Hill Jacqueline A, Cassano Jennifer M, Goodale Margaret B, Fortier Lisa A
Am J Vet Res. 2017 Jul;78(7):867-875. doi: 10.2460/ajvr.78.7.867.
OBJECTIVE To determine whether major histocompatability complex (MHC) class II expression in equine mesenchymal stem cells (MSCs) changes with exposure to a proinflammatory environment reflective of an inflamed joint. SAMPLE Cryopreserved bone marrow-derived MSCs from 12 horses and cartilage and synovium samples from 1 horse euthanized for reasons other than lameness. PROCEDURES In part 1 of a 3-part study, the suitability of a quantitative reverse transcriptase PCR (qRT-PCR) assay for measurement of MHC class II expression in MSCs following stimulation with interferon (IFN)-γ was assessed. In part 2, synoviocyte-cartilage cocultures were or were not stimulated with interleukin (IL)-1β (10 ng/mL) to generate conditioned media that did and did not (control) mimic an inflamed joint environment. In part 3, a qRT-PCR assay was used to measure MSC MHC class II expression after 96 hours of incubation with 1 of 6 treatments (control-conditioned medium, IL-1β-conditioned medium, and MSC medium alone [untreated control] or with IL-1β [10 ng/mL], tumor necrosis factor-α [10 ng/mL], or IFN-γ [100 ng/mL]). RESULTS The qRT-PCR assay accurately measured MHC class II expression. Compared with MHC class II expression for MSCs exposed to the untreated control medium, that for MSCs exposed to IL-1β was decreased, whereas that for MSCs exposed to IFN-γ was increased. Neither the control-conditioned nor tumor necrosis factor-α medium altered MHC class II expression. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that MSC exposure to proinflammatory cytokine IL-1β decreased MHC class II expression and antigenicity. Treatment of inflamed joints with allogeneic MSCs might not be contraindicated, but further investigation is warranted.
目的 确定马间充质干细胞（MSCs）中主要组织相容性复合体（MHC）II类分子的表达是否会因暴露于反映炎症关节的促炎环境而发生变化。样本 来自12匹马的冷冻保存的骨髓源性MSCs以及来自1匹因非跛行原因实施安乐死的马的软骨和滑膜样本。步骤 在一项3部分研究的第1部分中，评估了定量逆转录聚合酶链反应（qRT-PCR）检测法在测量干扰素（IFN）-γ刺激后MSCs中MHC II类分子表达方面的适用性。在第2部分中，滑膜细胞-软骨共培养物分别用白细胞介素（IL）-1β（10 ng/mL）刺激或不刺激，以产生模拟和未模拟（对照）炎症关节环境的条件培养基。在第3部分中，使用qRT-PCR检测法测量在6种处理之一（对照条件培养基、IL-1β条件培养基、单独的MSCs培养基[未处理对照]或与IL-1β[10 ng/mL]、肿瘤坏死因子-α[10 ng/mL]或IFN-γ[100 ng/mL]）孵育96小时后的MSCs MHC II类分子表达。结果 qRT-PCR检测法准确测量了MHC II类分子的表达。与暴露于未处理对照培养基的MSCs的MHC II类分子表达相比，暴露于IL-1β的MSCs的表达降低，而暴露于IFN-γ的MSCs的表达增加。对照条件培养基和肿瘤坏死因子-α培养基均未改变MHC II类分子的表达。结论及临床意义 结果表明，MSCs暴露于促炎细胞因子IL-1β会降低MHC II类分子的表达和抗原性。用同种异体MSCs治疗炎症关节可能并非禁忌，但有必要进一步研究。
