中国实用医刊
中國實用醫刊
중국실용의간
CENTRAL PLAINS MEDICAL JOURNAL
2013年
2期
3-6
,共4页
韩际奥%马应杰%杨丽%王志凌%韩莉%王郁杰
韓際奧%馬應傑%楊麗%王誌凌%韓莉%王鬱傑
한제오%마응걸%양려%왕지릉%한리%왕욱걸
慢性乙型病毒感染者%树突状细胞%细胞因子诱导的杀伤细胞%免疫效应
慢性乙型病毒感染者%樹突狀細胞%細胞因子誘導的殺傷細胞%免疫效應
만성을형병독감염자%수돌상세포%세포인자유도적살상세포%면역효응
Chronic hepatitis B virus infection%Dendritic cells%Cytokine-induced killer cells%Immune response
目的 研究慢性乙型病毒感染者与健康人外周血树突状细胞(DC)经HBsAg活化后的免疫功能的差异.方法 从慢性HBV感染者、健康人外周血中培养扩增DC和CIK,在DC成熟前,加入纯的HBsAg刺激,并再与同一来源的CIK共同培养.用流式细胞仪检测DC、CIK表型,用ELISA法检测DC、CIK共培养上清液中的IL-12浓度,用CCK-8比色法测定DC诱导CIK对HepG2.2.15细胞的杀伤活性.结果 未经HBsAg致敏的健康人DC表面标志CDla、CD80及CD83明显高于慢性HBV感染者(P＜0.05)；经HBsAg致敏的健康人DC表面标志均高于慢性HBV感染者,差异有统计学意义(P＜0.01)；慢性HBV感染者经HBsAg致敏的DC表面标志与未经HBsAg致敏者比较差异无统计学意义(P＞0.05).CIK细胞CD3 CD8、CD3 CD56的双阳性表达率健康人HBsAg致敏者明显高于慢性HBV感染者及未经HBsAg致敏的健康人(P＜0.01,P＜0.05)；慢性HBV感染组HBsAg致敏与未经HBsAg致敏者比较差异无统计学意义(P＞0.05).HBsAg致敏DC诱导CIK对HepG2.2.15细胞的杀伤率健康人显著高于慢性HBV感染者(P＜0.01).健康人HBsAg致敏DC、CIK共培养上清液中的IL-12浓度显著高于慢性HBV感染者(P＜0.01)；慢性HBV感染者HBsAg致敏与未经HBsAg致敏IL-12含量差异无统计学意义(P＞0.05).结论 慢性乙型病毒感染者、健康人DC经HBsAg活化后对HepG2.2.15细胞的免疫效应存在显著差异:健康人高于慢性HBV感染者.
目的 研究慢性乙型病毒感染者與健康人外週血樹突狀細胞(DC)經HBsAg活化後的免疫功能的差異.方法 從慢性HBV感染者、健康人外週血中培養擴增DC和CIK,在DC成熟前,加入純的HBsAg刺激,併再與同一來源的CIK共同培養.用流式細胞儀檢測DC、CIK錶型,用ELISA法檢測DC、CIK共培養上清液中的IL-12濃度,用CCK-8比色法測定DC誘導CIK對HepG2.2.15細胞的殺傷活性.結果 未經HBsAg緻敏的健康人DC錶麵標誌CDla、CD80及CD83明顯高于慢性HBV感染者(P＜0.05)；經HBsAg緻敏的健康人DC錶麵標誌均高于慢性HBV感染者,差異有統計學意義(P＜0.01)；慢性HBV感染者經HBsAg緻敏的DC錶麵標誌與未經HBsAg緻敏者比較差異無統計學意義(P＞0.05).CIK細胞CD3 CD8、CD3 CD56的雙暘性錶達率健康人HBsAg緻敏者明顯高于慢性HBV感染者及未經HBsAg緻敏的健康人(P＜0.01,P＜0.05)；慢性HBV感染組HBsAg緻敏與未經HBsAg緻敏者比較差異無統計學意義(P＞0.05).HBsAg緻敏DC誘導CIK對HepG2.2.15細胞的殺傷率健康人顯著高于慢性HBV感染者(P＜0.01).健康人HBsAg緻敏DC、CIK共培養上清液中的IL-12濃度顯著高于慢性HBV感染者(P＜0.01)；慢性HBV感染者HBsAg緻敏與未經HBsAg緻敏IL-12含量差異無統計學意義(P＞0.05).結論 慢性乙型病毒感染者、健康人DC經HBsAg活化後對HepG2.2.15細胞的免疫效應存在顯著差異:健康人高于慢性HBV感染者.
