Eukaryotic Kxpression of the Major Antigenic Determinants Evoking Neutralizing Antibodies in Human Cytomegalovirus ( HCMV ) Isolated in Korea

ABSTRACT

Human cytomegalovirus (HCMV) isolated from Korean patients is different in the antigenic and genomic structure of gB from the laboratory-adapted strain. To dissect the reactivity to HCMV glycoprotein B (gB) domains, each domain gene of gB of HCMV SNUCH1, Korean isolate, was amplified from the extracted DNA of the virus-infected fibroblasts with the specific primers by polymerase chain reaction (PCR). Amplified DNA was cloned into pcDNA3. Immunofluorescent staining and western blot analysis revealed that the expressed gB in mammalian cells was immunoreactive and equivalent to the naturally expressed gB in virus-infected fibroblasts. The antigenic component reactive with monoclonal antibodies, MCMVA 57, 88, and 98 appeared at the D3 domain of gB molecule, and that with MCMVA 66 and 135 at the D2b domain. Antibody titer was measured with HCMV-infected fibroblasts and the domains of gB expressed in mammalian cells. There was no correlation between the antibody titer to the whole HCMV and neutralizing antibody titer, and between the antibody titer to whole HCMV and whole gB. It was more reasonable to use whole gB than whole HCMV in the comparison with the neutralizing antibody titer. D3 was representative domain in gB molecule in the anti-gB reactivity. Conclusively it is highly recommendable to use the representing isolates in Korea and its domains for the detection of antibody or the analysis of antigen in the aspect of immunological properties and molecular structures.