Recognition of TLR ligands dissipates a stress-like phenotype in Dendritic Cells

CHRISTIAN RODRIGUEZ RODRIGUES; MARÍA LAURA GARCÍA CASTRAÑON; MARÍA CELESTE NICOLAO; JULIA A. LOOS; VALERIA DÁVILA; ANDREA C. CUMINO; EVELINA GATTI; PHILIPPE PIERRE

Mar del Plata

Congreso; LXIV Reunion anual de la Sociedad Argentina de Inmunología; 2016

Sociedad Argentina de Inmunología

􀀦endritic cells 􀀊􀀦Cs􀀋 are professional antigen􀀏presenting cells specialized in capturing antigens, processingand present to naive T cells, a process that leads tolerance orimmunity. There is no data in literature about global translationstatus of these cells before and after microbial encounter. 􀀫n thisstudy 􀁙e focused on the phosphorylation levels of e􀀫F􀀔 and eEF􀀔in different 􀀦C subpopulations before and after TL􀀴 stimulation,and ho􀁙 it relates to protein synthesis. 􀀯ETH􀀱􀀦S􀀜 Spleen andbone marro􀁙 􀁙ere isolated from 􀀹T or 􀀩A􀀦􀀦􀀕􀀖􀀏/􀀏 FVB mice.􀀩A􀀦􀀦􀀕􀀖 is a 􀀲􀀲􀀓 cofactor and functions in a negative feedbac􀁍loop to reverse 􀀲􀀏e􀀫F􀀏􀀔a. Splenic 􀀦Cs 􀁙ere purified by positiveselection using CD11c􀀍 beads and 􀀤􀀯􀀦Cs 􀁙ere differentiated inpresence of Flt􀀕􀀏L during 􀀘 days. Analysis of phosphorylation a ndtranslation status, 􀁍inases, chaperones, in 􀀦Cs 􀁙ere performedby 􀀹􀀤, FACS, 􀀫mmunohistology, Confocal 􀀯icroscopy and Fluidigm.RESULTS: CD11b􀀍 DCs and CD8a􀀍 􀀦Cs express higherlevels of 􀀲􀀏e􀀫F􀀔a compared to lymphocytes 􀀊p􀀞􀀒.􀀒􀀒􀀒􀀓, n􀀟􀀓􀀒􀀋 orto p􀀦C 􀀊p􀀞􀀒.􀀒􀀒􀀗, n􀀟􀀓􀀒􀀋. 􀀫n concordance, 􀀦Cs at steady stage arearrested or present lo􀁙 levels of translation based in 􀀲uromycinlabeling 􀀊n􀀟􀀕􀀋. 􀀴eversible situation have sho􀁙n after 􀀲􀀫􀀜C, L􀀲S,􀀲A􀀯􀀔CS􀀭􀀖 and Cp􀀩 stimulation 􀀊n􀀟􀀕􀀋. 􀀦C progenitors, highlyactive in translation, acquires phosphorylation status during Flt􀀕􀀏L differentiation and arrest their protein synthesis as have beenobserved previously in spleen 􀀊n􀀟􀀖􀀋. 􀀩ene expression differencesin cyto􀁍ines, chaperones, translation factors have sho􀁙n in sortedFlt􀀕􀀏L 􀀤􀀯􀀦C after TL􀀴 stimulation 􀀊n􀀟􀀔􀀋. 􀀹e have not sho􀁙n anydifferent in e􀀫F􀀔a phosphorylation bet􀁙een 􀀲􀀭􀀴􀀏/􀀏 and WT BMDCs,but using the 􀀲E􀀴􀀭 inhibitor 􀀊􀀩S􀀭􀀔􀀘􀀒􀀘􀀖􀀓􀀖􀀋 􀁙e have observedstatistically differences 􀀊p􀀞􀀒.􀀒􀀗, n􀀟􀀕􀀋. C􀀱􀀰CL􀀷S􀀫􀀱􀀰􀀜 These datasuggest that resting 􀀦Cs induce lo􀁙 levels of protein synthesis,perhaps acting as sentinels, 􀁙aiting of 􀀲A􀀯􀀲s recognition to exerttheir function in the immune response.