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Delinking CARD9 and IL-17: CARD9 Protects against Candida tropicalis Infection through a TNF-α-Dependent, IL-17-Independent Mechanism.

Whibley N, Jaycox JR, Reid D, Garg AV, Taylor JA, Clancy CJ, Nguyen MH, Biswas PS, McGeachy MJ, Brown GD, Gaffen SL - J. Immunol. (2015)

Bottom Line: Consistently, WT mice depleted of TNF-α were more susceptible to C. tropicalis, and CARD9-deficient neutrophils and monocytes failed to produce TNF-α following stimulation with C. tropicalis Ags.However, TNF-α treatment of neutrophils in vitro enhanced their ability to kill C. tropicalis.Moreover, CARD9-dependent production of TNF-α enhances the candidacidal capacity of neutrophils, limiting fungal disease during disseminated C. tropicalis infection.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology and Clinical Immunology, Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15261;

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Related in: MedlinePlus

Neutrophils and monocytes are required for protection against disseminated C. tropicalis. (A and B) WT mice were treated with anti-Ly6G or anti-Gr1 Ab or isotype-control Abs on days −1 and +1 and infected with 1 × 104 CFU/g C. tropicalis yeast cells on day 0. (A) CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6G−Ly6C+ monocyte numbers were measured by flow cytometry of blood samples at the time of C. tropicalis infection. Data are representative of two experiments (each data point represents an individual mouse). Cells were gated through leukocyte, single cell, and live cell gates. *p < 0.05, Mann–Whitney U test. (B) Survival of each cohort was monitored for 28 d. Data are pooled from six experiments (WT + PBS, n = 16; WT + IgG2a, n = 8; WT + IgG2b, n = 12; WT + anti-Ly6G, n = 10; WT + anti-Gr1, n = 18). ****p < 0.0001, log-rank (Mantel–Cox) test. (C and D) WT mice were treated with PBS, PBS liposomes (PBS-lip), or clodronate liposomes (Clodronate-lip) 24 h prior to infection with C. tropicalis yeast cells on day 0. (C) CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6G−Ly6C+ monocyte numbers were measured by flow cytometry of blood samples at the time of C. tropicalis infection. Data are representative of two experiments (each data point represents an individual mouse). Cells were gated through leukocyte, single cell, and live cell gates. *p < 0.05, Mann–Whitney U test. (D) Survival of each cohort was monitored for 28 d. Data are pooled from two experiments (WT + PBS, n = 5; WT + PBS-lip, n = 7; WT + clodronate-lip, n = 8). ****p < 0.0001, log-rank (Mantel–Cox) test. (E) WT mice were treated with PBS, PBS-lip, or clodronate-lip 3 d prior to infection with C. tropicalis yeast cells on day 0. Survival of each cohort was monitored for 28 d. Data are pooled from two experiments (WT + PBS, n = 10; WT + PBS-lip, n = 9; WT + clodronate-lip, n = 10). *p < 0.05, log-rank (Mantel–Cox) test.
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fig03: Neutrophils and monocytes are required for protection against disseminated C. tropicalis. (A and B) WT mice were treated with anti-Ly6G or anti-Gr1 Ab or isotype-control Abs on days −1 and +1 and infected with 1 × 104 CFU/g C. tropicalis yeast cells on day 0. (A) CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6G−Ly6C+ monocyte numbers were measured by flow cytometry of blood samples at the time of C. tropicalis infection. Data are representative of two experiments (each data point represents an individual mouse). Cells were gated through leukocyte, single cell, and live cell gates. *p < 0.05, Mann–Whitney U test. (B) Survival of each cohort was monitored for 28 d. Data are pooled from six experiments (WT + PBS, n = 16; WT + IgG2a, n = 8; WT + IgG2b, n = 12; WT + anti-Ly6G, n = 10; WT + anti-Gr1, n = 18). ****p < 0.0001, log-rank (Mantel–Cox) test. (C and D) WT mice were treated with PBS, PBS liposomes (PBS-lip), or clodronate liposomes (Clodronate-lip) 24 h prior to infection with C. tropicalis yeast cells on day 0. (C) CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6G−Ly6C+ monocyte numbers were measured by flow cytometry of blood samples at the time of C. tropicalis infection. Data are representative of two experiments (each data point represents an individual mouse). Cells were gated through leukocyte, single cell, and live cell gates. *p < 0.05, Mann–Whitney U test. (D) Survival of each cohort was monitored for 28 d. Data are pooled from two experiments (WT + PBS, n = 5; WT + PBS-lip, n = 7; WT + clodronate-lip, n = 8). ****p < 0.0001, log-rank (Mantel–Cox) test. (E) WT mice were treated with PBS, PBS-lip, or clodronate-lip 3 d prior to infection with C. tropicalis yeast cells on day 0. Survival of each cohort was monitored for 28 d. Data are pooled from two experiments (WT + PBS, n = 10; WT + PBS-lip, n = 9; WT + clodronate-lip, n = 10). *p < 0.05, log-rank (Mantel–Cox) test.

