• TL1A is a TNF superfamily ligand that binds to death receptor 3 (DR3) and delivers potent costimulatory signals, regulating effector cell proliferation, activation, apoptosis, and the production of cytokines and chemokines. Soluble decoy receptor 3 (DcR3) can neutralize sTL1A/DR3 signaling and inhibit apoptosis, dampen inflammation, and limit tissue damage by neutralizing LIGHT and FasL.
• IL-23 is a heterodimeric cytokine composed of p40 and p19 subunits, mainly secreted by macrophages and dendritic cells. By binding to its receptor (IL-23R), IL-23 drives downstream proinflammatory cytokine release and plays a central role in Th17-mediated pathology observed in multiple autoimmune diseases.
• In TL1A/IL23A/IL12B humanized mice, the extracellular domain–encoding region of the mouse Tl1a gene is replaced by its human TL1A counterpart, and the full-length mouse Il23a and Il12b coding regions are replaced with human IL23A and IL12B, respectively. Soluble human TL1A and human IL-23 are exclusively detectable in homozygous TL1A/IL23A/IL12B humanized mice but not in wild-type (WT) mice. Humanization of TL1A, IL23A, and IL12B does not alter immune-cell frequency or distribution in the spleen, blood, or lymph nodes, nor does it affect blood-cell composition, morphology, liver enzymes (ALT, AST), or overall liver health.

Key Advantages

• Triple humanized cytokine design enabling simultaneous evalsuation of TL1A-, IL-23-, and IL-12–targeting therapies.
• Physiological expression under endogenous mouse promoters ensures natural regulation and biological relevance.
• Normal immune cell homeostasis in spleen, blood, and lymph nodes.
• Active human cytokine signaling, producing soluble human TL1A and IL-23 from dendritic cells.
• Robust colitis phenotype induced by TNBS, including inflammation, cytokine elevation, and fibrosis.
• Ideal for IBD preclinical studies, colitis drug evalsuation, mechanistic cytokine pathway research, and combination antibody therapy testing.

Application

• In vivo evalsuation of anti-human TL1A, anti-human IL23p19, and combination biologics
• Mechanistic studies of the TL1A–IL23–IL12 cytokine axis
• TNBS-induced acute colitis and fibrosis
• IBD drug discovery and efficacy assessment
• Cytokine stimulation studies in dendritic cells
• Modeling TL1A/IL23/IL12–driven inflammation

TL1A

• The exons 1-4 of mouse Tl1a gene that encode extracellular domain were replaced by human counterparts in B-hTL1A/hIL23A/hIL12B mice.
• The genomic region of mouse Tl1a gene that encodes transmembrane domain and cytoplasmic portion was retained. The promoter, 5’UTR and 3’UTR region of the mouse gene were also retained. The TL1A expression was driven by endogenous mouse Tl1a promoter, while mouse Tl1a gene transcription and translation will be disrupted.

IL23A

• The exons 1-4 of mouse Il23a gene that encode the whole molecule (ATG to STOP codon) were replaced by human counterparts in B-hTL1A/hIL23A/hIL12B mice.
• The promoter, 5’UTR and 3’UTR region of the mouse gene were retained. The human IL23A expression was driven by endogenous mouse Il23a promoter, while mouse Il23a gene transcription and translation will be disrupted.

IL12B

• The exons 2-8 of mouse Il12b gene that encode the whole molecule (ATG to STOP codon), including 3’UTR were replaced by human counterparts in B-hTL1A/hIL23A/hIL12B mice.
• The promoter and 5’UTR region of the mouse gene were retained. The human IL12B expression was driven by endogenous mouse Il12b promoter, while mouse Il12b gene transcription and translation will be disrupted.

• Human TL1A, IL23A, and IL12B mRNA were specifically and correctly expressed in B-hTL1A/hIL23A/hIL12B mice.

