Journal: Advanced Science

Article Title: Genetically‐Programmed Hypervesiculation of Lactiplantibacillus Plantarum Increases Production of Bacterial Extracellular Vesicles with Therapeutic Efficacy in a Preclinical Inflammatory Bowel Disease Model

doi: 10.1002/advs.202512679

Figure Lengend Snippet: LAB BEVs exhibit anti‐inflammatory effects in vitro. A–D) Mouse RAW264.7 (A‐B) or human dTHP1 (C‐D) macrophages pretreated with BEVs display reduced inflammatory TNF‐α secretion following stimulation with 10 ng mL −1 LPS (for RAW264.7) or 250 ng mL −1 LPS + 20 ng mL −1 IFNγ (for dTHP1). Dosing (A–D): V.V. Low = 5E6 particles mL −1 , V. Low = 5E7 particles mL −1 , Low = 5E8 particles/ml, Med = 5E9 particles/ml, High = 5E10 particles mL −1 . E) RAW264.7 macrophages pretreated with BEVs from Gram‐negative probiotic EcN BEVs or Gram‐positive probiotic L. plantarum (Lp) BEVs followed by LPS stimulation; Dosing (E): Low = 1E9 particles mL −1 , High = 5E10 particles mL −1 . F) dTHP1 macrophages were treated with 5E9 particles/ml of BEVs that were previously covalently labeled with fluorescent Alexa Fluor 647;24 h later, cell uptake of fluorescently‐labeled BEVs was analyzed by flow cytometry. Controls included BEVs not labeled with Alexa Fluor 647 (unlabeled), PBS/Vehicle treated cells (No treat), or free dye (Alexa Fluor 647 carboxylate 10 um) G) dTHP1 viability 24 h after treatment with Lc, Lp, or EcN BEVs (2e11 particles mL −1 ) and LPS+IFNγ stimulation was assessed by CCK8 assay. Abbreviations in figures are as follows: Lp ( L. plantarum ), Lc ( L. paracasei ), Lre ( L. reuteri ), Lrh ( L. rhamnosus ), EcN ( E. coli Nissle 1917), DH5a ( E. coli DH5a), Dex (Dexamathosone). Statistical significance determined with one‐way ANOVA and Tukey post hoc test. If two groups share at least one letter, their means are not significantly different (α=0.05). If they share no letters, they are significantly different ( p < 0.05). For example, “a” versus “ab” = not significant; “a” versus “b” = significant. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Error bars ± SD.

Article Snippet: All lactic acid bacteria (LAB) strains were obtained through ATCC ( Lacticaseibacillus rhamnosus GG (ATCC 53 103), Lacticaseibacillus paracasei (ATCC 334), Limosilactobacillus reuteri F 275 (ATCC 23 272), L. plantarum WCFS1 (ATCC BAA‐793).

Techniques: In Vitro, Labeling, Flow Cytometry, CCK-8 Assay

Journal: Advanced Science

Article Title: Genetically‐Programmed Hypervesiculation of Lactiplantibacillus Plantarum Increases Production of Bacterial Extracellular Vesicles with Therapeutic Efficacy in a Preclinical Inflammatory Bowel Disease Model

doi: 10.1002/advs.202512679

Figure Lengend Snippet: LAB BEVs reduce severity in acute DSS‐induced colitis. Mice received BEVs (2.5E9 particles mouse −1 day −1 , oral gavage, Days 1–7). DSS (2.5%) was given in drinking water Days 0–6, then normal water Days 7–8 (washout). A) Body‐weight change relative to Day 0. B) Colon length at Day 8. C,D) Mesenteric lymph‐node CD4+ T‐cell populations/activation at endpoint: Treg (CD4+CD8−CD25+Foxp3+), Th17 (CD4+CD8−Foxp3−RORγt+), Naive (CD4+CD44−CD62L+), Effector (CD4+CD44+CD62L−), Central memory (CD4+CD44+CD62L+). E) qPCR array of human UC–associated genes in proximal colon, shown as log2 fold‐change versus sham (vehicle/PBS) colitis. F) Pearson correlation of colitis‐associated genes comparing treatment groups with naïve/healthy mice; R^2 closer to 1 indicates greater similarity to naive. G) RT‐qPCR of selected colitis genes in pooled, bulk colon RNA. Abbreviations: L. plantarum (Lp), L. paracasei (Lc), L. reuteri (Lre), L. rhamnosus (Lrh), E. coli Nissle 1917 (EcN), E. coli DH5a (DH5a). Statistics: one‐way ANOVA with Tukey post hoc; different letters p < 0.05. If two groups share at least one letter, their means are not significantly different (α = 0.05). If they share no letters, they are significantly different ( p < 0.05). For example, “a” versus “ab” = not significant; “a” versus “b” = significant.

Article Snippet: All lactic acid bacteria (LAB) strains were obtained through ATCC ( Lacticaseibacillus rhamnosus GG (ATCC 53 103), Lacticaseibacillus paracasei (ATCC 334), Limosilactobacillus reuteri F 275 (ATCC 23 272), L. plantarum WCFS1 (ATCC BAA‐793).

Techniques: Activation Assay, Quantitative RT-PCR