慢性乙型肝炎患者血清HBVDNA含量和IL-12对其PBMC产生Th1/Th2类细胞因子影响的相关性研究

Relationship between HIBVDNA in serum and effect of IL-12 on Th1/Th2-type cytokins poduction of PBMC in chronic hepatitis B virus infection patients

目的：研究慢性乙型肝炎患者血清ＨＢＶＤＮＡ含量对ＩＬ－１２诱导其ＰＢＭＣ产生Ｔｈ１／Ｔｈ２类细胞因子协同效应的影响。方法：分离 ５０例慢性乙型肝炎患者外周血单个核细胞，分别与 ＰＨＡ（１００ ｕｇ／ｍｌ）、ＨＢｃＡｇ（１ｕｇ／ｍｌ）、ＨＢｅＡｇ（ １ ｕｇ／ｍｌ）单独或联合ＩＬ－１２（１０ ｎｇ／ｍｌ）体外培养４８ ｈ，ＥＬＩＳＡ法检测培养上清液细胞因子 ＩＬ－２、ＩＦＮ－γ、ＩＬ－４、ＩＬ－１０。荧光定量ＰＣＲ检测患者血清 ＤＮＡ含量，并分成 ＨＢＶＤＮＡ小于１０３拷贝／ｍｌ、１０３～１０５拷贝／ｍｌ、１０５～１０７拷贝／ｍｌ、大于 １０７拷贝／ｍｌ４组。结果：以 ＨＢＶＤＮＡ小于１０３拷贝／ｍｌ组做对照组，比较发现无论抗原（ＰＨＡ、ＨＢｃＡｇ、ＨＢｅＡｇ）单独诱导还是联合ＩＬ－１２共同诱导，随着血清ＨＢＶＤＮＡ含量的增高，ＰＢＭＣ产生ＩＬ－２和ＩＦＮ－γ水平逐渐降低，产生ＩＬ－４和ＩＬ－１０水平逐渐升高，并且ＩＬ－１２对ＰＢＭＣ产生ＩＦＮ－γ的增殖效应逐渐减弱，特别是血清ＨＢＶＤＮＡ大于 １０７拷贝／ｍｌ患者几乎无明显增殖效应。结论：高水平血清ＨＢＶＤＮＡ含量对ＩＬ－１２诱导慢性乙型肝炎ＰＢＭＣ产生ＩＦＮ－γ协同效?

Objective:To examine the influence of the HBV virus load on co-stimulatory effect of rhIL-12 on Th1/Th2-type cytokines production of PBMC in chronic hepatitis B virus infection patients. Methods: PBMC of 50 chronic hepatitis B virus infection patients were stimulated with PHA(100 U/ml),XHBcAg(1 ug/ml),HBeAg(1 ug/ml) in the presence or obsence of IL-12(10ng/m1).IL-2,JFN-y,LL-4,IL-10 production were determined by EUSA after 48 hours. HBVDNA in serum were determined by fluorecence quantity PCR(FQ-PCR) . The patients were divided into 4 groups(HBVDNA< 103 copies/ml, 103 - 105 copies/ml, 105 - 107 copies/ml, >107 copies/ml). Results: After stimulation with IL-12 and HBcAg/HBeAg or PHA ,co-stimulatory effect on IFN-y production were decreased while level of HBVDNA in serum was high. Co-stimulatory effect on IFN-y production almost was not observed in HBVDNA> 107 copies/mi group. Conclusion: The presence of high amount of HBVDNA in serum is associated with suppressed co-stimulatory and regulatory effect of IL-12 to Th1/Th2 profilation