Species specific analysis of TL1A, IL23A and IL12B gene expression in wild-type C57BL/6 mice and homozygous B-hTL1A/hIL23A/hIL12B mice by RT-PCR. Colon and lung were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTL1A/hIL23A/hIL12B mice (H/H;H/H;H/H). Mouse Tl1a, Il23a and Il12b mRNA were detectable only in wild-type C57BL/6 mice. Human TL1A, IL23A, and IL12B mRNA were detectable only in homozygous B-hTL1A/hIL23A/hIL12B mice, but not in wild-type C57BL/6 mice.

• Soluble human TL1A was exclusively detectable in homozygous B-hTL1A/hIL23A/hIL12B mice but not wild-type C57BL/6 mice.

Soluble TL1A expression analysis in B-hTL1A/hIL23A/hIL12B mice by ELISA. Bone marrow derived dendritic cells were produced by culturing the bone marrow from wild-type C57BL/6 mice and homozygous B-hTL1A/hIL23A/hIL12B mice (female, 9-week-old, n=3), which were stimulated with LPS in vitro. After stimulation, the supernatants were collected and the level of soluble TL1A was analyzed by ELISA. Soluble human TL1A was exclusively detectable in homozygous B-hTL1A/hIL23A/hIL12B mice, but not in wild-type C57BL/6 mice. Values are expressed as mean ± SEM. ND: not detectable.

• Human IL23 was exclusively detectable in homozygous B-hTL1A/hIL23A/hIL12B mice but not wild-type C57BL/6 mice.

Strain specific IL23 expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hTL1A/hIL23A/hIL12B mice by ELISA. Bone marrow derived dendritic cells were produced by culturing the bone marrow from wild-type C57BL/6 mice and homozygous B-hTL1A/hIL23A/hIL12B mice (female, 6-week-old, n=3), which were stimulated with LPS in vitro. After stimulation, the supernatants were collected and the levels of mouse and human IL23 were analyzed by ELISA (R&D, M2300; R&D, D2300B). Mouse IL23 was only detectable in wild-type C57BL/6 mice. Human IL23 was exclusively detectable in homozygous B-hTL1A/hIL23A/hIL12B mice. Values are expressed as mean ± SEM. ND: not detectable.

• The percentages of T cells, B cells, NK cells, DCs, monocytes, macrophages, and neutrophils in homozygous B-hTL1A/hIL23A/hIL12B mice were similar to those in C57BL/6 mice.
• Humanization of TL1A, IL23A, and IL12B do not affect normal immune cell development or splenic distribution.

Analysis of leukocyte subpopulations by flow cytometry in immune organs and blood. Splenocytes, peripheral blood, and lymph nodes were isolated from female C57BL/6 and B-hTL1A/hIL23A/hIL12B mice (female, 7-week-old, n = 3). Single live cells were gated on the CD45⁺ population and analyzed by flow cytometry as indicated. Values are expressed as mean ± SEM.

• The proportions of CD4⁺ T cells, CD8⁺ T cells, and Tregs in homozygous B-hTL1A/hIL23A/hIL12B mice were comparable to those in C57BL/6 mice.
• Humanization of TL1A, IL23A, and IL12B do not affect normal T cell development, differentiation, or splenic distribution.

Analysis of T-cell subpopulations by flow cytometry in immune organs and blood. Splenocytes, peripheral blood, and lymph nodes were isolated from female C57BL/6 and B-hTL1A/hIL23A/hIL12B mice (female, 7-week-old, n = 3). Single live cells were gated on the CD3⁺ T-cell population and analyzed by flow cytometry as indicated. Values are expressed as mean ± SEM.

Growth curve of wild-type C57BL/6 and B-hTL1A/hIL23A/hIL12B mice. Eight-week-old mice were grouped by sex (10 males and 10 females). Body weight was measured on the same day of every two week, until 32 weeks. The minimum and maximum body weights shown in the table were calculated from the mean ± SD. The growth curve of the B-hTL1A/hIL23A/hIL12B mice was similar to the growth curve of C57BL/6 mice.

• No significant differences were observed compared with wild-type mice.

Complete blood count (CBC) of B-hTL1A/hIL23A/hIL12B mice. Values are expressed as mean ± SD.

• No significant differences were observed compared with wild-type mice.

