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Interferon-γ-producing immature myeloid cells confer protection against severe invasive group A Streptococcus infections.

Matsumura T, Ato M, Ikebe T, Ohnishi M, Watanabe H, Kobayashi K - Nat Commun (2012)

Bottom Line: Interferon-γ is necessary to protect mice, and is produced by a novel population of granulocyte-macrophage colony-stimulating factor-dependent immature myeloid cells with ring-shaped nuclei.These interferon-γ-producing immature myeloid cells express monocyte and granulocyte markers, and also produce nitric oxide.Our results indicate that interferon-γ-producing immature myeloid cells have a protective role during the early stage of severe invasive group A Streptococcus infections.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.

ABSTRACT
Cytokine-activated neutrophils are known to be essential for protection against group A Streptococcus infections. However, during severe invasive group A Streptococcus infections that are accompanied by neutropenia, it remains unclear which factors are protective against such infections, and which cell population is the source of them. Here we show that mice infected with severe invasive group A Streptococcus isolates, but not with non-invasive group A Streptococcus isolates, exhibit high concentrations of plasma interferon-γ during the early stage of infection. Interferon-γ is necessary to protect mice, and is produced by a novel population of granulocyte-macrophage colony-stimulating factor-dependent immature myeloid cells with ring-shaped nuclei. These interferon-γ-producing immature myeloid cells express monocyte and granulocyte markers, and also produce nitric oxide. The adoptive transfer of interferon-γ-producing immature myeloid cells ameliorates infection in wild-type and interferon-γ-deficient mice. Our results indicate that interferon-γ-producing immature myeloid cells have a protective role during the early stage of severe invasive group A Streptococcus infections.

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IFN-γ-producing cells are detected in the peripheral blood and the bone marrow of GAS-infected mice.(a–d) Non-infected C3H/HeN mice or (a–e) mice i.p. infected with NIH34 (3.0×107 CFU per mouse) for 42 h were i.v. injected with monensin. Six hours later, the mice were killed and their splenocytes (a–e), peripheral blood cells (a,b), and bone marrow cells (c,d) were immediately stained for the indicated markers, and analysed by ICS assay. (a,c) The numbers (%) in the plots represent the proportion of IFN-γ+ subsets to total cells. (b,d,e) Lower panels show the cells gated on the IFN-γ+ population. (d,e) Rectangle gates in the plot represent mature granulocytes (d) and Eos (e). The numbers (%) in the plots represent the proportion of mature granulocytes (d) or IFN-γ+ Eos (e) to total cells. Data are representative of three independent experiments. (f) The numbers of PMNs in the bone marrow, peripheral blood, and spleen from non-infected or NIH34-infected mice at 48 h. Data are expressed as mean±s.d. (n=3). The differences compared with non-infected mice were statistically significant (*P<0.05, **P<0.01) as determined by Student's t-test.
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f4: IFN-γ-producing cells are detected in the peripheral blood and the bone marrow of GAS-infected mice.(a–d) Non-infected C3H/HeN mice or (a–e) mice i.p. infected with NIH34 (3.0×107 CFU per mouse) for 42 h were i.v. injected with monensin. Six hours later, the mice were killed and their splenocytes (a–e), peripheral blood cells (a,b), and bone marrow cells (c,d) were immediately stained for the indicated markers, and analysed by ICS assay. (a,c) The numbers (%) in the plots represent the proportion of IFN-γ+ subsets to total cells. (b,d,e) Lower panels show the cells gated on the IFN-γ+ population. (d,e) Rectangle gates in the plot represent mature granulocytes (d) and Eos (e). The numbers (%) in the plots represent the proportion of mature granulocytes (d) or IFN-γ+ Eos (e) to total cells. Data are representative of three independent experiments. (f) The numbers of PMNs in the bone marrow, peripheral blood, and spleen from non-infected or NIH34-infected mice at 48 h. Data are expressed as mean±s.d. (n=3). The differences compared with non-infected mice were statistically significant (*P<0.05, **P<0.01) as determined by Student's t-test.

