Differential cytokines patterns of LPS-stimulated PBMC treated with lactic acid bacteria or its metabolic products: in vitro approach to select strains with specific probiotic properties

JUAREZ, GE; FONT DE VALDEZ, G; RODRIGUEZ, AV

Buenos Aires

Congreso; LVIII Reunión Anual de la Sociedad Argentina de Inmunología; 2010

Sociedad Argentina de Inmunología

<!-- /* Font Definitions */ @font-face {font-family:"Cambria Math"; panose-1:2 4 5 3 5 4 6 3 2 4; mso-font-charset:0; mso-generic-font-family:roman; mso-font-pitch:variable; mso-font-signature:-1610611985 1107304683 0 0 159 0;} @font-face {font-family:"Arial Narrow"; panose-1:2 11 6 6 2 2 2 3 2 4; mso-font-charset:0; mso-generic-font-family:swiss; mso-font-pitch:variable; mso-font-signature:647 2048 0 0 159 0;} /* Style Definitions */ p.MsoNormal, li.MsoNormal, div.MsoNormal {mso-style-unhide:no; mso-style-qformat:yes; mso-style-parent:""; margin:0cm; margin-bottom:.0001pt; mso-pagination:widow-orphan; font-size:12.0pt; font-family:"Times New Roman","serif"; mso-fareast-font-family:"Times New Roman";} .MsoChpDefault {mso-style-type:export-only; mso-default-props:yes; font-size:10.0pt; mso-ansi-font-size:10.0pt; mso-bidi-font-size:10.0pt;} @page WordSection1 {size:612.0pt 792.0pt; margin:70.85pt 3.0cm 70.85pt 3.0cm; mso-header-margin:36.0pt; mso-footer-margin:36.0pt; mso-paper-source:0;} div.WordSection1 {page:WordSection1;} --> Abstract Among the differential immunomodulatory properties observed in Lactic acid bacteria (LAB) are outlined their anti-inflammatory features with potential application in chronic inflammatory diseases. The aim of this work was employing a single in vitro technique to select between LAB with technological interest, those strains with potential anti-inflammatory properties to be further tested in animal models. Human peripheral blood mononuclear cells (PBMC) isolated from healthy donors (24-34 years old), using the Ficoll-Hypaque technique, were stimulated with E. coli O26:B6 LPS (0,5 µg/mL) as a pro-inflammatory challenge and then were incubated in presence of 5 viable LAB strains from CERELA culture collection or with its respective conditioned media (CM), on RPMI medium at 37ºC (5% CO2) at different incubation times. TNF-α and IL-10 were detected in the free cell supernatants by ELISA. Results show that Lactobacillus reuteri CRL 1101 CM reduces release of TNF-α by 46±27% (p= 0,01) and 54±17% (p=0,05) at 4 and 6 h incubation respectively compared whit control values, whereas increases IL-10 production 2,7±0,7 (p=0,01) and 2,9±0,6 (p=0,05) folds. CM from Lactobacillus rhamnosus CRL 1505 and Streptococcus thermophilus CRL 1190 strains increase IL-10 release 2,7±0,3 (p=0,05) and 1,9±0,3 (p=0,05) folds, but CRL 1505 strain CM reduces TNF-α release only by 15±13% (p=0,05) at 4 h incubation and CRL 1190 strain CM do not produce significant differences on TNF-α secretion at 4 h incubation (1,24±0,44 folds). Metabolic products present in the CM from L. reuteri CRL 1101 strain has suitable in vitro anti-inflammatory characteristics to be deeper studied and tested as experimental anti-inflammatory therapy on inflammation animal models. This single method could be appropriate to select LAB with potential immunomodulatory properties before animal investigations and could optimize the use of in vivo assays.