A reproducible immunopotency assay to measure mesenchymal stromal cell-mediated T-cell suppression.
A reproducible immunopotency assay to measure mesenchymal stromal cell-mediated T-cell suppression.
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DOI：
10.1016/j.jcyt.2014.10.002
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发表时间：
2015-02
期刊：
影响因子：
4.5
通讯作者：
Hematti, Peiman
中科院分区：
文献类型：
--
作者：
Bloom, Debra D.;Centanni, John M.;Bhatia, Neehar;Emler, Carol A.;Drier, Diana;Leverson, Glen E.;Mckenna, David H., Jr.;Gee, Adrian P.;Lindblad, Robert;Hei, Derek J.;Hematti, Peiman
骨髓间充质干细胞（MSCs）的T细胞抑制特性已被认为是其在许多人类临床试验中利用的主要作用模式和基础。然而，没有建立良好的可重复的测定来测量MSC介导的T细胞抑制。在威斯康星大学麦迪逊分校细胞治疗生产辅助（PACT）中心，我们开发了一种体外质量控制T细胞抑制免疫效力测定（IPA），其利用抗CD 3和抗CD 28抗体刺激T细胞增殖。我们测量了MSC诱导的抑制CD 4 + T细胞增殖在不同的效应细胞与靶细胞的比例，使用定义的外周血单核细胞，并平行比较参考标准MSC产品。我们计算了每种MSC产品抑制CD 4 + T细胞的IPA值。在三个独立的PACT中心产生的11个MSC产品的细胞表面表型标记和T细胞抑制特性进行了评价。流式细胞术结果显示了典型的MSC细胞表面标志物谱。T细胞增殖抑制水平存在显著差异，IPA值范围为27%至88%。然而，MSC抑制与HLA-DR表达无关。我们已经开发了一种可重复的免疫效价测定来测量同种异体MSC介导的CD 4 + T细胞抑制。可能需要进行额外的研究，以确定这些体外试验结果如何与MSC的其他免疫调节特性相关，以及评估该试验预测体内疗效的能力。
The T cell suppressive property of bone marrow derived mesenchymal stromal cells (MSCs) has been considered a major mode of action and basis for their utilization in a number of human clinical trials. However, there is no well-established reproducible assay to measure MSC-mediated T cell suppression. At the University of Wisconsin-Madison Production Assistance for Cellular Therapy (PACT) Center we developed an in vitro quality control T cell suppression immunopotency assay (IPA) which utilizes anti-CD3 and anti-CD28 antibodies to stimulate T cell proliferation. We measured MSC-induced suppression of CD4+ T cell proliferation at various effector to target cell ratios using defined peripheral blood mononuclear cells and in parallel compared to a reference standard MSC product. We calculated an IPA value for suppression of CD4+ T cells for each MSC product. Eleven MSC products generated at three independent PACT centers were evaluated for cell surface phenotypic markers and T cell suppressive properties. Flow cytometry results demonstrated typical MSC cell surface marker profiles. There was significant variability in the level of suppression of T cell proliferation with IPA values ranging from 27% to 88%. However, MSC suppression did not correlate with HLA-DR expression. We have developed a reproducible immunopotency assay to measure allogeneic MSC-mediated suppression of CD4+ T cells. Additional studies may be warranted to determine how these in vitro assay results may correlate with other immunomodulatory properties of MSCs, in addition to evaluating the ability of this assay to predict in vivo efficacy.
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