Mentions: Anti-Gr-1 mAb detects Ly-6C+ monocytes and Ly-6G+ PMNs. Therefore, to determine which subset of Gr-1+ CD11b+ cells is responsible for IFN-γ production during severe invasive GAS infections, we investigated the surface phenotype of IFN-γ-producing cells isolated from the spleens of mice at 48 h post-infection. The ICS assay revealed that IFN-γ-producing cells had the phenotype of monocytes (F4/80low CX3CR1+) and PMNs (Ly-6G+ Ly-6Clow) (Fig. 3). Additionally, they expressed no lymphoid (CD27, IL-7Rα) or granulocyte-lineage (CCR3, Siglec-F, c-Kit, IL-5Rα (H7)) markers, but exhibited particular profiles of CCR2− CD31+ CD34− CD38+ CD44high CD49d+ CD62L+ CD69+ IL-5Rα (T21)high Siglec-H− (Fig. 3; Supplementary Table S1). In this model, the most prominent GAS infection was present in the kidney1827. In accordance with the bacterial burden in the peritoneal cavity, spleen and kidney, a higher proportion of IFN-γ-producing cells accumulated in the kidney of C3H/HeN mice, and also C57BL/6 mice i.p. infected with NIH34 (Supplementary Fig. S2). By contrast, lower proportion of IFN-γ-producing cells existed at the sites of infection. A skin-infection model yielded similar results (Supplementary Fig. S2). Furthermore, IFN-γ-producing cells were detected in the spleens from mice infected with various STSS strains1827 (Supplementary Fig. S3). These cells were also detected in the peripheral blood and (in particularly high frequency) the bone marrow from NIH34-infected mice (Fig. 4a–d; Supplementary Fig. S4). Interestingly, Siglec-F+ eosinophils (Eos) stained with IL-5Rα (H7) and IL-5Rα (T21), whereas IFN-γ-producing cells stained well with T21, but not with H7 (Fig. 4e), suggesting that IFN-γ-producing cells were phenotypically distinct from Eos. IFN-γ-producing cells were also phenotypically distinct from Ly-6Clow CD31− PMNs (Fig. 4d). Moreover, the number and proportion of PMNs were markedly reduced in the spleen, peripheral blood, and bone marrow at 48 h post-infection (Fig. 4d and f), as reported in human STSS cases28.


Interferon-γ-producing immature myeloid cells confer protection against severe invasive group A Streptococcus infections.

Matsumura T, Ato M, Ikebe T, Ohnishi M, Watanabe H, Kobayashi K - Nat Commun (2012)

IFN-γ-producing cells are detected in the peripheral blood and the bone marrow of GAS-infected mice.(a–d) Non-infected C3H/HeN mice or (a–e) mice i.p. infected with NIH34 (3.0×107 CFU per mouse) for 42 h were i.v. injected with monensin. Six hours later, the mice were killed and their splenocytes (a–e), peripheral blood cells (a,b), and bone marrow cells (c,d) were immediately stained for the indicated markers, and analysed by ICS assay. (a,c) The numbers (%) in the plots represent the proportion of IFN-γ+ subsets to total cells. (b,d,e) Lower panels show the cells gated on the IFN-γ+ population. (d,e) Rectangle gates in the plot represent mature granulocytes (d) and Eos (e). The numbers (%) in the plots represent the proportion of mature granulocytes (d) or IFN-γ+ Eos (e) to total cells. Data are representative of three independent experiments. (f) The numbers of PMNs in the bone marrow, peripheral blood, and spleen from non-infected or NIH34-infected mice at 48 h. Data are expressed as mean±s.d. (n=3). The differences compared with non-infected mice were statistically significant (*P<0.05, **P<0.01) as determined by Student's t-test.
© Copyright Policy - open-access
Related In: Results  -  Collection

