期刊介绍

中华普通外科学文献简介:中华普通外科学文献简介,中华普通外科学文献简介[详细]

期刊信息

主管:
中华人民共和国国家卫生健康委员会
主办:
中华医学会
承办:
中山大学附属第一医院
中山大学附属中山医院
出版:
中华医学电子音像出版社有限责任公司
总编辑: 王深明
编辑部主任: 汪谦
刊 期: 双月刊
创刊时间: 2007年2月
国内刊号: 11-9148/R
国际刊号: 1674-0793
订 购:
中山大学附属第一医院期刊中心
510080, 广州市中山二路58号
电话:(020)87331056
Email:pwwxcma@126.com
定价: 每期28元,全年168元
全文收录:
中国科技核心期刊(中国科技论 文统计源期刊) 中国学术期刊网络出版总库 中文科技期刊数据库(全文版) 中国生物医学期刊引文数据库( CMCI) 美国化学文摘(CA) 首席医学网
首页 > 期刊列表 > 人肝癌浸润淋巴细胞的分离培养及其CD8阳性亚群特性研究
查看全文 下载全文

标题: 人肝癌浸润淋巴细胞的分离培养及其CD8阳性亚群特性研究
摘要:

目的:从原发性肝癌(HCC)患者术后的肿瘤组织中提取、培养肿瘤浸润性淋巴细胞(TILs),进一步分选、扩增其CD8阳性亚群(CD8+TILs),并研究其特性。方法:标本取自38例肝癌根治术后的肿瘤组织,免疫组织化学染色观察癌巢组织和癌旁组织TILs的分布,通过机械剪碎、混合酶消化和不连续密度梯度离心法分离提取TIL前体细胞。采用重组人白介素2进行激活,抗CD3、抗CD28抗体进行共刺激扩增,用磁珠分选出CD8+TILs,CCK8法观察CD8+TILs对肝癌细胞Hep3B、HepG2生长的影响。结果:TILs富集于癌旁组织,12例成功提取TILs,6例培养并实现大量扩增,扩增17~50倍后细胞数为(1.2~2.5)×108个,CD3+细胞比例为(77.53±16.37)%,CD3+CD4+比例为(27.08±21.56)%,CD3+CD8+比例(44.55±12.73)%,细胞活力为(90.5±3.0)%,CD8+TILs对肿瘤细胞系Hep3B、HepG2有较强的生长抑制作用。结论:本研究成功建立高效提取、培养CD8+TILs的方法,获得具有高抗肿瘤活性的CD8+TILs,为晚期肝癌等实体瘤的过继免疫治疗提供研究基础。

英文摘要: Objective:To investigate the methods of expansion of CD8 positive tumor-infiltrating lymphocytes (CD8+TILs) obtained from human hepatocellular carcinoma (HCC) and to study their immunological characteristics. Methods:Thirty-eight tumor tissue specimens were obtained from radical liver cancer surgery. Immunohistochemistry staining was performed on cancer nests and side tissues to study the distribution of TILs, and TIL precursor cells were extracted by mechanical shearing, mixed enzyme digestion and discontinuous density gradient centrifugal separation. Activated with high dose of recombinant human interleukin 2 at first, the isolated cells were co-stimulated of anti-CD3 and anti-CD28 antibody. The CD8+TILs were sorted by magnetic beads. The effect of CD8+TILs on the growth of Hep3B and HepG2 cells lines was observed in CCK8 cytotoxicity experiment in vitro. Results:TILs were enriched in paracancer tissues, 12 cases were successfully extracted, 6 cases were achieved a large amplification among them. The number of amplificated cells was (1.2-2.5)×108 after 17-50 times expansion fold, and the proportion of CD3+ cells in the paracancer tissues was (77.53±16.37)%, CD3+CD4+ ratio was (27.08±21.56)%, CD3+CD8+ ratio was (44.55±12.73)%, and the cell viability was (90.5±3.0)%. Moreover, CD8+TILs showed strong growth inhibition effect on Hep3B and HepG2 cell lines. Conclusions:This study successfully establishes a method for efficient extraction and cultivation of CD8 + TILs, and obtains CD8 +TILs with high antitumor activity, which provides a research basis for adoptive immunotherapy of solid tumors such as advanced liver cancer.
作者:

黄镇辉1,2;徐晓艳3;蒋小峰3;薛平3;曾嘉敏4;魏宜胜1,2

作者单位: 广州医科大学附属第二医院胃肠外科1,外科实验室2,肝胆外科3,病理科4
期刊: 中华普通外科学文献(电子版)
年.卷(期):页码 2020 .14(3):179-183
中图分类号:
文章编号:
引用格式: [1]黄镇辉1,2;徐晓艳3;蒋小峰3;薛平3;曾嘉敏4;魏宜胜1,2.人肝癌浸润淋巴细胞的分离培养及其CD8阳性亚群特性研究[J/CD].中华普通外科学文献(电子版),2020,14(3):179-183 .
关键词: 淋巴细胞,肿瘤浸润;癌,肝细胞;CD8阳性T淋巴细胞;免疫,细胞
英文关键词: Lymphocytes, tumor-infiltrating;Carcinoma, hepatocellular; CD8-positive T-lymphocytes;Immunity, cellular

欢迎阅读《中华普通外科学文献(电子版)》!您是该文第185位读者!

友情链接