Posaconazole in Vitro Action on Peripheral Blood Mononuclear Cells From Sarcoidosis Patients

Abstract

SESSION TITLE: Diffuse Lung Disease 1 SESSION TYPE: Original Investigation Poster PRESENTED ON: Wednesday, November 1, 2017 at 01:30 PM - 02:30 PM PURPOSE: Treatment with anti-fungal medication is as efficient as corticosteroids in sarcoidosis in many, albeit not all, patients.Insoluble β-glucan, a component in the fungal cell wall, induced the production of several cytokines from peripheral blood mononuclear cells (PBMC) “in vitro”. Against this background two hypotheses can be formulated: anti-fungal medication influences immune cell activity and decreases the secretion of cytokines after stimulation with insoluble β-glucan and anti-fungal medication acts on insoluble β-glucan and decreases its capacity to induce cytokine production from PBMC. METHODS: Human PBMCs from sarcoidosis (n=6) and healthy subjects (n=5) were stimulated for 4h with particulate (1→3)-β-D-glucan (BGP) after 24h preincubation with posaconazole, with methylprednisolone or with medium alone. We preincubated the BGP with or without the Posaconazole for 1h before adding the PBMCs from healthy subjects for additional 4h. The viability of the cells was determined, the concentrations of TNF-α in cell culture supernatants were measured and expression of NF-kB mRNA. RESULTS: When adding the PBMCs from healthy subjects for additional 4 h to BGP preincubated either with or without Posaconazole, there was no difference in TNF-α production (884 ± 23.9 pg/ml; 981 ± 70.2 pg/ml). Meanwhile, the secretion of TNF-α was significantly higher in sarcoidosis when PBMCs were stimulated with BGP (2162 ± 432.6 pg/ml; 1338 ± 116.7 pg/ml; p=0,032) than in healthy subjects. However, in sarcoidosis patients, preincubation of PBMCs with Posaconazole significantly reduced the secretion of TNF-α after stimulation with BGP (p=0,046), while having no effect on the healthy. On the other hand, Posaconazole significantly reduced the expression of NF-Kb in both groups and there was no significant difference in cell viability in cultures. Thus, the results demonstrate that the inhibitory effect of Posaconazole on PBMCs from sarcoidosis patients is neither a cytotoxic effect of Posaconazole on PBMCs nor a result of the interaction of Posaconazole with BGP. CONCLUSIONS: We have demonstrated that posaconazole “in vitro”significantly reduces the secretion of TNF-α from sarcoidosis patients. The results are consistent with our previous clinical research.There, we have already demonstrated that antifungal treatment is as efficient as corticosteroid treatment of granulomatous inflammation in many, albeit not all,sarcoidosis patients. CLINICAL IMPLICATIONS: Additional studies are required to define the phenotype where the antifungal treatment is not efficient. DISCLOSURE: The following authors have nothing to disclose: Marjeta Tercelj-Zorman, Sanja Stopinsek, Barbara Salobir, Alojzij Ihan, Sasa Simcic No Product/Research Disclosure Information