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探索HLA-E分子的表达对习惯性流产(recurrent spontaneous abortion,RSA)及妊高症(pregnancy-induced hypertension,PIH)患者外周血γδT细胞细胞毒效应的影响。通过固相抗体法体外分离并扩增外周γδT血细胞作为效应细胞,以HLA-E转染的LcL721.221细胞(.221E)及滋养层细胞JAR作为靶细胞,采用4 h乳酸脱氢酶(LDH)释放试验观察NK细胞对靶细胞的细胞毒效应。结果显示,来自RSA、PIH及正常对照组的γδ细胞均不能有效杀伤HLA-E转染的.221E细胞及JAR细胞,但未经HLA-E转染的LcL721.221细胞则被溶解;抗HLA-E单抗3D12及抗CD94单抗HP-3B1的阻断可以分别部分恢复效应细胞对靶细胞LcL721.221E的杀伤,但对JAR细胞没有影响;与正常对照组相比,RSA及PIH患者外周血γδT细胞对.221、.221E及JAR细胞的细胞毒活性没有显著性差异(P>0.05)。这说明HLA-E分子的体外表达可以保护靶细胞防止γδT细胞的杀伤,该保护机制主要是通过γδT细胞受体CD94/NKG2对靶细胞表面HLA-E分子的识别来实现的;滋养层细胞对γδT细胞杀伤的抵抗可能存在MHC I类非依赖的机制。
To investigate the effect of HLA-E expression on cytotoxicity of γδT cells in peripheral blood of patients with recurrent spontaneous abortion (RSA) and pregnancy-induced hypertension (PIH). The peripheral γδT blood cells were isolated and expanded as effector cells by solid-phase antibody in vitro. LcL721.221 cells (.221E) transfected with HLA-E and JARs of trophoblast cells were used as target cells. After 4 h of lactate dehydrogenase ) Release test to observe the cytotoxic effect of NK cells on target cells. The results showed that γδ cells from RSA, PIH and normal control group can not effectively kill HLA-E-transfected221E cells and JAR cells, but not LcL721.221 cells transfected with HLA-E were dissolved; anti-HLA -E monoclonal antibody 3D12 and anti-CD94 monoclonal antibody HP-3B1 can partially restore effector cell killing of target cell LcL721.221E, respectively, but have no effect on JAR cells. Compared with the normal control group, the peripheral There was no significant difference in the cytotoxic activity of γδT cells between the 221, 221 and JAR cells (P> 0.05). This indicates that the in vitro expression of HLA-E molecules can protect the target cells against γδT cell killing, which is mainly through the γδT cell receptor CD94 / NKG2 recognition of HLA-E molecules on the target cell surface to achieve; trophoblast cell pairs Resistance to γδT cell killing may be an MHC class I-independent mechanism.