Wang N, Wang Y, Jiang P, Lü M, Hu Z, Xu X
Institute of Basic Medicine, Xi'an Medical University, Xi'an 710021, China.
Department of General Practitioners, Xi'an Medical University, Xi'an 710021, China.
Nan Fang Yi Ke Da Xue Xue Bao. 2022 Sep 20;42(9):1288-1295. doi: 10.12122/j.issn.1673-4254.2022.09.03.
To explore the role of DNAM-1 in the activation, proliferation and function of type Ⅰ regulatory T cells (Tr1 cells).
Anti-CD3/CD28 antibodies were used to stimulate mouse T cells derived from the spleen of wild-type (WT) mice, and the expression level of DNAM-1 in resting and activated Tr1 cells was evaluated with flow cytometry. Na?ve CD4 T cells isolated by magnetic cell sorting from the spleens of WT mice and DNAM-1 knockout (KO) mice were cultured in Tr1 polarizing conditions for 3 days, after which CD25 and CD69 expressions were measured using flow cytometry. The induced Tr1 cells were labelled with CFSE and cultured in the presence of anti-CD/CD28 antibodies for 3 days, and their proliferative activity was analyzed. The expressions of IL-10 and p-STAT5 in DNAM-1-deficient Tr1 cells were detected before and after IL-2 stimulation.
The expression level of DNAM-1 was significantly upregulated in CD4 T cells and Tr1 cells after stimulation with anti-CD3/CD28 antibodies ( < 0.05). DNAM-1 knockout did not cause significant changes in the number or proportion of Tr1 cells, but but significantly increased the expression levels of the activation markers CD69 and CD25 ( < 0.05). Compared with WT Tr1 cells, DNAM-1-deficient Tr1 cells exhibited reduced proliferative activity ( < 0.05) with downregulated IL-10 production ( < 0.05) and decreased expressions of Il-10 and Gzmb mRNA ( < 0.05). In DNAM-1-deficient Tr1 cells, IL-2 stimulation significantly reduced IL-10 secretion level and the expression of p-STAT5 as compared with WT Tr1 cells.
DNAM-1 participate in the activation and proliferation of Tr1 cells and affect the biological functions of Tr1 cells through the IL-2/STAT5 pathway.
探讨DNAX辅助分子-1（DNAM-1）在Ⅰ型调节性T细胞（Tr1细胞）激活、增殖及功能中的作用。
用抗CD3/CD28抗体刺激野生型（WT）小鼠脾脏来源的小鼠T细胞，采用流式细胞术评估静息和激活的Tr1细胞中DNAM-1的表达水平。通过磁珠细胞分选从WT小鼠和DNAM-1基因敲除（KO）小鼠脾脏中分离出的初始CD4 T细胞，在Tr1极化条件下培养3天，之后用流式细胞术检测CD25和CD69的表达。将诱导的Tr1细胞用羧基荧光素二乙酸琥珀酰亚胺酯（CFSE）标记，并在抗CD3/CD28抗体存在的情况下培养3天，分析其增殖活性。在白细胞介素-2（IL-2）刺激前后，检测DNAM-1缺陷型Tr1细胞中白细胞介素-10（IL-10）和磷酸化信号转导子和转录激活子5（p-STAT5）的表达。
用抗CD3/CD28抗体刺激后，CD4 T细胞和Tr1细胞中DNAM-1的表达水平显著上调（P<0.05）。DNAM-1基因敲除未引起Tr1细胞数量或比例的显著变化，但显著增加了激活标志物CD69和CD25的表达水平（P<0.05）。与WT Tr1细胞相比，DNAM-1缺陷型Tr1细胞的增殖活性降低（P<0.05），IL-10产生下调（P<0.05），Il-10和颗粒酶B（Gzmb）mRNA的表达降低（P<0.05）。在DNAM-1缺陷型Tr1细胞中，与WT Tr1细胞相比，IL-2刺激显著降低了IL-10分泌水平和p-STAT5的表达。
DNAM-1参与Tr1细胞的激活和增殖，并通过IL-2/STAT5途径影响Tr1细胞的生物学功能。
