为了确保在国家过敏和传染病研究所赞助的多发性硬化症(HALT-MS)和系统性硬化症(SCOT)的研究新药方案中进行自体造血细胞移植(HCT)的可比移植物,实施了控制生产的药物主文件方法,包括通用的主生产批量记录和现场特定的标准操作程序以及“关键要素”。“我们使用健康供体的造血祖细胞(HPCs)评估了流式细胞术和受控速率冷冻保存位点之间的可比性以及冷冻保存移植物的稳定性。造血祖细胞,单采-CD34+富集,自体使用(Auto-CD34+HPC)移植规范包括冷冻保存前≥70%活CD34+细胞.对于两个协议,进行了110次单采;冷冻保存了121批Auto-CD34HPC,其中107个(88.4%)符合发布标准。移植物输注的中位数为25天(范围,17至68)用于HALT-MS的单采后(n=24),和25天(范围,14至78)为SCOT(n=33)。受试者接受的预保存剂量中位数为5.1×106活CD34+细胞/kg(范围,对于HALT-MS和5.6×106活CD34+细胞/kg(范围,2.6至10.2)对于SCOT。粒细胞恢复的中位数为11天(范围,9至15)HALT-MS和10天的HCT后(范围,8到12)对于SCOT,与CD34+细胞剂量无关。受试者接受最后一次血小板输注的中位数为9天(范围,6至16)用于HALT-MS和8天(范围,6至23)对于SCOT;较高的CD34/kg剂量与较快的血小板恢复有关。冷冻保存的健康供体CD34+HPC在6个月的气相液氮储存中的稳定性测试显示出一致的69%至73%的活CD34+细胞回收率。用于HALT-MS和SCOT方案的Auto-CD34+HPC的制造在所有位点上是可比较的,并且支持粒细胞和血小板的及时恢复。
To ensure comparable grafts for autologous hematopoietic cell transplantation (HCT) in the National Institute of Allergy and Infectious Diseases-sponsored Investigational New Drug protocols for multiple sclerosis (HALT-MS) and systemic sclerosis (SCOT), a Drug Master File approach to control manufacture was implemented, including a common Master Production Batch Record and site-specific standard operating procedures with \"Critical Elements.\" We assessed comparability of flow cytometry and controlled rate cryopreservation among sites and stability of cryopreserved grafts using hematopoietic progenitor cells (HPCs) from healthy donors. Hematopoietic Progenitor Cells, Apheresis-CD34+ Enriched, for Autologous Use (Auto-CD34+HPC) graft specifications included ≥70% viable CD34+ cells before cryopreservation. For the 2 protocols, 110 apheresis collections were performed; 121 lots of Auto-CD34+HPC were cryopreserved, and 107 of these (88.4%) met release criteria. Grafts were infused at a median of 25 days (range, 17 to 68) post-apheresis for HALT-MS (n = 24), and 25 days (range, 14 to 78) for SCOT (n = 33). Subjects received precryopreservation doses of a median 5.1 × 106 viable CD34+ cells/kg (range, 3.9 to 12.8) for HALT-MS and 5.6 × 106 viable CD34+ cells/kg (range, 2.6 to 10.2) for SCOT. Recovery of granulocytes occurred at a median of 11 days (range, 9 to 15) post-HCT for HALT-MS and 10 days (range, 8 to 12) for SCOT, independent of CD34+ cell dose. Subjects received their last platelet transfusion at a median of 9 days (range, 6 to 16) for HALT-MS and 8 days (range, 6 to 23) for SCOT; higher CD34+/kg doses were associated with faster platelet recovery. Stability testing of cryopreserved healthy donor CD34+ HPCs over 6 months of vapor phase liquid nitrogen storage demonstrated consistent 69% to 73% recovery of viable CD34+ cells. Manufacturing of Auto-CD34+HPC for the HALT-MS and SCOT protocols was comparable across all sites and supportive for timely recovery of granulocytes and platelets.