Deletion of the Candida albicans histidine kinase gene CHK1 improves recognition by phagocytes through an increased exposure of cell wall β-1,3-glucans
The pathogenic fungus Candida albicans is able to cover its most potent proinflammatory cell wall molecules, the β-glucans, underneath a dense mannan layer, so that the pathogen becomes partly invisible for immune cells such as phagocytes. As the C. albicans histidine kinases Chk1p, Cos1p and CaSln1p had been reported to be involved in virulence and cell wall biosynthesis, we investigated whether deletion of the respective genes influences the activity of phagocytes against C. albicans. We found that among all histidine kinase genes, CHK1 plays a prominent role in phagocyte activation. Uptake of the deletion mutant Δchk1 as well as the acidification of Δchk1-carrying phagosomes was significantly increased compared with the parental strain. These improved activities could be correlated with an enhanced accessibility of the mutant β-1,3-glucans for immunolabelling. In addition, any inhibition of β-1,3-glucan-mediated phagocytosis resulted in a reduced uptake of Δchk1, while ingestion of the parental strain was hardly affected. Moreover, deletion of CHK1 caused an enhanced release of interleukins 6 and 10, indicating a stronger activation of the β-1,3-glucan receptor dectin-1. In conclusion, the Chk1p protein is likely to be involved in masking β-1,3-glucans from immune recognition. As there are no homologues of fungal histidine kinases in mammals, Chk1p has to be considered as a promising target for new antifungal agents.
AdamsE. L.,
RiceP. J.,
GravesB.,
EnsleyH. E.,
YuH.,
BrownG. D.,
GordonS.,
MonteiroM. A.,
Papp-SzaboE.other authors2008; Differential high-affinity interaction of dectin-1 with natural or synthetic glucans is dependent upon primary structure and is influenced by polymer chain length and side-chain branching. J Pharmacol Exp Ther 325:115–123
AlexL. A.,
KorchC.,
SelitrennikoffC. P.,
SimonM. I.1998; COS1, a two-component histidine kinase that is involved in hyphal development in the opportunistic pathogen Candida albicans. Proc Natl Acad Sci U S A 95:7069–7073
BrachmannC. B.,
DaviesA.,
CostG. J.,
CaputoE.,
LiJ.,
HieterP.,
BoekeJ. D.1998; Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14:115–132
BrownG. D.,
TaylorP. R.,
ReidD. M.,
WillmentJ. A.,
WilliamsD. L.,
Martinez-PomaresL.,
WongS. Y.,
GordonS.2002; Dectin-1 is a major β-glucan receptor on macrophages. J Exp Med 196:407–412
CaleraJ. A.,
CalderoneR.1999a; Flocculation of hyphae is associated with a deletion in the putative CaHK1 two-component histidine kinase gene from Candida albicans. Microbiology 145:1431–1442
CaleraJ. A.,
CalderoneR. A.1999b; Identification of a putative response regulator two-component phosphorelay gene ( CaSSK1) from Candida albicans. Yeast 15:1243–1254
CaleraJ. A.,
ZhaoX. J.,
De BernardisF.,
SheridanM.,
CalderoneR.1999; Avirulence of Candida albicans CaHK1 mutants in a murine model of hematogenously disseminated candidiasis. Infect Immun 67:4280–4284
CaleraJ. A.,
ZhaoX. J.,
CalderoneR.2000; Defective hyphal development and avirulence caused by a deletion of the SSK1 response regulator gene in Candida albicans. Infect Immun 68:518–525
