Bifidobacteria from human origin: interaction with phagocytic cells.

AIM OF THE STUDY: Given that phagocytic cells are main players of the host immune response, we studied the interaction of bifidobacteria with monocytic THP-1 cells in nonopsonic conditions. METHODS AND RESULTS: Association/internalization, cell response (expression of HLA-DR and TLR2), M1/M2 macrophage polarization and colocalization of micro-organisms with Lysotracker or transferrin were evaluated. Screening with eight Bifidobacterium strains showed two patterns of interactions with THP-1 cells: high and low association and phagocytosis. Two strains with different surface properties were further studied: B. bifidum CIDCA 5310 and B. adolescentis CIDCA 5317. Strain CIDCA 5310 showed higher levels of colocalization in lysosome than strain CIDCA 5317. Both strains stimulated TLR2 expression. Strain CIDCA 5317 significantly increases HLA-DR expression, however, when cells are stimulated with IFN-γ, strain CIDCA 5310 induces the highest value of expression. Noteworthy, strain CIDCA 5310 was able to upregulate both M1 and M2 markers of macrophage polarization. CONCLUSIONS: Our results demonstrate that bifidobacteria from human origin show different patterns of interaction with phagocytic cells thus leading to different cell responses. These findings add further insight on the mechanisms involved in the biologic effects of probiotics. SIGNIFICANCE AND IMPACT OF THE STUDY: Knowledge of the interaction of bifidobacteria with key players of the host immune response is paramount for the understanding of the mechanisms involved in the beneficial effects.

Lactobacillus delbrueckii subsp lactis CIDCA 133 modulates response of human epithelial and dendritic cells infected with Bacillus cereus.

It is known that probiotic microorganisms are able to modulate pathogen virulence. This ability is strain dependent and involves multiple interactions between microorganisms and relevant host's cell populations. In the present work we focus on the effect of a potentially probiotic lactobacillus strain (Lactobacillus delbrueckii subsp. lactis CIDCA 133) in an in vitro model of Bacillus cereus infection. Our results showed that infection of intestinal epithelial HT-29 cells by B. cereus induces nuclear factor kappa B (NF-κB) pathway. Noteworthy, the presence of strain L. delbrueckii subsp.lactis CIDCA 133 increases stimulation. However, B. cereus-induced interleukin (IL)-8 production by epithelial cells is partially abrogated by L. delbrueckii subsp. lactis CIDCA 133. These findings suggest that signalling pathways other than that of NF-κB are involved. In a co-culture system (HT-29 and monocyte-derived dendritic cells), B. cereus was able to translocate from the epithelial (upper) to the dendritic cell compartment (lower). This translocation was partially abrogated by the presence of lactobacilli in the upper compartment. In addition, infection of epithelial cells in the co-culture model, led to an increase in the expression of CD86 by dendritic cells. This effect could not be modified in the presence of lactobacilli. Interestingly, infection of enterocytes with B. cereus triggers production of proinflammatory cytokines by dendritic cells (IL-8, IL-6 and tumour necrosis factor alpha (TNF-α)). The production of TNF-α (a protective cytokine in B. cereus infections) by dendritic cells was increased in the presence of lactobacilli. The present work demonstrates for the first time the effect of L. delbrueckii subsp. lactis CIDCA 133, a potentially probiotic strain, in an in vitro model of B. cereus infection. The presence of the probiotic strain modulates cell response both in infected epithelial and dendritic cells thus suggesting a possible beneficial effect of selected lactobacilli strains on the course of B. cereus infection.

Virulence of Bacillus cereus: a multivariate analysis.

Biological activity and presence of DNA sequences related to virulence genes were studied in 21 strains of the Bacillus cereus group. The activity of spent culture supernatants and the effect of infection by vegetative bacterial cells were assessed on cultured human enterocytes (Caco-2 cells). The effect of extracellular factors on the detachment, necrosis and mitochondrial dehydrogenase activity of cultured human enterocytes was studied. Hemolytic activity on rabbit red blood cells was also evaluated and the effect of direct procaryotic-eucaryotic interactions was assessed in infection assays with vegetative bacterial cells. Concerning virulence genes, presence of the DNA sequences corresponding to the genes entS, entFM, nhe (A, B and C), sph, hbl (A, B, C and D), piplC and bceT was assessed by PCR. Ribopatterns were determined by an automated riboprinting analysis after digestion of the DNA with EcoRI. Principal component analysis and biplots were used to address the relationship between variables. Results showed a wide range of biological activities: decrease in mitochondrial dehydrogenase activity, necrosis, cell detachment and hemolytic activity. These effects were strain-dependent. Concerning the occurrence of the DNA sequences tested, different patterns were found. In addition, ribotyping showed that strains under study grouped into two main clusters. One of these clusters includes all the strains that were positive for all the DNA sequences tested. Positive and negative correlations between variables under study were evidenced. Interestingly, high detaching strains were positively correlated with the presence of the sequences entS, nheC and sph. Within gene complexes, high correlation was found between sequences of the hbl complex. In contrast, sequences of the nhe complex were not correlated. Some strains clustered together in the biplots. These strains were positive for all the DNA sequences tested and they were able to detach enterocytes upon infection. Our results highlight the multifactorial character of the virulence of the B. cereus group and show the correlation between ribopatterns, occurrence of toxin genes and biological activity of the strains under study.