Session Type: ePosters
Session Title: ePosters
Authors(s): A. Vikelouda (1), M. Simitsopoulou (1), L. Skoura (2), C. Antachopoulos (1), E. Roilides (1)
Authors Affiliations(s): (1) Infectious Diseases Unit, 3rd Department of Pediatrics, Aristotle University School of Medicine, Greece, (2) Microbiology Departmen, AHEPA Hospital, Greece
Background:
Scedosporium apiospermum can cause a wide range of invasive infections depending on host immunity. Scedosporium spp. can grow in biofilms, extracellular structures resistant to antifungal therapy or host immune mechanisms. Knowledge on the interplay between pathogen, antimicrobial agent and innate immune cells is pivotal in the fight against drug-resistant infections. We investigated whether deoxycholate amphotericin-B (DAMB), liposomal amphotericin-B (LAMB) and voriconazole (VRC) modulate immune response of human neutrophils (PMN) against S. apiospermum biofilms (BF).
Methods:105 cfu/ml of a BF-producing strain were grown in RPMI at 37οC for 48h to form mature BF. MICs of DAMB, LAMB and VRC were assessed by the XTT reduction assay and MIC50s were determined as ≥50% BF damage. PMN’s from healthy volunteers (106/ml in RPMI/10% human serum) were co-incubated with BF at effector-to-target ratio 5:1 and 1 mg/l DAMB, 2mg/l LAMB or 32mg/l VRC for 3 hours at 37οC/5% CO2. The cytokines released in culture supernatants were determined via multiplex ELISA using chemiluminescence. Microplate image capture and data analysis were performed using Quansys Q-view software. Protein concentrations between drug-treated and untreated PMN’s were compared by ANOVA (P<0.05). Two independent experiments were performed.
Results:MIC50s of DAMB, LAMB and VRC for BF were 1, 2 and 32 mg/l, respectively. IL-1β, IL-8, IL-10, and IL-23 were detected in the culture supernates. Co-incubation of S. apiospermum BF with DAMB or VRC significantly increased IL-1β, IL-8 and IL-10 levels as compared to untreated controls (IL-1β: 9±0.4 and 8.6±0.2 vs 7.8±0.1 pg/ml; IL-8: 184±18 and 217±39 vs 102±4 pg/ml; IL-10: 9.5±1.6 vs 5.1±0.3 pg/ml;P<0.01). All antifungal agents caused significantly increased levels of IL-8, IL-10, and IL-23 compared to the combination treatment of PMN+BF+drug (P<0.01).
Conclusions:DAMB and VRC showed an immunomodulatory profile towards PMN against biofilm damage. However, apart from the robust production of IL-8, the remaining cytokines were differentially produced but at low amounts which could lead to a dampened immune response. This may constitute a mechanism for the prolongation of a BF life-style with serious complications to the patient’s health.
Session Type: ePosters
Session Title: ePosters
Authors(s): A. Vikelouda (1), M. Simitsopoulou (1), L. Skoura (2), C. Antachopoulos (1), E. Roilides (1)
Authors Affiliations(s): (1) Infectious Diseases Unit, 3rd Department of Pediatrics, Aristotle University School of Medicine, Greece, (2) Microbiology Departmen, AHEPA Hospital, Greece
Background:
Scedosporium apiospermum can cause a wide range of invasive infections depending on host immunity. Scedosporium spp. can grow in biofilms, extracellular structures resistant to antifungal therapy or host immune mechanisms. Knowledge on the interplay between pathogen, antimicrobial agent and innate immune cells is pivotal in the fight against drug-resistant infections. We investigated whether deoxycholate amphotericin-B (DAMB), liposomal amphotericin-B (LAMB) and voriconazole (VRC) modulate immune response of human neutrophils (PMN) against S. apiospermum biofilms (BF).
Methods:105 cfu/ml of a BF-producing strain were grown in RPMI at 37οC for 48h to form mature BF. MICs of DAMB, LAMB and VRC were assessed by the XTT reduction assay and MIC50s were determined as ≥50% BF damage. PMN’s from healthy volunteers (106/ml in RPMI/10% human serum) were co-incubated with BF at effector-to-target ratio 5:1 and 1 mg/l DAMB, 2mg/l LAMB or 32mg/l VRC for 3 hours at 37οC/5% CO2. The cytokines released in culture supernatants were determined via multiplex ELISA using chemiluminescence. Microplate image capture and data analysis were performed using Quansys Q-view software. Protein concentrations between drug-treated and untreated PMN’s were compared by ANOVA (P<0.05). Two independent experiments were performed.
Results:MIC50s of DAMB, LAMB and VRC for BF were 1, 2 and 32 mg/l, respectively. IL-1β, IL-8, IL-10, and IL-23 were detected in the culture supernates. Co-incubation of S. apiospermum BF with DAMB or VRC significantly increased IL-1β, IL-8 and IL-10 levels as compared to untreated controls (IL-1β: 9±0.4 and 8.6±0.2 vs 7.8±0.1 pg/ml; IL-8: 184±18 and 217±39 vs 102±4 pg/ml; IL-10: 9.5±1.6 vs 5.1±0.3 pg/ml;P<0.01). All antifungal agents caused significantly increased levels of IL-8, IL-10, and IL-23 compared to the combination treatment of PMN+BF+drug (P<0.01).
Conclusions:DAMB and VRC showed an immunomodulatory profile towards PMN against biofilm damage. However, apart from the robust production of IL-8, the remaining cytokines were differentially produced but at low amounts which could lead to a dampened immune response. This may constitute a mechanism for the prolongation of a BF life-style with serious complications to the patient’s health.
