白细胞介素-18基因修饰的胎肝细胞经脾移植对小鼠免疫功能的影响  

Effects of intrasplenic transplantation of IL-18 gene-modified fetal hepatocytes on mouse immune function

作　　者：姚航平[1] 张立煌[1] 孙文佶[1] 冷建杭[1] 

机构地区：[1]浙江大学医学院免疫学研究所,浙江杭州310031

出　　处：《浙江大学学报（医学版）》2002年第2期115-120,共6页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金重点资助项目 (39730 4 2 0 )

摘　　要：目的 :观察表达 m IL- 18的重组腺病毒基因修饰的胎肝细胞 (Adm IL- 18/ BNL.CL2 )经脾移植对正常小鼠免疫功能的影响。方法 :实验组小鼠经脾移植 Adm IL- 18/ BNL.CL2 ,同时设 Lac Z病毒对照组 (Ad- Lac Z/ BNL.CL2 ) ,BNL.CL2 细胞对照组及空白对照组。2周后处死 ,留取血清 ,制备腹腔巨噬细胞、脾淋巴细胞、肝组织匀浆液 ,提取肝组织总 RNA。采用 ELISA法检测各组小鼠血清、腹腔 Mφ和脾细胞培养上清、肝匀浆中细胞因子的含量 ;采用半定量 RT- PCR法 ,检测肝组织细胞因子 m RNA相对表达量 ;以 LDH释放法测定腹腔 Mφ杀伤活性和脾 NK细胞活性 ,用 MTT还原比色法测定脾淋巴细胞的增殖活性。结果 :实验组小鼠血清、细胞培养上清及肝匀浆中 ,IL- 18、IL - 2、IFN-γ、TNF-α含量均高于其它对照组 ,而 IL - 4、IL - 10水平则低于对照组 ;半定量 RT- PCR结果与EL ISA检测结果一致 ;同时 ,实验组腹腔 Mφ的杀伤活性和脾 NK细胞活性 ,及脾淋巴细胞增殖活性也明显高于对照组。结论 :Adm IL - 18能有效转染至胎肝细胞并稳定表达 m IL- 18;Adm IL - 18基因修饰的胎肝细胞经脾移植后 ,可显著提高肝脏、脾脏免疫细胞活性 ,活化腹腔 Mφ,促进 Th1类细胞因子表达 ,抑制 Th2类细胞因子的分泌。Objective: To investigate the effects of IL 18 gene modified fetal hepatocytes (AdmIL 18/BNL.CL 2) intrasplenic transplantation on mouse immune function. Methods: Forty mice were evenly divided into 4 groups of 10. Each group received an intrasplenic transplantation one of the following: AdmIL 18/BNL.CL 2, Ad LacZ/BNL.CL 2 (virus control), BNL.CL 2 (cell control) and PBS (blank control). After two weeks, the mice were sacrificed. Serum cytokine levels, Mφ and splenic cell culture supernatant and liver tissue extracts supernatants were measured using ELISA. Hepatic cytokines mRNA expression were determined by RT PCR. The cytotoxicity of peritoneal Mφ and NK activity of splenocytes were detected by LDH release assay. The proliferation of splenic lymphocytes was determined by MTT assay. Results: The IL 18, IL 2, IFN γ, TNF α levels of serum, Mφ and splenocyte culture supernatant, liver tissue extracts supernatants in mice transplanted with AdmIL 18/BNL.CL 2 were higher and the IL 4, IL 10 levels were lower compared to their levels in other 3 groups. The highest IL 18, IL 2, IFN γ, TNF α and the lowest IL 4, IL 10 mRNA expressions in the liver were observed in mice transplan ted with AdmIL 18/BNL.CL 2. The mice transplanted with AdmIL 18/BNL.CL 2 showed significantly increased cytotoxicity of Mφ, NK activity and splenic cell proliferation compared with the other 3 groups. Conclusion: AdmIL 18 can be effectively transfected into mice fetal heptocytes which subsequently IL 18. Intrasplenic transplantation of IL 18 gene modified fetal hepatocytes may augment mouse immune function and provide an useful basis for targeted gene therapy of liver disease.

关 键 词：白细胞介素-18 基因转移 重组腺病毒载体 胎肝细胞 脾内移植 免疫功能 动物实验 

分 类 号：R392.11[医药卫生—免疫学]