Potentially probiotic Lactobacillus strains with anti-proliferative activity induce cytokine/chemokine production and neutrophil recruitment in mice

Abstract

Lactobacillus pentosus B281 and Lactobacillus plantarum B282 are two Lactobacillus strains previously isolated from fermented table olives. Both strains were found to possess probiotic properties and displayed desirable technological characteristics for application as starters in novel functional food production. In the present study the anti-proliferative and immunostimulatory activities of the two strains were investigated. Firstly, we demonstrated that live L. pentosus B281 and L. plantarum B282 significantly inhibited the growth of human colon cancer cells (Caco-2) in a time- and dose-dependent manner. By employing the air pouch system in mice, we showed that administration of both strains led to a rapid and statistically significant infiltration of leukocytes in the air pouch exudates. The phenotypical characterisation of the recruited immune cells was performed by flow cytometry analysis. We demonstrated that the majority of the infiltrated leukocytes were neutrophils. Finally by using the Mouse Cytokine Array Panel A Detection Antibody cocktail, we showed that both strains induced the expression of granulocyte-colony stimulating factor, interleukin (IL)-1α, IL-1β, IL-6, chemokine (C-X-C motif) ligand (CXCL)-1, chemokine (C-C motif) ligand (CCL)-3, CCL-4, and CXCL-2 and diminished the expression levels of soluble intercellular adhesion molecule, macrophage colony-stimulating factor and metallopeptidase inhibitor 1. Our results showed that both strains display anti-proliferative and immunostimulatory properties equal or even better in some cases than those of established and commonly used probiotic strains. These findings further support the probiotic character of the two strains.

Assessment of the probiotic potential of lactic acid bacteria isolated from kefir grains: evaluation of adhesion and antiproliferative properties in in vitro experimental systems

“…Accordingly, it has been demonstrated that cell-free supernatants from probiotic L. casei ATCC 334 and L. rhamnosus GG attenuate cell invasion of the human colon carcinoma cell line HCT-116 by reducing the activity of matrix metalloproteinase-9 and the levels of the tight junction protein zona occludens-1 (Escamilla et al 2012). Similarly, we have shown recently, that CM of the probiotic strains L. pentosus B281 and L. plantarum B282, inhibited proliferation of Caco-2 cells through induction of a G1 arrest of the cell cycle and downregulation of specific cyclin genes (Saxami et al 2017). An elegant study demonstrated that ferrichrome isolated from the CM of L. casei ATCC 334 exerted anti-cancer properties both in vitro and in a mouse xenograft model (Konishi et al 2016).…”

“…The quantitative analysis of bacterial adhesion to HT-29 cells was performed as reported by Saxami et al (2016). Briefly, 3 × 10 5 cells/well were seeded in 24-well culture plates.…”

“…The anti-proliferative effects of lactobacilli were determined on HT-29 cells by sulforhodamine B (SRB) colorimetric assay as described previously (Saxami et al 2016). Briefly, HT-29 cells were seeded in 96-well microplates at a density of 6 × 10 3 cells per well.…”

Assessment of the Immunomodulatory Properties of the Probiotic Strain Lactobacillus paracasei K5 In Vitro and In Vivo

“…As shown in Figure 1B, the injection of L. paracasei K5 induced a rapid and strong recruitment of leukocytes in the exudates of air pouches, significantly higher than L. casei ATCC 393 and L. rhamnosus GG (p < 0.05). Specifically, the administration of L. paracasei K5 increased the infiltration of leukocytes in the exudates by eight-fold compared to control mice that received only the PBS solution, whereas the administration of L. casei ATCC 393 and L. rhamnosus GG enhanced the recruitment of leukocytes up to six-fold compared to the control, as recorded before [18].…”

“…Distinct modulation of the immune response in vivo following probiotic administration has been well documented. For example, we have showed previously that administration of L. pentosus B281 and L. plantarum B282, two recently characterized probiotic strains, induced significant recruitment of leukocytes in the air-pouch exudates of mice and differential expression of specific cytokines and chemokines [18]. Similarly, the probiotic strain Lactobacillus casei BL23 promoted a Th17-biased immune response, followed by the expression of proinflammatory cytokines (IL-6, IL-17, IL-10, and TGF-β), in a mouse model of intestinal inflammation [19].…”