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Open Access

Differential Sensitivity to IL-12 Drives Sex-Specific Differences in the CD8+ T Cell Response to Infection

Kristel Joy Yee Mon, Elizabeth Goldsmith, Neva B. Watson, Jocelyn Wang, Norah L. Smith and Brian D. Rudd
ImmunoHorizons April 1, 2019, 3 (4) 121-132; DOI: https://doi.org/10.4049/immunohorizons.1800066
Kristel Joy Yee Mon
*Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853; and
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Elizabeth Goldsmith
†College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO 80523
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Neva B. Watson
*Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853; and
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Jocelyn Wang
*Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853; and
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Norah L. Smith
*Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853; and
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Brian D. Rudd
*Department of Microbiology and Immunology, Cornell University, Ithaca, NY 14853; and
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  • FIGURE 1.
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    FIGURE 1.

    Sex-specific differences in the numbers and phenotype of Ag-specific CD8+ T cells postinfection.

    (A and E) Schematic of experimental design: female and male C57BL/6 mice were infected with either 5 × 103 CFU of LM-gB or 2 × 105 PFU of VACV-gB and serially bled to monitor gB-specific CD8+ T cell responses. (B and F) Relative numbers of female (red) and male (blue) gB-specific CD8+ T cells in the blood at various dpi. (C and G) Representative contour plots of gB-specific CD8+ T cells in male and female mice on day 7 postinfection. Asterisks indicate differences between male and female groups. (D and H) Percentage of female and male gB-specific CD8+ T cells at various dpi that exhibit a SLEC (KLRG1hiCD127lo) or MPEC (KLRG1loCD127hi) phenotype. Significance was determined by two-way ANOVA. Data are representative of two experiments (n = 8 mice per group overall). *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.

  • FIGURE 2.
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    FIGURE 2.

    Cell-intrinsic differences between male and female donor CD8+ T cells postinfection.

    (A) Schematic of experimental design: 1 × 104 gBT-I CD8+ T cells from adult female (Thy1.1, CD45.2) and male (Thy1.2, CD45.2) donors were cotransferred into congenic recipients (Thy1.2, CD45.1). The recipient mice were then infected with 5 × 103 CFU LM-gB and serially bled at days 5, 7, and 14. (B) Relative numbers of female (red) and male (blue) donor gBT-I CD8+ T cells per ml of blood at days 5, 7, and 14 postinfection. (C) Representative contour plots of female and male gB-specific CD8+ T cells at the peak of the primary response (day 7) of infection. Percentage of female and male donor gBT-I CD8+ T cells at various dpi that exhibit a SLEC (KLRG1hiCD127lo) or MPEC (KLRG1loCD127hi) phenotype. Significance was determined by two-way ANOVA. Data are representative of two experiments (n = 12 mice per group overall). (D) Representative histogram overlay and geometric mean fluorescence intensity of T-bet expression in male and female donor gB-specific CD8+ T cells from spleens of infected mice at 7 dpi. (E) Representative histogram overlays and geometric mean fluorescence intensity or percentage of positive gate of cytolytic molecules at 7 dpi. Significance was determined by unpaired t test. Data are representative of two experiments (n = 10 mice per group overall). **p < 0.01, ***p < 0.001, ****p < 0.0001. NS, p > 0.05.

  • FIGURE 3.
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    FIGURE 3.

    Comparison of male and female CD8+ T cells primed with different amounts of cognate Ag.

    (A) Schematic of experimental design: 1 × 104 gBT-I CD8+ T cells from female (Thy1.1, CD45.2) and male (Thy1.2, CD45.2) donor mice were cotransferred into Ly5.2 recipient mice (Thy1.2, CD45.1). These recipients were then infected with 5 × 103 CFU LM-gB mixed with 5 × 103 CFU L. monocytogenes at the indicated ratios and serially bled at days 5, 7, and 14. (B) Relative numbers of adult female (red) and male (blue) donor gBT-I CD8+ T cells per ml of blood in mice infected with different amounts of LM-gB at various dpi. (C) Representative contour plots of female and male gB-specific CD8+ T cells at the peak of the primary response (day 7) of infection. (D) Percentage of female and male donor gBT-I CD8+ T cells in different treatment groups at various dpi that exhibit a SLEC (KLRG1hiCD127lo) or MPEC (KLRG1loCD127hi) phenotype. Significance was determined by two-way ANOVA. Data are representative of two experiments (n = 15 mice per group). ***p < 0.001, ****p < 0.0001.

