解剖学报 ›› 2020, Vol. 51 ›› Issue (4): 548-552.doi: 10.16098/j.issn.0529-1356.2020.04.012

• 肿瘤生物学 • 上一篇    下一篇

胰岛素样生长因子2 mRNA结合蛋白3限制性表位肽诱导的细胞毒性T淋巴细胞的体外免疫应答

陈红莲 时冉冉 张丽 刘辉 马永超*   

  1. 漯河医学高等专科学校内科学教研室,河南 漯河 462000
  • 收稿日期:2019-02-27 修回日期:2019-09-04 出版日期:2020-08-06 发布日期:2020-08-06
  • 通讯作者: 马永超 E-mail:13839527381@163.com
  • 基金资助:
    河南省重点研发与推广专项(科技攻关)项目;河南省高等学校重点科研项目

In vitro immune response of insulin-like growth factor 2 mRNA-binding protein 3 restricted cytotoxic T lymphocyte epitope from lung cancer immunotherapy target

CHEN Hong-lian SHI Ran-ran ZHANG Li LIU Hui MA Yong-chao*   

  1. Department of Internal Medicine, Luohe Medical College, He’nan Luohe 462000, China
  • Received:2019-02-27 Revised:2019-09-04 Online:2020-08-06 Published:2020-08-06
  • Contact: MA Yong-chao E-mail:13839527381@163.com

摘要:

目的 探讨胰岛素样生长因子2 mRNA结合蛋白3(IGF2BP3)限制性细胞毒性T淋巴细胞(CTL)表位诱导的CTL对肺癌细胞的作用。 方法 使用软件NetCTL 1.2、SYFPEITHI和IEDB预测选取IGF2BP3的人类白细胞抗原A2(HLA-A2)限制性表位肽。T2A2与表位肽的亲和力实验检测候选表位与T2A2细胞表面HLA-A2分子的结合能力,酶联免疫斑点(ELISPOT)实验检测候选表位肽诱导CTL分泌γ干扰素(IFN-γ)的能力,羧基荧光素二醋酸盐琥珀酰亚胺酯(CFSE)荧光染色检测细胞诱导CTL的能力。 结果 P143、P199、P280、P409、P515具有较好的结合力。表位肽P409、P515诱导的CTL具有较好的分泌IFN-γ的能力,并且对靶细胞具有一定的杀伤作用。  结论 P409、P515有望用于肿瘤的过继免疫治疗,可以成为抗肿瘤多肽免疫治疗疫苗的候选表位。

关键词: 胰岛素样生长因子2 mRNA结合蛋白3, 人类白细胞抗原A2, 细胞毒性T淋巴细胞, 免疫印迹法, 人 

Abstract:

Objective Cancer immunotherapy is attractive for antigen-specific T cell-mediated antitumor therapy, especially in induction of cytotoxic T lymphocytes(CTL). In this report, we evaluated insulin-like growth factor 2 mRNA-binding protein 3(IGF2P3) restricted epitope-induced cytotoxic T lymphocytes effects in human lung cancer cells.   Methods The human leukocyte antigen A2(HLA-A2) restricted epitope peptides of IGF2P3 were selected by NetCTL 1.2, SYFPEITHI and IEDB software prediction. The binding affinity of the peptides to HLA-A02 molecules was evaluated by T2A2 cells binding assay. enzyme-linked immunosport(ELISPOT) assay was used to investigate the ability of the peptide to induce specific restricted cytotoxic T lymphocytes (CTL) and release of interferon γ(IFN-γ). The ability of the peptides to induce T cell response was investigated by carboxyfluorescein succinimidyl amino ester(CFSE) fluorescent staining.   Results The candidate peptide P143, P199, P280, P409 and P515 showed moderate affinity toward HLA-A2 molecule. ELISPOT assay showed P409, P515 were able to induce specific CTL and higher levels of IFN-γ were released. The CTL induced by P409, P515 lysed target cells.  Conclusion P409 and P515 have the potential for adoptive immunotherapy and can be used as candidate epitopes for new anti-tumor polypeptide immunotherapy vaccine. P409 and P515 can be used in peptide-based immunotherapy for lung cancer.

Key words: Insulin-like growth factor 2 mRNA-binding protein 3, Human leukocyte antigen A2, Cytotoxic T lymphocyte, Western blotting, Human

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