泡球蚴感染过程中宿主CD_4^+细胞凋亡和凋亡相关基因转录水平的研究  

CD_4^+cells apoptosis and apoptosis-associated gene expression in host infected with alveolar echinococcosis

作　　者：李富荣 石佑恩[2] 史大中[3] DA Vuitton PS Craig 

机构地区：[1]深圳市人民医院(暨南大学医学院第二附属医院)临床医学研究中心,深圳518020 [2]华中科技大学同济医学院寄生虫学教研室 [3]兰州医学院寄生虫学教研室 [4]Universite de Franche-comte,France [5]Division of Biological,the University of Salford,UK

出　　处：《中国人兽共患病杂志》2003年第4期59-62,65,共5页Chinese Journal of Zoonoses

基　　金：欧共体基金资助项目 (STD -2 )

摘　　要：目的　为了探讨泡球蚴感染宿主CD+ 4 淋巴细胞缺失的机制 ,以及泡球蚴感染与宿主CD+ 4 淋巴细胞凋亡及凋亡相关基因转录水平的相关性。方法　利用尼龙柱和补体法从泡球蚴感染 12周 ,2 5周和正常对照组BALB/c小鼠脾脏分离出纯CD+ 4 ,CD+ 8细胞 ,在体外分别经EmAg ,anti-CD3 ,IL - 2 ,TNFα ,PWM刺激培养 16h ;Tunel和PI双染后 ,FCM分析凋亡细胞数 ;并用C -myc ,TGF - β ,bcl- 2cDNA探针检测感染 2 5周EmAg诱导CD+ 4 细胞的凋亡相关基因转录水平。结果　FCM分析 ,感染 12周组CD+ 4 ,CD+ 8细胞凋亡数同正常对照组无明显差别 (P >0 0 5 ) ,感染 2 5周组CD+ 4 细胞凋亡数较正常对照组显著增高 (P <0 0 1) ,也显著性高于同组CD+ 8细胞 (P <0 0 1)。CD+ 4 细胞内C -myc,TGF - βmRNA水平较正常对照组显著升高 (P <0 0 5 ) ,而bcl- 2mRNA水平则减弱 (P <0 0 5 )。结论　泡球蚴寄生宿主后期 ,可诱导宿主成熟T细胞中CD+ 4 细胞发生凋亡 ,使宿主处于免疫抑制状态。凋亡的形成与凋亡信号增加 ,抑制信号减弱所引起。Aim To study the relationship between alveolar echinococcus infection and apoptosis of CD + 4lymphocyte in host Methods BALB/c mice were infected with alveolar echinococcus for 12 weeks and 25 weeks respectively as experiment group The uninfected mice were used as control group CD + 4T cells were purified Purified CD + 4cells was cultured in complete medium and were stimulated with EmAg,anti CD 3mAb,rIL 2,mouse rTNFα and PWM respectively After 16h,cells were collected and stained by TUNEL assays and PI Flow cytometry analysis Messenger RNA(mRNA) expression of bcl 2,C myc,TGF β in CD + 4stimulated with EmAg(25 weeks of post infection)was determined by the method of nucleic acid in situ hybridization Results Compared with control group,the apoptosis level of CD + 4cells in 12 weeks of post infection group was not significant change The apoptosis level of CD + 4T cells showed significantly increase in 25 weeks post infection group(P<0 01),and as well as CD + 8T cell increased But apoptosis of CD + 4T cell was higher than that of CD + 8T cell There was significantly difference about express level of C myc,TGF β,bcl 2 mRNA in CD + 4T cell between 25 weeks of post infection group and normal control Level of C myc mRNA and TGF βmRNA express increased,and bcl 2 mRNA express decreased in CD + 4T cell Conclusion CD + 4T cell apoptosis was a prominent feature in later period of alveolar echinococcus infection death promoter signals increased and death suppresser signals decreased may been reason responsible,in part,for the apoptosis in CD + 4lymphocytes of infected group

关 键 词：泡球蚴感染 宿主 CD4^+ 细胞凋亡 凋亡相关基因 转录 T细胞亚群 核酸原位杂交 

分 类 号：R383.3[医药卫生—医学寄生虫学]