T淋巴细胞乙肝病毒感染状态与临床意义的研究.pdfVIP

中国科技论文在线 T 淋巴细胞乙肝病毒感染状态与临床意义的 研究# 严勤，李用国** 5 （哈尔滨医科大学附属第一医院，感染性疾病科，哈尔滨 150001 ） 摘要：目的：利用细胞体外培养扩增技术检测外周血Ｔ淋巴细胞乙肝病毒DNA 载量与临床 转归的关系。方法：选取乙肝病毒携带者3 例，慢性乙型肝炎病人32 例，利用密度梯度离 心法，提取外周血单个核细胞，再利用免疫磁珠法，富集T 淋巴细胞，将T 细胞在淋巴因 子的诱导下体外培养扩增，于不同的时间点检测细胞及培养上清的 HBV-DNA 载量。利用 10 免疫组化，原位杂交及逆转录PCR 技术证明T 细胞是否可以感染乙肝病毒。结果：HBV 可 以感染T 细胞，并稳定复制，合成抗原。T 细胞在体外培养可扩增，同时感染的HBV 也复 制增加。血清病毒检测结果与T 细胞病毒检测结果呈正相关，T 细胞病毒的检出与肝组织活 检的炎症纤维化分级级别呈正相关。 关键词：T 淋巴细胞；乙肝病毒；临床预后 15 T lymphocytes in hepatitis B virus infection status and the clinical significance YAN Qin, LI Yongguo (The First Hospital of Harbin Medical University,Department of infection disease, Harbin 150001) 20 Abstract: IObjective Using cells cultured in vitro amplification technology to detect hepatitis B virus DNA load in peripheral blood T lymphocytes and explore the clinical significance.Method: Three cases of hepatitis B virus carriers, 32 cases of chronic hepatitis B patients were involved. Peripheral blood mononuclear cells were separated by density gradient centrifugation over Ficoll-Hypaque, And T cells were purified from the mononuclear cells fraction using human CD3 25 positive selection kit, The T cell were inoculated into 9/24-well plates, then the cells were cultured for 12 days. Intracellular HBV were examined by immunohistochemistry and situ hybridization, intracellular and supernatant HBV DNA levels were examined at day 0, 1, 6