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目的 探讨MHCⅡ类分子反式激活因子 (CⅡTA)突变体抑制HLAⅡ类分子表达的可能性。方法 构建不含起始密码子的pcDNA3mCⅡTA2、含起始密码子的pcDNA3mCⅡTA3和含起始密码子及NLS(nuclearlocalizationsignal)的pcDNA3mCⅡTA4突变体。用脂质体转染法将上述 3种突变体及pcDNA3空载体转入HeLa细胞和Raji细胞。用流式细胞术和RT PCR法观察它们对HeLa/Raji细胞HLA DR/DQ分子的诱导性和组成性表达的影响。结果 在细胞和基因水平证实pcDNA3mCⅡTA3和pcDNA3mCⅡTA4对HeLa/Raji细胞HLA DR/DQ的表达起明显抑制作用 ,而pcDNA3mCⅡTA2和pcDNA3空载体无此作用。结论 成功构建的pcDNA3mCⅡTA3和pcDNA3mCⅡTA4能抑制HLAⅡ类分子的表达
Objective To investigate the possibility of MHC class Ⅱ transactivator (CⅡTA) mutants inhibiting the expression of HLA class II molecules. Methods pcDNA3mCⅡTA2 without start codon, pcDNA3mCⅡTA3 with start codon and pcDNA3mCⅡTA4 mutant with start codon and NLS (nucleus localization signals) were constructed. The three kinds of mutants and pcDNA3 empty vector were transfected into HeLa cells and Raji cells by lipofectamine. Their effects on the induction and constitutive expression of HLA DR / DQ molecules in HeLa / Raji cells were observed by flow cytometry and RT PCR. Results The results showed that pcDNA3mCⅡTA3 and pcDNA3mCⅡTA4 significantly inhibited HLA-DR / DQ expression in HeLa / Raji cells at the cellular and genetic level, whereas pcDNA3mCⅡTA2 and pcDNA3 null vectors did not. Conclusion The constructed pcDNA3mCⅡTA3 and pcDNA3mCⅡTA4 can inhibit the expression of HLA class II molecules