Department of Virology, Statens Serum Institut, Denmark.
J Immunol Methods. 2010 Apr 30;356(1-2):70-8. doi: 10.1016/j.jim.2010.01.014. Epub 2010 Feb 13.
Immunotherapies based on reinfusion of autologous cells incubated ex vivo with peptides reconstituted in toxic solvents, such as DMSO, are now performed on a routine basis. However, the toxic effects of the most common solvent used, DMSO, on T cell responses from human PBMCs, have not previously been evaluated in detail. Here, in preparation for a first-in-man human phase I vaccine trial comprising reinfusion of autologous HIV peptide-pulsed PBMCs, human PBMCs from healthy and HIV-infected donors were exposed in vitro to a range of DMSO concentrations, and for a range of time periods. Polychromatic flow cytometry was used to evaluate the influence of DMSO on functional T cell responses. We report that high concentrations of up to 10% of DMSO for 1 hour do not affect the cell viability, the magnitude or the functional profile of CD4(+) and CD8(+) T cell responses, regardless of antigen specificity and HLA class I restriction. In contrast, >2% for >2 hours compromises these responses. These data are relevant in the design of immunotherapies based on pulsing a large number of peptides onto antigen presenting cells prior to reinfusion.
基于用有毒溶剂（如 DMSO）重建的自体细胞培养物的免疫疗法，目前已常规用于临床。然而，迄今为止，尚未详细评估最常用溶剂 DMSO 对来自人 PBMC 的 T 细胞反应的毒性作用。在这里，为了准备包含自体 HIV 肽脉冲 PBMC 再输注的首次人体 I 期疫苗试验，健康和 HIV 感染供体的人 PBMC 体外暴露于一系列 DMSO 浓度和一系列时间段。多色流式细胞术用于评估 DMSO 对功能性 T 细胞反应的影响。我们报告称，高达 10%的高浓度 DMSO 作用 1 小时不会影响细胞活力、CD4(+)和 CD8(+)T 细胞反应的强度或功能谱，无论抗原特异性和 HLA 类 I 限制如何。相比之下，超过 2%的 DMSO 作用超过 2 小时会损害这些反应。这些数据对于设计基于将大量肽脉冲到抗原呈递细胞上，然后再进行再输注的免疫疗法很重要。
