肿瘤抗原MUC4 HLA-A*0201限制性CTL表位的体外免疫效应研究
DOI:
作者:
作者单位:

作者简介:

通讯作者:

中图分类号:

基金项目:

国家自然科学基金资助项目(30471691)


In vitro immune response of cytotoxic T lymphocyte induced by dendritic cells pulsed with tumor associated antigen MUC4
Author:
Affiliation:

Fund Project:

  • 摘要
  • |
  • 图/表
  • |
  • 访问统计
  • |
  • 参考文献
  • |
  • 相似文献
  • |
  • 引证文献
  • |
  • 资源附件
  • |
  • 文章评论
    摘要:

    目的:探讨肿瘤抗原MUC4 HLA-A*0201限制性细胞毒性T淋巴细胞(CTL)表位肽负载树突状细胞诱导CTL的体外免疫效应-方法:从外周血单个核细胞(PBMC)中诱导树突状细胞(DC)-用之前表位预测法预测并合成的五种肽分子分别脉冲成熟的DC,并刺激HLA-A*0201+健康人自体CD8+T细胞成为CTL-用酶联免疫斑点法(ELISPOT),检测CTL IFN-γ的分泌-同时用乳酸脱氢酶(LDH)释放法检测其对靶细胞的杀伤作用-结果:人PBMC可在多种细胞因子的作用下被诱导称为DC-肽P01204诱导的CTL,在不同的效靶比可特异性杀伤P01204脉冲的T2细胞[5 ∶ 1时为(9.03 ± 0.24)%,10 ∶ 1时为(19.62 ± 0.44)%,20 ∶ 1时为(27.93 ± 2.22)%]和MUC4+-HLA-A2+的HCT-116细胞[5 ∶ 1时为(7.26 ± 0.18)%,10 ∶ 1时为(16.83 ± 0.40)%,20 ∶ 1时为(25.23 ± 1.35)%],均高于阴性对照组(P < 0.05),能够产生IFN-γ的细胞数(130.33 ± 6.58)明显高于阴性对照组(P < 0.05)-结论:肿瘤相关抗原MUC4的HLA-A*0201限制性CTL表位P01204,能诱导人外周血单核细胞的CTL反应,该活化的CTL能分泌免疫活性物质诱导特异性靶细胞的凋亡-

    Abstract:

    Objective:To explore the in vitro immune response to tumor associated antigen MUC4-derived cytotoxic T lymphocyte(CTL) epitope-pulsed dendritic cell(DC). Methods:DCs was induced from the HLA-A*0201-positive healthy individuals’ peripheral blood mononuclear cells(PBMC). DCs’ phenotype was identified by flow cytometry. CD8+ T cells were isolated and purified through MACS(magnetic activated cell sorting). Mature DCs were pulsed with each of the five synthesized peptides which were selected as possible CTL epitopes by software analysis previously. Autologous CD8+ T cells from the HLA-A*0201 healthy donor were stimulated with the peptide-pulsed DCs as CTL. CTL activity was assessed by lactate dehydrogenase(LDH) release assay and IFN-γ released by enzyme-linked immuno spot assay(ELISPOT). Results:Mature DCs could be induced from PBMCs through several cytokines’ action. CTL induced by peptide P01204 could lyse T2 cells pulsed with peptide P01204[(9.03 ± 0.24)%,(19.62 ± 0.44)%,(27.93 ± 2.22)% at ratios of 5 ∶ 1,10 ∶ 1,20 ∶ 1] and HCT-116 cells(MUC4+,HLA-A2+)[(7.26 ± 0.18)%,(16.83 ± 0.40)%,(25.23 ± 1.35)% at ratios of 5 ∶ 1,10 ∶ 1,20 ∶ 1]. The number of CTL induced by peptide P01204 which could secret IFN-γ(130.33 ± 6.58) was obviously higher than that in the negative group. Conclusion:Tumor associated antigen MUC4-derived HLA-A*0201-restrictive cytotoxic T lymphocyte(CTL) epitope P01204 could induce PBMCs’ CTL reaction. The CTL could secret immunologic active material to induce the specific target cells’ lysis.

    参考文献
    相似文献
    引证文献
引用本文

孟 凯,吴峻立,高文涛,苗 毅.肿瘤抗原MUC4 HLA-A*0201限制性CTL表位的体外免疫效应研究[J].南京医科大学学报(自然科学版),2007,(12):1353-1357

复制
分享
文章指标
  • 点击次数:
  • 下载次数:
  • HTML阅读次数:
  • 引用次数:
历史
  • 收稿日期:2007-09-12
  • 最后修改日期:
  • 录用日期:
  • 在线发布日期:
  • 出版日期: