Strain Name |
C57BL/6-Tnfsf15tm2(TNFSF15)Bcgen Itga4tm1(ITGA4)Bcgen Itgb7tm1(ITGB7)Bcgen/Bcgen |
Common Name |
B-hTL1A/hα4β7 mice |
Background |
C57BL/6 |
Catalog Number |
113037 |
Aliases |
TL1, TL1A, TNLG1B, VEGI, VEGI192A; CD49D, IA4; NA; |
||
NCBI Gene ID |
9966, 3676, 3695 |
Description
Targeting strategy
Gene targeting strategy for B-hTL1A/hα4β7 mice.
The exons 1-4 of mouse Tl1a gene that encode the extracellular domain were replaced by human TL1A exons 1-4 in B-hTL1A/hα4β7 mice.
The exons 2-27 of mouse Itga4 gene that encode the extracellular domain were replaced by human ITGA4 exons 2-27 in B-hTL1A/hα4β7 mice.
The exons 2-14 of mouse Itgb7 gene that encode the extracellular domain were replaced by human ITGB7 exons 3-15 in B-hTL1A/hα4β7 mice.
Protein expression analysis in spleen
Strain specific ITGA4 and ITGB7 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hTL1A/hα4β7 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice and homozygous B-hTL1A/hα4β7 mice. Protein expression was analyzed with anti-mouse ITGA4 antibody (Biolegend, 103705), anti-mouse ITGB7 antibody (Biolegend, 120607), anti-human ITGA4 antibody (Biolegend, 304307) anti-human ITGB7 antibody (Invitrogen, MA5-23541) by flow cytometry. Mouse ITGA4 and ITGB7 were only detectable in wild-type C57BL/6JNifdc mice. Human ITGA4 and ITGB7 were exclusively detectable in homozygous B-hTL1A/hα4β7 mice, but not in wild-type C57BL/6JNifdc mice.
Strain specific ITGA4 and ITGB7 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hTL1A/hα4β7 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice and homozygous B-hTL1A/hα4β7 mice. Protein expression was analyzed with anti-mouse ITGA4 antibody (Biolegend, 103705), anti-mouse ITGB7 antibody (Biolegend, 120607), anti-human ITGA4 antibody (Biolegend, 304307) anti-human ITGB7 antibody (Invitrogen, MA5-23541) by flow cytometry. Mouse ITGA4 and ITGB7 were only detectable in wild-type C57BL/6JNifdc mice. Human ITGA4 and ITGB7 were exclusively detectable in homozygous B-hTL1A/hα4β7 mice, but not in wild-type C57BL/6JNifdc mice.
Strain specific ITGA4 and ITGB7 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hTL1A/hα4β7 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice and homozygous B-hTL1A/hα4β7 mice. Protein expression was analyzed with anti-mouse ITGA4 antibody (Biolegend, 103705), anti-mouse ITGB7 antibody (Biolegend, 120607), anti-human ITGA4 antibody (Biolegend, 304307) anti-human ITGB7 antibody (Invitrogen, MA5-23541) by flow cytometry. Mouse ITGA4 and ITGB7 were only detectable in wild-type C57BL/6JNifdc mice. Human ITGA4 and ITGB7 were exclusively detectable in homozygous B-hTL1A/hα4β7 mice, but not in wild-type C57BL/6JNifdc mice.
Strain specific ITGA4 and ITGB7 expression analysis in wild-type C57BL/6JNifdc mice and homozygous humanized B-hTL1A/hα4β7 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6JNifdc mice and homozygous B-hTL1A/hα4β7 mice. Protein expression was analyzed with anti-mouse ITGA4 antibody (Biolegend, 103705), anti-mouse ITGB7 antibody (Biolegend, 120607), anti-human ITGA4 antibody (Biolegend, 304307) anti-human ITGB7 antibody (Invitrogen, MA5-23541) by flow cytometry. Mouse ITGA4 and ITGB7 were only detectable in wild-type C57BL/6JNifdc mice. Human ITGA4 and ITGB7 were exclusively detectable in homozygous B-hTL1A/hα4β7 mice, but not in wild-type C57BL/6JNifdc mice.
Protein expression analysis
Soluble TL1A expression analysis in B-hTL1A/hα4β7 mice by ELISA. Bone marrow derived dendritic cells (BMDCs) were produced by culturing the bone marrow from wild-type C57BL/6JNifdc mice and homozygous B-hTL1A/hα4β7 mice (female, 8 weeks-old, n=3), which were stimulated with 1 μg/mL LPS in vitro for 18h. 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/hα4β7 mice but not wild-type C57BL/6JNifdc mice. Values are expressed as mean ± SEM. ND: not detectable.
