The ability of intima smooth muscle cells (SMCs) and blood leukocytes to control the development of Herpes simplex type 1 virus (HSV-1) infection in human vascular endothelial cells (ECs) was studied under a co-culture conditions. It was shown that cultured ECs supported HSV-1 reproduction followed by cytopathogenic effect (CPE) and accumulation of infectious virus in the cell culture medium. At the multiplicity of infection (MOI) ranging from 1 to 0.1 TCID(50), co-culture of ECs with SMCs growing in TransWell inserts significantly delayed the development of signs of virus reproduction. Similar effect was found, when after infection ECs were cultured in SMC-conditioned medium. At a MOI 0.001, SMC-conditioned media completely abrogated both CPE and virus accumulation, and leaded to the establishment of latent infection: this infection could persist in a subculture of infected cells and be reactivated after substitution of conditioned medium by standard EC growing medium. The decrease of virus infection in ECs was also observed, when the cells were cultured in medium conditioned by human peripheral blood leukocytes, containing numerous cytokines (but not IFN). At a MOI 0.001 we have found neither visual signs of infection, nor virus reproduction; the PCR analyses were also negative. The results obtained suggest that resident cellular components of the vascular intima (SMCs) and leukocytes may be involved in the mechanism of endothelium-virus interaction and modify the pathogenesis of HSV-1 disease.