Generation and characterisation of therapeutic tolerogenic dendritic cells for rheumatoid arthritis
Generation and characterisation of therapeutic tolerogenic dendritic cells for rheumatoid arthritis
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DOI：
10.1136/ard.2009.126383
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发表时间：
2010-11-01
影响因子：
27.4
通讯作者：
Hilkens, Catharien M. U.
中科院分区：
文献类型：
--
作者：
Harry, Rachel A.;Anderson, Amy E.;Hilkens, Catharien M. U.
目的致耐受性树突状细胞（tolDCs）作为靶向自身反应性T细胞治疗类风湿性关节炎（RA）等自身免疫性疾病的一种有效方法。作者的目标是将类风湿关节炎的tolDC疗法引入临床。在这里，作者解决了与使用当前良好制造规范（cGMP）-符合试剂从RA患者制造tolDC相关的关键翻译问题，tolDC的稳定性，以及合适的质量控制标记物的选择。单磷酰脂质A，在符合cGMP的培养基CellGroDC中。通过流式细胞术、[H-3]胸苷掺入和ELISA.Results确定了tolDC和tolDC调节的自体CD 4 T细胞的功能性，从RA患者建立的临床级tolDC表现出典型的致耐受性表型，与HC tolDC相比，其具有减少的共刺激分子、促炎细胞因子的低产生和自体抗原特异性T细胞的刺激受损。Toll样受体2（TLR-2）由tolDC而不是成熟DC高度表达。此外，tolDC抑制成熟DC诱导的T细胞增殖、干扰素。和白细胞介素17的产生，并使T细胞对进一步的刺激反应迟钝。重要的是，tolDCs是表型稳定的免疫抑制药物的情况下，是难治性的进一步挑战与促炎mediators.Conclusions建立从RA患者的tolDCs来自健康供体。TLR-2被鉴定为用于tolDC的质量控制的理想标志物。这些tolDC具有潜在的致耐受性和高度稳定性，是治疗RA中定制免疫调节的有前景的细胞治疗剂。
Objectives Tolerogenic dendritic cells (tolDCs) constitute a promising experimental treatment for targeting autoreactive T cells in autoimmune diseases, including rheumatoid arthritis (RA). The authors' goal is to bring tolDC therapy for RA to the clinic. Here the authors address key translational issues related to the manufacturing of tolDCs from RA patients with current good manufacturing practice (cGMP)-compliant reagents, the stability of tolDCs, and the selection of suitable quality control markers.Methods Human monocyte-derived tolDCs were established from RA patients and healthy controls (HCs) using the immunosuppressive drugs dexamethasone and vitamin D 3, and the cGMP-grade immunomodulator, monophosphoryl lipid A, in the cGMP-compliant medium, CellGroDC. The functionality of tolDCs and tolDC-modulated autologous CD4 T cells was determined by flow cytometry, [H-3] thymidine incorporation and ELISA.Results Clinical-grade tolDCs established from patients with RA exhibit a typical tolerogenic phenotype of reduced costimulatory molecules, low production of proinflammatory cytokines and impaired stimulation of autologous antigen-specific T cells, comparable to HC tolDCs. Toll-like receptor 2 (TLR-2) was highly expressed by tolDCs but not mature DCs. Furthermore, tolDCs suppressed mature DC-induced T cell proliferation, interferon. and interleukin 17 production, and rendered T cells hyporesponsive to further stimulation. Importantly, tolDCs were phenotypically stable in the absence of immunosuppressive drugs and were refractory to further challenge with proinflammatory mediators.Conclusions tolDCs established from patients with RA are comparable to those derived from healthy donors. TLR-2 was identified as an ideal marker for quality control of tolDCs. Potently tolerogenic and highly stable, these tolDCs are a promising cellular therapeutic for tailored immunomodulation in the treatment of RA.