ChauhanN.,
CalderoneR.2008; Two-component signal transduction proteins as potential drug targets in medically important fungi. Infect Immun 76:4795–4803
ChauhanN.,
InglisD.,
RomanE.,
PlaJ.,
LiD.,
CaleraJ. A.,
CalderoneR.2003; Candida albicans response regulator gene SSK1 regulates a subset of genes whose functions are associated with cell wall biosynthesis and adaptation to oxidative stress. Eukaryot Cell 2:1018–1024
Galán-DiezM.,
AranaD. M.,
Serrano-GomezD.,
KremerL.,
CasasnovasJ. M.,
OrtegaM.,
Cuesta-DominguezA.,
CorbiA. L.,
PlaJ.other authors2010; Candida albicansβ-glucan exposure is controlled by the fungal CEK1-mediated mitogen-activated protein kinase pathway that modulates immune responses triggered through dectin-1. Infect Immun 78:1426–1436
GantnerB. N.,
SimmonsR. M.,
UnderhillD. M.2005; Dectin-1 mediates macrophage recognition of Candida albicans yeast but not filaments. EMBO J 24:1277–1286
GoodridgeH. S.,
SimmonsR. M.,
UnderhillD. M.2007; Dectin-1 stimulation by Candida albicans yeast or zymosan triggers NFAT activation in macrophages and dendritic cells. J Immunol 178:3107–3115
GowN. A.,
NeteaM. G.,
MunroC. A.,
FerwerdaG.,
BatesS.,
Mora-MontesH. M.,
WalkerL.,
JansenT.,
JacobsL.other authors2007; Immune recognition of Candida albicansβ-glucan by dectin-1. J Infect Dis 196:1565–1571
Ibata-OmbettaS.,
JouaultT.,
TrinelP. A.,
PoulainD.2001; Role of extracellular signal-regulated protein kinase cascade in macrophage killing of Candida albicans. J Leukoc Biol 70:149–154
JanuszM. J.,
AustenK. F.,
CzopJ. K.1988; Phagocytosis of heat-killed blastospores of Candida albicans by human monocyte beta-glucan receptors. Immunology 65:181–185
JouaultT.,
El Abed-El BehiM.,
Martinez-EsparzaM.,
BreuilhL.,
TrinelP. A.,
ChamaillardM.,
TrotteinF.,
PoulainD.2006; Specific recognition of Candida albicans by macrophages requires galectin-3 to discriminate Saccharomyces cerevisiae and needs association with TLR2 for signaling. J Immunol 177:4679–4687
KlippelN.,
BilitewskiU.2007; Phagocytosis assay based on living Candida albicans for the detection of effects of chemicals on macrophages function. Anal Lett 40:1400–1411
KruppaM.,
GoinsT.,
CutlerJ. E.,
LowmanD.,
WilliamsD.,
ChauhanN.,
MenonV.,
SinghP.,
LiD.other authors2003; The role of the Candida albicans histidine kinase ( CHK1) gene in the regulation of cell wall mannan and glucan biosynthesis. FEMS Yeast Res 3:289–299
KruppaM.,
Jabra-RizkM. A.,
MeillerT. F.,
CalderoneR.2004; The histidine kinases of Candida albicans: regulation of cell wall mannan biosynthesis. FEMS Yeast Res 4:409–416
KumagaiY.,
ChengZ.,
LinM.,
RikihisaY.2006; Biochemical activities of three pairs of Ehrlichia chaffeensis two-component regulatory system proteins involved in inhibition of lysosomal fusion. Infect Immun 74:5014–5022
Le CabecV.,
EmorineL. J.,
ToescaI.,
CougouleC.,
Maridonneau-PariniI.2005; The human macrophage mannose receptor is not a professional phagocytic receptor. J Leukoc Biol 77:934–943
LiD.,
BernhardtJ.,
CalderoneR.2002; Temporal expression of the Candida albicans genes CHK1 and CSSK1, adherence, and morphogenesis in a model of reconstituted human esophageal epithelial candidiasis. Infect Immun 70:1558–1565
