Journal: Nature microbiology

Article Title: Colonic goblet cell-associated antigen passages mediate physiologic and beneficial translocation of live gut bacteria in preweaning mice

doi: 10.1038/s41564-025-01965-1

Figure Lengend Snippet: (A) Representative images from the proximal colon of DOL17 mice with gut-resident bacteria (eubacterial FISH probe; red) identified within a goblet cell (UEA1+ green) and DAPI (in blue) visualized in XY, YZ and XZ plane. (B) CFU/organ of L. animalis WU recovered from intestinal and extraintestinal tissues of Math1 f/f (n=9) and goblet cell deficient Math1 f/f vil-Cre-ER T2 (n=13) DOL17 mice fed with L. animalis WU . (C) CFU/organ of L. animalis WU recovered from the intestine and extraintestinal tissues of nontreated (n=14), EGF treated (n=14) and pan-S1PR modulator (FTY720) treated (n=6) DOL17 mice fed with L. animalis WU . (D) Flow cytometry plots and (E) graph showing depletion of F4/80+ MHCII+ CD45+ splenic cells in Clodrosome treated preweaning mice (n=5) when compared to nontreated mice (n=4). (F) CFU/organ of L. animalis WU recovered intestinal and extraintestinal tissues of PBS (n=4) and Clodrosome (n=4) treated preweaning mice. (G) CFU of in vitro cultured L. animalis WU recovered after treatment of 10 3 L. animalis WU with gentamicin at the denoted concentrations for 2 hours. CFU/organ of L. animalis WU recovered from (H) MLNs (n=3) and (I) Spleens (n=4) of preweaning mice in presence or absence of gentamicin treatment before cell lysis. Image in panel A is representative of colon sections from unmanipulated DOL17 mice from 3 independent litters in which 13 of 125 goblet cells (UEA1+) and 1 of 553 (UEA1-) colonocytes imaged in 3 dimensions (z-stacks) contained bacteria (eubacterial FISH probe +). Bar graphs presented as mean+/− SEM. Data points in panels H and I represent paired observations of CFUs within one half of the same MLN or spleen. Two- tailed Student’s t-test was used in E, H and I and intestinal tissues in B and F. A one-sided cumulative binomial distribution probability test was used for extraintestinal tissues in B, C, and F. A one-way ANOVA with at Dunnett’s post test was used for intestinal tissues in C and G.

Article Snippet: Statistical analyses performed using DEseq2 with FDR<0.05 and >2 fold change for B in Partek ® FLOW ® and with one-way ANOVA with a Dunnett’s post test for C-J.

Techniques: Translocation Assay, Bacteria, Flow Cytometry, In Vitro, Cell Culture, Lysis, Two Tailed Test

Journal: Nature microbiology

Article Title: Colonic goblet cell-associated antigen passages mediate physiologic and beneficial translocation of live gut bacteria in preweaning mice

doi: 10.1038/s41564-025-01965-1

Figure Lengend Snippet: (A) Schematic representation of experimental setup used in investigating transcriptomic changes associated with L. animalis WU translocation in MLN of DOL17 mice. (B) Heatmap of 69 differentially expressed genes in sorted cellular populations defined as in (A) and confirmed by CFUs in the liver. (C) Peripheral blood neutrophils, and serum (D) IL6, (E) TNFα, (F) IL10, (G) CCL2, (H) CXCL9, (I) CXCL10 and (J) IFNα measured in DOL17 mice that are non-treated (n=6) or treated with L. animalis WU fed orally ( L. ani oral ) (n=6) or administered by i.p. injection ( L. ani i.p .) (n=6). Graphs presented as mean +/− SEM. Statistical analyses performed using DEseq2 with FDR<0.05 and >2 fold change for B in Partek ® FLOW ® and with one-way ANOVA with a Dunnett’s post test for C-J.

Article Snippet: Statistical analyses performed using DEseq2 with FDR<0.05 and >2 fold change for B in Partek ® FLOW ® and with one-way ANOVA with a Dunnett’s post test for C-J.

Techniques: Translocation Assay, Injection

Journal: Nature microbiology

Article Title: Colonic goblet cell-associated antigen passages mediate physiologic and beneficial translocation of live gut bacteria in preweaning mice

doi: 10.1038/s41564-025-01965-1

Figure Lengend Snippet: (A) Flow cytometry gating strategy for identifying Th1 cytokines (TNFα and IFNɣ) secreted by CD45 + CD3 + CD4 + T cells in MLNs and spleens. Frequency of TNFα+ and IFNɣ+ CD4+ T cells in (B-C) MLNs and (D-E) spleens of nontreated (control) (n=9), L. animalis WU fed (n=5) and L. animalis WU +EGF fed mice (n=6). (F) Flow cytometry gating strategy for identifying Th2 cytokines (IL4 and IL13) secreted by CD45 + CD3 + CD4 + T cells in MLNs and spleens. Frequency of IL13+ and IL4+ CD4+ T cells in (G-H) MLNs and (I-J) spleens of nontreated (control) (n=9), L. animalis WU fed (n=5) and L. animalis WU +EGF fed mice (n=6). (K) Flow cytometry gating strategy for identifying Th17 cytokines (IL17 and IL22) secreted by CD45 + CD3 + CD4 + T cells in MLNs and spleens. Frequency of IL17+ and IL22+ CD4+ T cells in (L-M) MLNs and (N-O) spleens of nontreated (control) (n=9), L. animalis WU fed (n=5) and L. animalis WU +EGF fed mice (n=6). Statistical analyses were performed by one-way ANOVA with Dunnett’s post test. Graphs represent mean+/− SEM. P values are as denoted.

Article Snippet: Statistical analyses performed using DEseq2 with FDR<0.05 and >2 fold change for B in Partek ® FLOW ® and with one-way ANOVA with a Dunnett’s post test for C-J.

Techniques: Flow Cytometry, Control

Journal: Nature microbiology

Article Title: Colonic goblet cell-associated antigen passages mediate physiologic and beneficial translocation of live gut bacteria in preweaning mice

doi: 10.1038/s41564-025-01965-1

Figure Lengend Snippet: A) Graphs and B) representative flow plots of Foxp3+ and RORγt+ Foxp3+ regulatory T cell populations in the MLN and spleen of unmanipulated mice (Control) (n=4 for MLNs and 3 for spleens), mice fed L. animalis WU from DOL10–20 ( L. animalis WU ) (n=4 for MLNs and 3 for spleens), and mice fed L. animalis WU given EGF gavage from DOL10–20 ( L. animalis WU +EGF ) (n=4 for MLNs and spleen). Graphs represent mean +/− SEM. Statistical analyses were performed by one way ANOVA with a Dunnett’s post test.

Article Snippet: Statistical analyses performed using DEseq2 with FDR<0.05 and >2 fold change for B in Partek ® FLOW ® and with one-way ANOVA with a Dunnett’s post test for C-J.

Techniques: Control