Effects of H2-B1 gene transfection on the immunogenicity of mouse vascular endothelial cells

Abstract

Objective The pEGFP-N1 -H2B1 plasmid vector was transfected to mouse vascular endothelial cells (VECs) , the immunogenicity effect of VECs was investigated, and the possible mechanism of immune tolerance after heart transplantation mirroring matemal-fetal immune tolerance was speculated. Methods The mouse VECs were transfected with pEGFP-Nl-H2Bl plasmid vector. The efficiency of transfection was tested by flow cytometry, the expression of H2-B1 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) , and the cytolysis of peripheral blood mononuclear cells (PBMC)was investigated at 24, 48 and 72 h, respectively. Results The expression levels of H2-B1 in 0. 5 mg/L and 1.0 mg/L pEGFP-N1-H2B1 plasmid vector groups were higher than in control group (P <0.05, P <0. 01, respectively). After 48 h, the cytolysis of PBMC to VECs in H2-B1-transfected groups was significantly attenuated as compared with control group ( P <0.05). Conclusion The H2-B1 gene may have a possible immunoregulative effect. Transfection of pEGFP-N1-H2B1 plasmid vector to mouse VECs can reduce the immunogenicity of VECs, attenuate the cytotoxicity of PBMC, and induce immune tolerance. Key words: H2-B1 gene; Immunogenicity; Vascular endothelial cell