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Reversible defects in natural killer and memory CD8 T cell lineages in interleukin 15-deficient mice.

Kennedy MK, Glaccum M, Brown SN, Butz EA, Viney JL, Embers M, Matsuki N, Charrier K, Sedger L, Willis CR, Brasel K, Morrissey PJ, Stocking K, Schuh JC, Joyce S, Peschon JJ - J. Exp. Med. (2000)

Bottom Line: IL-15(-/-) mice displayed marked reductions in numbers of thymic and peripheral natural killer (NK) T cells, memory phenotype CD8(+) T cells, and distinct subpopulations of intestinal intraepithelial lymphocytes (IELs).Specific defects associated with IL-15 deficiency were reversed by in vivo administration of exogenous IL-15.These data reveal critical roles for IL-15 in the development of specific lymphoid lineages.

View Article: PubMed Central - PubMed

Affiliation: Immunex Corporation, Seattle, Washington 98101, USA.

ABSTRACT
C57BL/6 mice genetically deficient in interleukin 15 (IL-15(-/-) mice) were generated by gene targeting. IL-15(-/-) mice displayed marked reductions in numbers of thymic and peripheral natural killer (NK) T cells, memory phenotype CD8(+) T cells, and distinct subpopulations of intestinal intraepithelial lymphocytes (IELs). The reduction but not absence of these populations in IL-15(-/-) mice likely reflects an important role for IL-15 for expansion and/or survival of these cells. IL-15(-/-) mice lacked NK cells, as assessed by both immunophenotyping and functional criteria, indicating an obligate role for IL-15 in the development and functional maturation of NK cells. Specific defects associated with IL-15 deficiency were reversed by in vivo administration of exogenous IL-15. Despite their immunological defects, IL-15(-/-) mice remained healthy when maintained under specific pathogen-free conditions. However, IL-15(-/-) mice are likely to have compromised host defense responses to various pathogens, as they were unable to mount a protective response to challenge with vaccinia virus. These data reveal critical roles for IL-15 in the development of specific lymphoid lineages. Moreover, the ability to rescue lymphoid defects in IL-15(-/-) mice by IL-15 administration represents a powerful means by which to further elucidate the biological roles of this cytokine.

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IL-15−/− mice have a deficiency in selective populations of IELs. IELs were isolated from individual 8-wk-old female C57BL/6 or IL-15−/− mice (n = 3/group). The isolated IELs were pooled after the individual cell yields were determined for each mouse. The pooled samples were incubated with the indicated mAb and analyzed using three-color flow cytometry. Viable lymphocytes were gated on the basis of forward and side scatter. A total of 200,000 events was collected for each sample. (A–D) IELs were incubated with mAb specific for CD3, Thy1.2, and TCR-α/β or TCR-γ/δ. Gated populations of CD3+ IELs were analyzed for expression of Thy1.2 and TCR-α/β (A and B) or Thy1.2 and TCR-γ/δ (C and D). The numbers shown represent the percentage of CD3+ IELs within each quadrant. (E–H) IELs were incubated with mAb specific for TCR-α/β, CD8α, and CD4 or CD8β. Gated populations of TCR-α/β+ IELs were analyzed for expression of CD4 and CD8α (E and F) or CD8β and CD8α (G and H). The numbers shown represent the percentage of TCR-α/β+ IELs within each quadrant.
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Figure 4: IL-15−/− mice have a deficiency in selective populations of IELs. IELs were isolated from individual 8-wk-old female C57BL/6 or IL-15−/− mice (n = 3/group). The isolated IELs were pooled after the individual cell yields were determined for each mouse. The pooled samples were incubated with the indicated mAb and analyzed using three-color flow cytometry. Viable lymphocytes were gated on the basis of forward and side scatter. A total of 200,000 events was collected for each sample. (A–D) IELs were incubated with mAb specific for CD3, Thy1.2, and TCR-α/β or TCR-γ/δ. Gated populations of CD3+ IELs were analyzed for expression of Thy1.2 and TCR-α/β (A and B) or Thy1.2 and TCR-γ/δ (C and D). The numbers shown represent the percentage of CD3+ IELs within each quadrant. (E–H) IELs were incubated with mAb specific for TCR-α/β, CD8α, and CD4 or CD8β. Gated populations of TCR-α/β+ IELs were analyzed for expression of CD4 and CD8α (E and F) or CD8β and CD8α (G and H). The numbers shown represent the percentage of TCR-α/β+ IELs within each quadrant.

