Increasing evidence suggests that human cytomegalovirus (HCMV) contributes to the development of both restenosis and atherosclerosis. We have worked on developing a DNA vaccination method to prevent CMV infection-caused increased neointimal response to balloon injury. Sixty young (3-week-old) rats were ordered; 30 rats were injected with RCMV (10 to 6th power TCID50) I.P.; 30 control rats were injected with conditioned media (same batch of RCMV contained media, but has been filtered through a 0.1 micron filter device to clear out virus, and the conditioned media has been tested for infectious virus for 3 weeks on rat fibroblast cell line, and showed no CPE). Ten latently infected rats will be vaccinated one month later with pUC18-IE1/IE2 125 micrograms each by using a Jet Gun; 10 latently infected rats will be vaccinated with pUC 18 only; 10 rats will be mock-vaccinated with PBS (DNA was diluted with PBS). The same strategy will be applied to 30 noninfected rats: 10 with IE1/IE2, 10 with pUC18 only, 10 with PBS. The vaccination procedures will be repeated twice at 3 weeks duration. Two weeks following the last injection, all rats will be operated on for left carotid balloon injury. Six weeks following surgery, all rats will be sacrificed for neointimal/medium ratio analysis. Upon sacrificing, blood will be collected from all rats for cytokine testing, and salivary gland will be collected for infectious virus testing. If no infectious virus are found, salivary glad will be tested for RCMV DNA sequence.
