Mucins and polysaccharides from Echinococcus granulosus laminar layer induce a mild maturation phenotype in dendritic cells and promote gene expression of pro-inflammatory cytokines.

PLA, NATALIA; CHOP, MAIA; NICOLAO MARÍA CELESTE; LOOS JULIA .A.; CUMINO ANDREA C.; RODRIGUEZ R CHRISTIAN

Buenos Aires

Congreso; LXV REUNIÓN ANUAL DE LA SOCIEDAD ARGENTINA DE INVESTIGACIÓN CLÍNICA (SAIC); 2020

Sociedad Argentina de Investigaciones Clínica y Sociedades Conjuntas

Background: The laminar layer is an acellular structure rich in glycansthat surrounds the metacestode, creating a mechanical andimmunological protective barrier, crucial in the Echinococcus?hostinterface. The aim of this work is to analyze if purified laminar layer(pLL) could trigger maturation and cytokine production in dendriticcells by activating mTOR pathway, a central regulator of cell metabolismand environmental signals. Methods: BMDCs were cultured incomplete RPMI supplemented with 100 ng/ml Flt3-L.The germinal layer was removed from pLL by washing with 2M NaCl.Flow cytometry: FITC or PE-conjugated mAbs directed to CD11c,CD40, CD80, CD86, MHC I and MHC II were used. Immunoblotingand confocal microscopy: mouse anti-puromycin and rabbit anti-Phospho-mTOR antibodies were used. Gene expression analysisof IL-10, IL-12p35, IL-6, TNF-a, IL-23p19, TGF-b was performedon a Real Time PCR System Results: The pLL of Echinococcus inducea non-statistically significant, but a trend in the up-regulationof CD86 and MHC II and that expression change was diminishedby the use of rapamycin (n=5). Conversely, no changes in the expressionof CD40, CD80 or MHC I were registered. Further studieswere done to analyze whether pLL was also able to stimulate theproduction of cytokines by BMDCs. When BMDCs were stimulatedwith pLL induce the expression of IL-6 and TNF-a (n=3 *p