Knock out of sHSP genes determines some modifications in the probiotic attitude of Lactiplantibacillus plantarum

Abstract

Objective We investigated whether the knock out of small heat shock protein (sHSP) genes (hsp1, hsp2 and hsp3) impact on probiotic features of Lactiplantibacillus plantarum WCFS1, aiming to find specific microbial effectors involved in microbe-host interplay. Results The probiotic properties of L. plantarum WCFS1 wild type, hsp1, hsp2 and hsp3 mutant clones were evaluated and compared through in vitro trials. Oro-gastro-intestinal assays pointed to significantly lower survival for hsp1 and hsp2 mutants under stomach-like conditions, and for hsp3 mutant under intestinal stress. Adhesion to human enterocyte-like cells was similar for all clones, though the hsp2 mutant exhibited higher adhesiveness. L. plantarum cells attenuated the transcriptional induction of pro-inflammatory cytokines on lipopolysaccharide-treated human macrophages, with some exception for the hsp1 mutant. Intriguingly, this clone also induced a higher IL10/IL12 ratio, which is assumed to indicate the anti-inflammatory potential of probiotics. Conclusions sHSP genes deletion determined some differences in gut stress resistance, cellular adhesion and immuno-modulation, also implying effects on in vivo interaction with the host. HSP1 might contribute to immunomodulatory mechanisms, though additional experiments are necessary to test this feature.

Probiotic Gastrointestinal Transit and Colonization After Oral Administration: A Long Journey

“…Nishiyama et al compared the adhesion ability of 31 L. pentosus strains to mucin and discovered a highly adhesive L. pentosus strain, which over-produced four moonlighting proteins including sHSPs (Peŕez Montoro et al, 2018). A recent study investigated the impact of knockout of the sHSP genes (including HSP1, HSP2, and HSP3) on adhesion of L. plantarum WCFS1 to human enterocyte-like cells, demonstrating that sHSP genes deletion lowered GIT stress resistance and adhesion capacity (Longo et al, 2020).…”

Immunomodulatory Activity on Human Macrophages by Cell-Free Supernatants to Explore the Probiotic and Postbiotic Potential of Lactiplantibacillus plantarum Strains of Plant Origin

“…Quantitative RT-PCR was performed for the transcriptional analysis of genes encoding tumour necrosis factor-alpha (TNF-α), interleukin 8 (IL-8), interleukin 10 (IL-10) and interleukin 12 (IL-12), by using QuantiFastSybr ® Green PCR kit (Qiagen) in a real-time instrument (ABI 7300, Applied Biosystem, Foster City, CA, USA), by applying the ΔΔCt method. β-Actin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) transcript levels were used as internal normaliser, as previously described [ 47 , 49 ].…”

Antimicrobial Properties, Functional Characterisation and Application of Fructobacillus fructosus and Lactiplantibacillus plantarum Isolated from Artisanal Honey

“…Human monocytoid leukaemia-derived cells, THP-1 (from Sigma-Aldrich), were cultivated in RPMI-1640 (Gibco, Carlsbad, CA) supplemented with 10% (v/v) fetal bovine serum (FBS), 2 mM L-glutamine, 50 U mL −1 penicillin and 50 μg mL −1 streptomycin, in 5% CO 2 , at 37 °C. Immunostimulation experiments were performed as previously reported [ 39 ]. Briefly, THP-1 cells were seeded (5 × 10 5 cells/well) in 24-well tissue culture-treated plates (EuroClone, Milan, Italy), and 100 ng mL −1 phorbol 12-myristate 13-acetate (PMA) (Sigma-Aldrich) was added to induce differentiation into macrophages.…”