尖锐湿疣病损组织粗提蛋白对混合淋巴细胞反应的影响

The effect on mixed lymphocyte reaction of crude extracts from condyloma acuminata lesions

目的 通过观察尖锐湿疣病损组织粗提蛋白刺激后的树突状细胞(DC)对混合淋巴细胞反应的影响,探讨尖锐湿疣病损粗提蛋白对DC免疫功能的影响。方法 用聚合酶联反应(PCR)鉴定尖锐湿疣病损组织中人乳头瘤病毒(HPV)6 DNA和HPV11 DNA。人脐带血和外周血分离培养10 d的DC分别用尖锐湿疣病损组织和健康儿童包皮组织粗提蛋白刺激2次。另设空白对照组磷酸盐缓冲液(PBS)刺激。用四甲基偶氮唑蓝(MTT)法检测DC刺激同种异体和自体混合淋巴细胞反应的能力。同种异体淋巴细胞来源于脐带血和外周血,同种自体淋巴细胞来源于外周血。结果 PCR结果显示尖锐湿疣病损组织中HPV6b DNA阳性。尖锐湿疣病损粗提蛋白刺激后的DC刺激来源于脐血的T淋巴细胞,其增殖指数(5.514)高于包皮对照组(2.645)和空白对照组(1.698,DC∶T均为1∶11)。尖锐湿疣病损粗提蛋白刺激后的DC刺激来源于外周血的T淋巴细胞的增殖指数(2.352)高于包皮对照组(1.083)和空白对照组(0.644,DC∶T均为1∶3)。结论 尖锐湿疣病损粗提蛋白可能因存在的HPV病毒抗原对DC的刺激和功能活化,进一步促进了T淋巴细胞的增殖活化,并进而推测可以提高机体抗病毒特异性细胞免疫。

Objective To investigate the effect of dendritic cells pulsed with the crude extracts from condyloma acuminata lesions on the mixed lymphocyte reaction. Methods HPV6b DNA and HPV11 DNA in condyloma acuminata lesions were detected by PCR. Plastic-adherent mononuclear cells (MNCs) were isolated from umbilical cord blood and peripheral blood, cultured with cytokines including granulocyte-macrophage-colony- stimulating factor (GM-CSF), interleukin-4 (IL-4) and Bacterial lipopolysaccharide (LPS) in fetal bovine serum-containing media. We incubated the dendritic cells with crude extracted proteins from the condyloma acuminata lesions or foreskins. The T lymphocytes were isolated from umbilical cord blood and peripheral blood. The mixed lymphocyte reaction induced by dendritic cells (DCs) was observed by MTT method. Results The PCR result showed that HPV6b DNA was positive in the condyloma acuminata lesions. After T lymphocytes were incubated with DCs which pulsed with crude extracted protein from the condyloma acuminata lesions, the values of the stimulation index (SI) of T lymphocytes from cord blood and peripheral blood were 5.514 (DC∶T ratio 1∶11) and 2.352 (DC∶T ratio 1∶3) respectively. While, the ones in the foreskin control group (2.645, DC∶T ratio 1∶11; 1.085, DC∶T ratio 1∶3) and in the PBS control group (1.698, DC∶T ratio 1∶11; 0.644, DC∶T ratio 1∶3) were relative lower. Conclusions The results suggest that the crude extracts from condyloma acuminata might enhance the antigen presenting capacity of dendritic cells which subsequently elicitated the T lymphocytes.