Page 17 - 南京医科大学学报自然科学版

第42卷第7期 南京医科大学学报（自然科学版）
2022年7月 Journal of Nanjing Medical University（Natural Sciences） ·913 ·

·基础研究·

负载HIP的耐受性树突状细胞对BDC2.5 T细胞的免疫调节作用

祁颜艳，顾 愹，杨 涛 *

南京医科大学第一附属医院内分泌科，江苏 南京 210029

［摘 要］ 目的：探究负载 HIP（hybrid insulin peptide）的耐受性树突状细胞（tolerogenic dendritic cell，tolDC）对致糖尿病性
BDC2.5 T细胞的免疫调节作用。方法：采用细胞因子诱导生成NOD（non⁃obese diabetic）小鼠骨髓细胞来源的未成熟树突状细
胞（immature dendritic cell，iDC），培养过程中额外加入维生素 D3 可生成 tolDC，脂多糖（lipopolysaccharide，LPS）刺激 24 h 后可
以分别获得 LPS⁃iDC、LPS⁃tolDC，收集其上清液检测细胞因子白介素（interleukin，IL）⁃12p70 及转化生长因子⁃β（transforming
growth factor⁃β，TGF⁃β），通过形态学及流式细胞术鉴定上述树突状细胞（dendritic cell，DC）的表型。将致糖尿病性 T 细胞即
BDC2.5 CD4 T 细胞与负载 HIP 的 4 组 DC 共孵育 72 h 后，检测 T 细胞增殖、活化以及调节性 T 细胞（regulatory T cell，Treg）的
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产生情况。结果：表型鉴定结果显示 tolDC 呈现低表达共刺激分子 CD80 和 CD86、高表达共抑制分子 PD⁃L1 的耐受性表型，
在脂多糖的刺激下仍保持稳定，且与LPS⁃iDC相比，LPS⁃tolDC 分泌低水平的IL⁃12p70、高水平的TGF⁃β。负载HIP的tolDC及
LPS⁃tolDC均可以抑制BDC2.5 T细胞的增殖和活化，诱导抗原特异性Treg的产生。结论：负载HIP的tolDC可以通过其稳定的
耐受表型及功能抑制致糖尿病性BDC2.5 T细胞的增殖活化，诱导抗原特异性Treg的产生。
［关键词］ 1型糖尿病；耐受性树突状细胞；免疫治疗；表位
［中图分类号］ R587.1 ［文献标志码］ A ［文章编号］ 1007⁃4368（2022）07⁃913⁃08
doi：10.7655/NYDXBNS20220702

The immunoregulatory effect of HIPs⁃loaded tolerogenic dendritic cells on BDC2.5 T cells

QI Yanyan，GU Yong，YANG Tao *
Department of Endocrinology，the First Affiliated Hospital of Nanjing Medical University，Nanjing 210029，China

［Abstract］ Objective：To investigate the immunoregulatory effect of tolerogenic dendritic cells（tolDCs）loaded with hybrid insulin
peptides（HIPs）on diabetogenic BDC2.5 T cells. Methods：Bone marrow derived imature DCs（iDCs）of non⁃obese diabetic（NOD）
mice were induced with cytokines，and additional vitamin D3 was added to generate tolDCs. After 24 h stimulation with
lipopolysaccharide（LPS），the supernatants of LPS⁃iDCs and LPS⁃tolDCs were collected to detect cytokines IL⁃12p70 and TGF⁃β，and
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the phenotype of the above DCs was identified by morphology and flow cytometry. Diabetogenic T cells，namely BDC2.5 CD4 T cells
were incubated with HIPs ⁃ loaded DCs for 3 days，and the proliferation，activation and regulatory T cells（Tregs）production were
detected. Results：Phenotypic identification results showed that tolDCs had low expression of costimulatory molecules CD80，CD86，
and high expression of coinhibitory molecule PD⁃L1. The phenotype of tolDCs remained stable under the stimulation of LPS，and the
secretion of IL⁃12p70 was low while the secretion of TGF⁃β was high compared with LPS⁃iDCs. Both tolDCs and LPS⁃tolDCs loaded
with HIPs could inhibit the proliferation and activation of BDC2.5 T cells and induce the production of antigen ⁃ specific Tregs.
Conclusion：HIPs loaded tolerogenic dendritic cells can inhibit the proliferation and activation of diabetogenic BDC2.5 T cells and
promote the production of Tregs through their stable tolerance phenotype and function.
［Key words］ type 1 diabetes mellitus；tolerogenic dendritic cell；immunotherapy；epitope
［J Nanjing Med Univ，2022，42（07）：913⁃920］

1 型糖尿病（type 1 diabetes mellitus，T1DM）是

［基金项目］ 国家自然科学基金（81770777，82170837） 一种 T 细胞介导的以胰岛β细胞靶向破坏为特征的
∗ 自身免疫疾病，自身反应性T细胞在β细胞破坏中起
通信作者（Corresponding author），E⁃mail：yangt@njmu.edu.cn