Liu J W, Lu X, Yang Z M, Deng L J, Yang L
Beijing Biohealthcare Biotechnology Co. Ltd, Beijing 101318, China.
National Cancer Center/Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2017 Oct 18;49(5):840-846.
To explore the potential of autologous dendritic cells (DCs) pulsed with caner/testis antigen NY-ESO-1 peptides in inducing specific cytotoxic T lymphocyte (CTLs) response and antineoplastic immune function of specific CTLs.
Fifteen patients with II to III stage positive HLA -A0201 and NY-ESO-1 were enrolled in the Cancer Hospital Chinese Academy of Medical Sciences on the basis of preclinical experiments from November 2014 to October 2015, and their peripheral blood mononuclear cells (PBMCs) and peripheral blood lymphocytes (PBLs) were isolated. The PBMCs were induced into DCs and pulsed with NY-ESO-1 peptide. The phenotypes of DCs were stained with antibodies against HLA-DRCD11c,CD80,CD83 and CD86, and subsequently analyzed by multichannel flow cytometry (FCM). The killing effects of CTLs pulsed with HLA-A0201-binding peptide NY-ESO-1 and the potential of autologous DCs pulsed with NY-ESO-1 peptides in inducing specific cytotoxic T lymphocytes (CTLs) responses were determined. The patients were administered two infusions of auto-logous CTLs for 1 time every two weeks. The total infusion was with 2 times. The immunological responses and clinical responses were examined in 1 week after the final administration.
The immunophenotype of DCs pulsed with NY-ESO-1 peptide was analyzed, HLA-DRCD11c cells (93.6%±1.2%), CD80 cells (87.3%±3.6%), CD83 cells (82.8%±2.5%) and CD86 cells (93.4%±6.4%). PBLs isolated from patients primed by DCs pulsed with NY-ESO-1 peptide proliferated continuously and the proliferation index (PI) of the PBLs were analyzed. There was significant difference between the DCs loaded with polypeptides and those unloaded, though it could promote the proliferation of PBLs, but the PI was significantly lower than that of the DCs loaded with NY-ESO-1 peptide (P<0.05). The average percentage of special CTLs primed by DCs pulsed with NY-ESO-1 peptides was significantly higher than that in the control group (5.2%±1.2% vs. 0.4%±0.1%). CTLs induced by NY-ESO-1 pulsed DCs exerted a stronger killing effect on T2 cell line pulsed with NY-ESO-1 peptide than that in the control group at the ratio of E (effect) to T (target) as 30:1, P<0.05. The cytokine levels in the patients'sera such as IFN-γ, IL-2 and IL-12 were increased after treatments [(132.9±10.2) μg/L vs. (46.4±3.1) μg/L; (101.3±6.4) μg/L vs. (26.7±1.2) μg/L; (51.3±2.6) μg/L vs. (26.4±1.1) μg/L; all P<0.05], and the percentages of antigen-specific CD8IFN-γ increased in these patients (P<0.01).
Auto-DCs pulsed with NY-ESO-1 peptides can induce the proliferation of allogenic CTLs, which elicit specific immune responses ex vivo or in vivo, and boost anticancer immunity markedly.
探讨用癌胚抗原NY-ESO-1肽脉冲处理的自体树突状细胞（DCs）诱导特异性细胞毒性T淋巴细胞（CTLs）应答及CTLs抗肿瘤免疫功能的潜力。
在2014年11月至2015年10月临床前实验的基础上，选取中国医学科学院肿瘤医院15例II至III期HLA -A0201阳性且NY-ESO-1阳性的患者，分离其外周血单个核细胞（PBMCs）和外周血淋巴细胞（PBLs）。将PBMCs诱导分化为DCs并用NY-ESO-1肽脉冲处理。用抗HLA-DR、CD11c、CD80、CD83和CD86的抗体对DCs的表型进行染色，随后通过多通道流式细胞术（FCM）进行分析。测定用HLA-A0201结合肽NY-ESO-1脉冲处理的CTLs的杀伤作用以及用NY-ESO-1肽脉冲处理的自体DCs诱导特异性细胞毒性T淋巴细胞（CTLs）应答的潜力。患者每两周接受1次自体CTLs输注，共输注2次。在最后一次给药后1周检查免疫应答和临床应答。
分析用NY-ESO-1肽脉冲处理的DCs的免疫表型，HLA-DR⁺CD11c⁺细胞（93.6%±1.2%）、CD80⁺细胞（87.3%±3.6%）、CD83⁺细胞（82.8%±2.5%）和CD86⁺细胞（93.4%±6.4%）。分析从用NY-ESO-1肽脉冲处理的DCs致敏的患者中分离的PBLs的增殖情况以及PBLs的增殖指数（PI）。负载多肽的DCs与未负载多肽的DCs之间存在显著差异，虽然后者可促进PBLs增殖，但其PI显著低于负载NY-ESO-1肽的DCs（P<0.05）。用NY-ESO-1肽脉冲处理的DCs致敏的特异性CTLs的平均百分比显著高于对照组（5.2%±1.2%对0.4%±0.1%）。在效靶比（E/T）为30:1时，NY-ESO-1脉冲DCs诱导的CTLs对用NY-ESO-1肽脉冲处理的T2细胞系的杀伤作用强于对照组，P<0.05。治疗后患者血清中IFN-γ、IL-2和IL-12等细胞因子水平升高[（132.9±10.2）μg/L对（46.4±3.1）μg/L；（101.3±6.4）μg/L对（26.7±1.2）μg/L；（51.3±2.6）μg/L对（26.4±1.1）μg/L；均P<0.05]，这些患者中抗原特异性CD8⁺IFN-γ⁺细胞百分比增加（P<0.01）。
用NY-ESO-1肽脉冲处理的自体DCs可诱导同种异体CTLs增殖，在体内外引发特异性免疫应答，并显著增强抗癌免疫力。
