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. 2004 Oct;72(10):5868-76.
doi: 10.1128/IAI.72.10.5868-5876.2004.

Dysregulated inflammatory response to Candida albicans in a C5-deficient mouse strain

Alaka Mullick  1 , Miria Elias, Serge Picard, Lucie Bourget, Orce Jovcevski, Susan Gauthier, Ashleigh Tuite, Penelope Harakidas, Craig Bihun, Bernard Massie, Philippe Gros
Affiliations

Dysregulated inflammatory response to Candida albicans in a C5-deficient mouse strain

Alaka Mullick et al. Infect Immun. 2004 Oct.

Abstract

Experimental infection of inbred mouse strains with Candida albicans provides a good model system to identify host genetic determinants that regulate onset of, response to, and ultimate outcome of disseminated candidiasis. The A/J mouse strain is exquisitely sensitive to infection with C. albicans, while the C57BL/6J strain is relatively resistant, as measured by survival following intravenous injection of Candida blastospores. This differential susceptibility is caused by an A/J-specific loss-of-function mutation in the C5 component of the complement pathway. C5 plays several critical roles in host response to infection, including target lysis and phagocyte recruitment. Therefore, to determine which of its functions were required for host resistance to candidiasis, a detailed comparative analysis of pathophysiology and host response to acute C. albicans infection was conducted in A/J and C57BL/6J mice. C5-sufficient C57BL/6J mice were found to succumb late in infection due to severe kidney pathology, typified by fungal replication and robust neutrophil-based inflammatory response associated with extensive tissue damage. In contrast, A/J mice were moribund within 24 h postinfection but displayed little if any kidney damage despite an inability to mobilize granulocytes and a high fungal load in the kidney. Rather, C5 deficiency in A/J mice was associated with higher levels of circulating cytokines tumor necrosis factor alpha, interleukin-6, monocyte chemotactic protein 1 (MCP-1), MCP-5, and eotaxin in response to C. albicans. Transfer of the C5-defective allele from A/J onto a C57BL/6J genetic background in recombinant congenic strain BcA17 recapitulated the phenotypic aspects of the susceptibility of A/J mice to C. albicans, confirming the causative role of C5 deficiency in the dysregulated cytokine response.

