负载HER-2/neu多肽的树突状细胞激发特异性CTL反应  

Specific CTL response induced by dendritic cells pulsed with HER-2/neu peptide

作　　者：孟东[1] 时伟锋[1] 孙春雷[1] 时宏珍 史央 朱晨瑶 唐金海[3] 

机构地区：[1]苏州大学附属第四医院乳腺外科,江苏无锡214035 [2]南京市免疫细胞工程技术研究中心,南京得康生物技术有限公司,江苏南京210019 [3]江苏省肿瘤医院普通外科,江苏南京210000

出　　处：《中国肿瘤生物治疗杂志》2012年第1期29-34,共6页Chinese Journal of Cancer Biotherapy

基　　金：南京市科学技术委员会高新技术产业化项目资助(No.201103058)~~

摘　　要：目的:探讨以HER-2/neu为靶抗原、树突状细胞(dendritic cell,DC)为抗原载体激发特异性细胞毒性T淋巴细胞(cytotoxic T lymphocytes,CTL)反应的能力及研制治疗型乳腺癌疫苗的可行性。方法:采集17例HLA-A201+HER-2/neu+乳腺癌患者外周血,分离单个核细胞与外周血淋巴细胞(peripheral blood lymphocyte,PBL),并诱导为成熟DC(mature dendriticcell,mDC);人工合成HER-2/neu多肽[E75(KIFGSLAFL)和GP2(IISAVVGIL)2条]负载mDC后体外反复致敏PBL(3次,每周1次),检测其激发HER-2/neu特异性CTL的能力与CTL的杀伤活性。同时于患者腹股沟淋巴结富集区皮内注射负载HER-2/neu多肽的DC,每周1次,共接种4次,检测接种前后患者外周血细胞因子和特异性的CTL水平变化,并进行DTH试验。结果:患者外周PBL经过负载HER-2/neu多肽DC共3轮致敏后,HER-2/neu多肽特异的CTL平均比例比对照组(未负载HER-2/neu多肽DC组)明显增高[(5.41±1.44)%vs(0.41±0.12)%,P<0.05];致敏后PBL对负载HER-2/neu多肽T2靶细胞的杀伤率明显高于对照组(未负载DC诱导的CTL)[效靶比为30∶1时,(35.5±4.7)%vs(11.2±1.4)%,P<0.05]。接种负载HER-2/neu多肽的DC后,患者体内血清中细胞因子IL-2、IL-12、IFN-γ水平较治疗前显著升高[(409.09±89.39)vs(148.79±28.32)ng/ml,(56.23±14.08)vs(24.49±56.23)ng/ml,(146.57±25.97)vs(67.77±39.35)ng/ml;均P<0.05],TNF-α和IL-10水平较治疗前变化不大(P>0.05)。患者DTH试验阳性率为47%(8/17),DTH阳性患者外周血中特异性CTL比例明显上升。结论:负载HER-2/neu多肽的DC体内、外均具有激发特异性CTL反应能力,可诱导Th1型细胞因子的分泌,未发生临床不良反应。Objective： To explore the potential of autologous dendritic cells（DCs） pulsed with HER-2/neu peptide in inducing specific cytotoxic T lymphocyte（CTL） response and feasibility of breast cancer vaccines.Methods： Seventeen breast cancer patients with positive HLA-A201 and HER-2/neu were enrolled and their peripheral blood mononuclear cells and lymphocytes were isolated and induced into DCs and pulsed with HER-2/neu peptide.The killing effect of CTLs against T2 cell line pulsed with HLA-A201-binding peptide HER-2/neu was determined.The patients were inoculated subcutaneously near the inguinal region with auto-DCs pulsed with HER-2/neu peptide for 4 times every week.The immunological responses and clinical responses were examined in 1 week after the final vaccination.Results： The average percentage of special CTLs primed by DCs pulsed with HER-2/neu peptide was significantly higher than that in the control group（CTLs primed by DCs unloaded with HER-2/neu peptide）（[5.41±1.44]% vs [0.41±0.12]%,P0.05）.CTLs induced by DCs exerted a stronger killing effect on T2 cell line pulsed with HER-2/neu peptide than that in control group（[35.5±4.7]% vs [11.2±1.4]% at the ratio of E [effect] to T [target] as 30∶1,P0.05）.Vaccination of DCs was well tolerated and no toxicity was observed.The cytokine levels in sera such as IL-2,IL-12 and IFN-γ were increased after vaccinations（[148.79±28.32] ng/ml vs [409.09±89.39] ng/ml,[24.49±56.23] ng/ml vs [56.23±14.08] ng/ml,[67.77±39.35] ng/ml vs [146.57±25.97] ng/ml,respectively,all P0.05）.The cytokine levels in sera such as TNF-α and IL-10 had no significant changes before and after vaccination.The results of DTH test were positive in 8 patients（8/17）,and the percentages of antigen-specific IFN-γ＋ CD8＋T increased in 8 patients（8/17）.Conclusion： Auto-DC vaccines pulsed with HER-2/neu peptide can elicit specific immune responses ex vivo and in vivo,and induce secretion of Th1 type cytokines from DCs and have no adverse reaction.

关 键 词：树突状细胞 乳腺癌 HER-2/neu多肽 细胞毒性T淋巴细胞 免疫治疗 

分 类 号：R737.9[医药卫生—肿瘤] R730.51[医药卫生—临床医学]