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f4: IFN-γ-producing cells are detected in the peripheral blood and the bone marrow of GAS-infected mice.(a–d) Non-infected C3H/HeN mice or (a–e) mice i.p. infected with NIH34 (3.0×107 CFU per mouse) for 42 h were i.v. injected with monensin. Six hours later, the mice were killed and their splenocytes (a–e), peripheral blood cells (a,b), and bone marrow cells (c,d) were immediately stained for the indicated markers, and analysed by ICS assay. (a,c) The numbers (%) in the plots represent the proportion of IFN-γ+ subsets to total cells. (b,d,e) Lower panels show the cells gated on the IFN-γ+ population. (d,e) Rectangle gates in the plot represent mature granulocytes (d) and Eos (e). The numbers (%) in the plots represent the proportion of mature granulocytes (d) or IFN-γ+ Eos (e) to total cells. Data are representative of three independent experiments. (f) The numbers of PMNs in the bone marrow, peripheral blood, and spleen from non-infected or NIH34-infected mice at 48 h. Data are expressed as mean±s.d. (n=3). The differences compared with non-infected mice were statistically significant (*P<0.05, **P<0.01) as determined by Student's t-test.
Mentions: Anti-Gr-1 mAb detects Ly-6C+ monocytes and Ly-6G+ PMNs. Therefore, to determine which subset of Gr-1+ CD11b+ cells is responsible for IFN-γ production during severe invasive GAS infections, we investigated the surface phenotype of IFN-γ-producing cells isolated from the spleens of mice at 48 h post-infection. The ICS assay revealed that IFN-γ-producing cells had the phenotype of monocytes (F4/80low CX3CR1+) and PMNs (Ly-6G+ Ly-6Clow) (Fig. 3). Additionally, they expressed no lymphoid (CD27, IL-7Rα) or granulocyte-lineage (CCR3, Siglec-F, c-Kit, IL-5Rα (H7)) markers, but exhibited particular profiles of CCR2− CD31+ CD34− CD38+ CD44high CD49d+ CD62L+ CD69+ IL-5Rα (T21)high Siglec-H− (Fig. 3; Supplementary Table S1). In this model, the most prominent GAS infection was present in the kidney1827. In accordance with the bacterial burden in the peritoneal cavity, spleen and kidney, a higher proportion of IFN-γ-producing cells accumulated in the kidney of C3H/HeN mice, and also C57BL/6 mice i.p. infected with NIH34 (Supplementary Fig. S2). By contrast, lower proportion of IFN-γ-producing cells existed at the sites of infection. A skin-infection model yielded similar results (Supplementary Fig. S2). Furthermore, IFN-γ-producing cells were detected in the spleens from mice infected with various STSS strains1827 (Supplementary Fig. S3). These cells were also detected in the peripheral blood and (in particularly high frequency) the bone marrow from NIH34-infected mice (Fig. 4a–d; Supplementary Fig. S4). Interestingly, Siglec-F+ eosinophils (Eos) stained with IL-5Rα (H7) and IL-5Rα (T21), whereas IFN-γ-producing cells stained well with T21, but not with H7 (Fig. 4e), suggesting that IFN-γ-producing cells were phenotypically distinct from Eos. IFN-γ-producing cells were also phenotypically distinct from Ly-6Clow CD31− PMNs (Fig. 4d). Moreover, the number and proportion of PMNs were markedly reduced in the spleen, peripheral blood, and bone marrow at 48 h post-infection (Fig. 4d and f), as reported in human STSS cases28.

Bottom Line: Interferon-γ is necessary to protect mice, and is produced by a novel population of granulocyte-macrophage colony-stimulating factor-dependent immature myeloid cells with ring-shaped nuclei.These interferon-γ-producing immature myeloid cells express monocyte and granulocyte markers, and also produce nitric oxide.Our results indicate that interferon-γ-producing immature myeloid cells have a protective role during the early stage of severe invasive group A Streptococcus infections.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, Japan.

ABSTRACT
Cytokine-activated neutrophils are known to be essential for protection against group A Streptococcus infections. However, during severe invasive group A Streptococcus infections that are accompanied by neutropenia, it remains unclear which factors are protective against such infections, and which cell population is the source of them. Here we show that mice infected with severe invasive group A Streptococcus isolates, but not with non-invasive group A Streptococcus isolates, exhibit high concentrations of plasma interferon-γ during the early stage of infection. Interferon-γ is necessary to protect mice, and is produced by a novel population of granulocyte-macrophage colony-stimulating factor-dependent immature myeloid cells with ring-shaped nuclei. These interferon-γ-producing immature myeloid cells express monocyte and granulocyte markers, and also produce nitric oxide. The adoptive transfer of interferon-γ-producing immature myeloid cells ameliorates infection in wild-type and interferon-γ-deficient mice. Our results indicate that interferon-γ-producing immature myeloid cells have a protective role during the early stage of severe invasive group A Streptococcus infections.

Show MeSH
Related in: MedlinePlus