Mentions: These results pointed to myeloid cells as likely mediators of immunity to C. tropicalis. Consistent with this idea is the fact that CARD9 is primarily expressed in myeloid lineage cells (12). Furthermore, neutropenia is a risk factor for disseminated C. tropicalis infection in humans, and mice deficient in neutrophils or monocytes/macrophages display increased susceptibility to disseminated C. albicans infection (25, 40–43). Accordingly, we first investigated a role for neutrophils in immunity to C. tropicalis by treating WT mice with anti-Ly6G Abs to deplete this population (Fig. 3A). Mice were given Abs on days −1 and +1 postinfection, and survival following C. tropicalis infection was assessed. As predicted, WT mice given anti-Ly6G Abs were significantly more susceptible to disseminated C. tropicalis infection compared with controls (Fig. 3B). All anti-Ly6G Ab–treated mice succumbed to infection by day 17 in contrast to only ∼10% mortality observed in controls at this time point (Fig. 3B). We also treated WT mice with anti-Gr1 Abs to deplete both neutrophils and monocytes (Fig. 3A). Anti-Gr1 Ab–treated mice were even more susceptible to C. tropicalis than were anti-Ly6G Ab–treated mice, with 100% mortality by day 8 (Fig. 3B). These results suggest that neutrophils are crucial for protective responses to C. tropicalis. Moreover, depletion of both neutrophils and monocytes leads to further enhanced susceptibility to disseminated C. tropicalis infection compared with depletion of neutrophils alone.


Delinking CARD9 and IL-17: CARD9 Protects against Candida tropicalis Infection through a TNF-α-Dependent, IL-17-Independent Mechanism.

Whibley N, Jaycox JR, Reid D, Garg AV, Taylor JA, Clancy CJ, Nguyen MH, Biswas PS, McGeachy MJ, Brown GD, Gaffen SL - J. Immunol. (2015)

