Journal: Cell

Article Title: Spatial proteogenomics reveals distinct and evolutionarily conserved hepatic macrophage niches

doi: 10.1016/j.cell.2021.12.018

Figure Lengend Snippet:

Article Snippet: Anti-Mouse CD5 (53-7.3) FITC , Miltenyi Biotec , 130-102-574; RRID: AB_2658608.

Techniques: Purification, Recombinant, Staining, cDNA Synthesis, Gene Expression, Software, Microscopy

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

Article Title: Donor B cells in Transplants Augment Clonal Expansion and Survival of Pathogenic CD4 + T cells That Mediate Autoimmune-like Chronic GVHD

doi: 10.4049/jimmunol.1200677

Figure Lengend Snippet: BALB/c recipients of SPL or B220−-SPL were sacrificed 5, 10, 15, 25, and 40 days post-HCT, and the donor CD5.1+CD4+ T cells from spleen, skin, and lung were evaluated for intracellular cytokine production. Mean±SE is shown for each time point (n=4-8 from 3-4 replicate experiments). A. Percentage of IFNγ+ cells among donor CD4+ T cells. No significant differences were observed (2-way ANOVA, p>.05). B. Percentage of IL-17+ cells among donor CD4+ T cells Although IL-17 was somewhat higher in the lung of recipients of B220−-SPL, no significant differences were observed (2-way ANOVA, p>.05). C. Percentage of IL-4+ cells among donor CD4+ T cells. A higher percentage of IL-4+ cells were observed in the SPL group compared with the B220−-SPL group (2-way ANOVA, p<0.01 for spleen and lung, p<0.05 for skin)

Article Snippet: Antibodies, flow cytometry analysis, and cell sorting FITC-CD5.1 (H11-86.1), APC-CD62L (MEL-14), PE-Cy7-Sca-1 (D7), PE-CD40 (3/23), PE-CD80 (16-10A1), PE-CD86 (GL1), PE-CD40L (MR1), PE-CD28 (37.51), and PE-streptavidin were purchased from BD Pharmingen (San Diego, CA). eFluor450-CD4 (RM4-5), PE-CD44 (IM7), eFluor780-TCRβ (H57-597), APC-IFNγ (XMG1.2), PE-IL-4 (11B11), PE-IL-5 (TRFK5), PE-IL-13 (eBio13A), PE-IL-17A (eBio17B7), APC-Foxp3 (FJK-16S), AlexaFluor780-B220 (RA3-6B2), PE-OX40 (OX-86), and Biotin-CD25 (eBio7D4) were purchased from eBioscience (San Diego, CA).

Techniques:

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

Article Title: Donor B cells in Transplants Augment Clonal Expansion and Survival of Pathogenic CD4 + T cells That Mediate Autoimmune-like Chronic GVHD

doi: 10.4049/jimmunol.1200677

Figure Lengend Snippet: A-C. BALB/c mice were irradiated (800 cGy) and injected with 5×106 CD4+CD25− splenoctyes from luciferase transgenic (luc+) DBA/2 mice and WT DBA/2 TBCD SPL (5×106), with or without WT DBA/2 B220+ B cells (10×106), then monitored for expansion of donor CD4+ T cells and signs of GVHD. A. CD4+ T cell expansion in mice was evaluated with bioluminescent imaging (BLI). One representative BLI pattern is shown per group per time point (n=8 from two replicate experiments). B. BLI intensity in terms of photons/s. A summary curve (mean±SE) is shown. Mice recieiving B cells had increased luminescence (2-way ANOVA, p<0.01). C. Recipieints of luc+CD4+ T cells with or without B cells were evaluated after HCT for GVHD-related skin damage and hair loss. Recipients of B cells had increased clinical cutaneous scores (2-way ANOVA, p<0.01). D-F. BALB/c mice were irradiated (800 cGy) and injected with either SPL or B220−-SPL and sacrificed 5, 10, 15, 25, and 40 days post-HCT, and their tissues were harvested and evaluated for the presence of infiltrating donor CD5.1+CD4+ T cells. Mean±SE is shown at each time point, n=4-8 from 2-3 replicate experiments. D. Donor CD4+ T cell yield in the spleen was evaluated, and was not significantly different between SPL and B220−-SPL groups (2-way ANOVA, p>0.1) E. Donor CD4+ T cell yield in the skin was evaluated. A higher yield was observed in SPL recipients (2-way ANOVA, p<0.01). F. Donor CD4+ T cell yield in the lung was evaluated. A higher yield was observed in SPL recipients (2-way ANOVA, p<0.01).

