMV IE, pp65 antigen by indirect immunofluorescent assay. (4) Observed the ultrastructural features of HF and the herpers virus-like particles through electron microscope.Results In the co-cultivation experiment of mice brain and lung with HCMV latency, the result as follows: (1) we observed the CPE in HF at the 7d, as well as the HCMV positive control, which show that HCMV recurrence from latency. The rate of...