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Extracellular Superoxide Dismutase Protects Histoplasma Yeast Cells from Host-Derived Oxidative Stress

DOI: 10.1371/journal.ppat.1002713

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Abstract:

In order to establish infections within the mammalian host, pathogens must protect themselves against toxic reactive oxygen species produced by phagocytes of the immune system. The fungal pathogen Histoplasma capsulatum infects both neutrophils and macrophages but the mechanisms enabling Histoplasma yeasts to survive in these phagocytes have not been fully elucidated. We show that Histoplasma yeasts produce a superoxide dismutase (Sod3) and direct it to the extracellular environment via N-terminal and C-terminal signals which promote its secretion and association with the yeast cell surface. This localization permits Sod3 to protect yeasts specifically from exogenous superoxide whereas amelioration of endogenous reactive oxygen depends on intracellular dismutases such as Sod1. While infection of resting macrophages by Histoplasma does not stimulate the phagocyte oxidative burst, interaction with polymorphonuclear leukocytes (PMNs) and cytokine-activated macrophages triggers production of reactive oxygen species (ROS). Histoplasma yeasts producing Sod3 survive co-incubation with these phagocytes but yeasts lacking Sod3 are rapidly eliminated through oxidative killing similar to the effect of phagocytes on Candida albicans yeasts. The protection provided by Sod3 against host-derived ROS extends in vivo. Without Sod3, Histoplasma yeasts are attenuated in their ability to establish respiratory infections and are rapidly cleared with the onset of adaptive immunity. The virulence of Sod3-deficient yeasts is restored in murine hosts unable to produce superoxide due to loss of the NADPH-oxidase function. These results demonstrate that phagocyte-produced ROS contributes to the immune response to Histoplasma and that Sod3 facilitates Histoplasma pathogenesis by detoxifying host-derived reactive oxygen thereby enabling Histoplasma survival.

References

[1]  Imlay JA (2003) Pathways of oxidative damage. Annu Rev Microbiol 57: 395–418.
[2]  Johnston RB Jr, Kitagawa S (1985) Molecular basis for the enhanced respiratory burst of activated macrophages. Fed Proc 44: 2927–2932.
[3]  Murray HW, Spitalny GL, Nathan CF (1985) Activation of mouse peritoneal macrophages in vitro and in vivo by interferon-gamma. J Immunol 134: 1619–1622.
[4]  Ajello L (1971) The medical mycological iceberg. HSMHA Health Rep 86: 437–448.
[5]  Fleischmann J, Wu-Hsieh B, Howard DH (1990) The intracellular fate of Histoplasma capsulatum in human macrophages is unaffected by recombinant human interferon-gamma. J Infect Dis 161: 143–145.
[6]  Brummer E, Stevens DA (1995) Antifungal mechanisms of activated murine bronchoalveolar or peritoneal macrophages for Histoplasma capsulatum. Clin Exp Immunol 102: 65–70.
[7]  Newman SL, Bucher C, Rhodes J, Bullock WE (1990) Phagocytosis of Histoplasma capsulatum yeasts and microconidia by human cultured macrophages and alveolar macrophages. Cellular cytoskeleton requirement for attachment and ingestion. J Clin Invest 85: 223–230.
[8]  Knox KS, Hage CA (2010) Histoplasmosis. Proc Am Thorac Soc 7: 169–172.
[9]  Kauffman CA (2001) Pulmonary histoplasmosis. Curr Infect Dis Rep 3: 279–285.
[10]  Eissenberg LG, Goldman WE (1987) Histoplasma capsulatum fails to trigger release of superoxide from macrophages. Infect Immun 55: 29–34.
[11]  Wolf JE, Kerchberger V, Kobayashi GS, Little JR (1987) Modulation of the macrophage oxidative burst by Histoplasma capsulatum. J Immunol 138: 582–586.
[12]  Wolf JE, Massof SE (1990) In vivo activation of macrophage oxidative burst activity by cytokines and amphotericin B. Infect Immun 58: 1296–1300.