목적 연구만성을형병독감염자여건강인외주혈수돌상세포(DC)경HBsAg활화후적면역공능적차이.방법 종만성HBV감염자、건강인외주혈중배양확증DC화CIK,재DC성숙전,가입순적HBsAg자격,병재여동일래원적CIK공동배양.용류식세포의검측DC、CIK표형,용ELISA법검측DC、CIK공배양상청액중적IL-12농도,용CCK-8비색법측정DC유도CIK대HepG2.2.15세포적살상활성.결과 미경HBsAg치민적건강인DC표면표지CDla、CD80급CD83명현고우만성HBV감염자(P＜0.05)；경HBsAg치민적건강인DC표면표지균고우만성HBV감염자,차이유통계학의의(P＜0.01)；만성HBV감염자경HBsAg치민적DC표면표지여미경HBsAg치민자비교차이무통계학의의(P＞0.05).CIK세포CD3 CD8、CD3 CD56적쌍양성표체솔건강인HBsAg치민자명현고우만성HBV감염자급미경HBsAg치민적건강인(P＜0.01,P＜0.05)；만성HBV감염조HBsAg치민여미경HBsAg치민자비교차이무통계학의의(P＞0.05).HBsAg치민DC유도CIK대HepG2.2.15세포적살상솔건강인현저고우만성HBV감염자(P＜0.01).건강인HBsAg치민DC、CIK공배양상청액중적IL-12농도현저고우만성HBV감염자(P＜0.01)；만성HBV감염자HBsAg치민여미경HBsAg치민IL-12함량차이무통계학의의(P＞0.05).결론 만성을형병독감염자、건강인DC경HBsAg활화후대HepG2.2.15세포적면역효응존재현저차이:건강인고우만성HBV감염자.
Objective To study differences of immune effect of dendritic cell activated by HBsAg between patients with chronic HBV and without HBV infective in vitro.Methods Dendritic cells (DC) and cytokine-induced killer(CIK) cells were cultured and amplified from chronic HBV infection and human peripheral blood.Before the DC mature,it was stimulation with adding pure HBsAg in cell culture medium.DC and CIK phenotype were detected by flow cytometry method.ELISA was used to detect the level of IL-12 in the upper liquid medium which DC and CIK cell mixed culture.The cell-killing activity of DC-induced CIK cell for HepG2.2.15 was measured by using CCK-8 colorimetric assay.Results Except HLA-DR,the positive rate of DC surface marker CDla,CD80 and CD83 in healthy people were higher than in chronic HBV infection (P ＜ 0.05),but healthy people DC activated by HBsAg,four surface markers were significantly higher than in chronic HBV infection (P ＜ 0.01).With or without HBsAg-pulsed,there was no significant difference in DC surface markers of chronic HBV infection.CD3 CD8,CD3 CD56 double positive expression rate of CIK cells in healthy people with HBsAg-pulsed were higher than in chronic HBV infection and those healthy without with HBsAg-pulsed.The difference statistically significant (P ＜0.01,P ＜ 0.05).There were not significantly differences chronic in HBV infection with or without HBsAg-pulsed (P ＞ 0.05) ; At healthy people,CIK cells HBsAg pulsed DC induced cell-killing rate of HepG2.2.15 cells was significantly higher than those in the chronic HBV infection (P ＜ 0.01).The level of IL-12 in HBsAg pulsed DC-CIK co-culture supermatants of healthy people was much higher than that of chronic HBV infection (P ＜ 0.001).There were not significant differences in chronic HBV infection with HBsAg vs without HBsAg pulsed (P ＞0.05).Conclusions The activation of the immune effect of HBsAg pulsed for HepG2.2.15 cells were significantly different:the healthy is stronger than that chronic HBV infection's.
