Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Blockade of CTLA-4 on CD4 + CD25 + regulatory T cells abrogates their function in vivo

doi:

Figure Lengend Snippet: Unfractionated splenocytes from WT, B7-1/B7-2 KO, and B7-1/B7-2/CTLA-4 KO mice were analysed by flow cytometry for the expression of CD4, CD25, and FoxP3. Mice were analysed at 6–8 wk of age. Representative plots show log10 fluorescence and are gated on CD4+ lymphocytes.

Article Snippet: FACS analysis of the sorted CD4 + CD45RB high population showed <1% FoxP3 + cells (anti-mouse FoxP3 staining set; eBioscience).

Techniques: Flow Cytometry, Expressing, Fluorescence

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Blockade of CTLA-4 on CD4 + CD25 + regulatory T cells abrogates their function in vivo

doi:

Figure Lengend Snippet: (A) WT and CTLA-4 KO TR were isolated from CTLA-4 KO mixed bone marrow chimeras by sorting for Ly9.1+CD4+CD25+ (CTLA-4 KO) and Ly9.1−CD4+CD25+ (CTLA-4-sufficient) populations as described above. The resulting populations were stained for expression of FoxP3 and analyzed by flow cytometry. Plots are gated on CD4+ lymphocytes, and percentages refer to the percentage of CD25+ FoxP3+ cells. (B) BALB/c RAG KO mice received WT CD4+CD45RBhigh cells alone or in combination with WT or CTLA-4 KO CD4+CD25+ cells. Some mice also received anti-IL10R mAb. Mice were sacrificed 8–10 wk after transfer and colons taken for histological analysis. Data are pooled from two independent experiments. In the presence of anti-IL-10R mAb, WT CD4+CD25+ TR mediated significant protection from colitis (p < 0.05), however, CTLA-4 KO CD4+CD25+ TR provided no significant protection from colitis (p > 0.05).

Article Snippet: FACS analysis of the sorted CD4 + CD45RB high population showed <1% FoxP3 + cells (anti-mouse FoxP3 staining set; eBioscience).

Techniques: Isolation, Staining, Expressing, Flow Cytometry

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Blockade of CTLA-4 on CD4 + CD25 + regulatory T cells abrogates their function in vivo

doi:

Figure Lengend Snippet: RAG2 KO mice received CD4+CD45RBhigh and CD4+CD25+ T cells together with anti-CTLA-4 mAb. Eight weeks later, mice were sacrificed and tissues taken for analysis. (A) Sections from mesenteric lymph nodes were stained for CD3 (red) and FoxP3 (green), or CD3 and a control rabbit Ig. Images are representative of the analysis of four mice from two independent experiments. Original magnification: ×630. (B) Representative analysis of FoxP3 expression by transferred CD4+ T cells from anti-CTLA-4-treated mice. Single-cell suspensions from spleen and mesenteric lymph nodes were stained for flow cytometric analysis. Plots show log10 fluorescence and are gated on CD3+CD4+ lymphocytes. (C) Proportion of CD4+ T cells expressing FoxP3 in the spleen and mesenteric lymph nodes of anti-CTLA-4 or control mice. Each point represents a single mouse analyzed by flow cytometry as in (B).

Article Snippet: FACS analysis of the sorted CD4 + CD45RB high population showed <1% FoxP3 + cells (anti-mouse FoxP3 staining set; eBioscience).

Techniques: Staining, Control, Expressing, Fluorescence, Flow Cytometry

Journal: Mucosal immunology

Article Title: miR-106a deficiency attenuates inflammation in murine IBD models.

doi: 10.1038/s41385-018-0091-7

Figure Lengend Snippet: ( A ) Histology of the colonic lamina propria demonstrates significant villus distortion in WT CD45RB High T cell recipients relative to miR-106a −/− T cell recipients following adoptive transfer ( B ) Adoptive transfer of CD4 + CD45RB High T cells from miR-106a −/− mice resulted in significantly attenuated weight loss, a surrogate marker of intestinal inflammation, relative to WT controls. This result coincided with a significant reduction in IFN + CD4 + T cells ( C ) and IL-17 + CD4 + T cells ( D ) in the colonic lamina propria as well as a reduction in IL-10-producing CD4 + T cells ( E ). Results represent mean ± SEM for 4–7 mice per group from two or more independent studies. *P<0.05, **P<0.01, ***P<0.001.

Article Snippet: Adoptive transfer colitis was induced by intraperitoneal injection of 1×10 6 cells/mouse of magnetically pre-enriched CD4 + T cells (EasySEP, STEMCELL Technologies, Vancouver, Ca) flow sorted into live CD4 + CD45RB High naïve T cells (BD FACSAria II, BD Biosciences, Franklin Lakes, NJ).

Techniques: Adoptive Transfer Assay, Marker