Neutrophils and monocytes are required for protection against disseminated C. tropicalis. (A and B) WT mice were treated with anti-Ly6G or anti-Gr1 Ab or isotype-control Abs on days −1 and +1 and infected with 1 × 104 CFU/g C. tropicalis yeast cells on day 0. (A) CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6G−Ly6C+ monocyte numbers were measured by flow cytometry of blood samples at the time of C. tropicalis infection. Data are representative of two experiments (each data point represents an individual mouse). Cells were gated through leukocyte, single cell, and live cell gates. *p < 0.05, Mann–Whitney U test. (B) Survival of each cohort was monitored for 28 d. Data are pooled from six experiments (WT + PBS, n = 16; WT + IgG2a, n = 8; WT + IgG2b, n = 12; WT + anti-Ly6G, n = 10; WT + anti-Gr1, n = 18). ****p < 0.0001, log-rank (Mantel–Cox) test. (C and D) WT mice were treated with PBS, PBS liposomes (PBS-lip), or clodronate liposomes (Clodronate-lip) 24 h prior to infection with C. tropicalis yeast cells on day 0. (C) CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6G−Ly6C+ monocyte numbers were measured by flow cytometry of blood samples at the time of C. tropicalis infection. Data are representative of two experiments (each data point represents an individual mouse). Cells were gated through leukocyte, single cell, and live cell gates. *p < 0.05, Mann–Whitney U test. (D) Survival of each cohort was monitored for 28 d. Data are pooled from two experiments (WT + PBS, n = 5; WT + PBS-lip, n = 7; WT + clodronate-lip, n = 8). ****p < 0.0001, log-rank (Mantel–Cox) test. (E) WT mice were treated with PBS, PBS-lip, or clodronate-lip 3 d prior to infection with C. tropicalis yeast cells on day 0. Survival of each cohort was monitored for 28 d. Data are pooled from two experiments (WT + PBS, n = 10; WT + PBS-lip, n = 9; WT + clodronate-lip, n = 10). *p < 0.05, log-rank (Mantel–Cox) test.
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fig03: Neutrophils and monocytes are required for protection against disseminated C. tropicalis. (A and B) WT mice were treated with anti-Ly6G or anti-Gr1 Ab or isotype-control Abs on days −1 and +1 and infected with 1 × 104 CFU/g C. tropicalis yeast cells on day 0. (A) CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6G−Ly6C+ monocyte numbers were measured by flow cytometry of blood samples at the time of C. tropicalis infection. Data are representative of two experiments (each data point represents an individual mouse). Cells were gated through leukocyte, single cell, and live cell gates. *p < 0.05, Mann–Whitney U test. (B) Survival of each cohort was monitored for 28 d. Data are pooled from six experiments (WT + PBS, n = 16; WT + IgG2a, n = 8; WT + IgG2b, n = 12; WT + anti-Ly6G, n = 10; WT + anti-Gr1, n = 18). ****p < 0.0001, log-rank (Mantel–Cox) test. (C and D) WT mice were treated with PBS, PBS liposomes (PBS-lip), or clodronate liposomes (Clodronate-lip) 24 h prior to infection with C. tropicalis yeast cells on day 0. (C) CD45+CD11b+Ly6G+ neutrophil and CD45+CD11b+Ly6G−Ly6C+ monocyte numbers were measured by flow cytometry of blood samples at the time of C. tropicalis infection. Data are representative of two experiments (each data point represents an individual mouse). Cells were gated through leukocyte, single cell, and live cell gates. *p < 0.05, Mann–Whitney U test. (D) Survival of each cohort was monitored for 28 d. Data are pooled from two experiments (WT + PBS, n = 5; WT + PBS-lip, n = 7; WT + clodronate-lip, n = 8). ****p < 0.0001, log-rank (Mantel–Cox) test. (E) WT mice were treated with PBS, PBS-lip, or clodronate-lip 3 d prior to infection with C. tropicalis yeast cells on day 0. Survival of each cohort was monitored for 28 d. Data are pooled from two experiments (WT + PBS, n = 10; WT + PBS-lip, n = 9; WT + clodronate-lip, n = 10). *p < 0.05, log-rank (Mantel–Cox) test.
Mentions: These results pointed to myeloid cells as likely mediators of immunity to C. tropicalis. Consistent with this idea is the fact that CARD9 is primarily expressed in myeloid lineage cells (12). Furthermore, neutropenia is a risk factor for disseminated C. tropicalis infection in humans, and mice deficient in neutrophils or monocytes/macrophages display increased susceptibility to disseminated C. albicans infection (25, 40–43). Accordingly, we first investigated a role for neutrophils in immunity to C. tropicalis by treating WT mice with anti-Ly6G Abs to deplete this population (Fig. 3A). Mice were given Abs on days −1 and +1 postinfection, and survival following C. tropicalis infection was assessed. As predicted, WT mice given anti-Ly6G Abs were significantly more susceptible to disseminated C. tropicalis infection compared with controls (Fig. 3B). All anti-Ly6G Ab–treated mice succumbed to infection by day 17 in contrast to only ∼10% mortality observed in controls at this time point (Fig. 3B). We also treated WT mice with anti-Gr1 Abs to deplete both neutrophils and monocytes (Fig. 3A). Anti-Gr1 Ab–treated mice were even more susceptible to C. tropicalis than were anti-Ly6G Ab–treated mice, with 100% mortality by day 8 (Fig. 3B). These results suggest that neutrophils are crucial for protective responses to C. tropicalis. Moreover, depletion of both neutrophils and monocytes leads to further enhanced susceptibility to disseminated C. tropicalis infection compared with depletion of neutrophils alone.

Bottom Line: Consistently, WT mice depleted of TNF-α were more susceptible to C. tropicalis, and CARD9-deficient neutrophils and monocytes failed to produce TNF-α following stimulation with C. tropicalis Ags.However, TNF-α treatment of neutrophils in vitro enhanced their ability to kill C. tropicalis.Moreover, CARD9-dependent production of TNF-α enhances the candidacidal capacity of neutrophils, limiting fungal disease during disseminated C. tropicalis infection.

View Article: PubMed Central - PubMed

Affiliation: Division of Rheumatology and Clinical Immunology, Department of Medicine, University of Pittsburgh, Pittsburgh, PA 15261;

Show MeSH
Related in: MedlinePlus