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B-hIL36R mice
Strain Name C57BL/6-Il1rl2tm1(IL1RL2)/Bcgen  Common Name  B-hIL36R mice
Background C57BL/6 Catalog number  110084
Aliases 
IL1RL2, IL-1Rrp2, IL-36R, IL1R-rp2, IL1RRP2, interleukin 1 receptor like 2
NCBI Gene ID
107527
Targeting strategy


Gene targeting strategy for B-hIL36R mice. 
The chimeric IL1RL2 CDS was inserted after the 5’UTR of mouse Il1rl2 gene in B-hIL36R mice. The insertion disrupts the endogenous murine Il1rl2 gene, resulting in the absence of mouse transcripts. 



mRNA expression analysis


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Strain specific analysis of IL36R gene expression in wild type (WT) mice and B-hIL36R mice by RT-PCR. 
Mouse Il1rl2 mRNA was detectable only in lung of WT mice (+/+). Human IL1RL2 mRNA was detectable only in homozygous B-hIL36R mice (H/H) but not in WT mice (+/+). 


mRNA expression profile of IL36R


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Quantitative PCR analysis of IL36R mRNA expression in wild-type (WT) mice and B-hIL36R mice. Total RNA was isolated from heart, liver, spleen, lung, kidney, brain, thymus, lymph node, uterus, ovary, testis, thyroid, skin, muscle, esophagus and small intestine of WT mice and homozygous B-hIL36R mice (n=3, 8 week-old), qPCR method was used to detect the RNA expression of IL36R. The results showed that the abundance of IL36R mRNA in spleen, lung, brain and thymus of B-hIL36R mice were significantly lower than that in WT mice, and the expression level of IL36R mRNA in other tissues of B-hIL36R mice were slightly lower than that in wild mice. The mean and standard deviation from triplicate samples are indicated.

IHC analysis of IL36R expression

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Immunohistochemical (IHC) analysis of IL36R expression in homozygous B-hIL36R mice. 

The small intestine and skin were collected from WT and homozygous B-hIL36R mice (H/H) that have established psoriasis models and prepared for optical coherence tomography (OCT). The tissues were analyzed by IHC with anti-IL36R antibody. (A), IL36R expression in the small intestine. (B), IL36R expression in the skin. Mouse IL36R was detectable in WT mice. Human IL36R was detectable in homozygous B-hIL36R mice due to the cross-reactivity of antibodies. The arrow indicates tissue cells with positive IL36R staining (brown). 

Analysis of spleen leukocytes cell subpopulations in B-hIL36R mice
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Analysis of spleen leukocyte subpopulations by FACS.
Splenocytes were isolated from female C57BL/6 and B-hIL36R mice (n=3, 7 week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for 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-hIL36R mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL36R in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in spleen. Values are expressed as mean ± SEM.


Analysis of spleen T cell subpopulations in B-hIL36R mice

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Analysis of spleen T cell subpopulations by FACS.
Splenocytes were isolated from female C57BL/6 and B-hIL36R mice (n=3, 7 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 CD3 T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8+ T cells, CD4+ T cells and Treg cells in homozygous B-hIL36R mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL36R in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in spleen. Values are expressed as mean ± SEM.



Analysis of lymph node leukocytes cell subpopulations in B-hIL36R mice
from clipboardAnalysis of lymph node leukocyte subpopulations by FACS.
Leukocytes were isolated from female C57BL/6 and B-hIL36R mice (n=3, 7 week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for CD45 population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells and NK cells in homozygous B-hIL36R mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL36R 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-hIL36R mice
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Analysis of lymph node T cell subpopulations by FACS.
Leukocytes were isolated from female C57BL/6 and B-hIL36R mice (n=3, 7 week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3 T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8+ T cells, CD4+ T cells and Treg cells in homozygous B-hIL36R mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL36R in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in lymph node. Values are expressed as mean ± SEM.

Analysis of blood leukocytes cell subpopulations in B-hIL36R mice


from clipboardAnalysis of blood leukocyte subpopulations by FACS.

Blood cells were isolated from female C57BL/6 and B-hIL36R mice (n=3, 7 week-old). Flow cytometry analysis of the blood leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for 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-hIL36R mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL36R 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 T cell subpopulations in B-hIL36R mice

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Analysis of blood T cell subpopulations by FACS.
Blood cells were isolated from female C57BL/6 and B-hIL36R mice (n=3, 7 week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3 T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8+ T cells, CD4+ T cells and Treg cells in homozygous B-hIL36R mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL36R in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in blood. Values are expressed as mean ± SEM.


In vivo efficacy of anti-human IL36R antibody in psoriasis model induced in B-hIL36R mice

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IMQ-induced skin inflammation in B-hIL36R mice phenotypically resembles psoriasis. Mice (female, 9 week-old, n=6) were scored daily for up to 7 days for body weight and clinical signs of skin inflammation following treatment with imiquimod (IMQ) cream. Mice in each group were treated with different doses of Spesolimab (in-house). (A) Experimental schedule for induction of psoriasis-like skin lesions in B-hIL36R mice. (B) Phenotypical presentation of mouse back skin at day 0 and day 6. (C) Body weight changes during treatment. (D-E) Erythema and scaling score of the back was scored daily. Additionally, the cumulative score (erythema plus scaling) is depicted. Values are expressed as mean ± SEM.


In vivo efficacy of anti-human IL36R antibody in psoriasis model induced in B-hIL36R mice

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Dose dependent effects of antibodies on keratinocyte proliferation and inflammatory cell infiltration in IMQ induced psoriasis-like skin lesions in B-hIL36R mice. Back skin was collected at the endpoint and stained with Hematoxylin and eosin (H&E). (A) H&E staining of the back skin. (B) Histological changes were scored. (C) Epidermal thickness of the mice. Results indicated that therapeutic effects of Spesolimab (in-house) on psoriasis-like skin lesions in B-hIL36R mice were dose dependent, confirming that B-hIL36R mice provide a powerful model for in vivo evaluation of anti-human IL36R antibody. Values are expressed as mean ± SEM.