[13]  Wolf JE, Abegg AL, Travis SJ, Kobayashi GS, Little JR (1989) Effects of Histoplasma capsulatum on murine macrophage functions: inhibition of macrophage priming, oxidative burst, and antifungal activities. Infect Immun 57: 513–519.
[14]  Baughman RP, Kim CK, Vinegar A, Hendricks DE, Schmidt DJ, et al. (1986) The pathogenesis of experimental pulmonary histoplasmosis. Correlative studies of histopathology, bronchoalveolar lavage, and respiratory function. Am Rev Respir Dis 134: 771–776.
[15]  Deepe GS Jr, Gibbons RS, Smulian AG (2008) Histoplasma capsulatum manifests preferential invasion of phagocytic subpopulations in murine lungs. J Leukocyte Biol 84: 669–678.
[16]  Schnur RA, Newman SL (1990) The respiratory burst response to Histoplasma capsulatum by human neutrophils. Evidence for intracellular trapping of superoxide anion. J Immunol 144: 4765–4772.
[17]  Kurita N, Brummer E, Yoshida S, Nishimura K, Miyaji M (1991) Antifungal activity of murine polymorphonuclear neutrophils against Histoplasma capsulatum. J Med Vet Mycol 29: 133–143.
[18]  Kurita N, Terao K, Brummer E, Ito E, Nishimura K, et al. (1991) Resistance of Histoplasma capsulatum to killing by human neutrophils. Evasion of oxidative burst and lysosomal-fusion products. Mycopathologia 115: 207–213.
[19]  Schaffner A, Davis CE, Schaffner T, Markert M, Douglas H, et al. (1986) In vitro susceptibility of fungi to killing by neutrophil granulocytes discriminates between primary pathogenicity and opportunism. J Clin Invest 78: 511–524.
[20]  Brummer E, Kurita N, Yosihida S, Nishimura K, Miyaji M (1991) Fungistatic activity of human neutrophils against Histoplasma capsulatum: correlation with phagocytosis. J Infect Dis 164: 158–162.
[21]  Holbrook ED, Edwards JA, Youseff BH, Rappleye CA (2011) Definition of the extracellular proteome of pathogenic-phase Histoplasma capsulatum. J Proteome Res 10: 1929–1943.
[22]  Sebghati TS, Engle JT, Goldman WE (2000) Intracellular parasitism by Histoplasma capsulatum: fungal virulence and calcium dependence. Science 290: 1368–1372.
[23]  Peskin AV, Winterbourn CC (2000) A microtiter plate assay for superoxide dismutase using a water-soluble tetrazolium salt (WST-1). Clin Chim Acta 293: 157–166.
[24]  De Groot PW, Ram AF, Klis FM (2005) Features and functions of covalently linked proteins in fungal cell walls. Fungal Genet Biol 42: 657–675.
[25]  Pittet M, Conzelmann A (2007) Biosynthesis and function of GPI proteins in the yeast Saccharomyces cerevisiae. Biochim Biophys Acta 1771: 405–420.
[26]  Edwards JA, Alore EA, Rappleye CA (2011) The yeast-phase virulence requirement for alpha-glucan synthase differs among Histoplasma capsulatum chemotypes. Eukaryot Cell 10: 87–97.
[27]  Bus JS, Gibson JE (1984) Paraquat: model for oxidant-initiated toxicity. Environ Health Perspect 55: 37–46.
[28]  Cocheme HM, Murphy MP (2008) Complex I is the major site of mitochondrial superoxide production by paraquat. J Biol Chem 283: 1786–1798.
[29]  Robertson AK, Cross AR, Jones OT, Andrew PW (1990) The use of diphenylene iodonium, an inhibitor of NADPH oxidase, to investigate the antimicrobial action of human monocyte derived macrophages. J Immunol Methods 133: 175–179.
[30]  Ellis JA, Mayer SJ, Jones OT (1988) The effect of the NADPH oxidase inhibitor diphenyleneiodonium on aerobic and anaerobic microbicidal activities of human neutrophils. Biochem J 251: 887–891.
