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Fig. 1. Level of Mac-1 expression on naive and recently activated LCMV GP33–41-specific T cells from TCR 318 transgenic mice. (A and B) Splenocytes from uninfected transgenic mice were incubated in vitro either with or without LCMV peptide GP33–41 for 5 h. Cells were surface stained with CyChrome-conjugated anti-CD8a, PE-conjugated anti-V<sub>α</sub>2 and FITC-conjugated anti-CD11b. Gates were set for CD8<sup>+</sup>V<sub>α</sub>2<sup>–</sup> and LCMV-specific CD8<sup>+</sup>V<sub>α</sub>2<sup>+</sup> cells, and the level of Mac-1 expression is depicted. (C and D) Splenocytes from transgenic mice infected i.v. with 10<sup>4</sup> p.f.u. of LCMV Armstrong 5 days earlier. Cells were incubated with GP33–41, surface stained with CyChrome-conjugated anti-CD8a, FITC-conjugated anti-V<sub>α</sub>2 (C) or FITC-conjugated anti-CD11b (D), permeabilized and stained with PE-conjugated anti-IFN-γ. Gates have been set for CD8<sup>+</sup> cells. CD11b expression as a marker to distinguish between recently activated effector CD8+ T cells and memory cells Int Immunol. 2001;13(4): doi: /intimm/ Int Immunol | © 2001 Japanese Society for Immunology
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Fig. 2. Mac-1 expression on primary LCMV-specific CD8<sup>+</sup> effector cells. Splenocytes from BALB/c mice infected i.v. with 4800 p.f.u. of LCMV Armstrong 8 days earlier and from matched controls were incubated with LCMV NP118–126, surface stained with CyChrome-conjugated anti-CD8a and FITC-conjugated anti-CD11b, permeabilized and stained with PE-conjugated anti-IFN-γ. Gates have been set for CD8<sup>+</sup> cells; LCMV-specific cells are defined as IFN-γ<sup>+</sup> (upper quadrants). CD11b expression as a marker to distinguish between recently activated effector CD8+ T cells and memory cells Int Immunol. 2001;13(4): doi: /intimm/ Int Immunol | © 2001 Japanese Society for Immunology
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Fig. 3. Level of Mac-1 expression on splenic LCMV specific effector and memory T cells (visualized through detection of IFN-γ intracellularly). C57BL/6 mice were infected with 200 p.f.u. LCMV Traub i.v., and on day 8, 50 and 110 p.i. splenocytes were stimulated with LCMV peptide GP33–41 for 5 h in vitro. Cells were surface stained with CyChrome-conjugated anti-CD8a and FITC-conjugated anti-CD11b, permeabilized and stained with PE-conjugated anti-IFN-γ. Gates have been set for CD8<sup>+</sup> T cells; LCMV-specific cells are defined as IFN-γ<sup>+</sup> (upper quadrants). LCMV-primed cells incubated without peptide gave results similar to naive cells (not shown). CD11b expression as a marker to distinguish between recently activated effector CD8+ T cells and memory cells Int Immunol. 2001;13(4): doi: /intimm/ Int Immunol | © 2001 Japanese Society for Immunology
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Fig. 4. Level of Mac-1 expression on splenic LCMV-specific effector and memory T cells (visualized through staining with tetramers). Splenocytes from C57BL/6 mice were infected with 200 p.f.u. LCMV Traub i.v., and on day 8 and 180 p.i. splenocytes were stained with PE-conjugated H-2D<sup>b</sup>–GP33–41 tetramers, FITC-conjugated anti-CD11b and CyChrome-conjugated anti-CD8a. Gates have been set for CD8<sup>+</sup> T cells; LCMV-specific cells are defined as tetramer<sup>+</sup> (upper quadrants) CD11b expression as a marker to distinguish between recently activated effector CD8+ T cells and memory cells Int Immunol. 2001;13(4): doi: /intimm/ Int Immunol | © 2001 Japanese Society for Immunology
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Fig. 5. Level of Mac-1 expression on LCMV-specific effector and memory T cells from lymph node. C57BL/6 mice were infected with 200 p.f.u. of LCMV Traub i.v., and on day 8 and 13 month p.i. lymph node cells were stimulated with LCMV peptide GP33–41 for 5 h in vitro. Cells were surface stained with CyChrome-conjugated anti-CD8a and FITC-conjugated anti-CD11b, permeabilized and stained with PE-conjugated anti-IFN-γ. Gates have been set for CD8<sup>+</sup> T cells; LCMV-specific cells are defined as IFN-γ<sup>+</sup> (upper quadrants). CD11b expression as a marker to distinguish between recently activated effector CD8+ T cells and memory cells Int Immunol. 2001;13(4): doi: /intimm/ Int Immunol | © 2001 Japanese Society for Immunology
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Fig. 6. Level of Mac-1 expression on VSV-specific effector and memory T cells. C57BL/6 were mice infected with 10<sup>6</sup> p.f.u. VSV i.v., and on day 6 and 40 p.i. splenocytes were stimulated with VSV NP 52–56 for 6 h in vitro. Cells were surface stained with CyChrome-conjugated anti-CD8a and FITC-conjugated anti-CD11b, permeabilized and stained with PE-conjugated anti-IFN-γ. Gates have been set for CD8<sup>+</sup> T cells; VSV-specific cells are defined as IFN-γ<sup>+</sup> (upper quadrants). VSV-primed cells incubated without peptide gave results similar to naive cells (not shown). CD11b expression as a marker to distinguish between recently activated effector CD8+ T cells and memory cells Int Immunol. 2001;13(4): doi: /intimm/ Int Immunol | © 2001 Japanese Society for Immunology
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Fig. 7. Mac-1 expression on secondary effectors generated in reinfected LCMV immune mice. BALB/c mice were infected with 200 p.f.u. of LCMV Armstrong and 6 months later half the mice were rechallenged with ~5×10<sup>5</sup> p.f.u. of LCMV Traub. On day 5 p.i., resting and re-stimulated splenocytes were cultured in vitro with LCMV peptide NP118–126 for 5 h. Splenocytes from acutely infected (day 8 p.i.) mice and naive controls were processed in parallel. Cells were surface stained with CyChrome-conjugated anti-CD8a and FITC-conjugated anti-CD11b, permeabilized and stained with PE-conjugated anti-IFN-γ. Gates have been set for CD8<sup>+</sup> T cells; LCMV-specific cells are defined as IFN-γ<sup>+</sup> (upper quadrants). CD11b expression as a marker to distinguish between recently activated effector CD8+ T cells and memory cells Int Immunol. 2001;13(4): doi: /intimm/ Int Immunol | © 2001 Japanese Society for Immunology
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