%0 Journal Article
%T Standardization and performance evaluation of mononuclear cell cytokine secretion assays in a multicenter study
%A Wayne G Shreffler
%A Cynthia M Visness
%A Melissa Burger
%A William W Cruikshank
%A Howard M Lederman
%A Maite de la Morena
%A Kristine Grindle
%A Agustin Calatroni
%A Hugh A Sampson
%A James E Gern
%J BMC Immunology
%D 2006
%I BioMed Central
%R 10.1186/1471-2172-7-29
%X Paired samples of PBMC were processed freshly, or after cryopreservation in a Nalgene container (NC) or a controlled-rate freezer (CRF). Although there were some differences between the methods, cryopreservation inhibited PHA-induced IL-10 secretion and Der f 1-induced IL-2 secretion, and augmented PHA-induced IL-2 secretion and spontaneous secretion of TNF-¦Á. In separate experiments, NC cryopreservation inhibited secretion of several cytokines (IL-13, IL-10, IFN-¦Ã, TNF-¦Á) by PHA-stimulated cord blood mononuclear cells. With the exception of PHA-induced IL-13, results from fresh and cryopreserved cord blood samples were not significantly correlated. Finally, in reproducibility studies involving processing of identical cell samples in up to 4 separate laboratories, variances in cytokine responses of fresh cells stimulated at separate sites did not exceed those in cryopreserved cells stimulated at a central site.Collectively, these studies indicate that cryopreservation can affect mononuclear cell cytokine response profiles, and that IL-10 secretion and antigen-induced responses may be especially vulnerable. These studies also demonstrate that mononuclear cell responses can be standardized for performance in a small number of laboratories for multicenter studies, and underscore the importance of measuring reproducibility and of testing whether cryopreservation techniques alter specific immunologic outcomes.With the increase in multi-center studies has come an increased need to perform cellular studies that can be standardized. The responses of blood mononuclear cells are commonly used as surrogates for in vivo immune responses to adaptive and innate immune stimuli. Performing cellular assays at a single central laboratory by the use of cryopreserved and shipped specimens is generally thought to improve their feasibility and uniformity. This approach has been used extensively in vaccine and tumor immunology research, and several methodological investigations on the use
%U http://www.biomedcentral.com/1471-2172/7/29