负载HIP的耐受性树突状细胞对BDC2.5T细胞的免疫调节作用
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E-mail:yangt@njmu.edu.cn

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R587.1

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国家自然科学基金(81770777,82170837)


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

    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 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.

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祁颜艳,顾愹,杨涛.负载HIP的耐受性树突状细胞对BDC2.5T细胞的免疫调节作用[J].南京医科大学学报(自然科学版),2022,42(7):913-920

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  • 在线发布日期: 2022-07-15
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