Dimethyl sulfoxide (DMSO) exposure to human peripheral blood mononuclear cells (PBMCs) abolish T cell responses only in high concentrations and following coincubation for more than two hours

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.