title: Comparison of cryopreservation methods on T-cell responses to islet and control antigens from type 1 diabetic patients and controls.
abstract: 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.
authors: University of Washington, Veterans Affairs Puget Sound Health Care System, Seattle, WA 98108, USA. bbrooks@u.washington.edu
authors: B
authors: B
authors: Brooks-Worrell
authors: T
authors: T
authors: Tree
authors: S I
authors: SI
authors: Mannering
authors: I
authors: I
authors: Durinovic-Bello
authors: E
authors: E
authors: James
authors: P
authors: P
authors: Gottlieb
authors: S
authors: S
authors: Wong
authors: Z
authors: Z
authors: Zhou
authors: L
authors: L
authors: Yang
authors: C M
authors: CM
authors: Cilio
authors: J
authors: J
authors: Reichow
authors: B
authors: B
authors: Menart
authors: R
authors: R
authors: Rutter
authors: R
authors: R
authors: Schreiner
authors: M
authors: M
authors: Pham
authors: L
authors: L
authors: Petrich de Marquesini
authors: O
authors: O
authors: Lou
authors: M
authors: M
authors: Scotto
authors: R
authors: R
authors: Mallone
authors: N C
authors: NC
authors: Schloot
mesh_terms: D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D002470:Cell Survival; D015925:Cryopreservation / Q000379:methods*; D003922:Diabetes Mellitus, Type 1 / Q000276:immunology*; D006801:Humans; D007515:Islets of Langerhans / Q000276:immunology*; D011384:Proinsulin / Q000276:immunology
chemical_list: D011384:Proinsulin