Zhang Le-Shi, Liu Qi-Fa, Huang Ke, Zhang Yu, Fan Zhi-Ping, Huang Shao-Liang
Department of Hematology, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China.
Zhonghua Nei Ke Za Zhi. 2009 Jul;48(7):542-6.
To assess whether treatment with mesenchymal stem cells (MSCs) is an effective adjunct therapy for refractory extensive chronic graft-versus-host disease (GVHD) resistant to conventional therapy.
12 patients with steroid-resistant extensive chronic GVHD were treated with MSCs. One patient received one dose, 10 received two doses, and the remaining three doses. The MSCs were obtained from HLA-identical sibling donors (n = 14), haploidentical donors (n = 2), unrelated mismatched donor (n = 1) and third-party HLA-mismatched donors (n = 7). Of the 11 patients treated with multiple infusions, 5 received cells derived from two donors. The median first dose of MSCs was 1.0 (0.4-2.1) x 10(6)/kg, the median second dose was 1.2 (0.8-1.9) x 10(6)/kg , and the third dose in one patient was 1.1 x 10(6)/kg. Meanwhile the proportion of CD3+, CD4+, CD8+, CD19+, CD4+, CD25+, FOXP3+, FOXP3+ CD4+ and FOXP3 + CD25+ was determined with double fluorescent-labeled antibodies and flow cytometry before and 4 weeks after the MSCs infusion.
No patients had side-effects during or immediately after the infusions of MSCs. After a treatment course of one to three doses, 3 patients had complete response( CR), 6 showed partial response (PR) and 3 did not respond; the total effective rate was 75% (9/12). Complete resolution was seen in the involvement of skin (3/12), lung (1/3), joints (1/5), liver (3/10), oral cavity (4/12) and eye (2/7). Response rate was not related to donor HLA-match. 3 CR patients discontinued all of the immunosuppressive agents without relapse 100 to 292 days after the MSC infusion and 6 PR patients taped all immunosuppressive agents after 60 to 79 days. Mean follow-up period was 1152 (795-1914) days, leukemia free survival rate was 91.7% (11/12) and the overall survival rate was 75% (9/12). The ratio of CD4/CD8 and the proportion of regulatory T cells were significantly higher than that before MSCs treatment.
Third-party MSCs were as effective as HLA-identical or haploidentical cells. This finding has practical implications and suggests that third-party cells can be prepared and stored frozen to be used for steroid-resistant extensive chronic GVHD therapy. It is concluded that MSCs may prevent the lethal cGVHD after allogeneic hematopoietic stem cell transplantation and raise the survival rate by increasing the ratio of CD4/CD8 and proportion of regulatory T cells in vivo.
评估间充质干细胞（MSCs）治疗是否为对传统治疗耐药的难治性广泛性慢性移植物抗宿主病（GVHD）的有效辅助治疗方法。
12例对类固醇耐药的广泛性慢性GVHD患者接受了MSCs治疗。1例患者接受1剂，10例接受2剂，其余3例接受3剂。MSCs取自HLA相同的同胞供者（n = 14）、单倍体供者（n = 2）、不相关的错配供者（n = 1）和第三方HLA错配供者（n = 7）。在接受多次输注治疗的11例患者中，5例接受了来自2个供者的细胞。MSCs的中位首剂为l.0（0.4 - 2.1）×10⁶/kg，中位第二剂为1.2（0.8 - 1.9）×10⁶/kg，1例患者的第三剂为1.1×10⁶/kg。同时，在输注MSCs前及输注后4周，用双荧光标记抗体和流式细胞术测定CD3⁺、CD4⁺、CD8⁺、CD19⁺、CD4⁺、CD25⁺、FOXP3⁺、FOXP3⁺CD4⁺和FOXP3⁺CD25⁺的比例。
在输注MSCs期间或输注后即刻，无患者出现副作用。经过1至3剂的一个疗程治疗后，3例患者完全缓解（CR），6例部分缓解（PR），3例无反应；总有效率为75%（9/12）。皮肤（3/12）、肺（1/3）、关节（1/5）、肝脏（3/10）、口腔（4/12）和眼睛（2/7）的受累情况完全缓解。缓解率与供者HLA配型无关。3例CR患者在输注MSCs后100至292天停用所有免疫抑制剂且未复发，6例PR患者在60至79天后减少了所有免疫抑制剂的用量。平均随访期为1152（795 - 1914）天，无白血病生存率为91.7%（11/12），总生存率为75%（9/12）。CD4/CD8比值和调节性T细胞比例显著高于MSCs治疗前。
第三方MSCs与HLA相同或单倍体细胞同样有效。这一发现具有实际意义，提示可以制备第三方细胞并冷冻保存，用于类固醇耐药的广泛性慢性GVHD治疗。结论是，MSCs可能预防异基因造血干细胞移植后的致死性慢性GVHD，并通过提高体内CD4/CD8比值和调节性T细胞比例提高生存率。