Complete blood count (CBC) of B-hTL1A/hIL23A/hIL12B mice. Values are expressed as mean ± SD.

• No significant differences were observed compared with wild-type mice.

Complete blood count (CBC) of B-hTL1A/hIL23A/hIL12B mice. Values are expressed as mean ± SD.

• No significant differences were observed compared with wild-type mice.

Complete blood count (CBC) of B-hTL1A/hIL23A/hIL12B mice. Values are expressed as mean ± SD.

• No significant differences were observed compared with wild-type mice.

Blood biochemical parameters of B-hTL1A/hIL23A/hIL12B mice are shown. Values are expressed as mean ± SD.

• No significant differences were observed compared with wild-type mice.

Blood biochemical parameters of B-hTL1A/hIL23A/hIL12B mice are shown. Values are expressed as mean ± SD.

• No significant differences were observed compared with wild-type mice.

Blood biochemical parameters of B-hTL1A/hIL23A/hIL12B mice are shown. Values are expressed as mean ± SD.

• No significant differences were observed compared with wild-type mice.

Blood biochemical parameters of B-hTL1A/hIL23A/hIL12B mice are shown. Values are expressed as mean ± SD.

• No obvious abnormalities were observed in all of the organs (brain, heart, lung, liver, spleen, stomach, small intestine, large intestine, kidney, uterus and ovary).

Histopathological analysis of organs in female B-hTL1A/hIL23A/hIL12B mice. Major organs from B-hTL1A/hIL23A/hIL12B mice were collected at 32 weeks of age and analyzed by H&E staining (female, n = 3).

• No obvious abnormalities were found in all of the organs (brain, heart, lung, liver, spleen, stomach, small intestine, large intestine, kidney, and testis).

Histopathological analysis of organs in male B-hTL1A/hIL23A/hIL12B mice. Major organs from B-hTL1A/hIL23A/hIL12B mice were collected at 32 weeks of age and analyzed by H&E staining (male, n = 3).

• Tulisokibart  and Risankizumab treatment efficiently improved TNBS-induced acute colitis.

The therapeutic efficacy of anti-human TL1A and anti-human IL23p19 antibodies on the TNBS-induced acute colitis model in B-hTL1A/hIL23A/hIL12B mice. TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice (female, 8-10 weeks-old, n=8). The control group (Sham) received intrarectal injections of 50% ethanol. The treatment groups received anti-human TL1A antibody Tulisokibart (10 mpk, provided by WuXi AppTec), anti-human IL23p19 antibody Risankizumab (10 mpk, provided by WuXi AppTec) alone or in combination. (A) Body weight change. (B) DAI score. (C) Colon Index. (D) Colon photo. An acute colitis disease model induced by TNBS was established in B-hTL1A/hIL23A/hIL12B mice, and administration of anti-human TL1A antibody Tulisokibart and anti-human IL23p19 antibody Risankizumab effectively improved TNBS-induced acute colitis, and their combination provided better efficacy. Values are expressed as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, versus Vehicle, ANOVA.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

• Tulisokibart  and Risankizumab treatment reduced inflammatory infiltration, epithelial damage and fibrotic progression in colon tissue.

H&E staining and Masson staining on the TNBS-induced acute colitis model in B-hTL1A/hIL23A/hIL12B mice. TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice (female, 8-10 weeks-old, n=8). The control group (Sham) received intrarectal injections of 50% ethanol. The treatment groups received anti-human TL1A antibody Tulisokibart (10 mpk, provided by WuXi AppTec), anti-human IL23p19 antibody Risankizumab (10 mpk, provided by WuXi AppTec) alone or in combination. (A) Pathological score. (B) Masson staining score. An acute colitis disease model induced by TNBS was established in B-hTL1A/hIL23A/hIL12B mice, and administration of anti-human TL1A antibody Tulisokibart and anti-human IL23p19 antibody Risankizumab effectively ameliorated colonic pathological damage and fibrotic progression in TNBS-induced acute colitis mice, and their combination provided better efficacy. Values are expressed as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, versus Vehicle, ANOVA.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

• Tulisokibart  and Risankizumab treatment reduced the production of inflammatory cytokines.

