Phenotypic characterisation of CD4CD25T cells from immunised IFN-γR KO and wild-type DBA/1 mice

Abstract

<p><b>Copyright information:</b></p><p>Taken from "Defective CD4CD25regulatory T cell functioning in collagen-induced arthritis: an important factor in pathogenesis, counter-regulated by endogenous IFN-γ"</p><p>Arthritis Research & Therapy 2005;7(2):R402-R415.</p><p>Published online 28 Jan 2005</p><p>PMCID:PMC1065335.</p><p>Copyright © 2005 Kelchtermans et al.; licensee BioMed Central Ltd.</p> CD4CD25T cells isolated from IFN-γR KO and wild-type mice show a similar expression pattern of activation markers, in naive and immunised conditions. CD4T cells were purified from the lymph node cells of eight IFN-γR KO and wild-type DBA/1 mice, either naive or having been immunised 21 days previously with collagen type II in complete Freund's adjuvant (purity more than 99%). CD4T cells were stained for CD25 in combination with CD69, CD62L, CD44 or cytolytic T lymphocyte-associated antigen-4 (CTLA-4). Dead cells were excluded by gating on propidium iodide-negative cells. The numbers represent the percentages of CD4CD25cells within the indicated marker. Decreased Foxp3 mRNA levels in CD4CD25Tcells from immunised mice. Lymph node cells were isolated from eight naive or immunised IFN-γR KO and wild-type DBA/1 mice. Purified CD4T cells were stained with anti-CD25-FITC and phycoerythrin-conjugated anti-CD4, and sorted. The purity of the sorted CD4CD25population was more than 97%. cDNA samples were prepared from 2 × 10cells of each population and were subjected to real-time quantitative PCR analyses. The relative quantity of Foxp3 in each sample was normalised to the quantity of β-actin. Error bars indicate standard error of the means of two (CD4CD24cells from naive mice) or three (CD4CD25cells from immunised mice) independent experiments. *< 0.05 for comparison with Foxp3 expression of cells isolated from immunised wild-type mice (Mann–Whitney -test)