Koehl Ulrike, Sörensen Jan, Esser Ruth, Zimmermann Stefanie, Grüttner Hans Peter, Tonn Torsten, Seidl Christian, Seifried Erhard, Klingebiel Thomas, Schwabe Dirk
University Hospital Frankfurt, Paediatric Haematology and Oncology, Frankfurt, Germany.
Blood Cells Mol Dis. 2004 Nov-Dec;33(3):261-6. doi: 10.1016/j.bcmd.2004.08.013.
Natural killer (NK) cells are thought to be of benefit in HLA-mismatched hematopoietic transplantation (H-SCT). Therefore, we developed a protocol for clinical-use expansion of highly enriched and IL-2-stimulated NK cells. Purification of unstimulated leukaphereses by a two-step T cell depletion with a final CD56 enrichment procedure leads to a mean purity of 95% CD56(+)CD3- NK cells with a four- to five-log depletion of T cells. So far, three pediatric patients with multiply relapsed acute lymphoblastic leukemia (ALL) or acute myelogenous leukemia (AML) were treated with repeated transfusions post-H-SCT. Directed killer immunoglobulin-like receptor (KIR) mismatches were demonstrated in all three cases. Although all patients showed blast persistence at the time of transplant, they reached complete remission and complete donor chimerism within 1 month post-H-SCT. NK cell therapy was tolerated well without graft-versus-host disease (GvHD) induction or other adverse events. The AML patient died of early relapse on day +80, while the ALL patients died of thrombotic-thrombocytopenic purpura and atypical viral pneumonia on days +45 and +152, respectively. This initial trial showed the feasibility of good manufacturing practice (GMP)-compliant NK cell isolation and expansion for clinical applications. We now launch a clinical phase I trial with activated NK cells post-H-SCT.
自然杀伤(NK)细胞被认为在人类白细胞抗原(HLA)不匹配的造血干细胞移植(H-SCT)中有益。因此,我们制定了一种用于临床应用的高度富集且经白细胞介素-2刺激的NK细胞扩增方案。通过两步T细胞去除和最终的CD56富集程序对未刺激的白细胞单采物进行纯化,可使CD56(+)CD3-NK细胞的平均纯度达到95%,T细胞减少四到五个对数级。到目前为止,三名多次复发的急性淋巴细胞白血病(ALL)或急性髓细胞白血病(AML)儿科患者在H-SCT后接受了重复输血治疗。在所有三例中均证实了定向杀伤细胞免疫球蛋白样受体(KIR)不匹配。尽管所有患者在移植时均表现出原始细胞持续存在,但他们在H-SCT后1个月内达到完全缓解并实现完全供体嵌合。NK细胞疗法耐受性良好,未诱发移植物抗宿主病(GvHD)或其他不良事件。AML患者在第80天死于早期复发,而ALL患者分别在第45天和第152天死于血栓性血小板减少性紫癜和非典型病毒性肺炎。这项初步试验表明了符合药品生产质量管理规范(GMP)的NK细胞分离和扩增用于临床应用的可行性。我们现在启动一项H-SCT后使用活化NK细胞的临床I期试验。