Figure 5
Figure 5. Role for endogenous STAT4 in the magnitude of CD8 T-cell expansion and proliferation. Responses in STAT4-deficient mice were compared with those in WT mice at different times after LCMV infection. (A) Total CD8 T-cell numbers and (B) BrdU incorporating CD8 T-cell numbers were measured in WT and STAT4-deficient mice that were uninfected (D0) or infected with LCMV for the indicated times. Individual symbols represent results from individual mice. Individual symbols show results from 2 to 8 mice collected from multiple experiments. Bars represent means. STAT4 effects on CD8 T-cell proliferation and STAT1 induction within total and antigen-specific subsets were evaluated. Responses in STAT4-deficient mice were compared with those in WT mice (C). The percentages of CD8 T cells were determined with flow cytometry to stain the subsets. (D) STAT1 and STAT4 expression levels were determined on D0 or D8 by Western blot analysis with the use of total, CD8, and non-CD8 cell preparations isolated from STAT4-deficient mice. Cell lysates were made and analyzed as indicated in Figure 4. Results are representative of 2 or more independent experiments with 3 animals individually tested. (E) To identify the proliferating populations, BrdU was administered in vivo, 2 hours before harvest as per samples in panel A, and CD8 T-cell subsets were examined for expression of STAT1 along with BrdU with the use of flow cytometry. Results are representative of 2 or more independent experiments, with percentages in flow panels showing means ± SEMs of 3 independent samples analyzed within 1 experiment. (F) The LCMV-specific CD8 T cells were separated from total and nonspecific CD8 T cells, on D8 of infection of STAT4-deficient mice, based on staining with pooled class 1 tetramers presenting the 3 immunodominant LCMV peptides for the B6 mice, NP396-404, GP267-286, and GP33-41. The total and sorted CD8 T-cell populations were analyzed for STAT1, STAT4, and IFNAR expression as described in Figure 4. Results are representative of 2 or more independent experiments.

Role for endogenous STAT4 in the magnitude of CD8 T-cell expansion and proliferation. Responses in STAT4-deficient mice were compared with those in WT mice at different times after LCMV infection. (A) Total CD8 T-cell numbers and (B) BrdU incorporating CD8 T-cell numbers were measured in WT and STAT4-deficient mice that were uninfected (D0) or infected with LCMV for the indicated times. Individual symbols represent results from individual mice. Individual symbols show results from 2 to 8 mice collected from multiple experiments. Bars represent means. STAT4 effects on CD8 T-cell proliferation and STAT1 induction within total and antigen-specific subsets were evaluated. Responses in STAT4-deficient mice were compared with those in WT mice (C). The percentages of CD8 T cells were determined with flow cytometry to stain the subsets. (D) STAT1 and STAT4 expression levels were determined on D0 or D8 by Western blot analysis with the use of total, CD8, and non-CD8 cell preparations isolated from STAT4-deficient mice. Cell lysates were made and analyzed as indicated in Figure 4. Results are representative of 2 or more independent experiments with 3 animals individually tested. (E) To identify the proliferating populations, BrdU was administered in vivo, 2 hours before harvest as per samples in panel A, and CD8 T-cell subsets were examined for expression of STAT1 along with BrdU with the use of flow cytometry. Results are representative of 2 or more independent experiments, with percentages in flow panels showing means ± SEMs of 3 independent samples analyzed within 1 experiment. (F) The LCMV-specific CD8 T cells were separated from total and nonspecific CD8 T cells, on D8 of infection of STAT4-deficient mice, based on staining with pooled class 1 tetramers presenting the 3 immunodominant LCMV peptides for the B6 mice, NP396-404, GP267-286, and GP33-41. The total and sorted CD8 T-cell populations were analyzed for STAT1, STAT4, and IFNAR expression as described in Figure 4. Results are representative of 2 or more independent experiments.

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