CAJ | 학술논문

目的研究免疫相关性血细胞减少症(IRP)患者外周血CD4+T淋巴细胞CD70 mRNA和蛋白的表达水平及CD70启动子甲基化水平,探讨CD70在IRP发病中的作用.方法选取35例IRP患者(初治组16例,恢复组19例)及15名正常对照,应用密度梯度离心法分离其外周血单个核细胞,免疫磁珠分选CD4+T淋巴细胞,应用实时定量PCR(RT-PCR)检测CD70 mRNA相对表达水平,采用流式细胞术检测CD4+CD70+/CD4+细胞比例,采用甲基化特异性PCR(MSP)法检测CD4+T淋巴细胞CD70基因启动子区域甲基化水平.结果 IRP患者外周血CD4+CD707CD4+细胞比例初治组[(7.46±1.51)％]高于恢复组[(5.95±1.34)％]和正常对照组[(1.83±0.60)％],恢复组高于正常对照组(P值均＜0.05);IRP初治组、恢复组及正常对照组CD70 mRNA的表达水分别为2.314 (0.200～6.084)、1.021(0.135～3.434)及0.353 (0.008～2.258),两两比较差异均有统计学意义(P＜0.05).IRP患者CD70启动子甲基化水平低于正常对照组(P＜0.05).IRP患者CD70表达率与CD5+B淋巴细胞比例呈正相关(r=0.533,P＜0.01).结论 CD4+T淋巴细胞CD70启动子区域低甲基化水平导致CD70的过表达可引起其免疫紊乱,可能在IRP的发病中发挥重要作用.
목적 연구면역상관성혈세포감소증(IRP)환자외주혈CD4+T림파세포CD70 mRNA화단백적표체수평급CD70계동자갑기화수평,탐토CD70재IRP발병중적작용.방법 선취35례IRP환자(초치조16례,회복조19례)급15명정상대조,응용밀도제도리심법분리기외주혈단개핵세포,면역자주분선CD4+T림파세포,응용실시정량PCR(RT-PCR)검측CD70 mRNA상대표체수평,채용류식세포술검측CD4+CD70+/CD4+세포비례,채용갑기화특이성PCR(MSP)법검측CD4+T림파세포CD70기인계동자구역갑기화수평.결과 IRP환자외주혈CD4+CD707CD4+세포비례초치조[(7.46±1.51)％]고우회복조[(5.95±1.34)％]화정상대조조[(1.83±0.60)％],회복조고우정상대조조(P치균＜0.05);IRP초치조、회복조급정상대조조CD70 mRNA적표체수분별위2.314 (0.200～6.084)、1.021(0.135～3.434)급0.353 (0.008～2.258),량량비교차이균유통계학의의(P＜0.05).IRP환자CD70계동자갑기화수평저우정상대조조(P＜0.05).IRP환자CD70표체솔여CD5+B림파세포비례정정상관(r=0.533,P＜0.01).결론 CD4+T림파세포CD70계동자구역저갑기화수평도치CD70적과표체가인기기면역문란,가능재IRP적발병중발휘중요작용.
Objective To study the expression of CD70 and the methylation level of CD70 promoter in immuno-related pancytopenia (IRP) patients,and explore the role of CD70 in the pathogenesis of IRP.Methods Thirty-five IRP patients and fifteen healthy donors were enrolled in this study.Peripheral blood mononuclear cells were isolated from venous blood by density gradient centrifugation,and CD4 + T cells were isolated by immunomagnetic beads.The mRNA level and the percentage of CD70 of CD4 + T cells were measured by real-time quantitative polymerase chain reaction (RT-PCR) and flow cytometry respectively.Methylation-Specific PCR (MSP) was performed to determine the methylation level of CD70 promoter.Results The percentage of CD70 expression on CD4+ T cells of untreated IRP patients [(7.46±1.51)％] was significantly higher than that of recovered ones [(5.95±1.34)％] and normal controls [(1.83±0.60)％],and that of recovered IRP patients was significantly higher than of normal controls (P＜0.05).The relative expressions of CD70 mRNA in CD4+ T cells were 2.314 (0.200-6.084),1.021 (0.135-3.434),0.353 (0.008-2.258) in three groups respectively.The differences among untreated IRP patients,recovered IRP patients and normal controls were significant (P＜0.05).The CD70 promoter methylation level in CD4+ T cells of all IRP patients was significantly lower than that of normal controls (P＜0.05).The expression of CD70 positively correlated to the ratio of CD5+ B cells (r=0.533,P＜0.01).Conclusions The overexpression of CD70 may lead to immunologic disarrangement of patients,which may play important role in the pathogenesis of IRP.