[31]  Jackson SH, Gallin JI, Holland SM (1995) The p47phox mouse knock-out model of chronic granulomatous disease. J Exp Med 182: 751–758.
[32]  Newman SL, Gootee L, Gabay JE, Selsted ME (2000) Identification of constituents of human neutrophil azurophil granules that mediate fungistasis against Histoplasma capsulatum. Infect Immun 68: 5668–5672.
[33]  Couto MA, Liu L, Lehrer RI, Ganz T (1994) Inhibition of intracellular Histoplasma capsulatum replication by murine macrophages that produce human defensin. Infect Immun 62: 2375–2378.
[34]  Lane TE, Wu-Hsieh BA, Howard DH (1994) Antihistoplasma effect of activated mouse splenic macrophages involves production of reactive nitrogen intermediates. Infect Immun 62: 1940–1945.
[35]  Tan AS, Berridge MV (2000) Superoxide produced by activated neutrophils efficiently reduces the tetrazolium salt, WST-1 to produce a soluble formazan: a simple colorimetric assay for measuring respiratory burst activation and for screening anti-inflammatory agents. J Immunol Methods 238: 59–68.
[36]  Hamilton AJ, Bartholomew MA, Figueroa J, Fenelon LE, Hay RJ (1990) Evidence that the M antigen of Histoplasma capsulatum var. capsulatum is a catalase which exhibits cross-reactivity with other dimorphic fungi. J Med Vet Mycol 28: 479–485.
[37]  Guimaraes AJ, Hamilton AJ, de MG, Nosanchuk JD, Zancope-Oliveira RM (2008) Biological function and molecular mapping of M antigen in yeast phase of Histoplasma capsulatum. PloS One 3: e3449.
[38]  Johnson CH, Klotz MG, York JL, Kruft V, McEwen JE (2002) Redundancy, phylogeny and differential expression of Histoplasma capsulatum catalases. Microbiology 148: 1129–1142.
[39]  Piddington DL, Fang FC, Laessig T, Cooper AM, Orme IM, et al. (2001) Cu,Zn superoxide dismutase of Mycobacterium tuberculosis contributes to survival in activated macrophages that are generating an oxidative burst. Infect Immun 69: 4980–4987.
[40]  De Groote MA, Ochsner UA, Shiloh MU, Nathan C, McCord JM, et al. (1997) Periplasmic superoxide dismutase protects Salmonella from products of phagocyte NADPH-oxidase and nitric oxide synthase. Proc Natl Acad Sci U S A 94: 13997–14001.
[41]  Fang FC, DeGroote MA, Foster JW, Baumler AJ, Ochsner U, et al. (1999) Virulent Salmonella typhimurium has two periplasmic Cu, Zn-superoxide dismutases. Proc Natl Acad Sci U S A 96: 7502–7507.
[42]  Melillo AA, Mahawar M, Sellati TJ, Malik M, Metzger DW, et al. (2009) Identification of Francisella tularensis live vaccine strain CuZn superoxide dismutase as critical for resistance to extracellularly generated reactive oxygen species. J Bacteriol 191: 6447–6456.
[43]  Giles SS, Batinic-Haberle I, Perfect JR, Cox GM (2005) Cryptococcus neoformans mitochondrial superoxide dismutase: an essential link between antioxidant function and high-temperature growth. Eukaryot Cell 4: 46–54.
[44]  Narasipura SD, Ault JG, Behr MJ, Chaturvedi V, Chaturvedi S (2003) Characterization of Cu,Zn superoxide dismutase (SOD1) gene knock-out mutant of Cryptococcus neoformans var. gattii: role in biology and virulence. Mol Microbiol 47: 1681–1694.
[45]  Narasipura SD, Chaturvedi V, Chaturvedi S (2005) Characterization of Cryptococcus neoformans variety gattii SOD2 reveals distinct roles of the two superoxide dismutases in fungal biology and virulence. Mol Microbiol 55: 1782–1800.
