摘要
目的 研究腺病毒为载体的基因治疗时淋巴细胞在肝组织免疫反应中的作用，探讨免疫抑制疗法在腺病毒载体基因治疗中的可行性。 方法 取8只恒河猴，经不同路径输入携带大肠杆菌lacZ基因或荧火虫荧光素酶基因luc的重组腺病毒6只。其中４只进行免疫抑制治疗。将含lacZ的质粒DNA注入2只动物作为对照。用免疫组织化学法检测β2-MG、HLA-DR、CD3、CD4、CD8及CD20。 结果 腺病毒介导的基因治疗时肝脏的β2-MG、HLA-DR、CD3、CD4及CD8阳性细胞明显增多。腺病毒载体和转基因均与肝损害有关, 表现为一过性, 呈轻、中度无黄疸性肝炎。免疫抑制的动物只要处于免疫抑制状态下就没有肝炎的表现，基因表达的时间延长。质粒介导的基因转导效果差, 无肝损害及免疫反应。所有动物B淋巴细胞抗原CD20始终阴性。 结论 腺病毒介导的基因治疗时肝脏的β2-MG、HLA-DR、CD3、CD4及CD8阳性细胞明显增多, 造成轻、中度一过性肝损害。使用免疫抑制药物可避免肝损害的发生并延长基因表达的时间。
Objective To define the role of lymphocyte subsets and investigate the efficiency of immunosuppression regimen in acute hepatitis in non-human primates after adenovirus mediated gene therapy. Methods Six rhesus monkeys were infused with E-1 deleted adenovirus (Ad) expressing E coli lacZ gene or luciferase by various routes. Four of 6 animals were immunosuppressed by cyclophosphamide and predenisone. Two monkeys were transfected with a lacZ containing plasmid with lipofectamine as the control. Lymphocyte subsets CD3, CD4, CD8, CD20 and MHC molecule β2-microglobulin(β2-MG) and HLA-DR in liver tissues were studied using immunohistochemical staining. Results Staining of β2-MG and HLA-DR on the membranes of hepatocyte and increased numbers of CD3+, CD4+ and CD8+ T-lymphocytes were detected in the livers after gene transfer, while B-lymphocytes were absent. The monkeys developed a mild to moderate transient hepatitis. This was accompanied by adenovirus-mediated T-cell proliferation and neutralizing antibodies to adenovirus. Drug-induced immune suppression enhanced the ability of adenovirus- mediated gene transfer. The development of acute hepatitis and the accompanying immune abnormalities were delayed in immunosuppressed monkeys until after discontinuation of immunosuppressive therapy. Lipofectamine-mediated gene transfer was inefficient and no immune response and liver damage were observed in the livers. Conclusions Increased numbers of β2-MG, HLA-DR, CD3, CD4 and CD8 antigen positive cells are presented in the monkey livers after adenovirus-mediated gene therapy and induce mild to moderate transient hepatic inflammation. Immunosuppression regimen may prolong transgene expression and delay the development of acute adenoviral hepatitis. [
出处
《中华肝脏病杂志》
CAS
CSCD
2001年第5期291-293,共3页
Chinese Journal of Hepatology