Mentions: IELs were isolated from individual C57BL/6 or IL-15−/− mice, and cell yields were determined. Once isolated, the IEL populations from individual mice were pooled and analyzed by three-color flow cytometry for expression of various cell surface markers. The absolute number of CD3+ IELs isolated from IL-15−/− mice was estimated to be approximately twofold lower than that of control mice. Within the CD3+ populations, the ratio of α/β to γ/δ IELs was ∼1:1 in control and 2:1 in IL-15−/− mice (data not shown). Thy1− IELs, which made up ∼50% of IEL populations in control mice (Fig. 4A and Fig. C), accounted for <5% of the total IELs in IL-15−/− mice (Fig. 4B and Fig. D). Within gated populations of TCR-α/β+ IELs, there were increases in the relative proportions of CD4+CD8− and CD4+CD8+ IELs in IL-15−/− mice compared with the controls (Fig. 4E and Fig. F). However, as the yield of CD3+ IELs from IL-15−/− mice was approximately twofold lower than that of control mice, the changes in the relative proportions of these IEL populations likely arise solely from a decrease in the absolute number of TCR-α/β+ CD4−CD8+ IELs in IL-15−/− mice compared with controls. The decrease in the TCR-α/β+CD4−CD8+ IEL population in IL-15−/− mice resulted from a selective decrease in the subset of IELs that express the CD8αα homodimer (Fig. 4G and Fig. H). Thus, IL-15−/− mice have a dramatic reduction in the specific IEL populations that are thought to arise or mature independently of the thymus.


Reversible defects in natural killer and memory CD8 T cell lineages in interleukin 15-deficient mice.

Kennedy MK, Glaccum M, Brown SN, Butz EA, Viney JL, Embers M, Matsuki N, Charrier K, Sedger L, Willis CR, Brasel K, Morrissey PJ, Stocking K, Schuh JC, Joyce S, Peschon JJ - J. Exp. Med. (2000)

