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Cryopreserved peripheral blood mononuclear cells are suitable for the assessment of immunological markers in type 1 diabetic children
Hälsouniversitet i Linköping.
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2008 (English)In: Cryobiology, ISSN 0011-2240, E-ISSN 1090-2392, Vol. 57, no 3, p. 201-208Article in journal (Refereed) Published
Abstract [en]

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-1 alpha, MIP-1 beta 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 GAD65 induced higher levels of IL-6, IFN-gamma, TNF-alpha and MIP-1 alpha, whereas lower secretion was found for IL-10 and IL-13 in cryopreserved PBMC. Stimulation with PHA induced lower secretion of IP-10, MCPA 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. (C) 2008 Elsevier Inc. All rights reserved.

Place, publisher, year, edition, pages
Elsevier, 2008. Vol. 57, no 3, p. 201-208
Keywords [en]
Cryopreservation; BMC; Cytokine; Chemokine; Multiplex fluorochrome technique; Real-time RT-PCR; Type 1 diabetes
National Category
Medical and Health Sciences
Identifiers
URN: urn:nbn:se:hj:diva-9407DOI: 10.1016/j.cryobiol.2008.08.001ISI: 000261665500003PubMedID: 18761006Scopus ID: 2-s2.0-56749180780OAI: oai:DiVA.org:hj-9407DiVA, id: diva2:223004
Available from: 2009-06-10 Created: 2009-06-10 Last updated: 2019-08-30Bibliographically approved

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