Article Snippet: Antibodies, flow cytometry analysis, and cell sorting FITC-CD5.1 (H11-86.1), APC-CD62L (MEL-14), PE-Cy7-Sca-1 (D7), PE-CD40 (3/23), PE-CD80 (16-10A1), PE-CD86 (GL1), PE-CD40L (MR1), PE-CD28 (37.51), and PE-streptavidin were purchased from BD Pharmingen (San Diego, CA). eFluor450-CD4 (RM4-5), PE-CD44 (IM7), eFluor780-TCRβ (H57-597), APC-IFNγ (XMG1.2), PE-IL-4 (11B11), PE-IL-5 (TRFK5), PE-IL-13 (eBio13A), PE-IL-17A (eBio17B7), APC-Foxp3 (FJK-16S), AlexaFluor780-B220 (RA3-6B2), PE-OX40 (OX-86), and Biotin-CD25 (eBio7D4) were purchased from eBioscience (San Diego, CA).

Techniques: Irradiation, Injection, Luciferase, Transgenic Assay, Imaging

Journal: PLoS Biology

Article Title: A Conserved CXXC Motif in CD3ε Is Critical for T Cell Development and TCR Signaling

doi: 10.1371/journal.pbio.1000253

Figure Lengend Snippet: (A) Total cell counts in various organs in the combined wt control (Cntl, open bars) versus the mutant transgenic mice (tgε Mut , filled bars). Cell counts were obtained after the lysis of red blood cells. Data from lymph nodes represent cell counts from inguinal, brachial, and axillary lymph nodes combined and are shown as the number per lymph node. Data shown are for 8 tgε Mut mice and 13 wt control mice (4 BL6, 1 tgε WT cd3ε +/− , 5 tgε Mut cd3ε +/− , and 3 tgε WT ). (B) Total cell counts in the thymus that were at the DN1, DN3, or DN4 stage. The stages are defined by CD44 and CD25 costaining, with a schematic view shown in the upper-left corner of the graph. Data shown are for 8 tgε Mut mice (circles) and 13 wt control mice (squares) as in (A) and 2 cd3ε −/− mice (diamonds). (C) Number of thymocytes that were at the double positive (DP) or single positive stages (CD4 SP and CD8 SP) in control (open bars) versus tgε Mut (filled bars) mice. The numbers of mice examined are as in (A). (D) The percentage of CD5 hi cells at the double positive stage of T cell development in the thymuses of control (open bars) versus tgε Mut (filled bars) mice. The numbers of mice examined are as in (A). (E) The percentage of CD69 hi cells at the double positive or single positive stages of T cell development in the thymuses of control (open bars) versus tgε Mut (filled bars) mice. Data shown are for 4 tgε Mut mice and 8 wt control (cntl) mice (3 BL6, 1 tgε WT cd3ε +/− , 1 tgε Mut cd3ε +/− , and 3 tgε WT ). For (B–E) cells from the thymus were costained with multiple antibodies against surface proteins (CD4, CD8, B220, CD44, CD25, CD5, and CD69). To avoid including non-T lineage cells and minimize nonspecific staining, only live B220 − cells were analyzed. The mean of the group and the SEM are indicated. p values were obtained from t tests. In some cases, fold decreases are indicated.

Article Snippet: Antibodies against CD4 (APC-Cy7-conjugated L3T4), CD5 (FITC- or PE-conjugated 53–7.3), CD8α (APC- or PerCp-conjugated 53–6.7), CD25 (PE- or PE-Cy7-conjugated PC61), CD62L (PE- or PE-Cy7-conjugated MEL-14), CD69 (PE-conjugated H1.2F3), and Thy1.2 (FITC-conjugated 53–2.1) were purchased from BD Pharmingen.

Techniques: Mutagenesis, Transgenic Assay, Lysis, Staining