Department of Biomedical Sciences, University of Minnesota-Duluth, 1035 University Drive, 341 SMED, Duluth, MN 55812, USA.
Department of Pathology, University of Iowa, Iowa City, IA 52242, USA.
Cells. 2023 Sep 28;12(19):2372. doi: 10.3390/cells12192372.
Acute graft versus host disease (GVHD) remains a significant complication following hematopoietic stem cell transplant (HSCT), despite improved human leukocyte antigen (HLA) matching and advances in prophylactic treatment regimens. Previous studies have shown promising results for future regulatory dendritic cell (DCreg) therapies in the amelioration of GVHD. This study evaluates the effects of cryopreservation on the generation of DCreg, the generation of young and older DCreg in serum-free media, and the feasibility of generating DCreg from young and older HSCT patient monocytes. DCregs were generated in X-vivo 15 serum-free media from donor or patient monocytes. This study includes the use of monocytes from young and older healthy, donor, and HSCT patients with varying hematological diseases. Phenotypic differences in cell populations were assessed via flow cytometry while pro-inflammatory and anti-inflammatory cytokine production was evaluated in culture medium. The number of DCreg generated from cryopreserved monocytes of healthy donors was not significantly different from freshly isolated monocytes. DCreg generated from cryopreserved monocytes had comparable levels of co-stimulatory molecule expression, inhibitory molecule expression, and cytokine production as freshly isolated monocytes. Young and older healthy donor monocytes generated similar numbers of DCreg with similar cytokine production and phenotype. Although monocytes from older HSCT patients generated significantly fewer DCreg, DCreg from young and older HSCT patients had comparable phenotypes and cytokine production. Monocytes from young and older myelodysplastic syndrome (MDS) patients generated reduced numbers of DCreg compared to non-MDS-derived DCreg. We demonstrate that the cryopreservation of monocytes from HSCT patients of varying hematological diseases allows for the cost-effective generation of DCreg on an as-needed basis. Although the generation of DCreg from MDS patients requires further assessment, these data support the possibility of -generated DCreg as a therapy to reduce GVHD-associated morbidity and mortality in young and older HSCT recipients.
急性移植物抗宿主病（GVHD）仍然是造血干细胞移植（HSCT）后的一个重大并发症，尽管人类白细胞抗原（HLA）匹配得到改善和预防性治疗方案有所进步。以前的研究表明，未来的调节树突状细胞（DCreg）疗法在改善 GVHD 方面有希望取得成果。本研究评估了冷冻保存对 DCreg 生成的影响、无血清培养基中年轻和年老 DCreg 的生成以及从年轻和年老 HSCT 患者单核细胞中生成 DCreg 的可行性。DCreg 是从供体或患者单核细胞在 X-vivo 15 无血清培养基中生成的。本研究包括使用来自年轻和年老健康、供体和患有各种血液疾病的 HSCT 患者的单核细胞。通过流式细胞术评估细胞群体的表型差异，同时评估培养物中促炎和抗炎细胞因子的产生。来自健康供体冷冻保存单核细胞的 DCreg 生成数量与新鲜分离的单核细胞无显著差异。来自冷冻保存单核细胞的 DCreg 具有与新鲜分离单核细胞相当的共刺激分子表达、抑制性分子表达和细胞因子产生水平。年轻和年老健康供体单核细胞生成的 DCreg 数量相似，细胞因子产生和表型相似。尽管来自老年 HSCT 患者的单核细胞生成的 DCreg 数量明显较少，但来自年轻和年老 HSCT 患者的 DCreg 具有相似的表型和细胞因子产生。与非 MDS 衍生的 DCreg 相比，来自年轻和年老骨髓增生异常综合征（MDS）患者的单核细胞生成的 DCreg 数量减少。我们证明，冷冻保存来自各种血液疾病 HSCT 患者的单核细胞允许在需要时以具有成本效益的方式生成 DCreg。尽管需要进一步评估从 MDS 患者生成 DCreg，但这些数据支持生成的 DCreg 作为降低年轻和年老 HSCT 受者 GVHD 相关发病率和死亡率的治疗方法的可能性。
