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

• Published in: Clinical and experimental immunology Vol. 163; no. 1; pp. 33 - 49
• Main Authors: Mallone, R, Mannering, S.I, Brooks-Worrell, B.M, Durinovic-Belló, I, Cilio, C.M, Wong, F.S, Schloot, N.C
• Format: Journal Article
• Language: English
• Published: Oxford, UK Blackwell Publishing Ltd 01.01.2011; Blackwell; Oxford University Press; Blackwell Science Inc
• Subjects: Analytical, structural and metabolic biochemistry; Animals; Antigens, Surface - immunology; Autoimmune diseases; Autoimmune Diseases - immunology; Basic Medicine; Biological and medical sciences; Blood Preservation - methods; Blood Preservation - standards; Cell Separation - methods; Cell Separation - standards; Clinical Trials as Topic; Cryopreservation - methods; Cryopreservation - standards; Diabetes; Diabetes Mellitus, Type 1 - immunology; Diabetes. Impaired glucose tolerance; ELISPOT; Endocrine pancreas. Apud cells (diseases); Endocrinopathies; Etiopathogenesis. Screening. Investigations. Target tissue resistance; freezing; Fundamental and applied biological sciences. Psychology; Humans; Immunologi inom det medicinska området; Immunology in the medical area; Indexing in process; Infections - immunology; Insulin-Secreting Cells - immunology; Lymphocytes; Medical and Health Sciences; Medical sciences; Medicin och hälsovetenskap; Medicinska och farmaceutiska grundvetenskaper; Mice; Monocytes - immunology; Neoplasms - immunology; Preservation; Review; shipping; Society; T cell; T cell receptors; T-Lymphocytes - immunology; tetramer; Endocrinopathy; Immune response; Preservation; Position; Diabetes mellitus; T cell; Conservation; Biochemistry; Enzyme immunoassay; ELISPOT; Freezing; Antigen; Blood cell; Immunology; Mononuclear cell; shipping; Tetramer; T-Lymphocyte; Isolation; Workshop
• ISSN: 0009-9104; 1365-2249
• DOI: 10.1111/j.1365-2249.2010.04272.x

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.