| Strain Name |
C57BL/6-Tnfrsf4tm1(TNFRSF4)Bcgen Tnfsf4tm1(TNFSF4)Bcgen/Bcgen
|
Common Name | B-hOX40/hOX40L mice |
| Background | C57BL/6 | Catalog number | 120543 |
|
Related Genes |
TNFRSF4（Tumor necrosis factor receptor superfamily, member 4, also known as OX40）; TNFSF4 ( tumor necrosis factor(TNF) superfamily member 4, also known as OX40L) |
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|
NCBI Gene ID |
22163,22164 | ||
Targeting strategy
Gene targeting strategy for B-hOX40/hOX40L mice. The exons 1-5 of mouse OX40 gene that encode the extracellular domain were replaced by human OX40 exons 1-5 in B-hOX40/hOX40L mice. The exons 2-3 of mouse Ox40l gene that encode the extracellular region were replaced by human OX40L exons 2-3 in B-hOX40/hOX40L mice .
Protein expression analysis
Strain specific analysis of OX40L gene expression in WT and B-hOX40/hOX40L mice by RT-PCR. (B)Mouse Ox40l mRNA was detectable in DC cell of wild-type (+/+) . Human OX40L mRNA was detectable only in B-hOX40/hOX40L (H/H) but not in +/+ mice.
Strain specific OX40 expression analysis in homozygous B-hOX40/hOX40L mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hOX40/hOX40L (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-OX40 antibody. Mouse OX40 was detectable in WT mice. Human OX40 was exclusively detectable in homozygous B-hOX40/hOX40L (H/H) but not WT mice.
Strain specific OX40L expression analysis in homozygous B-hOX40/hOX40L mice by flow cytometry. (A)Bone marrow cells were collected from WT and homozygous B-hOX40/hOX40L (H/H) mice. DCs were induced from bone marrow cells and stimulated with LPS. Then DCs were analyzed by flow cytometry with anti-OX40L antibodies. Mouse OX40L was detectable in WT mice. Human OX40L was exclusively detectable in homozygous B-hOX40/hOX40L (H/H) but not WT mice.
Analysis of spleen leukocytes cell subpopulations in B-hOX40/hOX40L mice
Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hOX40/hOX40L mice (n=3, 6-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hOX40/hOX40L mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hOX40/hOX40L in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in spleen. Values are expressed as mean ± SEM.
Analysis of blood leukocytes cell subpopulations in B-hOX40/hOX40L mice
Analysis of blood leukocyte subpopulations by FACS. Blood cells were isolated from female C57BL/6 and B-hOX40/hOX40L mice (n=5, 6-week-old). Flow cytometry analysis of the blood cell was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hOX40/hOX40L mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hOX40/hOX40L in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in blood. Values are expressed as mean ± SEM.
Analysis of blood leukocytes cell subpopulations in B-hOX40/hOX40L mice
Analysis of blood T cell subpopulations by FACS. Blood cells were isolated from female C57BL/6 and B-hOX40/hOX40L mice (n=3, 6-week-old). Flow cytometry analysis of the blood was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for TCRβ+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. The percent of CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hOX40/hOX40L mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hOX40/hOX40L in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in blood. Values are expressed as mean ± SEM.
Analysis of lymph node leukocytes cell subpopulations in B-hOX40/hOX40L mice
Analysis of lymph node leukocyte subpopulations by FACS. Lymph nodes were isolated from female C57BL/6 and B-hOX40/hOX40L mice (n=3, 6-week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for the CD45+ population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, NK cells in homozygous B-hOX40/hOX40L mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hOX40/hOX40L in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in lymph node. Values are expressed as mean ± SEM.
Analysis of lymph node T cell subpopulations in B-hOX40/hOX40L mice
In vivo efficacy of anti-human OX40L antibody with AD model
H&E staining of asthma-like model in B-hOX40/hOX40L mice. ear tissues were collected at the study endpoint and analyzed with H&E staining. The results showed that compared to the untreated group (G2), the group of mice treated with Amlitelimab (in-house) showed a significant reduction in histopathology score and score of eosinophil infiltration. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. AD: Atopic dermatitis.
In vivo efficacy of anti-human OX40L antibody with asthma model
H&E staining of asthma-like model in B-hOX40/hOX40L mice. Lung tissues were collected at the study endpoint and analyzed with H&E staining. The results showed that the group of mice treated with Amlitelimab (in-house) in inflammatory infiltration and mucus secretion in lung tissue was lower than that in untreated mice, indicating that B-hOX40/hOX40L mice provide a powerful preclinical model for in vivo evaluation of anti-human OX40L antibodies. Values are expressed as mean ± SEM. Significance was determined by unpaired t-test. *P < 0.05, **P < 0.01, ***P < 0.001.
In vivo efficacy of anti-human OX40L antibody with CIA model
Experimental schedule for Induction of CIA and in vivo efficacy of anti-human OX40L antibody. 50 μL CⅡ emulsion injection subcutaneously at 2 points-the base of the tail and buttocks on day 0 and day 21 respectively. Animals with disease onset were grouped individually into G1-G3 between day 22 to day 30 while ensuring similar average clinical scores on the day of grouping. Mouse body weight (A) and clinical score (B) post grouping were shown for selected timepoints.
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