Project description:Naïve CD44-lo/CD62L-hi/CD8+ T cells from C3H.SW mice were compared to CD44-hi/CD82L-lo/CD8+ effector memory T cells and CD44-lo/CD62L-hi/CD8+ postmitotic T cells, using 3 biological replicates of each type of sample. The later two cells types were highly purified at day 14 after transplantation from GVHD B6/SJL mice receiving donor C3H.SW mouse-derived naive CD44-lo/CD62L-hi/CD8+ T cells and T cell-depleted bone marrow. Recipient mice had first been lethally irradiated at a dose of 10Gy in two fractions. This is a MHC-identical minor histocompatibility antigen-mismatched mouse GVHD model of human allogeneic hematopoietic stem cell transplantation. Naive T cell samples were from pools of 2 mice each, while effector memory and postmitotic T cell samples were purified from pools of T cells from 4 mice each. After RNA extraction and cleanup, biotin labeled cRNA was prepared from 600 ng total RNA, using two rounds of in vitro transcription, and hybridized to Affymetrix Mouse Genome 430A 2.0 arrays using standard techniques. Keywords: Cell type comparison
Project description:Naïve CD44-lo/CD62L-hi/CD8+ T cells from C3H.SW mice were compared to CD44-hi/CD82L-lo/CD8+ effector memory T cells and CD44-lo/CD62L-hi/CD8+ postmitotic T cells, using 3 biological replicates of each type of sample. The later two cells types were highly purified at day 14 after transplantation from GVHD B6/SJL mice receiving donor C3H.SW mouse-derived naive CD44-lo/CD62L-hi/CD8+ T cells and T cell-depleted bone marrow. Recipient mice had first been lethally irradiated at a dose of 10Gy in two fractions. This is a MHC-identical minor histocompatibility antigen-mismatched mouse GVHD model of human allogeneic hematopoietic stem cell transplantation. Naive T cell samples were from pools of 2 mice each, while effector memory and postmitotic T cell samples were purified from pools of T cells from 4 mice each. After RNA extraction and cleanup, biotin labeled cRNA was prepared from 600 ng total RNA, using two rounds of in vitro transcription, and hybridized to Affymetrix Mouse Genome 430A 2.0 arrays using standard techniques. Keywords: Cell type comparison 9 samples were analyzed on 9 Affymetrix microarrays to assay mRNA levels. There were 3 biological replicates of each of 3 different cell types.
Project description:Naïve CD4+ T cells were isolated from spleen of AND TcR transgenic/green fluorescence protein (GFP) transgenic mice (Kaye et al., Nature 1989;341:746, Wright et al, Blood 2001;97:2278) that recognize a peptide of pigeon cytochrome C in the context of I-Ek and express CD44lo, CD62Lhi, CD45RBhi, and CD25-. After 4 days in vitro stimulation with antigen presenting cells (APC) under either Th1 or Th2 condition, naïve cells become Th1 or Th2 effector cells expressing CD44hi, CD62L lo, CD45RBhi, and CD25+. Additional 3 days culture in the absence of APC, those effector cells become rested expressing a phenotype similar to memory cells (CD44 hi, CD62L lo, CD45RB lo and CD25-). These rested effector cells were adaptively transferred into thymectomized, lethally irradiated, and T cell depleted bone marrow reconstituted mice and memory cells were isolated after 4-12 weeks by flow sort. Keywords: Microarray analysis of Th1 and Th2 effector and memory CD4+ T cells
Project description:CD4+ T cells are critical for defense against the Plasmodium parasites that cause malaria. To better understand CD4+ T cell effector mechanisms during malaria, we performed microarray analysis of CD4+ T cells from naïve and infected mice. Comparison of activated (CD44 hi CD62L lo) CD4+ T cells from infected mice to bulk CD4+ T cells from naïve mice revealed a subset of genes that were upregulated by infection with Plasmodium chabaudi. These results help generate a more complete picture of CD4+ T cell function in malaria.
