Limits...
Autoimmunity-associated protein tyrosine phosphatase PEP negatively regulates IFN-α receptor signaling.

Holmes DA, Suto E, Lee WP, Ou Q, Gong Q, Smith HR, Caplazi P, Chan AC - J. Exp. Med. (2015)

Bottom Line: Pep(-/-) hematopoietic progenitors demonstrate increased IFNAR signaling, increased IFN-inducible gene expression, and enhanced proliferation and activation compared to Pep(+/+) progenitors in response to IFN-α.In addition, Pep(-/-) mice treated with IFN-α display a profound defect in hematopoiesis, resulting in anemia, thrombocytopenia, and neutropenia when compared to IFN-α-treated Pep(+/+) mice.As SLE patients carrying the PTPN22(C1858T) risk variant have higher serum IFN-α activity, these data provide a molecular basis for how type I IFNs and PTPN22 may cooperate to contribute to lupus-associated cytopenias.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Immunology, Department of Translational Immunology, and Department of Pathology, Genentech, Inc., South San Francisco, CA 94080.

Show MeSH

Related in: MedlinePlus

PEP negatively regulates progenitor proliferation after administration of poly(I:C) or IFN-α in vivo. (A and B) Analysis of BM progenitor proliferation with poly(I:C) (A) or IFN-α (B) administration in vivo in mice. Pep+/+ and Pep−/− mice were injected i.p. with poly(I:C) (200 µg/mouse) for 23 h or infected with Ad5-IFN-α5 (107 PFU/mouse) for 36 h, followed by BrdU administration for 1 h. BrdU incorporation in BM progenitors was analyzed by FACS. Percentage of BrdU+ cells in Pep+/+ (open bar) and Pep−/− (grey bar) mice are depicted. n = 4 mice/genotype and data shown are representative of 3 independent experiments. (C–F) Analysis of cell cycle progression in poly(I:C) treated mice. Pep+/+ (top) and Pep−/− (bottom) mice were treated with poly(I:C) for 23 h, followed by BrdU administration for 1 h. Cell cycle progression in BM progenitors was analyzed by FACS for G0-G1, S, and G2-M in hematopoietic progenitors (LSK; C), LS−K cells (D), MEP (E), and CMP/GMP (F) from Pep+/+ (open bar) and Pep−/− (grey bar) mice. n = 4 mice/genotype and data shown are representative of 3 independent experiments. (G) Pep+/+ (n =15 mice, red) and Pep−/− (n = 15 mice, blue) mice were injected with poly(I:C) (200 µg/mouse). 48 h after poly(I:C) administration, mice were injected i.p. with 150 mg/kg 5-FU to determine intrinsic hematopoietic proliferation and exhaustion rates in vivo. Data are a compilation of three independent experiments. (H) Pep+/+ (n = 9 mice, red) and Pep−/− (n = 9 mice, blue) mice were injected once weekly for 5 wk (i.p.) with 150 mg/kg 5-FU to deplete proliferating BM progenitors to induce BM failure. Data shown are representative of three independent experiments unless otherwise stated. Values in graphs represent means ± SEM. Statistical analysis was done by two-tailed paired Student’s t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4493413&req=5

