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

被引:196
作者
Mallone, R. [1 ]
Mannering, S. I. [2 ]
Brooks-Worrell, B. M. [3 ]
Durinovic-Bello, I. [4 ]
Cilio, C. M. [5 ]
Wong, F. S. [6 ]
Schloot, N. C. [7 ]
机构
[1] Hop St Vincent de Paul, DeAR Lab Avenir, INSERM, U986, F-75674 Paris 14, France
[2] Univ Melbourne, St Vincents Inst Med Res, Dept Med, St Vincents Hosp, Fitzroy, Vic 3065, Australia
[3] Univ Washington, Vet Affairs Puget Sound Hlth Care Syst, Washington, DC USA
[4] Benaroya Res Inst, Seattle, WA USA
[5] Lund Univ, Dept Clin Sci, Cellular Autoimmun Unit, Malmo, Sweden
[6] Cardiff Univ, Ctr Endocrine & Diabet Sci, Cardiff, S Glam, Wales
[7] Univ Dusseldorf, Inst Clin Diabetol, German Diabet Ctr, Leibniz Inst Diabet Res,Dept Metab Dis,Univ Hosp, Dusseldorf, Germany
关键词
ELISPOT; freezing; shipping; T cell; tetramer; LEPTIN RECEPTOR MUTATION; WHOLE-BLOOD; AUTOIMMUNE ENCEPHALOMYELITIS; LYMPHOCYTE IMMUNOPHENOTYPE; ACETYLCHOLINE-RECEPTOR; CYTOKINE PRODUCTION; ELISPOT DETECTION; OXIDATIVE STRESS; CLINICAL-TRIALS; FLOW-CYTOMETRY;
D O I
10.1111/j.1365-2249.2010.04272.x
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
P>Autoimmune T cell responses directed against insulin-producing beta 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.
引用
收藏
页码:33 / 49
页数:17
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