MoranC.,
GrussemeyerC. A.,
SpaldingJ. R.,
BenjaminD. K.Jr,
ReedS. D.2009; Candida albicans and non- albicans bloodstream infections in adult and pediatric patients: comparison of mortality and costs. Pediatr Infect Dis J 28:433–435
NagahashiS.,
MioT.,
OnoN.,
Yamada-OkabeT.,
ArisawaM.,
BusseyH.,
Yamada-OkabeH.1998; Isolation of CaSLN1 and CaNIK1, the genes for osmosensing histidine kinase homologues, from the pathogenic fungus Candida albicans. Microbiology 144:425–432
NeteaM. G.,
BrownG. D.,
KullbergB. J.,
GowN. A.2008; An integrated model of the recognition of Candida albicans by the innate immune system. Nat Rev Microbiol 6:67–78
PoulainD.,
JouaultT.2004; Candida albicans cell wall glycans, host receptors and responses: elements for a decisive crosstalk. Curr Opin Microbiol 7:342–349
Ruiz-HerreraJ.,
ElorzaM. V.,
ValentinE.,
SentandreuR.2006; Molecular organization of the cell wall of Candida albicans and its relation to pathogenicity. FEMS Yeast Res 6:14–29
SinghS. D.,
RobbinsN.,
ZaasA. K.,
SchellW. A.,
PerfectJ. R.,
CowenL. E.2009; Hsp90 governs echinocandin resistance in the pathogenic yeast Candida albicans via calcineurin. PLoS Pathog 5:e1000532
SlackE. C.,
RobinsonM. J.,
Hernanz-FalconP.,
BrownG. D.,
WilliamsD. L.,
SchweighofferE.,
TybulewiczV. L.,
Reis e SousaC.2007; Syk-dependent ERK activation regulates IL-2 and IL-10 production by DC stimulated with zymosan. Eur J Immunol 37:1600–1612
SteeleC.,
MarreroL.,
SwainS.,
HarmsenA. G.,
ZhengM.,
BrownG. D.,
GordonS.,
ShellitoJ. E.,
KollsJ. K.2003; Alveolar macrophage-mediated killing of Pneumocystis carinii f. sp. muris involves molecular recognition by the dectin-1 β-glucan receptor. J Exp Med 198:1677–1688
TaylorP. R.,
BrownG. D.,
ReidD. M.,
WillmentJ. A.,
Martinez-PomaresL.,
GordonS.,
WongS. Y.2002; The β-glucan receptor, dectin-1, is predominantly expressed on the surface of cells of the monocyte/macrophage and neutrophil lineages. J Immunol 169:3876–3882
TaylorP. R.,
TsoniS. V.,
WillmentJ. A.,
DennehyK. M.,
RosasM.,
FindonH.,
HaynesK.,
SteeleC.,
BottoM.other authors2007; Dectin-1 is required for β-glucan recognition and control of fungal infection. Nat Immunol 8:31–38
TkaczJ. S.,
CybulskaE. B.,
LampenJ. O.1971; Specific staining of wall mannan in yeast cells with fluorescein-conjugated concanavalin A. J Bacteriol 105:1–5
TorosantucciA.,
ChianiP.,
De BernardisF.,
CassoneA.,
CaleraJ. A.,
CalderoneR.2002; Deletion of the two-component histidine kinase gene ( CHK1) of Candida albicans contributes to enhanced growth inhibition and killing by human neutrophils in vitro. Infect Immun 70:985–987
WheelerR. T.,
KombeD.,
AgarwalaS. D.,
FinkG. R.2008; Dynamic, morphotype-specific Candida albicansβ-glucan exposure during infection and drug treatment. PLoS Pathog 4:e1000227
Yamada-OkabeT.,
MioT.,
OnoN.,
KashimaY.,
MatsuiM.,
ArisawaM.,
Yamada-OkabeH.1999; Roles of three histidine kinase genes in hyphal development and virulence of the pathogenic fungus Candida albicans. J Bacteriol 181:7243–7247
Deletion of the Candida albicans histidine kinase gene CHK1 improves recognition by phagocytes through an increased exposure of cell wall β-1,3-glucans