Figure 6
AICD of lymph node T cells from B6.GT mice. Lymphocyte blasts were prepared from single cell suspensions of pooled axillary and brachial lymph node lymphocytes from B6.WT, B6.TRAIL−/−, B6.gld/gld, or B6.GT naive 5-week-old mice. Cells were cultured for 3 days on TCRVβ- and CD28-agonistic antibody-coated tissue culture plates and restimulated for 48 hours with isotype control, TCRVβ-, or TCRVβ- and CD28-agonistic antibodies in the presence of recombinant murine IL-2 or recombinant murine IFN-γ during the primary or secondary culture, as indicated. Cells were harvested and incubated with CD4-Pacific blue or CD8-PECy7-conjugated antibodies and stained with propidium iodide and annexin V-FITC and analyzed by flow cytometry. Data are the mean percentage of propidium iodide-negative annexin V-negative live cells plus or minus SD of triplicate cultures of (A) CD4+ T cells or (B) CD8+ T cells. AICD data were analyzed by 1-way analysis of variance, and significant differences (P < .05) relative to B6.WT cells (*) or relative to AICD of B6.gld/gld cells (**) are as indicated. Data shown are representative of repeated experiments on cells from independently generated mouse cohorts.

AICD of lymph node T cells from B6.GT mice. Lymphocyte blasts were prepared from single cell suspensions of pooled axillary and brachial lymph node lymphocytes from B6.WT, B6.TRAIL−/−, B6.gld/gld, or B6.GT naive 5-week-old mice. Cells were cultured for 3 days on TCRVβ- and CD28-agonistic antibody-coated tissue culture plates and restimulated for 48 hours with isotype control, TCRVβ-, or TCRVβ- and CD28-agonistic antibodies in the presence of recombinant murine IL-2 or recombinant murine IFN-γ during the primary or secondary culture, as indicated. Cells were harvested and incubated with CD4-Pacific blue or CD8-PECy7-conjugated antibodies and stained with propidium iodide and annexin V-FITC and analyzed by flow cytometry. Data are the mean percentage of propidium iodide-negative annexin V-negative live cells plus or minus SD of triplicate cultures of (A) CD4+ T cells or (B) CD8+ T cells. AICD data were analyzed by 1-way analysis of variance, and significant differences (P < .05) relative to B6.WT cells (*) or relative to AICD of B6.gld/gld cells (**) are as indicated. Data shown are representative of repeated experiments on cells from independently generated mouse cohorts.

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