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Cryopreserved peripheral blood mononuclear cells are suitable for the assessment of immunological markers in type 1 diabetic children

Axelsson, Stina (författare)
Linköpings universitet,Hälsouniversitetet,Pediatrik
Faresjö, Maria (författare)
Linköpings universitet,Hälsouniversitetet,Pediatrik,Hälsouniversitet i Linköping
Hedman, Maria (författare)
Linköpings universitet,Institutionen för klinisk och experimentell medicin,Hälsouniversitetet
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Ludvigsson, Johnny (författare)
Östergötlands Läns Landsting,Linköpings universitet,Hälsouniversitetet,Pediatrik,Barn- och ungdomskliniken i Linköping
Casas, Rosaura (författare)
Linköpings universitet,Hälsouniversitetet,Pediatrik
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 (creator_code:org_t)
Elsevier BV, 2008
2008
Engelska.
Ingår i: Cryobiology. - : Elsevier BV. - 0011-2240 .- 1090-2392. ; 57:3, s. 201-208
  • Tidskriftsartikel (refereegranskat)
Abstract Ämnesord
Stäng  
  • 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.

Nyckelord

Cryopreservation
BMC
Cytokine
Chemokine
Multiplex fluorochrome technique
Real-time RT-PCR
Type 1 diabetes
MEDICINE
MEDICIN

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