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目的探讨用纳米载体介导主要组织相容性复合物(majorhistocompatibilitycomplex,MHC)Ⅱ类抗原转录激活因子(MHCⅡtransactivator,CⅡTA)的反义RNA,抑制皮肤成纤维细胞表面MHCⅡ类抗原表达的可行性。方法将CⅡTA反义RNA(pDarⅡ质粒)稳定转染人类原代皮肤成纤维细胞(pDarⅡ-D),流式细胞仪检测pDarⅡ-D表面经典的MHCⅡ(HLA-DR、-DP、-DQ)类抗原表达,RT-PCR检测CⅡTA及经典的MHCⅡ类分子的mRNA水平。体外混合淋巴细胞反应检测pDarⅡ-D组刺激外周血T细胞反应的能力。结果与正义链组比较,在重组人干扰素(IFN)-γ诱导下,pDarⅡ-D组HLA-DR及-DP抗原诱导性表达分别降低了95.63%、87.89%;同时CⅡTA及经典的MHCⅡ类分子的mRNA含量明显减少(P<0.01);pDarⅡ-D组刺激T细胞分泌IL-2mRNA水平降低(P<0.05)。结论CⅡTA反义片段抑制了自身mRNA含量,从而阻止了其调控的MHCⅡ类分子的相应表达。
OBJECTIVE: To investigate the feasibility of inhibiting the expression of MHC class II antigen on the surface of human dermal fibroblasts by using antisense RNA of MHC Ⅱ transactivator (MHCⅡtransactivator, CⅡTA) mediated by nanocarriers. Methods C Ⅱ TA antisense RNA (pDar Ⅱ plasmid) was stably transfected into human primary dermal fibroblasts (pDarⅡ-D), and the classic MHCⅡ (HLA-DR, -DP, -DQ) The expression of antigen was detected by RT-PCR. The mRNA levels of CⅡTA and MHC class II molecules were detected by RT-PCR. In vitro mixed lymphocyte reaction assay pDar Ⅱ-D group to stimulate the peripheral blood T cell response ability. Results Compared with the normal control group, the induced expression of HLA-DR and -DP antigen in pDarⅡ-D group decreased by 95.63% and 87.89%, respectively, under the induction of recombinant human interferon (IFN) -γ. Meanwhile, CⅡTA and classical MHCⅡ (P <0.01). The mRNA level of IL-2 secreted by T cells in pDarⅡ-D group decreased (P <0.05). Conclusion CⅡTA antisense fragment inhibits the mRNA level of its own, thus preventing the corresponding expression of its regulated MHC class II molecules.