中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2012年
12期
2475-2478
,共4页
树突状细胞%转录因子%T淋巴细胞%免疫杀伤%干扰素-γ
樹突狀細胞%轉錄因子%T淋巴細胞%免疫殺傷%榦擾素-γ
수돌상세포%전록인자%T림파세포%면역살상%간우소-γ
Dendritic cells%Transcription factor%T-lyphocytes%Immune killer%Interferon-γ
目的 观察过表达细胞周期相关转录因子1(E2F-1)的MGC-803稳定株所制成的细胞抗原所引起的树突状细胞(DC)介导的细胞毒性T淋巴细胞(CTL)体外杀伤胃癌MGC-803细胞的作用.方法 用30μg/L重组人白细胞介素4(rhlL-4)、100 μg/L重组人粒细胞巨噬细胞刺激因子(rhGM-CSF)、50 μg/L重组人肿瘤坏死因子-α(rhTNF-α)和100 μg/L脂多糖(LPS)联合诱导健康人外周血单个核细胞成为DC,通过形态学及流式细胞仪对DC进行鉴定;实验分E2F-1组、NC组、MGC-803组和DC组,采用细胞计数试剂盒(CCK-8)法分别在DC:T为1:5、1:10、1:20、1:40时检测T淋巴细胞的增殖;采用乳酸脱氢酶(LDH)释放法在T淋巴细胞:MGC-803细胞分别为10:1、20:1、50:1时检测杀伤效果;用酶联免疫吸附试验(ELISA)法检测培养液中的干扰素(IFN)-γ的含量变化.结果 形态学和流式细胞仪检测结果显示经诱导的DC细胞成熟特征明显,过表达E2F-1组T淋巴细胞增殖能力刺激指数(SI)值(4.42 ±0.23)高于空载体组(2.95±0.53)和空白对照组(3.01±0.68),差异有统计学意义(P＜0.05);过表达E2F-1抗原致敏的DC诱导的CTL对胃癌MGC-803细胞株的杀伤率[(65.00±8.24)％]高于DC-CTL组[(45.25±7.89)％]、单纯MGC-803组[(41.83±4.79)％]、空载体组[(31.16±11.96)％]和空白对照组[(33.25±8.38)％],差异有统计学意义(P＜0.05);过表达E2F-1抗原组T淋巴细胞分泌IFN-γ的量[(571.03±5.96) ng/L]高于空载体组[(382.85±4.94) ng/L]、MGC-803组[(390.13±7.06) ng/L]和DC组[(356.03±5.36) ng/L],差异有统计学意义(P＜0.05).结论 过表达E2F-1的MGC-803抗原致敏的DC在体外可增加T淋巴细胞的增殖,加强T淋巴细胞对MGC-803细胞的杀伤力.
目的 觀察過錶達細胞週期相關轉錄因子1(E2F-1)的MGC-803穩定株所製成的細胞抗原所引起的樹突狀細胞(DC)介導的細胞毒性T淋巴細胞(CTL)體外殺傷胃癌MGC-803細胞的作用.方法 用30μg/L重組人白細胞介素4(rhlL-4)、100 μg/L重組人粒細胞巨噬細胞刺激因子(rhGM-CSF)、50 μg/L重組人腫瘤壞死因子-α(rhTNF-α)和100 μg/L脂多糖(LPS)聯閤誘導健康人外週血單箇覈細胞成為DC,通過形態學及流式細胞儀對DC進行鑒定;實驗分E2F-1組、NC組、MGC-803組和DC組,採用細胞計數試劑盒(CCK-8)法分彆在DC:T為1:5、1:10、1:20、1:40時檢測T淋巴細胞的增殖;採用乳痠脫氫酶(LDH)釋放法在T淋巴細胞:MGC-803細胞分彆為10:1、20:1、50:1時檢測殺傷效果;用酶聯免疫吸附試驗(ELISA)法檢測培養液中的榦擾素(IFN)-γ的含量變化.結果 形態學和流式細胞儀檢測結果顯示經誘導的DC細胞成熟特徵明顯,過錶達E2F-1組T淋巴細胞增殖能力刺激指數(SI)值(4.42 ±0.23)高于空載體組(2.95±0.53)和空白對照組(3.01±0.68),差異有統計學意義(P＜0.05);過錶達E2F-1抗原緻敏的DC誘導的CTL對胃癌MGC-803細胞株的殺傷率[(65.00±8.24)％]高于DC-CTL組[(45.25±7.89)％]、單純MGC-803組[(41.83±4.79)％]、空載體組[(31.16±11.96)％]和空白對照組[(33.25±8.38)％],差異有統計學意義(P＜0.05);過錶達E2F-1抗原組T淋巴細胞分泌IFN-γ的量[(571.03±5.96) ng/L]高于空載體組[(382.85±4.94) ng/L]、MGC-803組[(390.13±7.06) ng/L]和DC組[(356.03±5.36) ng/L],差異有統計學意義(P＜0.05).結論 過錶達E2F-1的MGC-803抗原緻敏的DC在體外可增加T淋巴細胞的增殖,加彊T淋巴細胞對MGC-803細胞的殺傷力.
