Propyzamide

AB wild-type and lck:gfp (7 days post-fertilization larvae)^[2]

0.2, 1, 5, and 20 μM

in E3 medium; co-administered with 25 μg/mL TNBS; 24-72 h starting at 7 d.p.f.

Boosted TNBS-induced intestinal inflammation in zebrafish, as indicated by intestinal scores and expression of pro-inflammatory genes.
In TNBS-exposed zebrafish, Propyzamide at 20 μM was tested in combination with TNBS; quantification of intestinal GFP-positive T cells in lck:gfp zebrafish was performed.
Did not induce intestinal inflammation in the absence of TNBS.
AHR activation by FICZ or NOS inhibition partially interfered with the worsening of TNBS-induced colitis by Propyzamide.

C57BL/6J (male, 8 weeks old)^[2]

100 mg/kg intraperitoneally; 100 mg/kg by oral gavage

intraperitoneally for 3 days until TNBS presensitization; then oral gavage for 6 consecutive days until TNBS rectal challenge

Worsened intestinal pathology as indicated by weight loss, colon shortening, and histological evaluation.
Increased IL-17+ CD4+ T cells, IL-17+ RORγt+ CD4+ T cells, IFNγ+ CD4+ T cells, and IFNγ+ CD8+ T cells in the colon, but did not modify IL-17 production by CD8+ T cells and γδ T cells.
Increased colonic CD45+CD3+ lymphocyte counts normalized by colon tissue length compared with vehicle control TNBS mice.
Also increased Ifng and Il17a expression in lamina propria mononuclear cells (LPMC) from TNBS mice compared with vehicle control TNBS mice, whereas naive mice showed lower expression.
RNA-seq detected upregulation of pro-inflammatory pathways, including leukocyte extravasation, integrin signalling, and NF-κB activation.
Upstream regulator analysis identified decreased AHR suppression of NF-κB-driven C/EBPβ.
qPCR validation detected increased Rela and Cebpb expression and increased Tnf, Il1b, Il23, and Il6 in lamina propria mononuclear cells; no changes were detected in Il10 or Tgfb expression.
scRNA-seq showed expansion of T cell and dendritic cell clusters, upregulation of NF-κB-driven C/EBPβ pro-inflammatory gene expression, and decreased AHR signalling.
Increased Vcam1 expression in the colon of TNBS mice and increased T cell expression of Itga4, but not Itga1.
Reduced microbiome diversity in the ileum and caecum and expanded Sutterellaceae; however, germ-free recipients of faecal microbiota from Propyzamide- or vehicle-exposed mice developed comparable intestinal inflammation after TNBS administration.

C57BL/6J (male, 8 weeks old)^[2]

100 mg/kg by gavage; anti-CD3 monoclonal antibody 200 μg intraperitoneally

by gavage for 7 days; then anti-CD3 monoclonal antibody intraperitoneally

Worsened intestinal pathology and T cell-driven inflammation in the anti-CD3 enteritis model, including worsened jejunal histopathology.
Transcriptomic analyses detected decreased AHR signalling, increased NF-κB activation, and increased expression of Cebpb and other pro-inflammatory molecules, as well as increased IL-17 and IFNγ production by CD4+ T cells.
Did not affect the number of small intestine immune cells or their transcriptional programs in the absence of anti-CD3.

C57BL/6J (male, 8 weeks old)^[2]

100 mg/kg intraperitoneally for 3 days and/or 100 mg/kg by oral gavage

intraperitoneally for 3 days and/or oral gavage

Did not alter weight, colon length, CD4+ T cells, CD8+ T cells, γδ T cells, or innate lymphoid cell numbers in the absence of TNBS.
Did not affect the number of small intestine immune cells or their transcriptional programs in the absence of anti-CD3.