• TL1A binds to death receptor 3 (DR3) to provide stimulatory signals for downstream signaling pathways, thereby regulating the proliferation, activation, apoptosis of effector cells, and the production of cytokines and chemokines. Soluble decoy receptor 3 (DcR3) may neutralize the effects of sTL1A/DR3. In addition, DcR3 can inhibit apoptosis, reduce inflammation, and prevent tissue damage by neutralizing LIGHT and FasL.
• TNF encodes a multifunctional proinflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. This cytokine is mainly secreted by macrophages. It can bind to its receptors TNFRSF1A and TNFRSF1B and exert its effects through these receptors.
• The genome of the mouse Tl1a gene encoding the extracellular domain was replaced with its human TL1A counterpart in in B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice. The genome of the mouse Tnf gene encoding the full-length protein was replaced with human TNF counterpart in B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice. The genome of the mouse Tnfr1 gene encoding the extracellular domain was replaced with its human TNFR1 counterpart in in B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice. The genome of the mouse Tnfr2 gene encoding the extracellular domain was replaced with its human TNFR2 counterpart in in B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice.
• Human TL1A, TNFA, TNFR1, and TNFR2 protein were only detectable in homozygous B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice.
• This product is used for the evaluation of the pharmacodynamics and safety of anti-human TL1A/TNFA antibodies in autoimmune diseases such as inflammatory bowel disease.

Gene targeting strategy for B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice.

The exons 1-4 of mouse Tl1a gene that encode extracellular domain were replaced by human counterparts in B-hTL1A/hTNFA/hTNFR1/hTNFR2 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 chimeric TL1A expression was driven by endogenous mouse Tl1a promoter, while mouse Tl1a gene transcription and translation will be disrupted.

The exons 1-4 of mouse Tnfa gene that encode signal peptide, extracellular domain, transmembrane domain, cytoplasmic region and 3’UTR are replaced by human counterparts in B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice. The promoter and 5’UTR region of the mouse gene are replaced by human counterparts. The human TNFA expression is driven by human TNFA promoter, while mouse Tnfa gene transcription and translation will be disrupted.

The exons 2-6 of mouse Tnfr1 gene that encode extracellular domain are replaced by human counterparts in B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice. The genomic region of mouse Tnfr1 gene that encodes transmembrane domain and cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric TNFR1 expression is driven by endogenous mouse Tnfr1 promoter, while mouse Tnfr1 gene transcription and translation will be disrupted.

The exons 2-6 of mouse Tnfr2 gene that encode extracellular domain are replaced by human counterparts in B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice. The genomic region of mouse Tnfr2 gene that encodes transmembrane domain and cytoplasmic portion is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric TNFR2 expression is driven by endogenous mouse Tnfr2 promoter, while mouse Tnfr2 gene transcription and translation will be disrupted.

Soluble TL1A expression analysis in B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice by ELISA. Bone marrow derived dendritic cells (BMDCs) were produced by culturing the bone marrow from wild-type C57BL/6 (+/+) and homozygous B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice (H/H;H/H;H/H;H/H), which were stimulated with LPS in vitro for 24 hrs. After stimulation, the supernatants were collected and the levels of soluble TL1A were measured using a species-specific human TL1A ELISA kit (R&D, DY1319-05). Soluble human TL1A was exclusively detectable in homozygous B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice but not in wild-type C57BL/6 mice. Values are expressed as mean ± SEM. ND: not detectable.

Strain specific TNFA expression analysis in wild-type C57BL/6 mice and B-hTL1A/hTNFA/hTNFR2/hTNFR1 mice by ELISA. Serum were collected from wild-type C57BL/6 (+/+) and homozygous B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice (H/H;H/H;H/H;H/H) stimulated with LPS in vivo for 24 hrs, and analyzed by ELISA with species-specific mouse TNFA ELISA kit (Biolegend, 430907) and human TNFA ELISA kit (Biolegend, 430207). Mouse TNFA was detectable in wild-type C57BL/6 mice. Human TNFA was exclusively detectable in homozygous B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice. Values are expressed as mean ± SEM. ND: not detectable.

Strain specific TNFR1 expression analysis in wild-type C57BL/6 mice and homozygous B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice (H/H;H/H;H/H;H/H), and analyzed by flow cytometry with species-specific anti-mouse CD120a antibody (Biolegend, 113005) and anti-human CD120a antibody (Biolegend, 369903). Mouse CD120a was only detectable in wild-type C57BL/6 mice. Human CD120a was exclusively detectable in homozygous B-hTL1A/hTNFA/hTNFR1/hTNFR2 mice.