摘要
目的研究热休克蛋白96(HSP-gp96)白血病抗原肽的抗白血病效应及相关机制。方法将不同的白血病细胞株(K562、HL-60、U937)分别分为A、B细胞株组,A组加预先构建的HSP-gp96重组腺病毒感染,B组不加以感染。将两组细胞株均制备为相应的抗原肽(分别设为A1、B1组),外周血单个核细胞诱导树突细胞(DC)后分别负载A1及B1组抗原肽以分别获取相应的DC复合物(分别设为A2、B2组)。用四甲基偶氮唑蓝反应比色法检测各组对T淋巴细胞增殖的影响,用乳酸脱氢酶释放法检测其在不同效靶比下的自然杀伤细胞(NK)活性和细胞毒性T淋巴细胞(CTL)效应,并比较K562来源的A2组复合物对3种白血病细胞株杀伤效应的不同。结果与A、B及B1组相比,3种不同细胞株的A1、A2和B2组均可刺激T淋巴细胞的增殖,同时也能增强NK细胞活性和CTL效应(P均<0.05)。其中A1、A2组对NK活性和CTL效应的增强尤为显著。且随着效靶比的增加,A1、A2和B2组NK细胞活性和CTL效应也明显增强。另外,K562细胞株制备的A2组复合物的CTL效应在K562细胞中明显增强,与HL-60及U937细胞相比,差异有统计学意义。结论 HSP-gp96白血病抗原肽抗白血病效应明显,经DC诱导后其作用增强,且后者对同源细胞株的杀伤效应具有特异性。
Objective To investigate the anti-leukemia effect of heat shock protein- peptide complex-96 （HSP-gp96）. Methods Human leukemia cell lines （K562, HL-60 and U937） were respectively divided into group A and group B. Only human leukemia cell lines in group A were infected with the constructed recombinant adenovirus Ad-gp96. The cell lines in groups A and B were prepared for antigen peptide and then categorized into group A1 and group B1. Peripheral blood mononuclear cell induced by dendritic ceils （DC） loading antigen peptide prepared the DC-complex （group A2 and group B2）. Of each group, the proliferation of T lymphocytes was detected by methyl thiazolyl tetrazolium cytotoxicity method, and cytotoxic lymphocyte （CTL） and natural killer （NK） cell activity at different effect- target ratio was measured by lactate dehydrogenase release assay. Results Compared with groups A, B and B~, the proliferation of T lymphocytes were stimulated more remarkably in groups A~, A2 and B~ （especially in ＇groups A1 and B2）, the activity of NK cell and CTL were enhanced （all P 〈 0.05）, and the cytotoxic effect in the leukemia cells as the effect- target ratio were higher. The gp96-peptide complex derived from K562 was more efficient in enhancing the activity of CTL than HL-60 and U937 cells. Conclusion Leukemia antigen gp96-peptide complex has a significant anti-leukemia effect, especially after stimulated by DCs. Furthermore, this anti-leukemia effect has a specificity.
出处
《中华危重症医学杂志（电子版）》
CAS
2013年第1期12-16,共5页
Chinese Journal of Critical Care Medicine:Electronic Edition
基金
国家自然科学基金(81172250)
浙江省自然科学基金(LY12H16015)
浙江省卫生厅(2011ZDA008
2012ZB070)
杭州市科技局项目(20120533Q02)
