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P860 Evaluation of the probiotic features of two different multi-strain probiotic preparations from two different manufacturers

S. Colombo*1, V. Sagheddu2, M. Elli2, D. Mora3

1Beingpharma, Milan, Italy, 2AAT-Advanced Analytical Technologies Srl, Fiorenzuola d’Arda, Italy, 3Università degli studi di Milano, Defens, Milano, Italy

Background

The aim of this work was to compare the probiotic adhesion features and immunomodulatory properties of 2 probiotic products characterised by the same bacterial content but manufactured in different places (Italy and USA).

Methods

The multi-strain probiotic products under investigation were composed by a mixture of 8 different strains. The two products were manufactured in Italy (VSL#3 - Lot 710061 expiry date 10/2019) and in US (Vivomixx - Lot 1708702 expiry date 31/03/2019). The cell viability in each lot was assessed by flow cytometry. The adhesive abilities of probiotics were evaluated on three different eukaryotic intestinal cell lines (Caco-2, HT-29 and mucus-producing HT29-MTX).

For the evaluation of immune-modulatory activity, human Dendritic Cells (hDCs) obtained from blood mononuclear cells were exposed for 6 days to cytokines cocktail (IL-4 20 ng/ml + GM-CSF 50 ng/ml) and then to both probiotic mixtures for 2 h with MOI 1:10. A positive inflammatory condition was obtained by incubating washed hDCs in fresh medium containing Salmonella sp. for additional two h. Finally, hDCs were then cultured for 23 h in complete medium with antibiotics for the final analysis of soluble and surface molecules with ELISA or cytofluorimetric analysis method, respectively.

Results

The two products showed a comparable amount of live cells, 2.2 1011 (FU/g) and 2.4 1011 (FU/g), respectively, for the VSL#3 and Vivomixx. The two tested products showed similar adhesive capacity to the 3 cell lines considered, with adhesion values in the range 67–71%. No significant differences were observed in the percentage of adhesiveness expressed by the two mixtures according to the type of cell line considered. Regarding the immune-modulatory ability of the two tested products, results showed that they have the same behaviours. The exposure of the probiotic mixtures to hDCs resulted in a meaningful reduction of the expression of IL-12 following Salmonella induction. We observed that both products slightly increase the proportion of HLA-DR+/CD11+ and CD80+/CD11+ hDCs. Moreover, both mixtures significantly reduce Salmonella-induced HLA-DR+/CD11+ and CD80+/CD11+ hDCs. Both probiotic products did not significantly induce TNF-α production by hDCs at basal condition, and they did not result in further TNF-α production if compared with that induced by Salmonella sp. alone even if the response of the two products was almost identical.

Conclusion

Investigative activities described in this work demonstrated a comparable adhesive performances and interesting immunomodulatory activity against inflammation induced by Salmonella sp. for both VSL#3 and Vivomixx showing same behaviours irrespective of the manufacturer.