Liang Chen, Chen Shu-lian, Wang Mei, Zhai Wen-jing, Zhou Zheng, Pang Ai-ming, Feng Si-zhou, Han Ming-zhe
Institute of Hematology and Blood Diseases Hospital, CAMS & PUMC, Tianjin,China.
Zhonghua Xue Ye Xue Za Zhi. 2013 Mar;34(3):213-6. doi: 10.3760/cma.j.issn.0253-2727.2013.03.007.
To investigate mesenchymal stem cells (MSCs) immunosuppressive activity in the presence of interferon-gamma (IFN-γ) to reveal synergistic immunomodulatory effects of IFN-γ and MSCs.
① MSCs were cultured in the presence or absence of IFN-γ（100 ng/ml）, the supernatants were collected for measurements of PGE2、HGF and TGF-β1 by ELISA kits. ② MSCs were cultured in the presence or absence of IFN-γ （100 ng/ml）for 48 h. The cDNA was analysed for the expression of human indoleamine 2, 3-dioxygenase（IDO）mRNA by semiquantitative RT-PCR. ③ Mononuclear cells (MNCs) were extracted from peripheral blood of healthy donors. The T cell proliferation was tested in the co-culture system added with MSCs, recombinant human IFN-γ (100 ng/ml) and anti-IFN-γ mAb (5 μg/ml) by BrdU ELISA kit.
①The immunosuppressive cytokines PGE2、HGF and TGF-β1 were detectable within 24-48 h in the supernatants. Their expressions were significantly up-regulated in the presence of IFN-γ. Concentrations of these cytokines were as of (1715.5±628.6) pg/ml vs (1344.5±709.4) pg/ml (P=0.001);(4031.8±1496.8) pg/ml vs (2452.4±1375.3) pg/ml（P=0.011）;(1753.5±413.8) pg/ml vs (1026.6±450.5) pg/ml（P<0.001）,respectively. ②The expression of IDO mRNA was undetectable when MSCs were cultured alone. In contrast, The IDO mRNA expression was remarkably enhanced in the presence of IFN-γ. ③Bone marrow-derived MSCs remarkably suppressed allogeneic T cell proliferation in vitro. Addition of exogenous IFN-γ had no significant effect on the inhibitory capacity of MSCs, the inhibitory ratios of T cell proliferation were （40.4±10.9）% vs（36.7±7.4）% (P=0.272). By contrast, the inhibitory ratio of T cell proliferation was significantly decreased in the presence of anti-IFN-γ mAb[(40.4±10.9)% vs (23.9±7.6)%,P=0.002].
①Human MSCs constitutively expressed immunosuppressive concentrations of PGE2, HGF and TGF-β1, and their expressions were significantly up-regulated by IFN-γ. ②IFN-γ-induced expression of IDO on MSCs involved in tryptophan catabolism. ③MSCs notably suppressed allogeneic T cell proliferation in vitro. IFN-γ promoted the immunosuppressive capacity of human MSCs, indicating the synergistic immunomodulatory effect of IFN-γ and MSCs.
研究在干扰素-γ（IFN-γ）存在的情况下间充质干细胞（MSCs）的免疫抑制活性，以揭示IFN-γ与MSCs的协同免疫调节作用。
①将MSCs在有或无IFN-γ（100 ng/ml）的条件下培养，收集上清液，用ELISA试剂盒检测PGE2、HGF和TGF-β1。②将MSCs在有或无IFN-γ（100 ng/ml）的条件下培养48小时。用半定量RT-PCR分析cDNA中人吲哚胺2,3-双加氧酶（IDO）mRNA的表达。③从健康供体的外周血中提取单个核细胞（MNCs）。用BrdU ELISA试剂盒在添加了MSCs、重组人IFN-γ（100 ng/ml）和抗IFN-γ单克隆抗体（5 μg/ml）的共培养体系中检测T细胞增殖。
①在24 - 48小时内可在上清液中检测到免疫抑制细胞因子PGE2、HGF和TGF-β1。在IFN-γ存在的情况下，它们的表达显著上调。这些细胞因子的浓度分别为（1715.5±628.6）pg/ml对（1344.5±709.4）pg/ml（P = 0.001）；（4031.8±1496.8）pg/ml对（2452.4±1375.3）pg/ml（P = 0.011）；（1753.5±413.8）pg/ml对（1026.6±450.5）pg/ml（P < 0.001）。②单独培养MSCs时未检测到IDO mRNA的表达。相反，在IFN-γ存在的情况下，IDO mRNA的表达显著增强。③骨髓来源的MSCs在体外显著抑制同种异体T细胞增殖。添加外源性IFN-γ对MSCs的抑制能力无显著影响，T细胞增殖抑制率分别为（40.4±10.9）%对（36.7±7.4）%（P = 0.272）。相比之下，在抗IFN-γ单克隆抗体存在的情况下，T细胞增殖抑制率显著降低[（40.4±10.9）%对（23.9±7.6）%，P = 0.002]。
①人MSCs组成性表达免疫抑制浓度的PGE2、HGF和TGF-β1，且它们的表达被IFN-γ显著上调。②IFN-γ诱导MSCs上IDO的表达参与色氨酸分解代谢。③MSCs在体外显著抑制同种异体T细胞增殖。IFN-γ促进人MSCs的免疫抑制能力，表明IFN-γ与MSCs具有协同免疫调节作用。
