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首页> 外文期刊>Cytotherapy >Levels of IL-17F and IL-33 correlate with HLA-DR activation in clinical-grade human bone marrow-derived multipotent mesenchymal stromal cell expansion cultures
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Levels of IL-17F and IL-33 correlate with HLA-DR activation in clinical-grade human bone marrow-derived multipotent mesenchymal stromal cell expansion cultures

机译：IL-17F和IL-33的水平与临床级人骨髓源性骨髓间充质细胞膨胀培养物中的HLA-DR活化相关

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Background aims: Multipotent mesenchymal stromal cell (MSC)-based medicines are extensively investigated for use in regenerative medicine and immunotherapy applications. The International Society for Cell and Gene Therapy (ISCT) proposed a panel of cell surface molecules for MSC identification that includes human leukocyte antigen (HLA)-DR as a negative marker. However, its expression is largely unpredictable despite production under tightly controlled conditions and compliance with current Good Manufacturing Practices. Herein, we report the frequency of HLA-DR expression in 81 batches of clinical grade bone marrow (BM)-derived MSCs and investigated its impact on cell attributes and culture environment. Methods: The levels of 15 cytokines (interleukin [IL]-1b, IL-4, IL-6, IL-10, IL-17A, IL-17F, IL-21, IL-22, IL-23, IL-25, IL-31, IL-33, interferon-gamma, soluble CD40 ligand and tumor necrosis factor-alpha) were determined in sera supplements and supernatants of BM-MSC cultures. Identity, multipotentiality and immunopotency assays were performed on high (20% of cells) and low (= 20% of cells) HLA-DR+ cultures. Results: A correlation was found between HLA-DR expression and levels of IL-17F and IL-33. Expression of HLA-DR did neither affect MSC identity, in vitro tri-lineage differentiation potential (into osteogenic, chondrogenic and adipogenic lineages), nor their ability to inhibit the proliferation of stimulated lymphocytes. Discussion: Out of 81 batches of BM-MSCs for autologous use analyzed, only three batches would have passed the ISCT criteria (2%), whereas 60.5% of batches were compliant with low HLA-DR values (= 20%). Although a cause-effect relationship cannot be drawn, we have provided a better understanding of signaling events and cellular responses in expansion culture conditions relating with HLA-DR expression.
机译：背景技术：随着再生医学和免疫疗法应用，广泛研究了多能间充质细胞（MSC）的药物。国际细胞和基因治疗学会（ISCT）提出了一种细胞表面分子的组，用于MSC鉴定，其包括人白细胞抗原（HLA）-DR作为阴性标记物。然而，尽管在紧密控制的条件下生产和遵守当前的良好制造实践，但其表达在很大程度上是不可预测的。在此，我们报告了81批次临床较骨髓（BM）的HLA-DR表达的频率，并研究了其对细胞属性和培养环境的影响。方法：15个细胞因子（白细胞介素[IL] -1B，IL-4，IL-6，IL-10，IL-17A，IL-17F，IL-21，IL-22，IL-23，IL-25的水平在BM-MSC培养物的血清补充剂和上清液中测定IL-31，IL-33，干扰素-γ-γ，可溶性CD40配体和肿瘤坏死因子-α。在高（＆ 20％的细胞）和低（＆ = 20％的细胞）HLA-DR +培养物中进行同一性，多电位性和免疫力测定。结果：在HLA-DR表达和IL-17F和IL-33的水平之间发现了相关性。 HLA-DR的表达既不影响MSC身份，体外三谱分化电位（成骨，软骨和脂肪和促进谱系），也不能抑制刺激淋巴细胞增殖的能力。讨论：出于81批BM-MSCs的BM-MSC分析，只有三个批次通过ISCT标准（＆lt 2％），而60.5％的批次符合低HLA-DR值（＆ = 20％） ）。虽然不能绘制致原效应关系，但我们提供了对与HLA-DR表达相关的扩增培养条件中的信令事件和细胞反应更好地理解。

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