  • FIGURE 4.
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    FIGURE 4.

    Analysis of male and female CD8+ T cells primed in vivo with an equivalent amount of IL-12.

    (A) Schematic of experimental design: 1 × 104 gBT-I CD8+ T cells from congenic adult female (Thy1.1, CD45.2) and male (Thy1.2, CD45.2) donors were cotransferred into Ly5.2 recipients (Thy1.2, CD45.1). These recipients were then immunized with 1 × 106 matured dendritic cells coated with gB peptide (i.v.). Recipients were injected with IL-12 or PBS (control) i.p. for three consecutive days and serially bled at days 5, 7, and 14. (B) Representative contour plots of female and male gB-specific CD8+ T cells at the peak of the primary response (day 7) of IL-12 immunization. (C) Relative numbers of adult female (red) and male (blue) donor gBT-I CD8+ T cells per milliliter of blood at days 5, 7, and 14 postimmunization. (D) Percentage of female and male donor gBT-I CD8+ T cells at various days postimmunization in the IL-12–treated group that exhibit a SLEC (KLRG1hiCD127lo) or MPEC (KLRG1loCD127hi) phenotype. Significance was determined by two-way ANOVA. Data are representative of two experiments (n = 16 mice per group). *p < 0.05, ****p < 0.0001.

  • FIGURE 5.
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    FIGURE 5.

    Comparison of male and female CD8+ T cells stimulated in vitro.

    (A) Schematic of experimental design: gBT-I CD8+ T cells from male and female mice were coated with CFSE and stimulated in vitro with plate-bound anti-CD3 and soluble CD28 in the absence or presence of IL-12. (B) Representative CFSE histograms of female and male CD8+ T cells at 72 h after TCR stimulation in vitro. (C) Representative contour plots of female and male gB-specific CD8+ T cells that are labeled in CFSE proliferation dye and express CD62L at 72 h after TCR stimulation in the absence or presence of IL-12. (D) Percentage of female and male CD8+ T cells that are CD62Llo in the absence or presence of IL-12 at 72 h after TCR stimulation. Significance was determined by two-way ANOVA. Data are representative of two experiments (n = 8 mice per group). Asterisks indicate differences between male and female groups. **p < 0.01, ***p < 0.001, ****p < 0.0001.

  • FIGURE 6.
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    FIGURE 6.

    Sex-specific differences in the phenotype of IL-12–deficient CD8+ T cells postinfection.

    (A) Schematic of experimental design: gBT-I CD8+ T cells from congenic female or male control mice (Thy1.1, CD45.2) and male or female IL-12Rβ KO (Thy1.2, CD45.2) donors were cotransferred into Ly5.2 recipients (Thy1.2, CD45.1). Recipient mice were then infected with 5 × 103 CFU LM-gB and serially bled at days 5, 7, and 14. (B) Representative contour plots of female and male donor gB-specific CD8+ T cells at the peak of the primary response (day 7) of infection. (C) Percentage of female and male donor gBT-I IL-12Rβ KO CD8+ T cells at various days postimmunization that exhibit a SLEC (KLRG1hiCD127lo) or MPEC (KLRG1loCD127hi) phenotype. Significance was determined by two-way ANOVA. Data are representative of two experiments (n = 20 mice per group). ***p < 0.001, ****p < 0.0001.

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Differential Sensitivity to IL-12 Drives Sex-Specific Differences in the CD8+ T Cell Response to Infection
Kristel Joy Yee Mon, Elizabeth Goldsmith, Neva B. Watson, Jocelyn Wang, Norah L. Smith, Brian D. Rudd
ImmunoHorizons April 1, 2019, 3 (4) 121-132; DOI: 10.4049/immunohorizons.1800066

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Differential Sensitivity to IL-12 Drives Sex-Specific Differences in the CD8+ T Cell Response to Infection
Kristel Joy Yee Mon, Elizabeth Goldsmith, Neva B. Watson, Jocelyn Wang, Norah L. Smith, Brian D. Rudd
ImmunoHorizons April 1, 2019, 3 (4) 121-132; DOI: 10.4049/immunohorizons.1800066
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