CAJ | 학술논문

目的转染pEGFP-N1-H2B1质粒载体至小鼠血管内皮细胞(VEC),观察导入H2-B1基因后VEC免疫原性的变化,探讨借鉴母胎免疫耐受模型诱导心脏移植术后免疫耐受的机制.方法以未处理的VEC为对照,采用流式细胞仪和逆转录-聚合酶链反应(RT-PCR)技术分别于转染H2-B1基因后24、48、72 h检测质粒转染效率以及H2-B1基因的mRNA相对表达量;将同源小鼠外周血单个核细胞(PBMC)和VEC按100:1和10:1两种不同效靶比混合,观察PBMC对靶细胞杀伤活性的变化.结果 H2-B1基因组H2-B1基因在VEC中的mRNA表达量明显高于对照组(0.5mg/L组,P<0.05;1.0 mg/L组,P<0.01).H2-B1基因转染48 h后,当效靶比为100:1时,与对照组细胞毒性14.29%比较,0.5 mg/L基因组细胞毒性为4.28%,提示转染H2-B1基因组PBMC对VEC的毒性作用明显降低(P<0.05).结论 H2-B1基因可能具有免疫调节功能,可降低VEC的免疫原性,抑制PBMC对VEC的杀伤活性,诱导免疫耐受.
목적 전염pEGFP-N1-H2B1질립재체지소서혈관내피세포(VEC),관찰도입H2-B1기인후VEC면역원성적변화,탐토차감모태면역내수모형유도심장이식술후면역내수적궤제.방법 이미처리적VEC위대조,채용류식세포의화역전록-취합매련반응(RT-PCR)기술분별우전염H2-B1기인후24、48、72 h검측질립전염효솔이급H2-B1기인적mRNA상대표체량;장동원소서외주혈단개핵세포(PBMC)화VEC안100:1화10:1량충불동효파비혼합,관찰PBMC대파세포살상활성적변화.결과 H2-B1기인조H2-B1기인재VEC중적mRNA표체량명현고우대조조(0.5mg/L조,P<0.05;1.0 mg/L조,P<0.01).H2-B1기인전염48 h후,당효파비위100:1시,여대조조세포독성14.29%비교,0.5 mg/L기인조세포독성위4.28%,제시전염H2-B1기인조PBMC대VEC적독성작용명현강저(P<0.05).결론 H2-B1기인가능구유면역조절공능,가강저VEC적면역원성,억제PBMC대VEC적살상활성,유도면역내수.
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.