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Endogenous IL-13 plays a crucial role in liver granuloma maturation during Leishmania donovani infection, independent of IL-4Rα-responsive macrophages and neutrophils.

McFarlane E, Carter KC, McKenzie AN, Kaye PM, Brombacher F, Alexander J - J. Infect. Dis. (2011)

Bottom Line: This correlated with significantly retarded granuloma maturation in IL-13(-/-) mice, defective interferon γ (IFN-γ) production, and elevated IL-4 and interleukin 10 (IL-10) levels.Because murine lymphocytes do not have IL-13 receptors, we examined the ability of macrophage/neutrophil-specific IL-4Rα(-/-) mice to control primary infection with L. donovani and to respond to chemotherapy.Consequently, in L. donovani infected BALB/c mice, IL-13 promotes hepatic granuloma formation and controls parasite burdens independently of direct effects on macrophages/neutrophils.

View Article: PubMed Central - PubMed

Affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK.

ABSTRACT
Previous studies comparing interleukin 4 receptor α (IL-4Rα)(-/-) and interleukin 4 (IL-4)(-/-) BALB/c mice have indicated that interleukin 13 (IL-13), whose receptor shares the IL-4Rα subunit with IL-4, plays a protective role during visceral leishmaniasis. We demonstrate that IL-13(-/-) BALB/c mice were less able to control hepatic growth of Leishmania donovani compared with wild-type mice. This correlated with significantly retarded granuloma maturation in IL-13(-/-) mice, defective interferon γ (IFN-γ) production, and elevated IL-4 and interleukin 10 (IL-10) levels. L. donovani-infected IL-13(-/-) mice also responded poorly to sodium stibogluconate-mediated chemotherapy compared with wild-type BALB/c mice. Because murine lymphocytes do not have IL-13 receptors, we examined the ability of macrophage/neutrophil-specific IL-4Rα(-/-) mice to control primary infection with L. donovani and to respond to chemotherapy. Macrophage/neutrophil-specific IL-4Rα(-/-) mice were as resistant to leishmaniasis as wild-type mice, and chemotherapy retained its efficacy. Consequently, in L. donovani infected BALB/c mice, IL-13 promotes hepatic granuloma formation and controls parasite burdens independently of direct effects on macrophages/neutrophils.

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Mean serum IFN-γ (A), IL-13 (B), IL-10 (C), and IL-4 (D) levels ± standard errors in wild-type and IL-13-/- BALB/c mice (n = 4) at day 35, and IL-12 p40/70 (E) at days 14 and 35 postinfection with L. donovani as measured in individual serum samples by enzyme-linked immunosorbent assay. Representative data from 4 independent experiments are shown. No measurable cytokine levels were observed in sera from uninfected wild-type and IL-13-/-mice. Analysis of intracellular IFN-γ expression (F) was performed on day 35 postinfection on splenocytes stimulated in the presence of 50 ng/mL phorbol myristate acetate (Sigma-Aldrich) and 500 ng/mL of ionomycin (Sigma-Aldrich) for 4 hours, with the addition of Brefeldin A during the last 2 hours as previously described. The antimouse antibody PerCP-labeled anti-CD4+, activated protein C–labeled anti-CD8+, peridinin chlorophyll protein–labeled, and activated protein C–labeled IgG isotype controls were used to stain the cell surfaces, and the protein E–labeled anti-mouse IFN-γ and protein E–labeled IgG isotype control were used for intracellular staining. All antibodies were obtained from BD Biosciences. Cells were analyzed with the FACSCanto. The FACsDiva software was used to analyze results. Representative of 2 separate experiments. *P < .05, **P < .01 wild-type versus IL-13-/-. ND = not detected.
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fig3: Mean serum IFN-γ (A), IL-13 (B), IL-10 (C), and IL-4 (D) levels ± standard errors in wild-type and IL-13-/- BALB/c mice (n = 4) at day 35, and IL-12 p40/70 (E) at days 14 and 35 postinfection with L. donovani as measured in individual serum samples by enzyme-linked immunosorbent assay. Representative data from 4 independent experiments are shown. No measurable cytokine levels were observed in sera from uninfected wild-type and IL-13-/-mice. Analysis of intracellular IFN-γ expression (F) was performed on day 35 postinfection on splenocytes stimulated in the presence of 50 ng/mL phorbol myristate acetate (Sigma-Aldrich) and 500 ng/mL of ionomycin (Sigma-Aldrich) for 4 hours, with the addition of Brefeldin A during the last 2 hours as previously described. The antimouse antibody PerCP-labeled anti-CD4+, activated protein C–labeled anti-CD8+, peridinin chlorophyll protein–labeled, and activated protein C–labeled IgG isotype controls were used to stain the cell surfaces, and the protein E–labeled anti-mouse IFN-γ and protein E–labeled IgG isotype control were used for intracellular staining. All antibodies were obtained from BD Biosciences. Cells were analyzed with the FACSCanto. The FACsDiva software was used to analyze results. Representative of 2 separate experiments. *P < .05, **P < .01 wild-type versus IL-13-/-. ND = not detected.

