CAJ | 학술논문

目的探讨CD4-CD8-双阴性T淋巴细胞(DNT细胞)的致病作用及其表面标志分子.方法取30只C3H/Hej小鼠,腹腔注射10 pfu 3型鼠肝炎病毒(MHV-3),采用磁珠分选和Cytotox96非放射性细胞杀伤活性测定方法,检测MHV-3感染后0、4、15、30、40 d脾脏DNT细胞分别对肝细胞、脾脏CD8+T淋巴细胞以及鼠巨细胞病毒感染后的脾脏CD8+T淋巴细胞的杀伤效应.免疫荧光抗体标记后,用流式细胞技术三色荧光分析法对脾脏内DNT细胞的表型进行检测.数据进行t检验或单因素方差分析,并用S-N-K法进行差异显著性检验. 结果 MHV-3感染后的DNT细胞对CD8+T淋巴细胞具有明显的杀伤作用,其杀伤作用随着效应细胞与靶细胞的比例增加而增强.感染15 d时,当效靶比为1:1,2.5:1,5:1,10:1时,其杀伤效率分别为8.1%,38.6%,62.4%,90.3%.而对感染后的肝细胞以及非相关病毒感染的CD8+T淋巴细胞无明显杀伤效应.表型分析提示此群DNT细胞为一群全新的细胞群(TCR αβ+CD3+CD4-CD8 CD25 CD28-CD30-CD44+).结论 C3H小鼠感染MHV-3后的TCR αβ+CD3+CD4-CD8 CD25-CD28-CD30 CD44+细胞对同种病毒感染后的CD8+T淋巴细胞有特异性杀伤作用,提示该群细胞在慢性病毒性肝炎的发生和发展中起着一定的负性免疫调节作用,导致病毒感染慢性迁延.
목적 탐토CD4-CD8-쌍음성T림파세포(DNT세포)적치병작용급기표면표지분자.방법 취30지C3H/Hej소서,복강주사10 pfu 3형서간염병독(MHV-3),채용자주분선화Cytotox96비방사성세포살상활성측정방법,검측MHV-3감염후0、4、15、30、40 d비장DNT세포분별대간세포、비장CD8+T림파세포이급서거세포병독감염후적비장CD8+T림파세포적살상효응.면역형광항체표기후,용류식세포기술삼색형광분석법대비장내DNT세포적표형진행검측.수거진행t검험혹단인소방차분석,병용S-N-K법진행차이현저성검험. 결과 MHV-3감염후적DNT세포대CD8+T림파세포구유명현적살상작용,기살상작용수착효응세포여파세포적비례증가이증강.감염15 d시,당효파비위1:1,2.5:1,5:1,10:1시,기살상효솔분별위8.1%,38.6%,62.4%,90.3%.이대감염후적간세포이급비상관병독감염적CD8+T림파세포무명현살상효응.표형분석제시차군DNT세포위일군전신적세포군(TCR αβ+CD3+CD4-CD8 CD25 CD28-CD30-CD44+).결론 C3H소서감염MHV-3후적TCR αβ+CD3+CD4-CD8 CD25-CD28-CD30 CD44+세포대동충병독감염후적CD8+T림파세포유특이성살상작용,제시해군세포재만성병독성간염적발생화발전중기착일정적부성면역조절작용,도치병독감염만성천연.
Objective To investigate role of CD4-CD8-T cells in murine hepatitis virus type 3 (MHV-3) induced chronic viral hepatitis in C3H/Hej mice and to identify their surface markers. Methods Thirty C3H/Hej mice received 10 Pfu MHV-3 intraperitoneally, the CD4-CD8-T cells were isolated using magnetic bead sorting on 0, 4, 15, 30, 40 days post MHV-3 infection. The cytotoxie effects of CD4-CD8-T cells on normal and infected hepatocytes, CD8+T cells and unrelated-virus (murine cytomegalovirus, MCMV) infected CD8+ T cells were examined by non-radioactive cytotoxicity assay. The surface markers of CD4- CD8- T cells were determined by flow cytometry. Results MHV-3 infected CD4-CD8-T cells showed significant cytotoxic effect on CD8+ T cells, but not on infected hepatocytes or MCMV infected CD8+ T cells. The analysis of cell surface markers demonstrated that the CD4-CD8-T cells are a completely new T cell subset. Conclusions CD4-CD8-T cells have significant cytotoxic effect on virus specific CD8+T cells in MHV-3 infected C3H/Hej mice, which suggests that CD4-CD8-T cells have immune modulatory functions in the development of chronic viral hepatitis. The phenotype of these CD4-CD8-T ceils detected by flow cytometry is TCR αβ+CD3+CD4-CD8-CD25-CD28-CD30-CD44+.