Hemato-Oncology Research Group, Biomedical Research Center, Pamplona, Spain.
Bone Marrow Transplant. 2013 Oct;48(10):1265-70. doi: 10.1038/bmt.2012.262. Epub 2013 Jan 14.
Recipients of hematopoietic SCT undergo a period of profound immunosuppression due to the chemotherapy and/or radiotherapy used for the conditioning and to the graft versus host reaction. SCT patients are highly susceptible to the development of viral infections such as CMV or EBV. The achievement of a competent immunological response, such as viral-specific T cells, is associated with a lower incidence of viral infections. Methods for direct identification of antigen-specific T cells have been based on the functional characteristics of these T cells. Techniques such as proliferation and ELISPOT assays, intracellular cytokine staining and IFN-γ capture have been used to quantitate and obtain viral-specific T cells. Multimers are composed of several MHC molecules loaded with immunodominant peptides joined to a fluorescent molecule, which signal can be quantified by a flow cytometer. Multimer technology together with recent advances in flow cytometry, have facilitated the monitoring and selection of antigen-specific T cells without the need for in vitro cultures and manipulation. This has resulted in a better characterization of the function and phenotype of the different subpopulations of T cells involved in the immune recovery post allogeneic SCT. It is becoming a distinct possibility to isolate individual antigen-specific T cells, without long-term culture techniques, and potentially use them as adoptive immunotherapy in the SCT setting.
接受造血干细胞移植(HSCT)的患者由于预处理过程中使用的化疗和/或放疗以及移植物抗宿主反应而经历了一段严重的免疫抑制期。HSCT 患者极易发生病毒感染,如 CMV 或 EBV。获得有效的免疫应答,如病毒特异性 T 细胞,与较低的病毒感染发生率相关。直接鉴定抗原特异性 T 细胞的方法基于这些 T 细胞的功能特征。增殖和 ELISPOT 测定、细胞内细胞因子染色和 IFN-γ 捕获等技术已被用于定量和获得病毒特异性 T 细胞。多聚体由与荧光分子连接的几个负载免疫优势肽的 MHC 分子组成,其信号可以通过流式细胞仪进行定量。多聚体技术与流式细胞术的最新进展相结合,促进了抗原特异性 T 细胞的监测和选择,而无需体外培养和操作。这使得对同种异体 HSCT 后免疫恢复过程中涉及的不同 T 细胞亚群的功能和表型进行更好的描述成为可能。有可能在不使用长期培养技术的情况下分离单个抗原特异性 T 细胞,并有可能将其用作 HSCT 中的过继免疫疗法。