Macrophage Migration Is Impaired within Candida albicans Biofilms
Abstract
:1. Introduction
2. Materials and Methods
2.1. C. albicans Strains and Growth Conditions
2.2. Preparation of J774.1 Murine Macrophage Cell Line
2.3. Macrophage Migration Assays
2.4. Analysis of Live Cell Microscopy Movies
2.5. Statistical Analyses
3. Results
3.1. Macrophage Migration towards C. albicans Is Not Altered by Multiplicity of Infection
3.2. Macrophage Migration Dynamics towards C. albicans Is Affected by Fungal Cell Wall Composition
3.3. Macrophage Migration is Diminished within a 3D C. albicans Biofilm
3.4. Macrophage Migration Is Diminished within 3D C. albicans Biofilms Formed from Glycosylation Defective Mutants
4. Discussion
Supplementary Materials
Acknowledgments
Author Contributions
Conflicts of Interest
References
- Drummond, R.A.; Gaffen, S.L.; Hise, A.G.; Brown, G.D. Innate defense against fungal pathogens. Cold Spring Harb. Perspect. Med. 2014, 5. [Google Scholar] [CrossRef] [PubMed]
- Erwig, L.P.; Gow, N.A. Interactions of fungal pathogens with phagocytes. Nat. Rev. Microbiol. 2016, 14, 163–176. [Google Scholar] [CrossRef] [PubMed]
- Ballou, E.R.; Avelar, G.M.; Childers, D.S.; Mackie, J.; Bain, J.M.; Wagener, J.; Kastora, S.L.; Panea, M.D.; Hardison, S.E.; Walker, L.A.; et al. Lactate signalling regulates fungal β-glucan masking and immune evasion. Nat. Microbiol. 2016, 2, 16238. [Google Scholar] [CrossRef] [PubMed]
- Ene, I.V.; Adya, A.K.; Wehmeier, S.; Brand, A.C.; MacCallum, D.M.; Gow, N.A.; Brown, A.J. Host carbon sources modulate cell wall architecture, drug resistance and virulence in a fungal pathogen. Cell. Microbiol. 2012, 14, 1319–1335. [Google Scholar] [CrossRef] [PubMed]
- Ene, I.V.; Heilmann, C.J.; Sorgo, A.G.; Walker, L.A.; de Koster, C.G.; Munro, C.A.; Klis, F.M.; Brown, A.J. Carbon source-induced reprogramming of the cell wall proteome and secretome modulates the adherence and drug resistance of the fungal pathogen Candida albicans. Proteomics 2012, 12, 3164–3179. [Google Scholar] [CrossRef] [PubMed]
- Ene, I.V.; Cheng, S.C.; Netea, M.G.; Brown, A.J.P. Growth of Candida albicans cells on the physiologically relevant carbon source lactate affects their recognition and phagocytosis by immune cells. Infect. Immun. 2013, 81, 238–248. [Google Scholar] [CrossRef] [PubMed]
- Pierce, C.G.; Vila, T.; Romo, J.A.; Montelongo-Jauregui, D.; Wall, G.; Ramasubramanian, A.; Lopez-Ribot, J.L. The Candida albicans Biofilm matrix: Composition, structure and function. J. Fungi 2017, 3, 14. [Google Scholar] [CrossRef] [PubMed]
- Ramage, G.; Saville, S.P.; Thomas, D.P.; Lopez-Ribot, J.L. Candida biofilms: An update. Eukaryot. Cell 2005, 4, 633–638. [Google Scholar] [CrossRef] [PubMed]
- Rajendran, R.; Sherry, L.; Nile, C.J.; Sherriff, A.; Johnson, E.M.; Hanson, M.F.; Williams, C.; Munro, C.A.; Jones, B.J.; Ramage, G. Biofilm formation is a risk factor for mortality in patients with Candida albicans bloodstream infection-scotland, 2012–2013. Clin. Microbiol. Infect. 2016, 22, 87–93. [Google Scholar] [CrossRef] [PubMed]
- Lewis, L.E.; Bain, J.M.; Lowes, C.; Gillespie, C.; Rudkin, F.M.; Gow, N.A.; Erwig, L.P. Stage specific assessment of Candida albicans phagocytosis by macrophages identifies cell wall composition and morphogenesis as key determinants. PLoS Pathog. 2012, 8, e1002578. [Google Scholar] [CrossRef] [PubMed]
