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Role of Toll-Like Receptors in the recognition of probiotics by monocyte-derived dendritic cells

AuthorsMartínez-Abad, Beatriz; Garrote, José Antonio ; Vallejo-Díez, Sara; Montalvillo, Enrique; Escudero-Hernández, Celia; Bernardo, David; Vázquez, Enrique ; Rueda, Ricardo; Arranz, Eduardo
Issue Date2012
CitationEMIG 2012
AbstractTo determine the recognition of probiotics and pathogen bacteria by TLRs in monocyte-derived dendritic cells through a PCR array. Monocytes were cultured with GM-CSF and IL-4 for 5 days. Escherichia coli, Salmonella typhimurium and Clostridium perfringens were used as positive bacterial control. Four species of probiotics were used from the lactobacillus genus: L1, L2, L3 and L4; and two strains from Bifidobacterium genus: B1 and B2. Bacteria were collected in the exponential phase of growth and monocyte-derived dendritic cells (moDCS) were stimulated in a ratio 50 bacteria: 1 moDC for 3 hours. Stimulated moDCs were harvested and RNAs extracted. moDCs were obtained from the collection of six blood donors. We studied the TLR pathway performing the RT2 Profiles TM PCR Array Human Toll-Like Receptor Signalling Pathway. Changes in the transcriptional expression of ACTB were estimated by the ΔΔCT method using basal condition as reference. The expression of TLR2 and molecules from the TLR signalling pathway was triggered by E. coli, S. typhimurium and L3. With the remainding probiotics, we observed a lower expression of TLR pathway molecules such as MyD88, IRAK2, MAPK and NFKB. Among probiotics, Bifidobacteria induced the lowest expression in our assay. CXCL10 and IFNB1 expression was decreased in a greater degree by Bifidobacteria. L3 produced an increase of IL10 and CSF3 expression which are involved in regulation. There are genes whose expression were up-regulated in all culture conditions such as IL1B, IL8, PTGS2, TNF, MAP2K3, CSF2 and IL1A. However, IFNG expression is specific for positive controls.
DescriptionPóster presentado al 8th European Mucosal Immunology Meeting celebrado en Dublín (Irlanda) del 10 al 13 de octubre de 2012.
Appears in Collections:(IBGM) Comunicaciones congresos
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