Abstract
The incidence of invasive fungal diseases has increased over the past decades, particularly in relation with the increase of immunocompromised patient cohorts (e.g., HIV-infected patients, transplant recipients, immunosuppressed patients with cancer). Opportunistic fungal pathogens such as Candida spp. are most often associated with serious systemic infections. Currently available antifungal drugs are rather unspecific, often with severe side effects. In some cases, their prophylactic use has favored emergence of resistant fungal strains. Major antifungal drugs target the biosynthesis of lipid components of the fungal plasma membrane or the assembly of the cell wall. For a more specific and efficient treatment and prevention of fungal infection, new therapeutic strategies are needed, including strengthening or stimulation of the residual host immune response. Achieving such a goal requires a better understanding of factors important for the defense and the survival of the host combating Candida spp. Where possible, primary cultures of mammalian immune cells of the innate immune system constitute a better suited model than transformed cell lines to study host-pathogen response and virulence. Hence, in vitro primary cell culture systems are a good strategy for a first screening of mutant strains of Candida spp. to identify virulence traits with regard to host cell response and pathogen invasion.
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Roeder A, Kirschning CJ, Rupec RA, Schaller M, Korting HC. Toll-like receptors and innate antifungal responses, Trends Microbiol 2004;12:44–9.
Netea MG, Van der Graaf C, Van der Meer JW, Kullberg BJ, Recognition of fungal pathogens by Toll-like receptors: Toll-like receptors and the host defense against microbial pathogens: bringing specificity to the innate-immune system, Eur J Clin Microbiol Infect Dis 2004;23:672–6.
Mencacci A, Cenci E, Del Sero G, et al, Defective co-stimulation and impaired Th1 development in tumor necrosis factor/lymphotoxin-alpha double-deficient mice infected with Candida albicans. Int Immunol 1998;10:37–48.
Vazquez-Torres A, Jones-Carson J, Wagner RD, Warner T, Balish E, Early resistance of interleukin-10 knock-out mice to acute systemic candidiasis, Infect Immun 1999;67:670–4.
Farah CS, Hu Y, Riminton S, Ashman RB, Distinct roles for interleukin-12p40 and tumour necrosis factor in resistance to oral candidiasis defined by gene-targeting, Oral Microbiol Immunol 2006;21: 252–5.
Vonk AG, Netea MG, van Krieken JH, van der Meer JW, Kullberg BJ, Delayed clearance of intraabdominal abscesses caused by Candida albicans in tumor necrosis factor-alpha- and lymphotoxin-alpha-deficient mice, J Infect Dis 2002;186:1815–22.
Ibata-Ombetta S, Jouault T, Trinel PA, Poulain D, Role of extracellular signal-regulated protein kinase cascade in macrophage killing of Candida albicans. J Leukoc Biol 2001;70:149–54.
Zhong B, Jiang K, Gilvary DL, et al, Human neutrophils utilize a Rac/Cdc42-dependent MAPK pathway to direct intracellular granule mobilization toward ingested microbial pathogens, Blood 2003;101:3240–8.
Choi JH, Choi EK, Park SJ, et al, Impairment of p38 MAPK-mediated cytosolic phospholipase A(2) activation in the kidneys is associated with pathogenicity of Candida albicans. Immunology 2007; 120:173–81.
Zal T, Volkmann A, Stockinger B, Mechanisms of tolerance induction in major histocompatibility complex class II-restricted T cells specific for a blood-borne self-antigen, J Exp Med 1994;180:2089–99.
Gillum AM, Tsay EY, Kirsch DR, Isolation of the Candida albicans gene for orotidine-5´-phosphate decarboxylase by complementation of S. cerevisiae ura3 and E. coli pyrF mutations, Mol Gen Genet 1984;198:179–82.
Hull CM, Johnson AD, Identification of a mating type-like locus in the asexual pathogenic yeast Candida albicans. Science 1999;285:1271–5.
Kovarik P, Stoiber D, Novy M, Decker T, Stat1 combines signals derived from IFN-gamma and LPS receptors during macrophage activation, EMBO J 1998;17:3660–8.
Overbergh L, Giulietti A, Valckx D, Decallonne R, Bouillon R, Mathieu C, The use of real-time reverse transcriptase PCR for the quantification of cytokine gene expression, J Biomol Tech 2003;14:33–43.
Pattyn F, Speleman F, De Paepe A, Vandesompele J, RTPrimerDB: the Real-Time PCR primer and probe database, Nucl Acids Res 2003;31:122–3.
Pattyn F, Robbrecht P, De Paepe A, Speleman F, Vandesompele J, RTPrimerDB: the real-time PCR primer and probe database, major update 2006. Nucl Acids Res 2006;34:D684–8.
Hume DA, Gordon S, Optimal conditions for proliferation of bone marrow-derived mouse macrophages in culture: the roles of CSF-1, serum, Ca2+, and adherence, J Cell Physiol 1983;117:189–94.
Inaba K, Inaba M, Romani N, et al, Generation of large numbers of dendritic cells from mouse bone marrow cultures supplemented with granulocyte/macrophage colony-stimulating factor, J Exp Med 1992;176:1693–702.
Herth W, Calcofluor white and Congo red inhibit chitin microfibril assembly of Poterioochromonas: evidence for a gap between polymerization and microfibril formation, J Cell Biol 1980;87:442–50.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2009 Humana Press, a part of Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Bourgeois, C., Majer, O., Frohner, I., Kuchler, K. (2009). In Vitro Systems for Studying the Interaction of Fungal Pathogens with Primary Cells from the Mammalian Innate Immune System. In: Rupp, S., Sohn, K. (eds) Host-Pathogen Interactions. Methods in Molecular Biology, vol 470. Humana Press. https://doi.org/10.1007/978-1-59745-204-5_11
Download citation
DOI: https://doi.org/10.1007/978-1-59745-204-5_11
Publisher Name: Humana Press
Print ISBN: 978-1-58829-886-7
Online ISBN: 978-1-59745-204-5
eBook Packages: Springer Protocols