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dc.contributor.authorShreffler, Wayne Gen_US
dc.contributor.authorVisness, Cynthia Men_US
dc.contributor.authorBurger, Melissaen_US
dc.contributor.authorCruikshank, William Wen_US
dc.contributor.authorLederman, Howard Men_US
dc.contributor.authorde la Morena, Maiteen_US
dc.contributor.authorGrindle, Kristineen_US
dc.contributor.authorCalatroni, Agustinen_US
dc.contributor.authorSampson, Hugh Aen_US
dc.contributor.authorGern, James Een_US
dc.date.accessioned2011-12-30T00:09:36Z
dc.date.available2011-12-30T00:09:36Z
dc.date.copyright2006en_US
dc.date.issued2006-12-12en_US
dc.identifier.citationShreffler, 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.issn1471-2172en_US
dc.identifier.urihttp://hdl.handle.net/2144/2683
dc.description.abstractBACKGROUND: 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.sponsorshipNational Institute of Allergy and Infectious Diseases; National Institutes of Health (N01-A1-25496, N01-A1-254082)en_US
dc.language.isoenen_US
dc.publisherBioMed Centralen_US
dc.rightsCopyright 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 (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.en_US
dc.rights.urihttp://creativecommons.org/licenses/by/2.0en_US
dc.titleStandardization and Performance Evaluation of Mononuclear Cell Cytokine Secretion Assays in a Multicenter Studyen_US
dc.typearticleen_US
dc.identifier.doi10.1186/1471-2172-7-29en_US
dc.identifier.pubmedid17156490en_US
dc.identifier.pmcid1762025en_US


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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 (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Except where otherwise noted, this item's license is described as 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 (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.