调节性（或“容忍性”）树突状细胞（DCregs）是诱导或恢复特定抗原耐受性的高度有前途的创新细胞疗法，适用于免疫介导的炎症性疾病，包括器官移植物排斥、骨髓移植后的移植物抗宿主病和各种自身免疫性疾病。适用于移植的DCregs具有潜力降低患者对非特异性免疫抑制药物的依赖，该类药物可能会引起严重的副作用，增加感染和某些癌症的风险。本研究目的是提供我们制备和验证大量符合良好制造规范的DCregs用于静脉注射给28例器官（肝脏）移植受者的经验，并讨论影响满足释放标准和达到目标细胞数的因素。DCregs是通过从同意的健康成年预定肝脏移植供体的非移动的白细胞分离产物中精选单核细胞分数，并在粒细胞-巨噬细胞集落刺激因子和白细胞介素（IL）-4中培养7天生成的。在培养期间的第0天和第4天加入维生素D3，第4天加入IL-10。释放和后释放标准包括细胞活力、纯度、表型、无菌和功能评估。单核细胞转化为DCregs的整体转化率为28±8.2％，产品存活率为94±5.1％。细胞表面T细胞共抑制：共刺激分子（程序性死亡配体-1：CD86）平均荧光强度比为3.9±2.2，CD40配体刺激后分泌的抗炎细胞因子：促炎细胞因子比（IL-10：IL-12p70）的平均比值为60±63（中位数=40）。从单个白细胞分离产物（n=25个供体）和两个白细胞分离产物（n=3个供体）生成的DCregs总数平均为489±223×106（n=28）。平均注射的DCregs总数为5.9±2.8×106/kg体重。对于27个产品中的25个（92.6％），达到了2.5-10×106/kg的目标细胞范围的DCreg数目。在良好制造规范条件下，从循环血单核细胞中很容易生成高纯度、符合多种质量标准的DCregs，以满足注入潜在器官移植受者的目标细胞数。版权所有©2022 年国际细胞和基因治疗协会。由Elsevier Inc.发表。保留所有权利。

Regulatory (or "tolerogenic") dendritic cells (DCregs) are a highly promising, innovative cell therapy for the induction or restoration of antigen-specific tolerance in immune-mediated inflammatory disorders. These conditions include organ allograft rejection, graft-versus-host disease following bone marrow transplantation and various autoimmune disorders. DCregs generated for adoptive transfer have potential to reduce patients' dependence on non-specific immunosuppressive drugs that can induce serious side effects and enhance the risk of infection and certain types of cancer. Here, our aim was to provide a detailed account of our experience manufacturing and validating comparatively large numbers of Good Manufacturing Practice-grade DCregs for systemic (intravenous) infusion into 28 organ (liver) transplant recipients and to discuss factors that influence the satisfaction of release criteria and attainment of target cell numbers.DCregs were generated in granulocyte-macrophage colony stimulating factor and interleukin (IL)-4 from elutriated monocyte fractions isolated from non-mobilized leukapheresis products of consenting healthy adult prospective liver transplant donors. Vitamin D3 was added on day 0 and 4 and IL-10 on day 4 during the 7-day culture period. Release and post-release criteria included cell viability, purity, phenotype, sterility and functional assessment. The overall conversion rate of monocytes to DCregs was 28 ± 8.2%, with 94 ± 5.1% product viability. The mean cell surface T-cell co-inhibitory to co-stimulatory molecule (programmed death ligand-1:CD86) mean fluorescence intensity ratio was 3.9 ± 2.2, and the mean ratio of anti-inflammatory:pro-inflammatory cytokine product (IL-10:IL-12p70) secreted upon CD40 ligation was 60 ± 63 (median = 40). The mean total number of DCregs generated from a single leukapheresis product (n = 25 donors) and from two leukapheresis products (n = 3 donors) was 489 ± 223 × 106 (n = 28). The mean total number of DCregs infused was 5.9 ± 2.8 × 106 per kg body weight. DCreg numbers within a target cell range of 2.5-10 × 106/kg were achieved for 25 of 27 (92.6%) of products generated.High-purity DCregs meeting a range of quality criteria were readily generated from circulating blood monocytes under Good Manufacturing Practice conditions to meet target cell numbers for infusion into prospective organ transplant recipients.Copyright © 2022 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.