Analysis of inflammatory cytokines on the TNBS-induced acute colitis model in B-hTL1A/hIL23A/hIL12B mice. TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice (female, 8-10 weeks-old, n=8). The control group (Sham) received intrarectal injections of 50% ethanol. The treatment groups received anti-human TL1A antibody Tulisokibart (10 mpk, provided by WuXi AppTec), anti-human IL23p19 antibody Risankizumab (10 mpk, provided by WuXi AppTec) alone or in combination. (A) The concentrations of IL-1β, IL-6, IL-17A, IFN-γ, TNF-α in intestinal mucosa. (B) The concentrations of IL-1β, IL-6, IL-17A, IFN-γ, TNF-α in serum. Administration of anti-human TL1A antibody Tulisokibart and anti-human IL23p19 antibody Risankizumab markedly reduced the production of inflammatory cytokines in TNBS-induced acute colitis mice, and their combination provided better efficacy. The results indicate that B-hTL1A/hIL23A/hIL12B mice are a powerful tool for evalsuating in vivo efficacy of the combination of anti-human TL1A antibody and anti-human IL23p19 antibody. Values are expressed as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, versus Vehicle, ANOVA.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

Experimental schedule for TNBS induced chronic colitis and in vivo efficacy of anti-TL1A and anti-IL23 antibodies in B-hTL1A/hIL23A/hIL12B mice. TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice on day0, day7, day14, and day21. The control group (Sham) received intrarectal injections of PBS. The treatment groups received anti-TL1A antibody RVT-3101 (8 mpk, provided by WuXi AppTec), anti-human TL1A antibody TEV-48574 (20 mpk, provided by WuXi AppTec), anti-human IL23p19 antibody Risankizumab (8 mpk, provided by WuXi AppTec) alone or in combination. This animal model can be used to evalsuate the research of anti-fibrotic IBD drugs.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

• Anti-TL1A and anti-Human IL23p19 antibodies treatment efficiently improved TNBS-induced chronic colitis

The therapeutic efficacy of anti-human TL1A and anti-human IL23p19 antibodies on the TNBS-induced chronic colitis model in B-hTL1A/hIL23A/hIL12B mice. TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice on day0, day7, day14, and day21. The control group (Sham) received intrarectal injections of PBS. The treatment groups received anti-TL1A antibody RVT-3101 (8 mpk, provided by WuXi AppTec), anti-human TL1A antibody TEV-48574 (20 mpk, provided by WuXi AppTec), anti-human IL23p19 antibody Risankizumab (8 mpk, provided by WuXi AppTec) alone or in combination. (A) Body weight change. (B) DAI score. A chronic colitis disease model induced by TNBS was established in B-hTL1A/hIL23A/hIL12B mice, and administration of anti-TL1A antibodies RVT-3101 and TEV-48574, and anti-human IL23p19 antibody Risankizumab effectively improved TNBS-induced chronic colitis, and combination therapy with RVT-3101 and Risankizumab provided better efficacy. Values are expressed as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, versus Vehicle, ANOVA.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

• Anti-TL1A and anti-Human IL23p19 antibodies treatment reduced inflammatory infiltration, epithelial damage and fibrotic progression in colon tissue.

H&E staining and Masson staining on the TNBS-induced chronic colitis model in B-hTL1A/hIL23A/hIL12B mice. TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice on day0, day7, day14, and day21. The control group (Sham) received intrarectal injections of PBS. The treatment groups received anti-TL1A antibody RVT-3101 (8 mpk, provided by WuXi AppTec), anti-human TL1A antibody TEV-48574 (20 mpk, provided by WuXi AppTec), anti-human IL23p19 antibody Risankizumab (8 mpk, provided by WuXi AppTec) alone or in combination. (A) Pathological score. (B) Masson staining score. A chronic colitis disease model induced by TNBS was established in B-hTL1A/hIL23A/hIL12B mice, and administration of anti-TL1A antibodies RVT-3101 and TEV-48574, and anti-human IL23p19 antibody Risankizumab effectively ameliorated colonic pathological damage and fibrotic progression. Values are expressed as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, versus Vehicle, ANOVA.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