Project description:Naïve CD4+ T cells were isolated from spleen of AND TcR transgenic/green fluorescence protein (GFP) transgenic mice (Kaye et al., Nature 1989;341:746, Wright et al, Blood 2001;97:2278) that recognize a peptide of pigeon cytochrome C in the context of I-Ek and express CD44lo, CD62Lhi, CD45RBhi, and CD25-. After 4 days in vitro stimulation with antigen presenting cells (APC) under either Th1 or Th2 condition, naïve cells become Th1 or Th2 effector cells expressing CD44hi, CD62L lo, CD45RBhi, and CD25+. Additional 3 days culture in the absence of APC, those effector cells become rested expressing a phenotype similar to memory cells (CD44 hi, CD62L lo, CD45RB lo and CD25-). These rested effector cells were adaptively transferred into thymectomized, lethally irradiated, and T cell depleted bone marrow reconstituted mice and memory cells were isolated after 4-12 weeks by flow sort. Generation and purification of Th1 and Th2 effector and memory CD4+ T cells of 42 samples.
Project description:To identify the role of chemokine receptor in inflammation of colon, we isolated CD3+CD4+ helper T cells harboring CXCR6 from colonic lamina propria of mice We used microarrays to identify the differentially expressed genes between CXCR6Hi Tcells and CXCR6Lo Tcells CXCR6 Hi or CXCR6 Lo T cells were isolated from colonic lamina propria for RNA extraction and hybridization on Affymetrix microarrays.
Project description:This study reports the evolution of the donor CD8 effector T cell transcriptomes at multiple sites and time points within the same host following allo-SCT. Donor derived CD8+ T cells were flow sorted from multiple organs in mice developing GVHD in two clinically relevant murine models of H-2b MHC-matched minor antigen-mismatched BMT: (1) B6>129 model – transfer of C57BL/6 polyclonal CD4+ and CD8+ T cells into 129/Sv BMT recipients; (2) F>M – transfer of monoclonal HY-specific TCR-transgenic MataHari CD8+ T cells into C57BL/6 male BMT recipients. To test the effect of early lymph node egress on CD8 effector T cell transcription profile, BMT recipients were treatment with FTY720 (1.0mg/kg/day, from D+3 to D+7).
Project description:This study reports the evolution of the donor CD8 effector T cell transcriptomes at multiple sites and time points within the same host following allo-SCT. Donor derived CD8+ T cells were flow sorted from multiple organs in mice developing GVHD in two clinically relevant murine models of H-2b MHC-matched minor antigen-mismatched BMT: (1) B6>129 model – transfer of C57BL/6 polyclonal CD4+ and CD8+ T cells into 129/Sv BMT recipients; (2) F>M – transfer of monoclonal HY-specific TCR-transgenic MataHari CD8+ T cells into C57BL/6 male BMT recipients.
Project description:This study reports the evolution of the donor CD8 effector T cell transcriptomes at multiple sites and time points within the same host following allo-SCT. Donor derived CD8+ T cells were flow sorted from multiple organs in mice developing GVHD in two clinically relevant murine models of H-2b MHC-matched minor antigen-mismatched BMT: (1) B6>129 model – transfer of C57BL/6 polyclonal CD4+ and CD8+ T cells into 129/Sv BMT recipients; (2) F>M – transfer of monoclonal HY-specific TCR-transgenic MataHari CD8+ T cells into C57BL/6 male BMT recipients.
Project description:Trascriptional analysis of CD2 hi and CD25 lo CD4+ effector T cells during acute viral infection. SMARTA cells were transferred into B6 mice, followed by infection with LCMV. At day 5 post-infection, CD25 hi and CD25 lo SMARTA cells were isolated from the spleen by FACS. Consistent with our prior studies showing that CD25 lo early effector cells give rise to both Tfh effector cells and memory T cells, we observed gene expression in the CD25 lo population consistent with Tfh differentiation. Conversely, CD25 hi effector cells expressed markers consistent with Th1 differentiation and short-term survival.