IL-15−/− mice have a deficiency in selective populations of IELs. IELs were isolated from individual 8-wk-old female C57BL/6 or IL-15−/− mice (n = 3/group). The isolated IELs were pooled after the individual cell yields were determined for each mouse. The pooled samples were incubated with the indicated mAb and analyzed using three-color flow cytometry. Viable lymphocytes were gated on the basis of forward and side scatter. A total of 200,000 events was collected for each sample. (A–D) IELs were incubated with mAb specific for CD3, Thy1.2, and TCR-α/β or TCR-γ/δ. Gated populations of CD3+ IELs were analyzed for expression of Thy1.2 and TCR-α/β (A and B) or Thy1.2 and TCR-γ/δ (C and D). The numbers shown represent the percentage of CD3+ IELs within each quadrant. (E–H) IELs were incubated with mAb specific for TCR-α/β, CD8α, and CD4 or CD8β. Gated populations of TCR-α/β+ IELs were analyzed for expression of CD4 and CD8α (E and F) or CD8β and CD8α (G and H). The numbers shown represent the percentage of TCR-α/β+ IELs within each quadrant.
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Figure 4: IL-15−/− mice have a deficiency in selective populations of IELs. IELs were isolated from individual 8-wk-old female C57BL/6 or IL-15−/− mice (n = 3/group). The isolated IELs were pooled after the individual cell yields were determined for each mouse. The pooled samples were incubated with the indicated mAb and analyzed using three-color flow cytometry. Viable lymphocytes were gated on the basis of forward and side scatter. A total of 200,000 events was collected for each sample. (A–D) IELs were incubated with mAb specific for CD3, Thy1.2, and TCR-α/β or TCR-γ/δ. Gated populations of CD3+ IELs were analyzed for expression of Thy1.2 and TCR-α/β (A and B) or Thy1.2 and TCR-γ/δ (C and D). The numbers shown represent the percentage of CD3+ IELs within each quadrant. (E–H) IELs were incubated with mAb specific for TCR-α/β, CD8α, and CD4 or CD8β. Gated populations of TCR-α/β+ IELs were analyzed for expression of CD4 and CD8α (E and F) or CD8β and CD8α (G and H). The numbers shown represent the percentage of TCR-α/β+ IELs within each quadrant.
Mentions: IELs were isolated from individual C57BL/6 or IL-15−/− mice, and cell yields were determined. Once isolated, the IEL populations from individual mice were pooled and analyzed by three-color flow cytometry for expression of various cell surface markers. The absolute number of CD3+ IELs isolated from IL-15−/− mice was estimated to be approximately twofold lower than that of control mice. Within the CD3+ populations, the ratio of α/β to γ/δ IELs was ∼1:1 in control and 2:1 in IL-15−/− mice (data not shown). Thy1− IELs, which made up ∼50% of IEL populations in control mice (Fig. 4A and Fig. C), accounted for <5% of the total IELs in IL-15−/− mice (Fig. 4B and Fig. D). Within gated populations of TCR-α/β+ IELs, there were increases in the relative proportions of CD4+CD8− and CD4+CD8+ IELs in IL-15−/− mice compared with the controls (Fig. 4E and Fig. F). However, as the yield of CD3+ IELs from IL-15−/− mice was approximately twofold lower than that of control mice, the changes in the relative proportions of these IEL populations likely arise solely from a decrease in the absolute number of TCR-α/β+ CD4−CD8+ IELs in IL-15−/− mice compared with controls. The decrease in the TCR-α/β+CD4−CD8+ IEL population in IL-15−/− mice resulted from a selective decrease in the subset of IELs that express the CD8αα homodimer (Fig. 4G and Fig. H). Thus, IL-15−/− mice have a dramatic reduction in the specific IEL populations that are thought to arise or mature independently of the thymus.

Bottom Line: IL-15(-/-) mice displayed marked reductions in numbers of thymic and peripheral natural killer (NK) T cells, memory phenotype CD8(+) T cells, and distinct subpopulations of intestinal intraepithelial lymphocytes (IELs).Specific defects associated with IL-15 deficiency were reversed by in vivo administration of exogenous IL-15.These data reveal critical roles for IL-15 in the development of specific lymphoid lineages.

View Article: PubMed Central - PubMed

Affiliation: Immunex Corporation, Seattle, Washington 98101, USA.

ABSTRACT
C57BL/6 mice genetically deficient in interleukin 15 (IL-15(-/-) mice) were generated by gene targeting. IL-15(-/-) mice displayed marked reductions in numbers of thymic and peripheral natural killer (NK) T cells, memory phenotype CD8(+) T cells, and distinct subpopulations of intestinal intraepithelial lymphocytes (IELs). The reduction but not absence of these populations in IL-15(-/-) mice likely reflects an important role for IL-15 for expansion and/or survival of these cells. IL-15(-/-) mice lacked NK cells, as assessed by both immunophenotyping and functional criteria, indicating an obligate role for IL-15 in the development and functional maturation of NK cells. Specific defects associated with IL-15 deficiency were reversed by in vivo administration of exogenous IL-15. Despite their immunological defects, IL-15(-/-) mice remained healthy when maintained under specific pathogen-free conditions. However, IL-15(-/-) mice are likely to have compromised host defense responses to various pathogens, as they were unable to mount a protective response to challenge with vaccinia virus. These data reveal critical roles for IL-15 in the development of specific lymphoid lineages. Moreover, the ability to rescue lymphoid defects in IL-15(-/-) mice by IL-15 administration represents a powerful means by which to further elucidate the biological roles of this cytokine.

Show MeSH
Related in: MedlinePlus