Comparison of herpes simplex virus replication in tlymphocyte and epithellal cell

Abstract

HSV can cause infection at various parts of the human body such as skin, mouth, eyes, genital area and brain. HSV usually causes localized infection at skin and mucous tissue which are known to be their tissue tropism. It has been reported that HSV can cause systemic infection in the immunocompromised host. Several studies of HSV replication in many cell types including peripheral blood mononuclear cells especially T lymphocyte have been reported. However, until now the mechanism of HSV replication in T lymphocyte has not been defined. In this study, the comparison of the ability of HSV-1 and HSV-2 replication in Vero, HEp-2, Jurkat and PHA-activated Jurkat cells at 12, 24, 48 hours post infection (h.p.i.) was done. The viral antigen detection by Indirect immonufluorescenc assay was determined at two, four, six, nine and 24 h.p.i. by using polyclonal anti-HSV-1 or HSV-2, anti-ICP0, anti-ICP22 and anti-ICP47 antibodies. The number of infected cells at 24 h.p.i. were detected by Flow cytometry. Moreover, the adsorption ability of virus to each cell type and HveA mRNA expression in Jurkat and PHA-activated Jurkat cells by RT-PCR were studies. The results showed that HSV-1 and HSV-2 can replicate in Jurkat cell, human leukemia T cell line. Although the yield production in T lymphocyte was very low when it was compared to the other epithelial cells, Vero and HEP-2 cells, the HSV-1 yield production could be enhanced after PHA activation while yield production of HSV-2 was not increased. The specific HSV proteins synthesis in Jurkat cells was delayed compared to that in HEp-2 cells. The kinetics study of IE proteins expression demonstrated that the expression of ICP22 in both HSV-1 and HSV-2 infected Jurkat and PHA-activated Jurkat cells and ICP0 in HSV-2 infected PHA-activated Jurkat cells were delayed 2 and 4 hours, compared to those HEp-2 cells. Detection of infected cells by using Flow cytometry showed that the number of HSV-1 infected cells between HEp-2 and Jurkat cells were different (71.58% vs 22.71%) and increased in PHA-activated Jurkat cells (40.07%). In the case of HSV-2 infected cells, the number of positive cells in HEp-2 and Jurkat cells were 57.01% and 47.15% whereas in PHA-activated Jurkat cells, the number of infected cell was declined (29.31%). The results from the adsorption experiment demonstrated that HEp-2 and PHA-activated Jurkat cells did absorb HSV-1 better than HSV-2 (93% vs 64% and 97% vs 47%) but the ability of Jurkat cells to adsorb HSV-1 was equal to HSV-2 (89%). Moreover, it was found that activation of Jurkat cells by PHA increased HSV receptor (HveA) mRNA expression in these cells. This study revealed that the replication of HSV in HEp-2 and T lymphocyte are different. The duration of successive steps in the replication cycle depends upon the types of cells, the virus strain and the multiplicity of infection. Increasing of HveA receptor in PHA-activated Jurkat cells could be demonstrated. This might be a reason for increasing HSV-1 production occurred after PHA activation.