Document Detail


Cryopreserved peripheral blood mononuclear cells are suitable for the assessment of immunological markers in type 1 diabetic children.
MedLine Citation:
PMID:  18761006     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cryopreserved peripheral blood mononuclear cells (PBMC) are commonly used when assessing immune responses in clinical trials, both for practical reasons and to minimize interassay variation, as samples are often collected and studied over time. This study investigated the effect of cryopreservation on cytokine and chemokine secretion, and on expression of regulatory T-cell associated markers, in samples from children with type 1 diabetes. PBMC were cultured before and after cryopreservation either with GAD(65) or PHA. Secretion of cytokines (IL-5, -6, -10, -12, -13 -17, IFN-gamma and TNF-alpha) and chemokines (IP-10, MCP-1, MIP-1alpha, MIP-1beta and RANTES) was analysed in cell supernatants using multiplex fluorochrome technique (Luminex). Expression of FOXP3 and TGF-beta mRNA was detected by multiplex real-time RT-PCR. Increased spontaneous secretion of IL-6, -10, -12, -13, IFN-gamma and MCP-1, and mRNA expression of FOXP3 and TGF-beta, was detected after cryopreservation. Stimulation with GAD(65) induced higher levels of IL-6, IFN-gamma, TNF-alpha and MIP-1alpha, whereas lower secretion was found for IL-10 and IL-13 in cryopreserved PBMC. Stimulation with PHA induced lower secretion of IP-10, MCP-1 and RANTES and FOXP3 mRNA expression after cryopreservation. Thus, cryopreserved PBMC were suitable to assess the immunological markers included in this study, even though their expression could differ from freshly handled cells.
Authors:
Stina Axelsson; Maria Faresjö; Maria Hedman; Johnny Ludvigsson; Rosaura Casas
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2008-08-12
Journal Detail:
Title:  Cryobiology     Volume:  57     ISSN:  1090-2392     ISO Abbreviation:  Cryobiology     Publication Date:  2008 Dec 
Date Detail:
Created Date:  2008-12-01     Completed Date:  2009-01-22     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  0006252     Medline TA:  Cryobiology     Country:  United States    
Other Details:
Languages:  eng     Pagination:  201-8     Citation Subset:  IM    
Affiliation:
Division of Paediatrics and Diabetes Research Centre, Department of Clinical and Experimental Medicine, Faculty of Health Sciences, Linköping University, S-581 85 Linköping, Sweden. stiax@imk.liu.se
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MeSH Terms
Descriptor/Qualifier:
Adolescent
Cells, Cultured
Chemokines / immunology,  secretion
Child
Cryopreservation / methods*
Cytokines / immunology,  secretion
Diabetes Mellitus, Type 1 / immunology
Female
Forkhead Transcription Factors / genetics,  immunology
Humans
Leukocytes, Mononuclear / immunology*,  metabolism,  secretion
Male
RNA, Messenger / metabolism
Transforming Growth Factor beta / genetics,  immunology
Chemical
Reg. No./Substance:
0/Chemokines; 0/Cytokines; 0/FOXP3 protein, human; 0/Forkhead Transcription Factors; 0/RNA, Messenger; 0/Transforming Growth Factor beta

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