- Rudkin, F.M.; Bain, J.M.; Walls, C.; Lewis, L.E.; Gow, N.A.R.; Erwig, L.P. Altered dynamics of Candida albicans phagocytosis by macrophages and PMNs when both phagocyte subsets are present. MBio 2013, 46, e00810–e00813. [Google Scholar] [CrossRef] [PubMed]
- Okai, B.; Lyall, N.; Gow, N.A.R.; Bain, J.M.; Erwig, L.P. Rab14 regulates maturation of macrophage phagosomes containing the fungal pathogen Candida albicans and outcome of the host-pathogen interaction. Infect. Immun. 2015, 83, 1523–1535. [Google Scholar] [CrossRef] [PubMed]
- Murad, A.M.; Lee, P.R.; Broadbent, I.D.; Barelle, C.J.; Brown, A.J.P. CIp10, an efficient and convenient integrating vector for Candida albicans. Yeast 2000, 16, 325–327. [Google Scholar] [CrossRef]
- Munro, C.A.; Bates, S.; Buurman, E.T.; Hughes, H.B.; Maccallum, D.M.; Bertram, G.; Atrih, A.; Ferguson, M.A.; Bain, J.M.; Brand, A.; et al. Mnt1p and Mnt2p of Candida albicans are partially redundant α-1,2-mannosyltransferases that participate in O-linked mannosylation and are required for adhesion and virulence. J. Biol. Chem. 2005, 280, 1051–1060. [Google Scholar] [CrossRef] [PubMed]
- Bates, S.; MacCallum, D.M.; Bertram, G.; Munro, C.A.; Hughes, H.B.; Buurman, E.T.; Brown, A.J.; Odds, F.C.; Gow, N.A. Candida albicans Pmr1p a secretory pathway P-type Ca2+/Mn2+-ATPase is required for glycosylation and virulence. J. Biol. Chem. 2005, 280, 23408–23415. [Google Scholar] [CrossRef] [PubMed]
- Mora-Montes, H.M.; Bates, S.; Netea, M.G.; Castillo, L.; Brand, A.; Buurman, E.T.; Díaz-Jiménez, D.F.; Jan Kullberg, B.; Brown, A.J.; Odds, F.C.; et al. A multifunctional mannosyltransferase family in Candida albicans determines cell wall mannan structure and host-fungus interactions. J. Biol. Chem. 2010, 285, 12087–12095. [Google Scholar] [CrossRef] [PubMed]
- Hobson, R.P.; Munro, C.A.; Bates, S.; MacCallum, D.M.; Cutler, J.E.; Heinsbroek, S.E.; Brown, G.D.; Odds, F.C.; Gow, N.A. Loss of cell wall mannosylphosphate in Candida albicans does not influence macrophage recognition. J. Biol. Chem. 2004, 279, 39628–39635. [Google Scholar] [CrossRef] [PubMed]
- Fonzi, W.A.; Irwin, M.Y. Isogenic strain construction and gene mapping in Candida albicans. Genetics 1993, 134, 717–728. [Google Scholar] [PubMed]
- Brand, A.; MacCallum, D.M.; Brown, A.J.; Gow, N.A.R.; Odds, F.C. Ectopic expression of URA3 can influence the virulence phenotypes and proteome of Candida albicans but can be overcome by targeted reintegration of URA3 at the RPS10 locus. Eukaryot. Cell 2004, 3, 900–909. [Google Scholar] [CrossRef] [PubMed]
- Lewis, L.E.; Bain, J.M.; Okai, B.; Gow, N.A.R.; Erwig, L.P. Live-cell video microscopy of fungal pathogen phagocytosis. J. Vis. Exp. 2013, 71, e50196. [Google Scholar] [CrossRef] [PubMed]
- Bain, J.M.; Louw, J.; Lewis, L.E.; Okai, B.; Walls, C.A.; Ballou, E.R.; Walker, L.A.; Reid, D.; Munro, C.A.; Brown, A.J.; et al. Candida albicans hypha formation and mannan masking of β-glucan inhibit macrophage phagosome maturation. MBio 2014, 5, e01874-14. [Google Scholar] [CrossRef] [PubMed]
- McKenzie, C.G.; Koser, U.; Lewis, L.E.; Bain, J.M.; Mora-Montes, H.M.; Barker, R.N.; Gow, N.A.; Erwig, L.P. Contribution of Candida albicans cell wall components to recognition by and escape from murine macrophages. Infect. Immun. 2010, 78, 1650–1658. [Google Scholar] [CrossRef] [PubMed]
- Dutton, L.C.; Nobbs, A.H.; Jepson, K.; Jepson, M.A.; Vickerman, M.M.; Aqeel Alawfi, S.; Munro, C.A.; Lamont, R.J.; Jenkinson, H.F. O-mannosylation in Candida albicans enables development of interkingdom biofilm communities. MBio 2014, 5, e00911–e00914. [Google Scholar] [CrossRef] [PubMed]