[46]  Hwang CS, Rhie GE, Oh JH, Huh WK, Yim HS, et al. (2002) Copper- and zinc-containing superoxide dismutase (Cu/ZnSOD) is required for the protection of Candida albicans against oxidative stresses and the expression of its full virulence. Microbiology 148: 3705–3713.
[47]  Cox GM, Harrison TS, McDade HC, Taborda CP, Heinrich G, et al. (2003) Superoxide dismutase influences the virulence of Cryptococcus neoformans by affecting growth within macrophages. Infect Immun 71: 173–180.
[48]  Gonzalez-Guerrero M, Oger E, Benabdellah K, Azcon-Aguilar C, Lanfranco L, et al. (2010) Characterization of a CuZn superoxide dismutase gene in the arbuscular mycorrhizal fungus Glomus intraradices. Curr Genet 56: 265–274.
[49]  Lanfranco L, Novero M, Bonfante P (2005) The mycorrhizal fungus Gigaspora margarita possesses a CuZn superoxide dismutase that is up-regulated during symbiosis with legume hosts. Plant Physiol 137: 1319–1330.
[50]  Lin CH, Yang SL, Chung KR (2009) The YAP1 homolog-mediated oxidative stress tolerance is crucial for pathogenicity of the necrotrophic fungus Alternaria alternata in citrus. Mol Plant Microbe Interact 22: 942–952.
[51]  Zhang N, Zhang S, Borchert S, Richardson K, Schmid J (2011) High levels of a fungal superoxide dismutase and increased concentration of a PR-10 plant protein in associations between the endophytic fungus Neotyphodium lolii and ryegrass. Mol Plant Microbe Interact 24: 984–992.
[52]  Patel RM, van Kan JA, Bailey AM, Foster GD (2008) RNA-mediated gene silencing of superoxide dismutase (bcsod1) in Botrytis cinerea. Phytopathology 98: 1334–1339.
[53]  Rolke Y, Liu S, Quidde T, Williamson B, Schouten A, et al. (2004) Functional analysis of H(2)O(2)-generating systems in Botrytis cinerea: the major Cu-Zn-superoxide dismutase (BCSOD1) contributes to virulence on French bean, whereas a glucose oxidase (BCGOD1) is dispensable. Mol Plant Pathol 5: 17–27.
[54]  Fradin C, De Groot P, MacCallum D, Schaller M, Klis F, et al. (2005) Granulocytes govern the transcriptional response, morphology and proliferation of Candida albicans in human blood. Mol Microbiol 56: 397–415.
[55]  Martchenko M, Alarco AM, Harcus D, Whiteway M (2004) Superoxide dismutases in Candida albicans: transcriptional regulation and functional characterization of the hyphal-induced SOD5 gene. Mol Biol Cell 15: 456–467.
[56]  Frohner IE, Bourgeois C, Yatsyk K, Majer O, Kuchler K (2009) Candida albicans cell surface superoxide dismutases degrade host-derived reactive oxygen species to escape innate immune surveillance. Mol Microbiol 71: 240–252.
[57]  Bink A, Vandenbosch D, Coenye T, Nelis H, Cammue BP, et al. (2011) Superoxide dismutases are involved in Candida albicans biofilm persistence against miconazole. Antimicrob Agents Chemother 55: 4033–4037.
[58]  Rappleye CA, Eissenberg LG, Goldman WE (2007) Histoplasma capsulatum alpha-(1,3)-glucan blocks innate immune recognition by the beta-glucan receptor. Proc Natl Acad Sci U S A 104: 1366–1370.
[59]  Netea MG, Gow NA, Munro CA, Bates S, Collins C, et al. (2006) Immune sensing of Candida albicans requires cooperative recognition of mannans and glucans by lectin and Toll-like receptors. J Clin Invest 116: 1642–1650.
[60]  Skrzypek F, Cenci E, Pietrella D, Rachini A, Bistoni F, et al. (2009) Dectin-1 is required for human dendritic cells to initiate immune response to Candida albicans through Syk activation. Microbes Infect 11: 661–670.
