不同浓度IL-2对体外诱导肽特异性细胞毒性T淋巴细胞培养体系的影响

李桉琪; 祁元明; 周哲骏

doi:10.3969/j.issn.1007-3969.2016.09.006

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不同浓度IL-2对体外诱导肽特异性细胞毒性T淋巴细胞培养体系的影响

更新时间：2025-12-31

- 不同浓度IL-2对体外诱导肽特异性细胞毒性T淋巴细胞培养体系的影响
- The influence of different concentrations of IL-2 on the cultivation system for peptide-specific CTL inductionin vitro
- 中国癌症杂志 2016年26卷第9期页码：756-762
- 作者机构：
  
  1. 郑州大学第一附属医院生物治疗中心,河南,郑州,450052
  2. 郑州大学第一附属医院肿瘤科,河南,郑州,450052
  3. 郑州大学生命科学学院,河南,郑州,450001
- 作者简介：
- 基金信息：
  
  国家自然科学基金面上项目(81171986);国家自然科学基金国际(地区)合作与交流项目(81261120402)。
- DOI：10.3969/j.issn.1007-3969.2016.09.006
  中图分类号：
- 网络出版：2016-10-26，
  
  纸质出版：2016-10-26
- 稿件说明：
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李桉琪,祁元明,周哲骏,等. 不同浓度IL-2对体外诱导肽特异性细胞毒性T淋巴细胞培养体系的影响[J]. 中国癌症杂志, 2016, 26(9): 756-762.

李桉琪, 祁元明, 周哲骏. The influence of different concentrations of IL-2 on the cultivation system for peptide-specific CTL inductionin vitro[J]. China Oncology, 2016, 26(9): 756-762.
李桉琪,祁元明,周哲骏,等. 不同浓度IL-2对体外诱导肽特异性细胞毒性T淋巴细胞培养体系的影响[J]. 中国癌症杂志, 2016, 26(9): 756-762. DOI： 10.3969/j.issn.1007-3969.2016.09.006.

李桉琪, 祁元明, 周哲骏. The influence of different concentrations of IL-2 on the cultivation system for peptide-specific CTL inductionin vitro[J]. China Oncology, 2016, 26(9): 756-762. DOI： 10.3969/j.issn.1007-3969.2016.09.006.

摘要

背景与目的：细胞毒性T淋巴细胞(cytotoxic T lymphocyte，CTL)介导的特异性细胞免疫在抗肿瘤免疫过程中发挥着主要作用。该研究探讨不同浓度IL-2(50、200和1 000 U/mL)对体外诱导表位肽特异性CTL培养体系培养的细胞亚群比例和功能的影响，以及高剂量IL-2是否会诱导该体系中的调节性T细胞(regulatory cell，Treg)的富集。方法：选取HLA-A2超型的肿瘤患者和健康供者各10例，取外周血分离外周血单核细胞，使用环氧合酶-2(Cox-2)来源的CTL表位肽P321(ILIGETIKI)与不同浓度IL-2体外诱导肽特异性CTL。运用流式细胞仪检测细胞的增殖能力、CD4

T淋巴细胞和CD8

T淋巴细胞亚群的比例、Treg细胞亚群的比例，以及CD8

T淋巴细胞分泌穿孔素(perforin)、颗粒酶B(granzyme-B)、干扰素IFN-γ的能力。使用Elispot实验检测实验组细胞分泌IFN-γ的能力。结果：高浓度的IL-2有利于细胞的增殖。肿瘤患者组的CD4

T淋巴细胞比例高于健康供者组，而CD8

T淋巴细胞比例较健康供者组低；不同浓度IL-2对CD4

T淋巴细胞、CD8

T淋巴细胞和Treg细胞亚群比例以及CD8

T淋巴细胞分泌穿孔素、颗粒酶B、IFN-γ的能力没有影响。随着IL-2浓度的增高，Elispot实验出现的阳性斑点数越多。结论：不同浓度IL-2条件对体外诱导表位肽特异性CTL培养体系培养出的细胞亚群比例和功能没有影响，在50~1 000 U/mL IL-2浓度范围内，高剂量的IL-2不会诱导该体系中的Treg的富集。但高浓度的IL-2可以提高细胞的增殖能力，并且促进细胞分泌IFN-γ，而高浓度的IL-2可以使培养体系中存在的NK细胞或NKT细胞能够非特异性产生IFN-γ，从而对Elispot实验产生干扰。因此，在体外诱导肽特异性CTL时选用50 U/mL的IL-2浓度可以很好地维持T细胞的增殖和存活，并且最大程度地降低对Elispot实验的干扰。

Abstract

Background and purpose: Cytotoxic T lymphocyte (CTL) plays a vital role in the process of antitumor immunology. The aim of this study was to investigate whether changes in concentration of IL-2 (50

200 and 1 000 U/mL) would affect the sub-population and cytotoxic function of cells cultivated by peptide-specific CTL induction system in vitro and also observe whether using the concentration of IL-2 at a range of 50-1 000 U/mL is beneficial to regulatory cells (Tregs) enrichment. Methods: Peripheral blood from 10 healthy donors and 10 cancer patients that were HLA-A2 positive

were collected in the study. HLA-A2 restricted CTL epitope P321 (ILIGETIKI) derived from COX-2 pulsed with different concentrations of IL-2 were used to induce peptides-specific CTL in vitro. Flow cytometry was performed to analyze the proliferative capability

the proportion of different T-cell subsets

and secretion of perforin

granzyme B and IFN-γ. IFN-γ secretion was assessed by ELISpot assay. Results: High concentration of IL-2 increased the proliferative activity. The percentage of CD4+ T cells of cancer patient group was significantly higher than that of healthy donor group

while the percentage of CD8+ T cells of cancer patient group was significantly lower than that of healthy donor group. And there was no significant difference in the percentages of CD4+ T cells

CD8+ T cells and Tregs among groups with different IL-2 concentrations. No difference was seen in cytokine (perforin

granzyme B

IFN-γ) secretion capacity of CD8+ T cells. ELISpot study revealed that high-dose IL-2 resulted in the increasing of IFN-γ secretion. Conclusion: The sub-population and the function of cells cultured by peptide-specific CTL induction system in vitro are not affected by different concentrations of IL-2. Furthermore

high concentrations of IL-2 (50-1 000 U/mL) do not provide the enrichment for Tregs. Higher concentration of IL-2 is likely to cause high secretion of IFN-γ in ELISpot assay. In order to exclude the distraction of NK cells or NKT cells

the concentration of 50 U/mL is better choice.

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