Session Type: ePosters
Session Title: ePosters
Authors(s): A. Vikelouda (1), M. Simitsopoulou (1), L. Skoura (2), C. Antachopoulos (1), E. Roilides (1)
Authors Affiliations(s): (1) Infectious Diseases Unit, 3rd Department of Pediatrics, Aristotle University School of Medicine, Greece, (2) Microbiology Departmen, AHEPA Hospital, Greece
Background:
Scedosporium apiospermum can cause a wide range of invasive infections depending on host immunity. Scedosporium spp. can grow in biofilms, extracellular structures resistant to antifungal therapy or host immune mechanisms. Knowledge on the interplay between pathogen, antimicrobial agent and innate immune cells is pivotal in the fight against drug-resistant infections. We investigated whether deoxycholate amphotericin-B (DAMB), liposomal amphotericin-B (LAMB) and voriconazole (VRC) modulate immune response of human neutrophils (PMN) against S. apiospermum biofilms (BF).
Methods:105 cfu/ml of a BF-producing strain were grown in RPMI at 37οC for 48h to form mature BF. MICs of DAMB, LAMB and VRC were assessed by the XTT reduction assay and MIC50s were determined as ≥50% BF damage. PMN’s from healthy volunteers (106/ml in RPMI/10% human serum) were co-incubated with BF at effector-to-target ratio 5:1 and 1 mg/l DAMB, 2mg/l LAMB or 32mg/l VRC for 3 hours at 37οC/5% CO2. The cytokines released in culture supernatants were determined via multiplex ELISA using chemiluminescence. Microplate image capture and data analysis were performed using Quansys Q-view software. Protein concentrations between drug-treated and untreated PMN’s were compared by ANOVA (P<0.05). Two independent experiments were performed.
Results:MIC50s of DAMB, LAMB and VRC for BF were 1, 2 and 32 mg/l, respectively. IL-1β, IL-8, IL-10, and IL-23 were detected in the culture supernates. Co-incubation of S. apiospermum BF with DAMB or VRC significantly increased IL-1β, IL-8 and IL-10 levels as compared to untreated controls (IL-1β: 9±0.4 and 8.6±0.2 vs 7.8±0.1 pg/ml; IL-8: 184±18 and 217±39 vs 102±4 pg/ml; IL-10: 9.5±1.6 vs 5.1±0.3 pg/ml;P<0.01). All antifungal agents caused significantly increased levels of IL-8, IL-10, and IL-23 compared to the combination treatment of PMN+BF+drug (P<0.01).
Conclusions:DAMB and VRC showed an immunomodulatory profile towards PMN against biofilm damage. However, apart from the robust production of IL-8, the remaining cytokines were differentially produced but at low amounts which could lead to a dampened immune response. This may constitute a mechanism for the prolongation of a BF life-style with serious complications to the patient’s health.
Session Type: ePosters
Session Title: ePosters
Authors(s): A. Vikelouda (1), M. Simitsopoulou (1), L. Skoura (2), C. Antachopoulos (1), E. Roilides (1)
Authors Affiliations(s): (1) Infectious Diseases Unit, 3rd Department of Pediatrics, Aristotle University School of Medicine, Greece, (2) Microbiology Departmen, AHEPA Hospital, Greece
Background:
Scedosporium apiospermum can cause a wide range of invasive infections depending on host immunity. Scedosporium spp. can grow in biofilms, extracellular structures resistant to antifungal therapy or host immune mechanisms. Knowledge on the interplay between pathogen, antimicrobial agent and innate immune cells is pivotal in the fight against drug-resistant infections. We investigated whether deoxycholate amphotericin-B (DAMB), liposomal amphotericin-B (LAMB) and voriconazole (VRC) modulate immune response of human neutrophils (PMN) against S. apiospermum biofilms (BF).
Methods:105 cfu/ml of a BF-producing strain were grown in RPMI at 37οC for 48h to form mature BF. MICs of DAMB, LAMB and VRC were assessed by the XTT reduction assay and MIC50s were determined as ≥50% BF damage. PMN’s from healthy volunteers (106/ml in RPMI/10% human serum) were co-incubated with BF at effector-to-target ratio 5:1 and 1 mg/l DAMB, 2mg/l LAMB or 32mg/l VRC for 3 hours at 37οC/5% CO2. The cytokines released in culture supernatants were determined via multiplex ELISA using chemiluminescence. Microplate image capture and data analysis were performed using Quansys Q-view software. Protein concentrations between drug-treated and untreated PMN’s were compared by ANOVA (P<0.05). Two independent experiments were performed.
Results:MIC50s of DAMB, LAMB and VRC for BF were 1, 2 and 32 mg/l, respectively. IL-1β, IL-8, IL-10, and IL-23 were detected in the culture supernates. Co-incubation of S. apiospermum BF with DAMB or VRC significantly increased IL-1β, IL-8 and IL-10 levels as compared to untreated controls (IL-1β: 9±0.4 and 8.6±0.2 vs 7.8±0.1 pg/ml; IL-8: 184±18 and 217±39 vs 102±4 pg/ml; IL-10: 9.5±1.6 vs 5.1±0.3 pg/ml;P<0.01). All antifungal agents caused significantly increased levels of IL-8, IL-10, and IL-23 compared to the combination treatment of PMN+BF+drug (P<0.01).
Conclusions:DAMB and VRC showed an immunomodulatory profile towards PMN against biofilm damage. However, apart from the robust production of IL-8, the remaining cytokines were differentially produced but at low amounts which could lead to a dampened immune response. This may constitute a mechanism for the prolongation of a BF life-style with serious complications to the patient’s health.