Genotypic Analysis of Human Cytomegalovirus Resistance to Ganciclovir Using Aqueous Humor of Immunocompetent Patients with Cytomegalovirus Anterior Uveitis

学位記番号：医博甲168

CMV infection of trabecular meshwork cells

Purpose: Human cytomegalovirus (HCMV) infections can cause endotheliitis which is associated with an elevation of the intraocular pressure (IOP). However, the mechanism of the IOP elevation has not been determined. The purpose of this study was to determine whether HCMV strains which are capable of infecting corneal endothelial cells can also replicate, induce anti-viral responses, and can reorganize the actin cytoskeleton in trabecular meshwork cells. Study design: Experimental study design Methods: Cultured primary human trabecular meshwork cells (HTMCs) were infected with the Towne or TB40/E strains of HCMV. TB40/E is trophic for vascular endothelial and corneal endothelial cells. Real-time PCR, western blot, and fluorescent immunostaining have been used to determine whether HCMV-infected HTMCs will support the expression of viral mRNA and protein, allow viral replication, and elicit anti-viral host responses. We also determined whether lytic replication was present after an HCMV infection. Results: HCMV infection led to the expression of viral mRNA and proteins of IE1, glycoprotein B(gB), and pp65. TB40/E infection induced interferon-β, a sign of host anti-viral immune response and MCP-1. Together with the induction of the regulators of actin cytoskeleton, myosin phosphatase Rho interacting protein (MPRIP) and monocyte chemotactic protein-1 (MCP-1), TB40/E induced a high level of expression of viral proteins, including IE1, gB, and pp65 as well as actin stress fiber formation, and achieved pathogenically high viral titers. Conclusions: Human trabecular meshwork cells support the replication of endotheliotropic TB40/E strain of HCMV which indicates that this strain may have high virulence for trabecular meshwork