- Johnson, C.J.; Cabezas-Olcoz, J.; Kernien, J.F.; Wang, S.X.; Beebe, D.J.; Huttenlocher, A.; Ansari, H.; Nett, J.E. The extracellular matrix of Candida albicans biofilms impairs formation of neutrophil extracellular traps. PLoS Pathog. 2016, 12, e1005884. [Google Scholar] [CrossRef] [PubMed]
- Mitchell, K.F.; Zarnowski, R.; Sanchez, H.; Edward, J.A.; Reinicke, E.L.; Nett, J.E.; Mitchell, A.P.; Andes, D.R. Community participation in biofilm matrix assembly and function. Proc. Natl. Acad. Sci. USA 2015, 112, 4092–4097. [Google Scholar] [CrossRef] [PubMed]
- Maridonneau-Parini, I. Control of macrophage 3D migration: A therapeutic challenge to limit tissue infiltration. Immunol. Rev. 2014, 262, 216–231. [Google Scholar] [CrossRef] [PubMed]
- Nobile, C.J.; Johnson, A.D. Candida albicans biofilms and human disease. Annu. Rev. Microbiol. 2015, 69, 71–92. [Google Scholar] [CrossRef] [PubMed]
- Thomas, D.P.; Pitarch, A.; Monteoliva, L.; Gil, C.; Lopez-Ribot, J.L. Proteomics to study Candida albicans biology and pathogenicity. Infect. Disord. Drug Targets 2006, 6, 335–341. [Google Scholar] [CrossRef] [PubMed]
- Katragkou, A.; Kruhlak, M.J.; Simitsopoulou, M.; Chatzimoschou, A.; Taparkou, A.; Cotton, C.J.; Paliogianni, F.; Diza-Mataftsi, E.; Tsantali, C.; Walsh, T.J. Interactions between human phagocytes and Candida albicans biofilms alone and in combination with antifungal agents. J. Infect. Dis. 2010, 201, 1941–1949. [Google Scholar] [CrossRef] [PubMed]
- Xie, Z.; Thompson, A.; Sobue, T.; Kashleva, H.; Xu, H.; Vasilakos, J.; Dongari-Bagtzoglou, A. Candida albicans biofilms do not trigger reactive oxygen species and evade neutrophil killing. J. Infect. Dis. 2012, 206, 1936–1945. [Google Scholar] [CrossRef] [PubMed]
- Chandra, J.; McCormick, T.S.; Imamura, Y.; Mukherjee, P.K.; Ghannoum, M.A. Interaction of Candida albicans with adherent human peripheral blood mononuclear cells increases C. albicans biofilm formation and results in differential expression of pro- and anti-inflammatory cytokines. Infect. Immun. 2007, 75, 2612–2620. [Google Scholar] [CrossRef] [PubMed]
Strain Name | Strain Number | Disrupted Target | Phenotype | Reference |
---|---|---|---|---|
CAI4-CIp10 | NGY152 | Not applicable (n/a) | Wild type | [13] |
mnt1Δ/mnt2Δ | NGY111 | α-(1,2)-mannosyltransferases | O-mannan defect | [14] |
mnt1Δ/mnt2Δ+MNT1 | NGY335 | n/a | Complemented mutant | [14] |
pmr1Δ | NGY355 | P-type Ca2+/Mns2+-ATPase | N-linked mannan and O-linked mannan and phosphomannan defect | [15] |
pmr1Δ+PMR1 | NGY356 | n/a | Complemented mutant | [15] |
mns1Δ | HMY5 | α-(1,2)-mannosidase | Terminal stage N-linked core mannan defect | [16] |
mnn4Δ | CDH15 | mannosylphosphate transferase | Phosphomannan incorporation defect | [17] |
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Alonso, M.F.; Gow, N.A.R.; Erwig, L.P.; Bain, J.M. Macrophage Migration Is Impaired within Candida albicans Biofilms. J. Fungi 2017, 3, 31. https://doi.org/10.3390/jof3030031
Alonso MF, Gow NAR, Erwig LP, Bain JM. Macrophage Migration Is Impaired within Candida albicans Biofilms. Journal of Fungi. 2017; 3(3):31. https://doi.org/10.3390/jof3030031
Chicago/Turabian StyleAlonso, Maria F., Neil A. R. Gow, Lars P. Erwig, and Judith M. Bain. 2017. "Macrophage Migration Is Impaired within Candida albicans Biofilms" Journal of Fungi 3, no. 3: 31. https://doi.org/10.3390/jof3030031