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Figures

FIG. 1.
FIG. 1.
Differential susceptibility of A/J and C57BL/6J mice to C. albicans infection. (A) Survival of A/J and C57BL/6J mice after intravenous infection with C. albicans. Twelve A/J and 25 C57BL/6J (B6) mice were injected intravenously with a C. albicans suspension and subsequently monitored for clinical signs as described in Materials and Methods. The figure shows the percent survival at various intervals postinfection. The figure presents data from one of three independent experiments. All three experiments gave comparable results. (B) Tissue distribution of C. albicans in A/J and C57BL/6J mice 24 h postinfection. Six mice each of the two strains were injected intravenously with 3 × 105 Candida blastospores, and tissue fungal load was determined 24 h postinfection as described in Materials and Methods. Results represent a compilation of two independent experiments. The standard error of the mean is indicated for each sample. Asterisks indicate statistically significant differences (P = 0.001).
FIG. 2.
FIG. 2.
Kidney pathophysiology in C. albicans-infected A/J and C57BL/6J mice. (A) Kidney function. BUN levels were determined in serum from uninfected and C. albicans-infected A/J and C57BL/6J mice (A/Jinf and B6inf., respectively) collected 24 h postinfection and in serum from C57BL/6J mice collected between days 7 and 30 postinfection (d7-30), when they were moribund (B6m). Isolation of serum and BUN determination were carried out as described in Materials and Methods. The standard error of the mean is indicated for each sample. Numbers in parentheses represent the number of mice analyzed in each experimental group. The results represent a compilation of two independent experiments. Asterisks indicate statistically significant differences (P = 0.001). (B) Kidney fungal load. Mice of the two strains were infected with 3 × 105 Candida blastospores, and kidney fungal load was determined 24 h postinfection (day 1) in both A/J and C57BL/6J mice. Between days 7 and 10 postinfection (days 7 to 10), moribund C57BL/6J (B6m) and healthy C57BL/6J mice were euthanized, and the fungal load was determined as described in Materials and Methods. The standard error of the mean is indicated for each sample. Numbers in parentheses represent the number of mice analyzed in each experimental group. Asterisks indicate statistically significant differences (P = 0.005).
FIG. 3.
FIG. 3.
Flow cytometric analysis of Gr-1+ cells in the spleens of A/J and C57BL/6J mice during C. albicans infection. Spleens were removed from control and C. albicans-infected A/J and C57BL/6J (A/Jinf. And B6inf., respectively) mice 24 h postinfection. Spleen cell preparations were stained with R-phycoerythrin-conjugated rat anti-mouse Gr-1 antibody. The percentage of cells positive for R-phycoerythrin fluorescence was determined by flow cytometry. The standard error of the mean is indicated for each sample. Numbers in parentheses indicate the number of mice analyzed per experimental group. Asterisks indicate statistically significant differences (P = 0.005).
FIG. 4.
FIG. 4.
TNF-α levels in the circulation of C. albicans-infected A/J and C57BL/6J mice. Mice of the two strains were infected with 3 × 105 Candida blastospores and exsanguinated at the indicated times. TNF-α levels were determined in serum from control and C. albicans-infected mice as described in Materials and Methods. The standard error of the mean is indicated for each sample. Numbers in parentheses represent the number of mice analyzed in each experimental group.
FIG. 5.
FIG. 5.
Cytokine profile in the circulation of C. albicans-infected A/J and C57BL/6J mice. Four mice each of the two strains were injected intravenously with 3 × 105 Candida blastospores and exsanguinated 24 h postinfection. Serum was isolated and pooled for each of the four experimental groups, control and C. albicans-infected A/J and C57BL/6J mice. Four hundred microliters of serum was used to determine the levels of cytokines and chemokines with the RayBio mouse cytokine array II membranes as described in Materials and Methods. The figure presents one of three experiments, all of which gave similar results. GCFS, granulocyte colony-stimulating factor.
FIG. 6.
FIG. 6.
Cytokine profile in the peritoneal exudate of A/J and C57BL/6J mice injected intraperitoneally with heat-killed C. albicans blastospores. Four mice each of the two strains were injected intraperitoneally with 108 heat-killed Candida blastospores in 1 ml of PBS. Sixteen hours postinjection, a peritoneal lavage was performed with 5 ml of PBS. Four hundred microliters of peritoneal fluid was used to determine the levels of cytokines and chemokines with the RayBio mouse cytokine array II membranes as described in Materials and Methods. The figure presents results from one of two experiments, both of which gave similar results.
FIG. 7.
FIG. 7.
C. albicans infection of BcA17 mice. A/J, C57BL/6J, and BcA17 mice were injected with 3 × 105 C. albicans organisms intravenously. Twenty-four hours later, the C. albicans-infected and uninfected A/J, C57BL/6J, and BcA17 mice were euthanized by exsanguination, and the kidneys were harvested. Kidney fungal load (A) and serum cytokine levels (B and C) were determined as described in Materials and Methods. The standard error of the mean is indicated for each sample. Numbers in parentheses indicate the number of mice analyzed in each experimental group. Asterisks indicate statistically significant differences (P = 0.005).
FIG. 8.
FIG. 8.
Circulating cytokines in BcA17 mice. Six A/J, C57BL/6J, and BcA17 mice each were injected with 3 × 105 C. albicans intravenously (A/J inf., C57BL/6J inf, and BcA17 inf., respectively). Twenty-four hours later, the C. albicans-infected and six uninfected BcA17 mice were euthanized by exsanguination. Serum cytokine profiles were determined with the RayBio mouse cytokine array II membranes as described in Materials and Methods. The figure presents one of two experiments, both of which gave similar results.
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References

    1. Ashman, R. B. 1998. Candida albicans: pathogenesis, immunity and host defense. Res. Immunol. 149:281-288. - PubMed
    1. Ashman, R. B. 1998. A gene (Carg1) that regulates tissue resistance to Candida albicans maps to chromosome 14 of the mouse. Microb. Pathog. 25:333-335. - PubMed
    1. Ashman, R. B., E. M. Bolitho, and J. M. Papadimitriou. 1993. Patterns of resistance to Candida albicans in inbred mouse strains. Immunol. Cell Biol. 71:221-225. - PubMed
    1. Ashman, R. B., A. Fulurija, and J. M. Papadimitriou. 1997. Evidence that two independent host genes influence the severity of tissue damage and susceptibility to acute pyelonephritis in murine candidiasis. Microb. Pathog. 22:187-192. - PubMed
    1. Ashman, R. B., A. Fulurija, and J. M. Papadimitriou. 1998. A second Candida albicans resistance gene (Carg2) regulates tissue damage, but not fungal clearance, in sub-lethal murine systemic infection. Microb. Pathog. 25:349-352. - PubMed

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