Mentions: IFN-γ is important in controlling granuloma maturation; Murray et al have previously shown that IFN-γ-deficient mice are unable to control parasite burdens and granuloma formation [2]. We found serum IFN-γ levels at day 35 postinfection to be significantly greater (P < .0001) in L. donovani–infected wild-type mice compared with levels in IL-13-/- BALB/c mice (Figure 3A). We also found L. donovani to induce high serum IL-13 levels in L. donovani–infected wild-type mice (Figure 3B). Conversely, both IL-4 and IL-10 serum levels were greater in IL-13-/- mice compared with levels in wild-type mice (Figure 3C and D respectively). Together these results show a positive association of IFN-γ and IL-13 production with granuloma maturation and a negative association with IL-4 and IL-10 levels. Consistent with these observations, IL-12 serum levels peaked earlier in infection in wild-type mice compared with levels in IL-13-/- mice (Figure 3E). As well as systemic cytokine levels, the percentage of IFN-γ-secreting CD4+ T cells and CD8+ T cells was significantly greater in the spleens of wild-type mice than in IL-13-/- mice (Figure 3F; wild-type mice, 7.9 ± 0.265% CD4+ IFN-γ+ T cells and 15.5 ± 0.354% CD8+ IFN-γ+ T cells; IL-13-/- mice, 6.1 ± 0.24% CD4+ IFN-γ+ T cells and 12.0 ± 0.841% CD8+ IFN-γ+ T cells).


Endogenous IL-13 plays a crucial role in liver granuloma maturation during Leishmania donovani infection, independent of IL-4Rα-responsive macrophages and neutrophils.

McFarlane E, Carter KC, McKenzie AN, Kaye PM, Brombacher F, Alexander J - J. Infect. Dis. (2011)

Mean serum IFN-γ (A), IL-13 (B), IL-10 (C), and IL-4 (D) levels ± standard errors in wild-type and IL-13-/- BALB/c mice (n = 4) at day 35, and IL-12 p40/70 (E) at days 14 and 35 postinfection with L. donovani as measured in individual serum samples by enzyme-linked immunosorbent assay. Representative data from 4 independent experiments are shown. No measurable cytokine levels were observed in sera from uninfected wild-type and IL-13-/-mice. Analysis of intracellular IFN-γ expression (F) was performed on day 35 postinfection on splenocytes stimulated in the presence of 50 ng/mL phorbol myristate acetate (Sigma-Aldrich) and 500 ng/mL of ionomycin (Sigma-Aldrich) for 4 hours, with the addition of Brefeldin A during the last 2 hours as previously described. The antimouse antibody PerCP-labeled anti-CD4+, activated protein C–labeled anti-CD8+, peridinin chlorophyll protein–labeled, and activated protein C–labeled IgG isotype controls were used to stain the cell surfaces, and the protein E–labeled anti-mouse IFN-γ and protein E–labeled IgG isotype control were used for intracellular staining. All antibodies were obtained from BD Biosciences. Cells were analyzed with the FACSCanto. The FACsDiva software was used to analyze results. Representative of 2 separate experiments. *P < .05, **P < .01 wild-type versus IL-13-/-. ND = not detected.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3105032&req=5

