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Comparison of cryopreservation methods on T‐cell responses to islet and control antigens from type 1 diabetic patients and controls
Author(s) -
BrooksWorrell B.,
Tree T.,
Mannering S. I.,
DurinovicBello I.,
James E.,
Gottlieb P.,
Wong S.,
Zhou Z.,
Yang L.,
Cilio C. M.,
Reichow J.,
Menart B.,
Rutter R.,
Schreiner R.,
Pham M.,
Petrich de Marquesini L.,
Lou O.,
Scotto M.,
Mallone R.,
Schloot N. C.
Publication year - 2011
Publication title -
diabetes/metabolism research and reviews
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.307
H-Index - 110
eISSN - 1520-7560
pISSN - 1520-7552
DOI - 10.1002/dmrr.1245
Subject(s) - peripheral blood mononuclear cell , cryopreservation , islet , cd8 , type 1 diabetes , andrology , antigen , immunology , viability assay , cell , t cell , biology , medicine , diabetes mellitus , immune system , endocrinology , in vitro , microbiology and biotechnology , biochemistry , embryo
Background Type 1 diabetes (T1D) is a cell‐mediated autoimmune disease characterized by destruction of the pancreatic islet cells. The use of cryopreserved cells is preferable to the use of freshly isolated cells to monitor clinical trials to decrease assay and laboratory variability. Methods The T‐Cell Workshop Committee of the Immunology of Diabetes Society compared two widely accepted T‐cell freezing protocols (warm and cold) to freshly isolated peripheral blood mononuclear cells from patients with T1D and controls in terms of recovery, viability, cell subset composition, and performance in functional assays currently in use in T1D‐related research. Nine laboratories participated in the study with four different functional assays included. Results The cold freezing method yielded higher recovery and viability compared with the warm freezing method. Irrespective of freezing protocol, B cells and CD8+ T cells were enriched, monocyte fraction decreased, and islet antigen‐reactive responses were lower in frozen versus fresh cells. However, these results need to take in to account that the overall response to islet autoantigens was low in some assays. Conclusions In the current study, none of the tested T‐cell functional assays performed well using frozen samples. More research is required to identify a freezing method and a T‐cell functional assay that will produce responses in patients with T1D comparable to responses using fresh peripheral blood mononuclear cells. Copyright © 2011 John Wiley & Sons, Ltd.