tudied. We found a role for dectin-1 and -2 that engage -glucan and mannans on the C. albicans cell wall that, together with a MyD88-dependent pathway, promote cPLA2 activation and eicosanoid production. Although C. albicans is a normal commensal organism, it is an opportunistic pathogen that is a leading cause of mycoses particularly in the immunocompromised and critically ill. There has been considerable interest in elucidating the mechanisms regulating SB366791 web immune responses to C. albicans because of the prevalence of fungal infections. Eicosanoids affect immune regulation by modulating cellular differentiation, phagocytic potential, migration and cytokine/ chemokine production. The types and balance of cytokines produced during the early responses of innate immune cells to infection influence the macrophage phenotype, differentiation of lymphocytes and adaptive immune responses. In this study, we compared cPLA2+/+ and cPLA2-/RPM to investigate the functional consequences of cPLA2 activation and the effect of endogenously produced eicosanoids on gene expression in response to C. albicans. Our results demonstrate that C. albicans-stimulated cPLA2 activation and the early production of prostanoids promotes an autocrine pathway in RPM that affects the expression of genes involved in host defense and to dampen inflammation. Mouse Strains Pathogen-free Balb/c mice were obtained from Harlan Sprague Dawley. cPLA2-/- mice were generated as previously described and backcrossed onto a Balb/c background for 10 generations. The TLR4 mutant mouse strain C3H/HeJ and control strain C3H/HeOuJ were obtained from The Jackson Laboratory. TLR2-/- and MyD88-/- mice were generated as previously described. MyD88+/C57BL/6/129 mice were crossed to generate MyD88-/- mice and MyD88+/+ littermate controls. C57BL/6 control mice were obtained from The Jackson Laboratory. Dectin-1-/- mice were produced as described previously, and age and strain matched controls obtained from Taconic. Mice were used for macrophage isolation at 7-12 wk of age. C. albicans Strains and Culture C. albicans was used for experiments unless otherwise indicated. The C. albicans Capmr1 null mutant defective in glycosylation, the re-integrant strain and parental wild-type control were generated as previously described. C. albicans strains were grown on Sabouraud dextrose agar plates and maintained at 4. RPM Infection The day before the experiment, the strains were streaked onto fresh Sabouraud dextrose agar plates and incubated overnight at 37. C. albicans was scraped from the plate and washed twice in endotoxin-free PBS. Live C. 26506265 albicans at a multiplicity of infection of 2 was used for all experiments. RPM Isolation RPM were obtained by peritoneal lavage as previously described. Cells were plated at a density of 5 x 105/cm2 and incubated for 2 h at 37 in a humidified atmosphere of 5% CO2 in air. After washing the cultures to remove non-adherent cells, the adherent macrophages were incubated in DMEM containing 10% heat inactivated FBS, 100 /ml 12419798 streptomycin sulfate, 100 units/ml penicillin G, 0.29 mg/ml glutamine for 16-18 h at 37. The cells were washed twice with serum-free DMEM containing 0.1% human serum albumin and then infected with 
C. albicans. Materials and Methods Ethics Statement The work with mice in this study was approved by the National Jewish Health Institutional Animal Care and Use Committee and conducted in accordance with their guidelines. Materials DMEM was from Cambrex BioScienc
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