Fig 1.
Phenotypic effects of Bin1 mAb treatment in DSS-treated mice.
There was a change in (A) weight of mice, (B) length of colon, (C) the amount of fecal pellets, and (D) urine drops after treatment with Bin1 mAb after subjecting to DSS treatment (*P<0.05, **P<0.01, ***P<0.001, ****P<0.0001, as determined by t test). All results are expressed as Mean ± SD; (n = 5 mice per treatment). Fig 1A. Day 7: Control and DSS *p<0.0038. Day 15: DSS and Bin1 mAb *p<0.0285, IgG and Bin1mAb *p<0.0156. Fig 1B. Day 7: Control and DSS *p<0.0002, Day 15: Control and DSS *p<0.0005, DSS and Bin1 mAb *p<0.0057, IgG and Bin1 mAb *p< 0.0015. Fig 1C. Day 7: Control and DSS *p<0.0008. Day 15: Control and DSS *p<0.0116, Control and IgG *p<0.0265. Fig 1D. Day 7: Control and DSS *p<0.0076. Bin1, bridging integrator 1; DSS, dextran sodium sulfate; mAb, monoclonal antibodies.
Fig 2.
Bin1 mAb treatment improved NeuN expressing enteric neurons and GFAP expressing glial cells after DSS-induced colitis.
Fig 2A. Effect of Bin1 mAb treatment on neuronal and glial abundance in the gut of DSS-induced colitis mice as determined by confocal microscopy. Bin1 mAb treated mice had high expression of NeuN and low expression of GFAP (n = 5). Fig 2B. Bin1 mAb treated mice had high levels of NeuN expressing neuronal cells in the muscularis of the colon as quantitated by CTCF analysis (*P<0.05, **P<0.01, as determined by t test). Control and DSS *p<0.0060, Control and IgG *p<0.0030, DSS and Bin1 mAb *p<0.0414, IgG and Bin1 mAb *p< 0.0234. Fig 2C. Bin1 mAb treated mice had low levels of GFAP expressing glial cells in the colon as quantitated by CTCF analysis (CTCF analysis, n = 3 mice tissues per treatment) (*P<0.05, **P<0.01, as determined by t test). Control and DSS *p<0.0006, Control and IgG *p<0.0002, DSS and Bin1 mAb *p<0.0007, IgG and Bin1 mAb *p<0.0003.
Fig 3.
The muscularis of the colon is not a continuous entity.
Fig 3A. The muscularis of the colon has openings (anigma) that transport glial cells (stained with GFAP), which protect the enteric neurons (n = 5). Fig 3B. H and E staining of the mouse colon showing that muscularis is not a continuous entity. The muscularis of the colon is punctuated with openings (anigma) (n = 5; representative image). Inset in figure B shows an intact serosa in the muscularis (large arrow) and anigma in the muscularis (long arrow).
Fig 4.
Treatment with Bin1 mAb improved PGP9.5 expressing enteric neurons after DSS-induced colitis.
Fig 4A. Effect of Bin1 mAb treatment on the expression of PGP9.5 neuronal cells in the colon of DSS-induced colitis mice as determined by confocal microscopy. Representative image from two independent experiments shown. Fig 4B. Bin1 mAb treated mice had high levels of PGP9.5 expressing neuronal cells in the muscularis of the colon as quantitated by CTCF analysis (CTCF analysis, n = 3 mice tissues per treatment) (*P<0.05, **P<0.01, as determined by t test). Control and DSS *p<0.0063, Control and IgG *p<0.0259, DSS and Bin1 mAb *p<0.0197.
Fig 5.
Bin1 mAb treatment improved MAP2 expressing enteric neurons after DSS-induced colitis.
Fig 5A. Effect of Bin1 mAb treatment on the expression of MAP2 and TuJ1 neuronal cells in the colon of DSS-induced colitis mice as determined by confocal microscopy. Representative image from two independent experiments shown. Fig 5B. Bin1 mAb treated mice had high levels of MAP2 expressing neuronal cells in the muscularis of the colon as quantitated by CTCF analysis (CTCF analysis, n = 3 mice tissues per treatment) (*P<0.05 as determined by t test). Control and DSS *p<0.0112, Control and IgG *p<0.0124, DSS and Bin1 mAb *p<0.0144, IgG and Bin1 mAb *p<0.0152.
Fig 6.
Morphological changes in bacteria of the gut after the treatment with Bin1 mAb.
The fecal pellets were collected from untreated control mice, DSS-induced colitis mice, IgG treated mice and Bin 1 mAb treated mice; they were cultured aerobically and anaerobically and the morphology observed under phase contrast microscopy. Fig 6A. Aerobic culture of microorganisms show that the phenotype of Bin1 mAb treated mice has bacteria similar to the control (Phase contrast, 500X) (n = 5; representative image). Fig 6B. Anaerobic culture of microorganisms from fecal pellets were slow growing. There was no change in phenotype of the microorganisms between Bin 1 mAb treated and control groups (Phase contrast, 500X) (n = 5; representative image).
Fig 7.
Microbiome sequencing by 16S rRNA showed lack of Firmicutes in the DSS-induced colitis mice in the fecal pellet or when cultured in a biosimulator under aerobic and anaerobic conditions.
The fecal pellets were collected from untreated control mice, DSS-induced colitis mice (D7, Day 7), IgG treated mice (D15, Day 15) and Bin 1 mAb (D15, Day 15) treated mice; they were directly analyzed or cultured aerobically and anaerobically in a biosimulator. The level of Firmicutes improved during treatment with the Bin1 mAb (n = 3 per treatment).
Fig 8.
The relative abundance of the microbiome of the fecal pellets of mice from different treatment conditions.
The bacteria of the genus Lactobacillus is totally absent in the UC mice. The UC mice is enriched in Enterobacteriaceae. (n = 3 mice per treatment).