从 G-CSF 动员的干细胞移植物中临床级生成肽刺激的 CMV/EBV 特异性 T 细胞。

Clinical-grade generation of peptide-stimulated CMV/EBV-specific T cells from G-CSF mobilized stem cell grafts.

机构信息

Dept. of Hematology/Oncology, University Hospital of Erlangen, Ulmenweg 18, 91054, Erlangen, Germany.

Center for Clinical Studies CCS, University Hospital of Erlangen, Krankenhausstr. 12, 91054, Erlangen, Germany.

出版信息

J Transl Med. 2018 May 9;16(1):124. doi: 10.1186/s12967-018-1498-3.

BACKGROUND

A major complication after allogeneic hematopoietic stem cell transplantation (aSCT) is the reactivation of herpesviruses such as cytomegalovirus (CMV) and Epstein-Barr virus (EBV). Both viruses cause significant mortality and compromise quality of life after aSCT. Preventive transfer of virus-specific T cells can suppress reactivation by re-establishing functional antiviral immune responses in immunocompromised hosts.

METHODS

We have developed a good manufacturing practice protocol to generate CMV/EBV-peptide-stimulated T cells from leukapheresis products of G-CSF mobilized and non-mobilized donors. Our procedure selectively expands virus-specific CD8+ und CD4+ T cells over 9 days using a generic pool of 34 CMV and EBV peptides that represent well-defined dominant T-cell epitopes with various HLA restrictions. For HLA class I, this set of peptides covers at least 80% of the European population.

RESULTS

CMV/EBV-specific T cells were successfully expanded from leukapheresis material of both G-CSF mobilized and non-mobilized donors. The protocol allows administration shortly after stem cell transplantation (d30+), storage over liquid nitrogen for iterated applications, and protection of the stem cell donor by avoiding a second leukapheresis.

CONCLUSION

Our protocol allows for rapid and cost-efficient production of T cells for early transfusion after aSCT as a preventive approach. It is currently evaluated in a phase I/IIa clinical trial.

背景

异基因造血干细胞移植（aSCT）后，巨细胞病毒（CMV）和 EBV 等疱疹病毒的再激活是一个主要并发症。这两种病毒都会导致 aSCT 后死亡率显著增加，并降低生活质量。通过在免疫功能低下的宿主中重新建立功能性抗病毒免疫反应，可以转移病毒特异性 T 细胞来预防病毒再激活。

方法

我们制定了一项良好生产规范（GMP）方案，从 G-CSF 动员和非动员供体的白细胞分离物中产生 CMV/EBV 肽刺激的 T 细胞。我们的方案使用通用的 34 种 CMV 和 EBV 肽池选择性地在 9 天内扩增病毒特异性 CD8+和 CD4+T 细胞，这些肽代表具有不同 HLA 限制的明确主导 T 细胞表位。对于 HLA Ⅰ类，这组肽至少覆盖了 80%的欧洲人群。