CAJ | 학술논문

目的探讨CD80、CD86和CD137L基因联合表达增强宿主细胞毒性T淋巴细胞(CTL)杀伤活性的机制.方法 BAL B/c小鼠随机分为5组,A组接种H22-Wt细胞,B组接种H22-neo细胞,C组接种H22-CD80/CD86+细胞,D组接种H22-CD137L+细胞,E组接种H22-CD80/CD86/CD137L+细胞.分别于第14、35、56和84天,每组每次随机选取2只小鼠处死取材.原位末端标记法(TUNEL)和DNA ladder法检测脾淋巴细胞凋亡.电泳迁移率法(EMSA)检测T细胞核因子KB(NF-KB)活性.结果 TUNEL检测结果显示,接种后第14天,A、B组脾淋巴细胞即出现大量凋亡,D组凋亡虽然较A、B组明显减少,但仍远高于C、E组.随接种时间推移,C组脾淋巴细胞凋亡逐渐增多,而E组这一趋势不明显,至第84天,C组和E组的脾淋巴细胞凋亡指数分别为30.8±9.2和14.4±4.5.DNA ladder检测结果显示,接种后第14、35、56和84天,C组和E组均检测出典型阳性结果,其中以C组第35、56和84天凋亡最明显.EMSA检测结果显示,A、B组T细胞NF-KB活性很低,D组显著高于A、B组,C、E组则显著高于A、B和D组.随着接种时间推移,E组NF-KB活性一直维持较高水平,而C组呈现逐渐下降趋势,至第84天,C组和E组的T细胞NF-KB活性分别为14.01±1.04和41.16±5.78.结论 H22-CD80/CD86/CD137L+变异株接种荷瘤鼠后,CD28信号和CD137信号的协同作用可显著增强活化T细胞NF-KB活性,这可能是CD80、CD86和CD137L基因联合表达显著增强宿主CTL杀伤活性的机制之一.
목적 탐토CD80、CD86화CD137L기인연합표체증강숙주세포독성T림파세포(CTL)살상활성적궤제.방법 BAL B/c소서수궤분위5조,A조접충H22-Wt세포,B조접충H22-neo세포,C조접충H22-CD80/CD86+세포,D조접충H22-CD137L+세포,E조접충H22-CD80/CD86/CD137L+세포.분별우제14、35、56화84천,매조매차수궤선취2지소서처사취재.원위말단표기법(TUNEL)화DNA ladder법검측비림파세포조망.전영천이솔법(EMSA)검측T세포핵인자KB(NF-KB)활성.결과 TUNEL검측결과현시,접충후제14천,A、B조비림파세포즉출현대량조망,D조조망수연교A、B조명현감소,단잉원고우C、E조.수접충시간추이,C조비림파세포조망축점증다,이E조저일추세불명현,지제84천,C조화E조적비림파세포조망지수분별위30.8±9.2화14.4±4.5.DNA ladder검측결과현시,접충후제14、35、56화84천,C조화E조균검측출전형양성결과,기중이C조제35、56화84천조망최명현.EMSA검측결과현시,A、B조T세포NF-KB활성흔저,D조현저고우A、B조,C、E조칙현저고우A、B화D조.수착접충시간추이,E조NF-KB활성일직유지교고수평,이C조정현축점하강추세,지제84천,C조화E조적T세포NF-KB활성분별위14.01±1.04화41.16±5.78.결론 H22-CD80/CD86/CD137L+변이주접충하류서후,CD28신호화CD137신호적협동작용가현저증강활화T세포NF-KB활성,저가능시CD80、CD86화CD137L기인연합표체현저증강숙주CTL살상활성적궤제지일.
Objective To study the mechanism of enhancement of the CTL activity in mice co-expressing of CD80, CD86 and CD137L genes. Methods The mice were randomly divided into five group, named A, B, C, D and E. The group A and B were control group (CG). H22-BAL B/c HCC mouse model was established by subcutaneous injection with hepatoceUular carcinoma cells of cell line H22-Wt (group A), H22-neo (group B), H22-CD80/CD86+ (group C), H22-CD137L+ (group D) and H22-CD80/CD86/CD137L+ (group E), respectively. On the 14th, 35th, 56th and 84th day after the first inoculation of tumor cells, TUNEL staining and DNA ladder examination were used to detect apoptosis of splenic T lymphocytes in all groups at each post-inoculation time point. Electroplioretie mobility-shift assay (EMSA) method was used to detect the activity of nuclear factor KB (NF-KB) in splenic T lymphocytes in each group at each time point post-inoculation. Results Apoptosis was found in a great number of T lymphecytes in CG on the 14th day, much more than that in group C and E. The number of apoptotic T cells in group C had a significant difference compared with that in the group E from 14th to 84th day (P=0.003). DNA ladder analysis showed typical positive results in group C and E. The significant apoptosia fragments were found in group C on 21st, 35th and 84th days. NF-KB activity ofT cells in groups C and E was remarkably higher than that of groups CG and D, with higher in group D than that of CG (P=0.002), and with no significant difference between group C and E on 14th day. The activity in group E was stable and remarkably higher than that of group C on 56th and 84th days after the first inoculation. Conclusion H22-CD80/CD86/CD137L+ induces higher NF-KB activity of the host T ceils by synergistic action of CD28 and CD137, which may be one of the mechanisms of enhancement of the host CTL activity induced by co-expression of CDS0, CD86 and CD137L genes.