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目的 了解CD+ 4、CD+ 8自然杀伤 (NK)T细胞在慢性HBV感染者外周血中的分布情况 ,并对其细胞毒性进行分析 ,阐明其在慢性HBV感染中的作用。方法 常规分离外周血单个核细胞(PBMCs) ,用重组人白细胞介素 12 / 2 (rhIL 12 /IL 2 )诱导 14d ,以鼠抗人CD4 单克隆抗体 (mAb)或抗人CD8mAb与抗人CD56mAb分别标记细胞样品 ,流式细胞术 (FCM)分析 ,CD+ 4CD+ 56同时阳性的细胞 ,即为CD+ 4NK T细胞 ;CD+ 8CD+ 56同时阳性的细胞为CD+ 8NK T细胞 ,因此获得CD+ 4、CD+ 8NK T细胞的百分比。并对细胞样品同时进行细胞毒性试验 ;分别以鼠抗人CD4 mAb、抗人CD8mAb及抗人CD56mAb和新鲜兔血清处理细胞 ,以鉴定CD+ 3 CD+ 4NK T、CD+ 3 CD+ 8NK T细胞的细胞毒活性。结果 rhIL 12 /IL 2诱导后 ,慢性HBV感染者外周血CD+ 4NK T细胞的含量远低于正常对照组。CD+ 4NK T细胞的比例(% )在正常对照组、慢性乙型肝炎 (CH)组和慢性无症状携带者 (AsC)组分别为 18 10± 4 2 0 ,6 95±2 85和 1 5 0± 1 30 ,经方差分析证实 3组之间差异具有显著性 (P <0 0 5 ) ;CD+ 8NK T细胞的比例 (% )在正常对照组、CH组及AsC组分别为 2 70± 1 10 ,2 2 0± 1 40和 3 10± 0 70 ,经方差分析显示 3组之间无统计学差异 (P >0 0 5 )。51Cr释放试验
Objective To investigate the distribution of CD + 4, CD + 8 natural killer (NK) T cells in peripheral blood of patients with chronic HBV infection and to analyze their cytotoxicity to clarify their role in chronic HBV infection. Methods Peripheral blood mononuclear cells (PBMCs) were isolated routinely and induced with rhIL 12 / IL 2 for 14 days. The anti - human CD56 mAb (anti - human CD4 mAb) or anti - human CD8 mAb The results of flow cytometry (FCM) analysis showed that CD + 4CD + 56 positive cells were CD + 4NK T cells. CD + 8CD + 56 positive CD + 8NK T cells were CD + 4, CD + 8NK T The percentage of cells. The cytotoxicity of CD + 3 CD + 4NK T, CD + 3 CD + 8NK T cells was evaluated by cytotoxicity assay simultaneously. The cells were treated with mouse anti-human CD4 mAb, anti-human CD8 mAb and anti-human CD56 mAb and fresh rabbit serum respectively . Results After rhIL 12 / IL 2 induction, the levels of CD + 4NK T cells in peripheral blood of patients with chronic HBV infection were much lower than those of normal controls. The percentage (%) of CD + 4NK T cells was 18 10 ± 4 2 0, 6 95 ± 2 85 and 1 0 0 in the normal control group, the chronic hepatitis group (CH) group and the chronic asymptomatic carrier group ± 30, and the difference between the three groups was significant (P <0.05) by analysis of variance. The percentage of CD + 8NK T cells in normal control group, CH group and AsC group was 2 70 ± 1 10 , 220 ± 1 40 and 3 10 ± 0 70 respectively. There was no significant difference between the three groups (P> 0.05) by ANOVA. 51Cr release test