• Anti-TL1A antibodies treatment reduced COL1A1 expression in smooth muscular tissue

COL1A1 expression analysis on the TNBS-induced chronic colitis model in B-hTL1A/hIL23A/hIL12B mice by Immunohistochemistry (IHC). TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice on day0, day7, day14, and day21. The control group (Sham) received intrarectal injections of PBS. The treatment groups received anti-TL1A antibody RVT-3101 (8 mpk, provided by WuXi AppTec), anti-human TL1A antibody TEV-48574 (20 mpk, provided by WuXi AppTec), anti-human IL23p19 antibody Risankizumab (8 mpk, provided by WuXi AppTec) alone or in combination. A chronic colitis disease model induced by TNBS was established in B-hTL1A/hIL23A/hIL12B mice, and administration of anti-TL1A antibodies RVT-3101 and TEV-48574 effectively reduced COL1A1 expression in smooth muscular tissue. Values are expressed as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, versus Vehicle, ANOVA.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

• Targeting the TL1A/IL-23 pathway synergistically improves the progression of intestinal diseases through a triple mechanism.

Anti-TL1A and anti-human IL23 antibodies reversed fibrosis decoded by integrating RNA-seq and Olink analysis on the TNBS-induced chronic colitis model in B-hTL1A/hIL23A/hIL12B mice. Colon tissues were collected at the study endpoint and analyzed by RNA-seq and Olink analysis. (A) PCA analysis. The treatment groups (mono- and combo-therapies) significantly separated from the vehicle group, confirming that drugs targeting the TL1A/IL-23 pathway systemically restructured the transcriptomic expression of the intestine. (B) GSEA analysis. The treatment groups (mono- and combo-therapies) significantly downregulated the fibrosis pathways (such as the TGF-β signaling pathway) and the inflammatory pathways (such as TNF-α, IL-6 signaling), achieving a "fibrosis-inhibition + anti-inflammatory" dual reversal at the genetic level. (C) Proteomics research. The TL1A/IL-23 targeted therapy significantly inhibits the key factors of the intestinal Th1/Th17 pathway (such as IL-17a, IFN-γ, TNF), precisely blocking the immune storm.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

Experimental schedule for TNBS induced chronic colitis and in vivo efficacy of anti-TL1A antibodies in B-hTL1A/hIL23A/hIL12B mice. TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice on day0, day7, day14, day21, day28, and day35. The mice were grouped on Day 27, the control group (Sham) received intrarectal injections of PBS. The treatment groups received anti-TL1A antibody RVT-3101 (20 mpk, provided by WuXi AppTec) and anti-human TL1A antibody TEV-48574 (20 mpk, provided by WuXi AppTec). This animal model can be used to evalsuate the research of anti-fibrotic IBD drugs.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.

• Anti-TL1A antibodies treatment efficiently improved TNBS-induced chronic colitis.

The therapeutic efficacy of anti-human TL1A and anti-human IL23p19 antibodies on the TNBS-induced chronic colitis model in B-hTL1A/hIL23A/hIL12B mice. TNBS solution was instilled into the colon lumen of B-hTL1A/hIL23A/hIL12B mice on day0, day7, day14, day21, day28, and day35. The mice were grouped on Day 27, the control group (Sham) received intrarectal injections of PBS. The treatment groups received anti-TL1A antibody RVT-3101 (20 mpk, provided by WuXi AppTec) and anti-human TL1A antibody TEV-48574 (20 mpk, provided by WuXi AppTec). (A) Body weight change. (B) DAI score. A chronic colitis disease model induced by TNBS was established in B-hTL1A/hIL23A/hIL12B mice, and administration of anti-TL1A antibodies RVT-3101 and TEV-48574 effectively improved TNBS-induced chronic colitis. Values are expressed as mean ± SEM. *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, versus Vehicle, ANOVA.

Note: This experiment was conducted by WuXi AppTec using B-hTL1A/hIL23A/hIL12B mice.