ABSTRACT
Objective To study the function of 4 different haplotypes cDNA which are constructed by two non-homonymy single nueleotide polymorphism (SNP) sites C19170G (Leu45Val) and C30799G (Ala500Gly) in the coding region of human CⅡTA gene. Methods HeLa cells were transfeeted with eu-karyotic expression vectors containing four different haplotypes cDNA. C Ⅱ TA mRNA and HLA classⅡanti-gen (HLA-DR, DP, DQ) were respectively detected by RT-PCR and indirect cell immunofluoreseence tech-nique in the untransfected and transfeeted with four eukaryotic expression vectors and empty vectors HeLa cells. The quantity of HLA classⅡ antigen were analyzed by flow eytometry. Results No expression of CⅡTA mRNA and HLA class Ⅱ antigen were observed on original HeLa cells and empty vector transfected cells. CⅡTA mRNA expression was emerged, and the expression of HLA class Ⅱ antigen were observed in the HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA. And there were not significantly different with the levels of HLA class Ⅱ antigen expression among HeLa cells transfected with eukaryotic expression vectors containing four different haplotypes cDNA ( P > 0.05 ). Con-dusion The SNP of Chinese at the sites C19170G(Leu45Val) and C30799G(Ala500Gly) in the coding site of C Ⅱ TA gene did not influence capability of CⅡTA trans-aetivating HLA class Ⅱgene expression.
ABSTRACT
Objective To investigate the role of MHC class Ⅱ transactivator(CⅡTA) in upregulating the expression of HLA class Ⅱ antigen in HepG2 cells.Methods Eukaryotic expression vector EBS-NPL-CⅡTA containing CⅡTA cDNA or the empty vector EBS-NPL was transfected into HepG2 cells respectively.CⅡTA mRNA and HLA class Ⅱ antigen(HLA-DR,DP,DQ) were respectively detecued by RT-PCR,indirect cell immunofluorescence technique and flow cytometry in original HepG2 cells,HepG2 cells transfected with EBS-NPL-CⅡTA or the empty vector EBS-NPL.Results The expression of CⅡTA mRNA and HLA class Ⅱ antigen(HLA-DR,DP,DQ) were not observed in original HepG2 cells and HepG2 cells transfected with empty vector,but in the HepG2 cells transfected with EBS-NPL-CⅡTA.Conclusion CⅡTA is a switching factor of mastering the expression of HLA class Ⅱ antigen in HepG2 cells.The lack of CⅡTA expression in HepG2 cells contributes to no expression of HLAⅡ antigen.
ABSTRACT
Objective To investigate the role of HLA class II antigen of human monocytes in pathogenesis of herpes simplex virus-2 (HSV-2) infection. Methods Human monocytes were infected by HSV-2 (333 Strain). The cultured cells were collected at the first, third, 5th, and 7th days of post-infection. The expression of HLA class II antigen of the infected monocytes was measured by alkaline phosphotase anti-alkaline phosphotase method (APAAP). Results The expression of HLA class II antigen on the monocytes significantly decreased at the first day of post-infection,and then gradually increased and reached to the normal control level at the 7th day of post-infection. Conclusion HLA class Ⅱ antigen expression of monocytes was inhibited by HSV-2 infection at early stage, which may be one of means of virus escaping host immunity. The expression of HLA class Ⅱ antigen may play an important role in HSV-2 pathogenicity and immunity.