fig3: Mean serum IFN-γ (A), IL-13 (B), IL-10 (C), and IL-4 (D) levels ± standard errors in wild-type and IL-13-/- BALB/c mice (n = 4) at day 35, and IL-12 p40/70 (E) at days 14 and 35 postinfection with L. donovani as measured in individual serum samples by enzyme-linked immunosorbent assay. Representative data from 4 independent experiments are shown. No measurable cytokine levels were observed in sera from uninfected wild-type and IL-13-/-mice. Analysis of intracellular IFN-γ expression (F) was performed on day 35 postinfection on splenocytes stimulated in the presence of 50 ng/mL phorbol myristate acetate (Sigma-Aldrich) and 500 ng/mL of ionomycin (Sigma-Aldrich) for 4 hours, with the addition of Brefeldin A during the last 2 hours as previously described. The antimouse antibody PerCP-labeled anti-CD4+, activated protein C–labeled anti-CD8+, peridinin chlorophyll protein–labeled, and activated protein C–labeled IgG isotype controls were used to stain the cell surfaces, and the protein E–labeled anti-mouse IFN-γ and protein E–labeled IgG isotype control were used for intracellular staining. All antibodies were obtained from BD Biosciences. Cells were analyzed with the FACSCanto. The FACsDiva software was used to analyze results. Representative of 2 separate experiments. *P < .05, **P < .01 wild-type versus IL-13-/-. ND = not detected.
Mentions: IFN-γ is important in controlling granuloma maturation; Murray et al have previously shown that IFN-γ-deficient mice are unable to control parasite burdens and granuloma formation [2]. We found serum IFN-γ levels at day 35 postinfection to be significantly greater (P < .0001) in L. donovani–infected wild-type mice compared with levels in IL-13-/- BALB/c mice (Figure 3A). We also found L. donovani to induce high serum IL-13 levels in L. donovani–infected wild-type mice (Figure 3B). Conversely, both IL-4 and IL-10 serum levels were greater in IL-13-/- mice compared with levels in wild-type mice (Figure 3C and D respectively). Together these results show a positive association of IFN-γ and IL-13 production with granuloma maturation and a negative association with IL-4 and IL-10 levels. Consistent with these observations, IL-12 serum levels peaked earlier in infection in wild-type mice compared with levels in IL-13-/- mice (Figure 3E). As well as systemic cytokine levels, the percentage of IFN-γ-secreting CD4+ T cells and CD8+ T cells was significantly greater in the spleens of wild-type mice than in IL-13-/- mice (Figure 3F; wild-type mice, 7.9 ± 0.265% CD4+ IFN-γ+ T cells and 15.5 ± 0.354% CD8+ IFN-γ+ T cells; IL-13-/- mice, 6.1 ± 0.24% CD4+ IFN-γ+ T cells and 12.0 ± 0.841% CD8+ IFN-γ+ T cells).

Bottom Line: This correlated with significantly retarded granuloma maturation in IL-13(-/-) mice, defective interferon γ (IFN-γ) production, and elevated IL-4 and interleukin 10 (IL-10) levels.Because murine lymphocytes do not have IL-13 receptors, we examined the ability of macrophage/neutrophil-specific IL-4Rα(-/-) mice to control primary infection with L. donovani and to respond to chemotherapy.Consequently, in L. donovani infected BALB/c mice, IL-13 promotes hepatic granuloma formation and controls parasite burdens independently of direct effects on macrophages/neutrophils.

View Article: PubMed Central - PubMed

Affiliation: Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, Glasgow, UK.

ABSTRACT
Previous studies comparing interleukin 4 receptor α (IL-4Rα)(-/-) and interleukin 4 (IL-4)(-/-) BALB/c mice have indicated that interleukin 13 (IL-13), whose receptor shares the IL-4Rα subunit with IL-4, plays a protective role during visceral leishmaniasis. We demonstrate that IL-13(-/-) BALB/c mice were less able to control hepatic growth of Leishmania donovani compared with wild-type mice. This correlated with significantly retarded granuloma maturation in IL-13(-/-) mice, defective interferon γ (IFN-γ) production, and elevated IL-4 and interleukin 10 (IL-10) levels. L. donovani-infected IL-13(-/-) mice also responded poorly to sodium stibogluconate-mediated chemotherapy compared with wild-type BALB/c mice. Because murine lymphocytes do not have IL-13 receptors, we examined the ability of macrophage/neutrophil-specific IL-4Rα(-/-) mice to control primary infection with L. donovani and to respond to chemotherapy. Macrophage/neutrophil-specific IL-4Rα(-/-) mice were as resistant to leishmaniasis as wild-type mice, and chemotherapy retained its efficacy. Consequently, in L. donovani infected BALB/c mice, IL-13 promotes hepatic granuloma formation and controls parasite burdens independently of direct effects on macrophages/neutrophils.

Show MeSH
Related in: MedlinePlus