Project description:Single cell transcriptomic analysis of human CD25+ CD127- CD4+ Treg cells and CD25- CD127+ CD4+ Tconv cells isolated from peripheral blood from two different donors
Project description:Comparison of gene expression profile of CD4+ CD25+, CD4+ CD25- CD45RBlow LAG3+ and CD4+ CD25- CD45RBlow LAG3- T cells. Naive CD4+CD25-CD45RBhigh T cells were used as a reference for pair comparison with values from the three other subsets.
Project description:The objective of the present study was to characterize the phenotype of CD4+CD25+Foxp3+ regulatory T cells (Tregs) in the course of parasitic Plasmodium yoelii (P .yoelii) infection of BALB/c mice. Therefore we performed microarray expression analysis of CD4+CD25+Foxp3+ Tregs isolated by FACS from spleens of non-infected mice and from spleens of mice infected with P. yoelii 3 days and 5 days post infection. By comparing the gene expression profiles, we were able to identify molecules which were differentially expressed by Tregs during parasitic infection and thereby might be involved in their immune-suppressive function. Moreover, we included CD4+CD25-Foxp3- T cells from spleens of non-infected and P. yoelii-infected mice in our analysis. It was proposed that immune-suppressive CD4+CD25-Foxp3- T cells might be induced during Plasmodium infection of mice. Thus, detailed gene expression data of these cells in comparison to CD4+CD25+Foxp3+ Tregs would contribute a better understanding in the phenotype. FACS sorted CD4+CD25+Foxp3+ Tregs and CD4+CD25-Foxp3- T cells from pooled spleens of non-infected Foxp3/ eGFP mice (served as reference) and from pooled spleens of P. yoelii infected Foxp3/ eGFP mice 3 days and 5 days post infection were analyzed as single probes.
Project description:A comparison of CD4+CD25+ and CD4+CD25- splenic T cells. Although both subsets have regulatory T cell activity in an induced transplantation tolerance model, the CD4+CD25+ subsets are apparently 10 fold enriched in regulatory T cells. SAGE analysis is performed and compared both on resting and CD3 activated populations. Keywords: other
Project description:Here we compare the effects of stimulation on cord blood derived CD4+ CD25+ (Treg) and CD4+ CD25- (Thelper) cells, isolated by MACS protocols & expanded in vitro using dynabeads. Expansion was carried out at a ratio of 3 beads/cell in the presence of 1000units/ml of recombinant human IL2 for 8 days, followed by 3 days of culture without beads. RNA was extracted from resting cells on day 4 after expansion. Stimulated cells were treated with Ionomycin for 2hrs prior to RNA extraction. Four way comparison of resting and stimulated Tregs versus Thelpers. A total of 18 arrays were analysed containing RNA from 5 individual cord blood donors.
Project description:Here we compare the effects of stimulation on cord blood derived CD4+ CD25+ (Treg) and CD4+ CD25- (Thelper) cells, isolated by MACS protocols & expanded in vitro using dynabeads. Expansion was carried out at a ratio of 3 beads/cell in the presence of 1000units/ml of recombinant human IL2 for 8 days, followed by 3 days of culture without beads. RNA was extracted from resting cells on day 4 after expansion. Stimulated cells were treated with Ionomycin for 2hrs prior to RNA extraction.