fig3: PEP negatively regulates progenitor proliferation after administration of poly(I:C) or IFN-α in vivo. (A and B) Analysis of BM progenitor proliferation with poly(I:C) (A) or IFN-α (B) administration in vivo in mice. Pep+/+ and Pep−/− mice were injected i.p. with poly(I:C) (200 µg/mouse) for 23 h or infected with Ad5-IFN-α5 (107 PFU/mouse) for 36 h, followed by BrdU administration for 1 h. BrdU incorporation in BM progenitors was analyzed by FACS. Percentage of BrdU+ cells in Pep+/+ (open bar) and Pep−/− (grey bar) mice are depicted. n = 4 mice/genotype and data shown are representative of 3 independent experiments. (C–F) Analysis of cell cycle progression in poly(I:C) treated mice. Pep+/+ (top) and Pep−/− (bottom) mice were treated with poly(I:C) for 23 h, followed by BrdU administration for 1 h. Cell cycle progression in BM progenitors was analyzed by FACS for G0-G1, S, and G2-M in hematopoietic progenitors (LSK; C), LS−K cells (D), MEP (E), and CMP/GMP (F) from Pep+/+ (open bar) and Pep−/− (grey bar) mice. n = 4 mice/genotype and data shown are representative of 3 independent experiments. (G) Pep+/+ (n =15 mice, red) and Pep−/− (n = 15 mice, blue) mice were injected with poly(I:C) (200 µg/mouse). 48 h after poly(I:C) administration, mice were injected i.p. with 150 mg/kg 5-FU to determine intrinsic hematopoietic proliferation and exhaustion rates in vivo. Data are a compilation of three independent experiments. (H) Pep+/+ (n = 9 mice, red) and Pep−/− (n = 9 mice, blue) mice were injected once weekly for 5 wk (i.p.) with 150 mg/kg 5-FU to deplete proliferating BM progenitors to induce BM failure. Data shown are representative of three independent experiments unless otherwise stated. Values in graphs represent means ± SEM. Statistical analysis was done by two-tailed paired Student’s t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Mentions: Previous studies have shown that chronic type I IFN stimulation induces hematopoietic progenitors to proliferate and lose self-renewal capacity, resulting in exhaustion and HSC dysfunction (Essers et al., 2009). To investigate the mechanism leading to decreased numbers of progenitor cells in Pep−/− BM after IFN-α treatment, we assessed BrdU incorporation by progenitor cells as a measure of progenitor proliferation. Consistent with increased activation observed in poly(I:C)-treated mice (Fig. 2 G), Pep−/− myeloid progenitors incorporated BrdU at a higher rate compared to Pep+/+ progenitors in poly(I:C)-treated mice (Fig. 3 A). We observed a similar increase in BrdU in Pep−/− compared to Pep+/+ myeloid progenitors after low-dose Ad5-IFN-α5 (107 PFU/mouse) administration (Fig. 3 B). This increased proliferation in Pep−/− LS−K, MEP, and CMP/GMP progenitors were associated with increased entry into the cell cycle after poly(I:C) treatment (Fig. 3, D–F). In contrast, no differences in BrdU incorporation or cell cycling were detected between Pep+/+ and Pep−/− HSC progenitors (LSK) after poly(I:C) treatment (Fig. 3 C).


Autoimmunity-associated protein tyrosine phosphatase PEP negatively regulates IFN-α receptor signaling.

Holmes DA, Suto E, Lee WP, Ou Q, Gong Q, Smith HR, Caplazi P, Chan AC - J. Exp. Med. (2015)

PEP negatively regulates progenitor proliferation after administration of poly(I:C) or IFN-α in vivo. (A and B) Analysis of BM progenitor proliferation with poly(I:C) (A) or IFN-α (B) administration in vivo in mice. Pep+/+ and Pep−/− mice were injected i.p. with poly(I:C) (200 µg/mouse) for 23 h or infected with Ad5-IFN-α5 (107 PFU/mouse) for 36 h, followed by BrdU administration for 1 h. BrdU incorporation in BM progenitors was analyzed by FACS. Percentage of BrdU+ cells in Pep+/+ (open bar) and Pep−/− (grey bar) mice are depicted. n = 4 mice/genotype and data shown are representative of 3 independent experiments. (C–F) Analysis of cell cycle progression in poly(I:C) treated mice. Pep+/+ (top) and Pep−/− (bottom) mice were treated with poly(I:C) for 23 h, followed by BrdU administration for 1 h. Cell cycle progression in BM progenitors was analyzed by FACS for G0-G1, S, and G2-M in hematopoietic progenitors (LSK; C), LS−K cells (D), MEP (E), and CMP/GMP (F) from Pep+/+ (open bar) and Pep−/− (grey bar) mice. n = 4 mice/genotype and data shown are representative of 3 independent experiments. (G) Pep+/+ (n =15 mice, red) and Pep−/− (n = 15 mice, blue) mice were injected with poly(I:C) (200 µg/mouse). 48 h after poly(I:C) administration, mice were injected i.p. with 150 mg/kg 5-FU to determine intrinsic hematopoietic proliferation and exhaustion rates in vivo. Data are a compilation of three independent experiments. (H) Pep+/+ (n = 9 mice, red) and Pep−/− (n = 9 mice, blue) mice were injected once weekly for 5 wk (i.p.) with 150 mg/kg 5-FU to deplete proliferating BM progenitors to induce BM failure. Data shown are representative of three independent experiments unless otherwise stated. Values in graphs represent means ± SEM. Statistical analysis was done by two-tailed paired Student’s t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
© Copyright Policy - openaccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4493413&req=5

