Isolation and preservation of peripheral blood mononuclear cells for analysis of islet antigen-reactive T cell responses: position statement of the T-Cell Workshop Committee of the Immunology of Diabetes Society

R Mallone, S I Mannering, B M Brooks-Worrell, I Durinovic-Belló, C M Cilio, F S Wong, N C Schloot, T-Cell Workshop Committee, Immunology of Diabetes Society, Barbara M Brooks-Worrell, Corrado M Cilio, Ivana Durinovic-Bellò, Peter A Gottlieb, Roberto Mallone, Stuart I Mannering, Nanette C Schloot, Tim I Tree, Susan Wong, R Mallone, S I Mannering, B M Brooks-Worrell, I Durinovic-Belló, C M Cilio, F S Wong, N C Schloot, T-Cell Workshop Committee, Immunology of Diabetes Society, Barbara M Brooks-Worrell, Corrado M Cilio, Ivana Durinovic-Bellò, Peter A Gottlieb, Roberto Mallone, Stuart I Mannering, Nanette C Schloot, Tim I Tree, Susan Wong

Abstract

Autoimmune T cell responses directed against insulin-producing β cells are central to the pathogenesis of type 1 diabetes (T1D). Detection of such responses is therefore critical to provide novel biomarkers for T1D 'immune staging' and to understand the mechanisms underlying the disease. While different T cell assays are being developed for these purposes, it is important to optimize and standardize methods for processing human blood samples for these assays. To this end, we review data relevant to critical parameters in peripheral blood mononuclear cell (PBMC) isolation, (cryo)preservation, distribution and usage for detecting antigen-specific T cell responses. Based on these data, we propose recommendations on processing blood samples for T cell assays and identify gaps in knowledge that need to be addressed. These recommendations may be relevant not only for the analysis of T cell responses in autoimmune disease, but also in cancer and infectious disease, particularly in the context of clinical trials.

© 2010 The Authors. Clinical and Experimental Immunology © 2010 British Society for Immunology.

Figures

Fig. 1
Fig. 1
Critical steps in processing blood samples for T cell assays.

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