Standardization and Performance Evaluation of Mononuclear Cell Cytokine Secretion Assays in a Multicenter Study


Show simple item record Shreffler, Wayne G en_US Visness, Cynthia M en_US Burger, Melissa en_US Cruikshank, William W en_US Lederman, Howard M en_US de la Morena, Maite en_US Grindle, Kristine en_US Calatroni, Agustin en_US Sampson, Hugh A en_US Gern, James E en_US 2011-12-30T00:09:36Z 2011-12-30T00:09:36Z 2006 en_US 2006-12-12 en_US
dc.identifier.citation Shreffler, Wayne G, Cynthia M Visness, Melissa Burger, William W Cruikshank, Howard M Lederman, Maite de la Morena, Kristine Grindle, Agustin Calatroni, Hugh A Sampson, James E Gern. "Standardization and performance evaluation of mononuclear cell cytokine secretion assays in a multicenter study." BMC Immunology 7:29. (2006) en_US
dc.identifier.issn 1471-2172 en_US
dc.description.abstract BACKGROUND: Cryopreservation of peripheral blood mononuclear cells has been used to preserve and standardize immunologic measurements for multicenter studies, however, effects of cryopreservation on cytokine responses are incompletely understood. In designing immunologic studies for a new multicenter birth cohort study of childhood asthma, we performed a series of experiments to determine the effects of two different methods of cryopreservation on the cytokine responses of cord and peripheral blood mononuclear cells. RESULTS: 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-α. In separate experiments, NC cryopreservation inhibited secretion of several cytokines (IL-13, IL-10, IFN-γ, TNF-α) 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. CONCLUSION: 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. en_US
dc.description.sponsorship National Institute of Allergy and Infectious Diseases; National Institutes of Health (N01-A1-25496, N01-A1-254082) en_US
dc.language.iso en en_US
dc.publisher BioMed Central en_US
dc.rights Copyright 2006 Shreffler et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution 2.0 License (, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. en_US
dc.rights.uri en_US
dc.title Standardization and Performance Evaluation of Mononuclear Cell Cytokine Secretion Assays in a Multicenter Study en_US
dc.type article en_US
dc.identifier.doi 10.1186/1471-2172-7-29 en_US
dc.identifier.pubmedid 17156490 en_US
dc.identifier.pmcid 1762025 en_US

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