fig3: PEP negatively regulates progenitor proliferation after administration of poly(I:C) or IFN-α in vivo. (A and B) Analysis of BM progenitor proliferation with poly(I:C) (A) or IFN-α (B) administration in vivo in mice. Pep+/+ and Pep−/− mice were injected i.p. with poly(I:C) (200 µg/mouse) for 23 h or infected with Ad5-IFN-α5 (107 PFU/mouse) for 36 h, followed by BrdU administration for 1 h. BrdU incorporation in BM progenitors was analyzed by FACS. Percentage of BrdU+ cells in Pep+/+ (open bar) and Pep−/− (grey bar) mice are depicted. n = 4 mice/genotype and data shown are representative of 3 independent experiments. (C–F) Analysis of cell cycle progression in poly(I:C) treated mice. Pep+/+ (top) and Pep−/− (bottom) mice were treated with poly(I:C) for 23 h, followed by BrdU administration for 1 h. Cell cycle progression in BM progenitors was analyzed by FACS for G0-G1, S, and G2-M in hematopoietic progenitors (LSK; C), LS−K cells (D), MEP (E), and CMP/GMP (F) from Pep+/+ (open bar) and Pep−/− (grey bar) mice. n = 4 mice/genotype and data shown are representative of 3 independent experiments. (G) Pep+/+ (n =15 mice, red) and Pep−/− (n = 15 mice, blue) mice were injected with poly(I:C) (200 µg/mouse). 48 h after poly(I:C) administration, mice were injected i.p. with 150 mg/kg 5-FU to determine intrinsic hematopoietic proliferation and exhaustion rates in vivo. Data are a compilation of three independent experiments. (H) Pep+/+ (n = 9 mice, red) and Pep−/− (n = 9 mice, blue) mice were injected once weekly for 5 wk (i.p.) with 150 mg/kg 5-FU to deplete proliferating BM progenitors to induce BM failure. Data shown are representative of three independent experiments unless otherwise stated. Values in graphs represent means ± SEM. Statistical analysis was done by two-tailed paired Student’s t test; *, P < 0.05; **, P < 0.01; ***, P < 0.001.
Mentions: Previous studies have shown that chronic type I IFN stimulation induces hematopoietic progenitors to proliferate and lose self-renewal capacity, resulting in exhaustion and HSC dysfunction (Essers et al., 2009). To investigate the mechanism leading to decreased numbers of progenitor cells in Pep−/− BM after IFN-α treatment, we assessed BrdU incorporation by progenitor cells as a measure of progenitor proliferation. Consistent with increased activation observed in poly(I:C)-treated mice (Fig. 2 G), Pep−/− myeloid progenitors incorporated BrdU at a higher rate compared to Pep+/+ progenitors in poly(I:C)-treated mice (Fig. 3 A). We observed a similar increase in BrdU in Pep−/− compared to Pep+/+ myeloid progenitors after low-dose Ad5-IFN-α5 (107 PFU/mouse) administration (Fig. 3 B). This increased proliferation in Pep−/− LS−K, MEP, and CMP/GMP progenitors were associated with increased entry into the cell cycle after poly(I:C) treatment (Fig. 3, D–F). In contrast, no differences in BrdU incorporation or cell cycling were detected between Pep+/+ and Pep−/− HSC progenitors (LSK) after poly(I:C) treatment (Fig. 3 C).

Bottom Line: Pep(-/-) hematopoietic progenitors demonstrate increased IFNAR signaling, increased IFN-inducible gene expression, and enhanced proliferation and activation compared to Pep(+/+) progenitors in response to IFN-α.In addition, Pep(-/-) mice treated with IFN-α display a profound defect in hematopoiesis, resulting in anemia, thrombocytopenia, and neutropenia when compared to IFN-α-treated Pep(+/+) mice.As SLE patients carrying the PTPN22(C1858T) risk variant have higher serum IFN-α activity, these data provide a molecular basis for how type I IFNs and PTPN22 may cooperate to contribute to lupus-associated cytopenias.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Immunology, Department of Translational Immunology, and Department of Pathology, Genentech, Inc., South San Francisco, CA 94080.

Show MeSH
Related in: MedlinePlus