Frequency of leukocyte subpopulations in spleen
Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6JNifdc mice and homozygous B-hTL1A/hα4β7 mice (female, 8-week-old, n=3). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequencies of T cell subpopulations. Percentages of T cells, B cells, NK cells, DCs, monocytes, macrophages, neutrophils, CD4+ T cells, CD8+ T cells and Tregs in B-hTL1A/hα4β7 mice were similar to those in C57BL/6JNifdc mice, demonstrating that humanization of TL1A, ITGA4 and ITGB7 do not change the frequency or distribution of these cell types in spleen. Values are expressed as mean ± SEM.
Frequency of leukocyte subpopulations in blood
Frequency of leukocyte subpopulations in blood by flow cytometry. Blood cells were isolated from wild-type C57BL/6JNifdc mice and homozygous B-hTL1A/hα4β7 mice (female, 8-week-old, n=3). A. Flow cytometry analysis of the blood cells was performed to assess the frequency of leukocyte subpopulations. B. Frequencies of T cell subpopulations. Percentages of T cells, B cells, NK cells, DCs, monocytes, macrophages, neutrophils, CD4+ T cells, CD8+ T cells and Tregs in B-hTL1A/hα4β7 mice were similar to those in C57BL/6JNifdc mice, demonstrating that humanization of TL1A, ITGA4 and ITGB7 do not change the frequency or distribution of these cell types in blood. Values are expressed as mean ± SEM.
Frequency of leukocyte subpopulations in lymph nodes
Frequency of leukocyte subpopulations in lymph node by flow cytometry. Leukocytes were isolated from wild-type C57BL/6JNifdc mice and homozygous B-hTL1A/hα4β7 mice (female, 8-week-old, n=3). A. Flow cytometry analysis of the leukocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequencies of T cell subpopulations. Percentages of T cells, B cells, NK cells, CD4+ T cells, CD8+ T cells and Tregs in B-hTL1A/hα4β7 mice were similar to those in C57BL/6JNifdc mice, demonstrating that humanization of TL1A, ITGA4 and ITGB7 do not change the frequency or distribution of these cell types in lymph nodes. Values are expressed as mean ± SEM.
TNBS induced acute colitis in B-hTL1A/hα4β7 mice
TNBS solution was instilled into the colon lumen of B-hTL1A/hα4β7 mice (female, 8-10 weeks-old, n=10). The control group (Sham) received intrarectal injections of 50% ethanol. The treatment groups received anti-human TL1A antibody RVT-3101 (10 mpk, provided by WuXi AppTec), anti-human α4β7 antibody Vedolizumab (10 mpk, provided by WuXi AppTec) alone or in combination. Body weight and DAI score were recorded daily. On day 5, the mice were sacrificed, and colon length and weight were recorded. (A) Body weight change. (B) DAI score. (C) Colon Index. (D) Colon photo. A TNBS-induced acute colitis model was established in B-hTL1A/hα4β7 mice. Administration of anti-human TL1A antibody RVT-3101 and anti-human α4β7 antibody Vedolizumab effectively improved TNBS-induced acute colitis, and their combination provided better efficacy. The results indicate that B-hTL1A/hα4β7 mice are a powerful tool for evaluating in vivo efficacy of the combination of anti-human TL1A antibody and anti-human α4β7 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 in B-hTL1A/hα4β7 mice.
TNBS solution was instilled into the colon lumen of B-hTL1A/hα4β7 mice (female, 8-10 weeks-old, n=10). The control group (Sham) received intrarectal injections of 50% ethanol. The treatment groups received anti-human TL1A antibody RVT-3101 (10 mpk, provided by WuXi AppTec), anti-human α4β7 antibody Vedolizumab (10 mpk, provided by WuXi AppTec) alone or in combination. Body weight and DAI score were recorded daily. On day 5, the mice were sacrificed, and the intestinal mucosa and serum were collected for cytokine analysis. (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. A TNBS-induced acute colitis model was established in B-hTL1A/hα4β7 mice. Administration of anti-human TL1A antibody RVT-3101 and anti-human α4β7 antibody Vedolizumab effectively improved TNBS-induced acute colitis, and their combination provided better efficacy. The results indicate that B-hTL1A/hα4β7 mice are a powerful tool for evaluating in vivo efficacy of the combination of anti-human TL1A antibody and anti-human α4β7 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 in B-hTL1A/hα4β7 mice.