Probiotic bacterial strains differentially modulate macrophage cytokine production in a strain-dependent and cell subset-specific manner

Abstract

Gut mucosal macrophages play a pivotal role in driving mucosal immune responses, resulting in either activation of inflammatory immune responses to pathogenic challenge or tolerance to beneficial luminal contents such as food and commensal bacteria. Macrophage responses elicited are dependent on tissue environment and the resulting cell subset, where homeostatic macrophages resemble the M2 macrophage subset and inflammatory macrophages resemble M1s. Probiotics can modulate macrophage function with outcome dependent on subset present. Using a THP-1 monocyte cell line-derived model of CD14high/low M1 and M2 macrophages, the aim of this study was to investigate the immunomodulatory effects of a panel of heat-killed probiotic bacteria and their secreted proteins on the subset-specific inflammatory marker profile of TNFa, IL-6 and NF?B. M1 and M2 cells were generated by differentiation of monocyte stable transfectants for high and low CD14 expression with phorbol 12-myristate 13-acetate and vitamin D3, respectively, where the resulting CD14lo M2 and CD14hi M1s mimicked homeostatic and inflammatory mucosal macrophages. Subsets were stimulated by enteropathic lipopolysaccharides in the presence or absence of heat-killed (HK) or secreted proteins (SP) from a panel of probiotic bacteria. Regulation of cytokine expression was measured by ELISA and NF?B activity by reporter assay. HK probiotics suppress CD14lo and augment CD14hi M1 and M2 production of TNFa whereas SPs augmented CD14hi M1 TNFa and were generally suppressive in the other subtypes. M2 macrophage IL-6 production was suppressed by both HK and SPs and differentially regulated in CD14lo and CD14hi M1s. NF?B activation failed to parallel the regulatory profiles for TNFa and IL-6 which is suggestive of probiotic bacteria exerting their regulatory effects on these cytokines in an NF?B-independent manner. In conclusion, HK and SP probiotics differentially regulate macrophage cytokines and NF?B activation in a subset-dependent manner and suggest a cautionary approach to probiotic treatment of mucosal inflammation. © 2011 Wageningen Academic Publishers.