목적 관찰과표체세포주기상관전록인자1(E2F-1)적MGC-803은정주소제성적세포항원소인기적수돌상세포(DC)개도적세포독성T림파세포(CTL)체외살상위암MGC-803세포적작용.방법 용30μg/L중조인백세포개소4(rhlL-4)、100 μg/L중조인립세포거서세포자격인자(rhGM-CSF)、50 μg/L중조인종류배사인자-α(rhTNF-α)화100 μg/L지다당(LPS)연합유도건강인외주혈단개핵세포성위DC,통과형태학급류식세포의대DC진행감정;실험분E2F-1조、NC조、MGC-803조화DC조,채용세포계수시제합(CCK-8)법분별재DC:T위1:5、1:10、1:20、1:40시검측T림파세포적증식;채용유산탈경매(LDH)석방법재T림파세포:MGC-803세포분별위10:1、20:1、50:1시검측살상효과;용매련면역흡부시험(ELISA)법검측배양액중적간우소(IFN)-γ적함량변화.결과 형태학화류식세포의검측결과현시경유도적DC세포성숙특정명현,과표체E2F-1조T림파세포증식능력자격지수(SI)치(4.42 ±0.23)고우공재체조(2.95±0.53)화공백대조조(3.01±0.68),차이유통계학의의(P＜0.05);과표체E2F-1항원치민적DC유도적CTL대위암MGC-803세포주적살상솔[(65.00±8.24)％]고우DC-CTL조[(45.25±7.89)％]、단순MGC-803조[(41.83±4.79)％]、공재체조[(31.16±11.96)％]화공백대조조[(33.25±8.38)％],차이유통계학의의(P＜0.05);과표체E2F-1항원조T림파세포분비IFN-γ적량[(571.03±5.96) ng/L]고우공재체조[(382.85±4.94) ng/L]、MGC-803조[(390.13±7.06) ng/L]화DC조[(356.03±5.36) ng/L],차이유통계학의의(P＜0.05).결론 과표체E2F-1적MGC-803항원치민적DC재체외가증가T림파세포적증식,가강T림파세포대MGC-803세포적살상력.
Objective To study the methods of induction of dendritic cells (DC) which derived from the peripheral blood of health adults and further to study the anti-tumor of kidney effect activated by DC.Methods With 30 μg/L recombinant human interleukin-4 (rhlL-4),100 pg/L recombined human GM-CSF (rhGM-CSF),50 μg/L recombinant human tumor necrosis factor-α (rhTNF-α) andl00 μg/L lipopolysaccharide (LPS) make nuclear cell become the DC,Through the morphological and flow cytometric identification of DC,cell cycle related transcription factor 1 (E2F-1) MGC-803 made antigen sensitization stable strains after DC with cell counting kit-8 (CCK-8) method to detect T lymphocyte proliferation in DC:T (1:5 ; 1:10 ; 1:20 ; 1:40).The killing effect of cytotoxic T lymphocyte (CTL) by DC loaded detect the Absorbance of different group by methyl thiazol tetrazolium (MTT) assay and calculate T lymphocytes stimulating index in T cell:MGC-803 (10:1,20:1,50:1).Results Morphology and Flow cytometric show the induction of DC cells mature features obvious and overexpressed E2F-1 antigen sensitization DC can promote T lymphocyte proliferation and express E2F-1 group T lymphocytes SI value (4.42 ± 0.23)was significantly higher than the two other groups [empty carrier group (2.95 ± 0.53) and blank control group (3.01 ±0.68),P ＜0.05].A E2F-1 antigen expression sensitization of DC induction to cancer of the stomach CTL MGC-803 cell damage rate (65.00 ± 8.24) ％ was significantly higher than other groups,[DC-CTL (45.25 ±7.8) ％,MGC-803 group (41.83 ±4.79)％,empty carrier group (31.16 ±11.96) ％,blank control group (33.25 ± 8.38) ％,P ＜ 0.05],express E2F-1 antigen group T lymphocytes secretion interferon (IFN)-γ amount (571.03 ± 5.96) ng/L is much higher than in other groups [(382.85 ±4.942) ng/L,(390.13 ±7.06) ng/L,(356.03 ±5.36) ng/L,P＜0.05].Conclusion Over expression E2F-1 MGC-803 antigen sensitization to DC in vitro can increase T lymphocytes added value and strengthen the T lymphocytes to kill MGC-803 cell.
