Basic Information
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Gene Targeting Strategy
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The chimeric IL1RL2 CDS was inserted after the 5’UTR of the murine Il1rl2 gene in B-hIL36R mice. This insertion disrupts the endogenous murine Il1rl2 gene, resulting in loss of murine Il1rl2 transcripts.
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mRNA Expression Analysis
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Species-specific IL36R gene expression analysis in wild-type and humanized B-hIL36R mice by RT-PCR. Murine Il36r mRNA was detected in lung isolated from wild-type (+/+) mice, while human IL36R mRNA was exclusively detected in homozygous B-hIL36R (H/H) mice.
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Analysis of Splenic Leukocytes in Humanized B-hIL36R Mice
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Analysis of splenic leukocyte subpopulations by FACS. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL36R (H/H) mice (female, 7 week-old, n=3) and analyzed by flow cytometry to assess leukocyte subpopulations. A. Single live cells were gated on CD45+ cells 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 wild-type mice, demonstrating that introduction of hIL36R in place of its mouse counterpart does not change the overall development, differentiation or distribution of these splenic cell types. Values are expressed as mean ± SEM.
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Analysis of Splenic T Cells in Humanized B-hIL36R Mice
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Analysis of splenic T cell subpopulations in humanized B-hIL36R mice. Splenocytes were isolated from wild-type C57BL/6 and B-hIL36R mice (female, n=3, 7-week-old), and analyzed by flow cytometry to assess splenic leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on CD3+ T cells and used for further analysis as indicated here. B. Percent of CD8+ T cells, CD4+ T cells and Tregs in homozygous B-hIL36R mice were comparable to those in wild-type mice, demonstrating that introduction of human IL36R in place of its mouse counterpart does not change the overall development, differentiation or distribution of these splenic T cell subtypes. Values are expressed as mean ± SEM.
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In vivo Efficacy of an Anti-Human IL36R Antibody in a Psoriasis Model
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IMQ-induced skin inflammation in B-hIL36R mice phenotypically resembles psoriasis. Humanized B-hIL36R 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) Phenotypic presentation of mouse back skin at day 0 and day 6. (C) Body weight changes during treatment. (D-E) Erythema, scaling and cumulative (erythema plus scaling) score of the back was scored daily. Values are expressed as mean ± SEM.
Dose-dependent effects of anti-human IL36R 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, and (C) epidermal thickness of the mice were scored. 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 antibodies. Values are expressed as mean ± SEM.
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Poster