[61]  Underhill DM, Rossnagle E, Lowell CA, Simmons RM (2005) Dectin-1 activates Syk tyrosine kinase in a dynamic subset of macrophages for reactive oxygen production. Blood 106: 2543–2550.
[62]  Gantner BN, Simmons RM, Underhill DM (2005) Dectin-1 mediates macrophage recognition of Candida albicans yeast but not filaments. EMBO J 24: 1277–1286.
[63]  Kennedy AD, Willment JA, Dorward DW, Williams DL, Brown GD, et al. (2007) Dectin-1 promotes fungicidal activity of human neutrophils. Eur J Immunol 37: 467–478.
[64]  Brummer E, Kurita N, Yoshida S, Nishimura K, Miyaji M (1991) Killing of Histoplasma capsulatum by gamma-interferon-activated human monocyte-derived macrophages: evidence for a superoxide anion-dependent mechanism. J Med Microbiol 35: 29–34.
[65]  Ding AH, Nathan CF, Stuehr DJ (1988) Release of reactive nitrogen intermediates and reactive oxygen intermediates from mouse peritoneal macrophages. Comparison of activating cytokines and evidence for independent production. J Immunol 141: 2407–2412.
[66]  Desai G, Nassar F, Brummer E, Stevens DA (1995) Killing of Histoplasma capsulatum by macrophage colony stimulating factor-treated human monocyte-derived macrophages: role for reactive oxygen intermediates. J Med Microbiol 43: 224–229.
[67]  Wu-Hsieh B, Howard DH (1984) Inhibition of growth of Histoplasma capsulatum by lymphokine-stimulated macrophages. J Immunol 132: 2593–2597.
[68]  Wu-Hsieh BA, Howard DH (1987) Inhibition of the intracellular growth of Histoplasma capsulatum by recombinant murine gamma interferon. Infect Immun 55: 1014–1016.
[69]  Cain JA, Deepe GS Jr (1998) Evolution of the primary immune response to Histoplasma capsulatum in murine lung. Infect Immun 66: 1473–1481.
[70]  Worsham PL, Goldman WE (1988) Quantitative plating of Histoplasma capsulatum without addition of conditioned medium or siderophores. J Med Vet Mycol 26: 137–143.
[71]  Woods JP, Heinecke EL, Goldman WE (1998) Electrotransformation and expression of bacterial genes encoding hygromycin phosphotransferase and beta-galactosidase in the pathogenic fungus Histoplasma capsulatum. Infect Immun 66: 1697–1707.
[72]  Bahn YS, Sundstrom P (2001) CAP1, an adenylate cyclase-associated protein gene, regulates bud-hypha transitions, filamentous growth, and cyclic AMP levels and is required for virulence of Candida albicans. J Bacteriol 183: 3211–3223.
[73]  Youseff BH, Rappleye CA (2012) RNAi-based gene silencing using a GFP sentinel system in Histoplasma capsulatum. In: Brand A, MacCallum D, editors. Host-Fungal Interactions: Manipulation of Fungal Gene Expression. New York: Humana Press.
[74]  Beers RF Jr, Sizer IW (1952) A spectrophotometric method for measuring the breakdown of hydrogen peroxide by catalase. J Biol Chem 195: 133–140.
[75]  Kouoh F, Gressier B, Luyckx M, Brunet C, Dine T, et al. (2000) A simple method for isolating human and rabbit polymorphonuclear neutrophils (PMNs). Biol Pharm Bull 23: 1382–1383.
[76]  Nauseef WM (2007) Isolation of human neutrophils from venous blood. Methods Mol Biol 412: 15–20.
[77]  Zhang X, Goncalves R, Mosser DM (2008) The isolation and characterization of murine macrophages. Curr Protoc Immunol Chapter 14: Unit 14 11.
[78]  Marion CL, Rappleye CA, Engle JT, Goldman WE (2006) An alpha-(1,4)-amylase is essential for alpha-(1,3)-glucan production and virulence in Histoplasma capsulatum. Mol Microbiol 62: 970–983.

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