自体DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞亚群表面分子的表达
投稿时间:2015-07-22  修订日期:2015-08-20  点此下载全文
引用本文:何剑营,续力云,陈冬冬,竺王玉,方可欣,周吉航,刘晓光,乐涵波,陈志军,吴国清,张永奎.自体DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞亚群表面分子的表达[J].医学研究杂志,2016,45(3):92-97
DOI: 10.11969/j.issn.1673-548X.2016.03.024
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作者单位E-mail
何剑营 316021 舟山医院细胞分子生物学实验室  
续力云 316021 舟山医院细胞分子生物学实验室  
陈冬冬 316021 舟山医院细胞分子生物学实验室  
竺王玉 316021 舟山医院细胞分子生物学实验室  
方可欣 316021 舟山医院细胞分子生物学实验室  
周吉航 316021 舟山医院细胞分子生物学实验室  
刘晓光 316021 舟山医院细胞分子生物学实验室  
乐涵波 舟山医院细胞分子生物学实验室, 胸心外科  
陈志军 舟山医院细胞分子生物学实验室, 胸心外科  
吴国清 310014 杭州, 浙江省人民医院肿瘤科  
张永奎 舟山医院细胞分子生物学实验室, 胸心外科 zhoushanzhang1@126.com 
基金项目:浙江省重大科技专项重点社会发展项目(2012C13015-1, 2012C13015-3)
中文摘要:目的 探讨自体DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞亚群表面分子表达情况.方法 抽取63例肺癌晚期患者的外周血分离获得外周血单个核细胞(PBMC),将与患者病理分类相同的肺癌细胞株经反复冻融处理获得可溶性抗原,制备肿瘤相关抗原肽负载DC-CIK细胞,并进行质量控制后回输.应用流式细胞术检测DC-CIK细胞治疗晚期肺癌患者前后T淋巴细胞表面CD3+HLA-DR+、CD3+CD25+、CD3+CD4+CD25+、CD3+CD8+HLA-DR+、CD3+CD8+CD38+细胞百分比及对比治疗前后表达情况.结果 成功获得成熟DC细胞,流式细胞术检测证实DC细胞成熟的标志分子CD11c、CD83、CD86及HLA-DR的表达均较高,分别为51.6%±10.3%、50.8%±9.7%、48.9%±11.4%、61.2%±6.5%;成功获得CIK细胞细胞,流式细胞术结果显示,CD3+CD56+双阳性细胞比例占21.3%±7.6%.与治疗前相比,DC-CIK细胞治疗后CD3+HLA-DR+(12.71±1.54 vs 11.44±4.13,P<0.05)、CD3+CD8+HLA-DR+(7.48±1.01 vs 5.20±1.01,P<0.01)、CD3+CD8+CD38+(8.27±1.71 vs 6.47±1.99,P<0.01)表达明显升高,CD3+CD4+CD25+(5.38±1.47 vs 6.12±0.67,P<0.05)表达明显降低.结论 采用自体DC-CIK免疫细胞治疗晚期肺癌可以明显提高患者免疫力.
中文关键词:DC细胞  CIK细胞  肺癌  T淋巴细胞亚群  免疫治疗
 
Surface Molecules Expressions on T Lymphocyte Subsets before and after Treatment with Autologus DC-CIK Cells in Patients with Advanced Lung Cancer.
Abstract:Objective To investigate the surface molecules expressions on T lymphocyte subsets before and after treatment with autologus DC-CIK cells in patients with advanced lung cancer. Methods Peripheral blood mononuclear cells(PBMCs) were derived from 63 patients with advanced lung cancer. And the tumor soluble antigen were derived from repeatedly frozenthawed lung cancer cell line with the corresponding pathological subtype. The DC-CIK cells were induced by the tumor-associated antigen peptide in vitro, and quality of the induced cells was assured for back transfusion. CD3+HLA-DR+, CD3+CD25+, CD3+CD4+CD25+, CD3+CD8+HLA-DR+ and CD3+CD8+CD38+ expressions on T lymphocyte subsets before and after treatment with autologus DC-CIK cells were analyzed by flow cytometry in advanced lung cancer patients. Results After 2 weeks of in vitro inducing PBMCs, mature DC cells and CIK cells were established. The expressions of CD11c, CD83,CD86 and HLA-DR, the molecular markers for DC cells maturation proved to be high expressed by flow cytometry, were 51.6%±10.3%,50.8%±9.7%, 48.9%±11.4%,61.2%±6.5%, respectively. And CD3+CD56+ double positive cells were showed to be 21.3%±7.6% among mature CIK cells by flow cytometry. Compared with pretreatment with DC-CIK cells, CD3+HLA-DR+(12.71±1.54 vs 11.44±4.13,P<0.05),CD3+CD8+HLA-DR+(7.48±1.01 vs 5.20±1.01,P<0.01)、CD3+CD8+CD38+(8.27±1.71 vs 6.47±1.99,P<0.01)expressions on T cells were significantly higher after treatment, and CD3+CD4+CD25+(5.38±1.47 vs 6.12±0.67,P<0.05)expression was significantly lower after treatment with DC-CIK cells. Conclusion After treatment with autologus DC-CIK cells, immunity of patients with advanced lung cancer was greatly improved.
keywords:DC cells  CIK cells  Lung cancer  T lymphocyte subsets  Immunotherapy
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