Basic Information
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Gene Targeting Strategy
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Gene targeting strategy for B-hTSLP/hTSLPR mice. The exons 1-5 of mouse Tslp gene that encode the full-length protein were replaced by human TSLP exons 1-4 in B-hTSLP/hTSLPR mice. The extracellular and transmembrane region of human TSLPR gene and the cytoplasmic region of mouse Tslpr gene were constructed into a chimeric CDS vector and inserted into the mouse exon 2. The targeted mice will express the chimeric TSLPR protein, while mouse TSLPR will no longer express.
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mRNA expression analysis
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Strain specific analysis of TSLP and TSLPR gene expression in wild type C57BL/6 mice and homozygous B-hTSLP/hTSLPR mice by RT-PCR. Mouse mRNA was prepared from ear grinding supernatant of wild type C57BL/6 mice (+/+;+/+) and homozygous B-hTSLP/hTSLPR mice (H/H;H/H) stimulated with calcipotriol (MC903). Mouse Tslp mRNA was only detectable in wild type C57BL/6 mice (+/+;+/+). Human TSLP mRNA was only detectable in homozygous B-hTSLP/hTSLPR mice but not in wild type mice. Mouse mRNA was prepared from splenocytes of wild type C57BL/6 mice and homozygous B-hTSLP/hTSLPR mice. Mouse Tslpr mRNA was only detectable in wild type C57BL/6 mice. Human TSLPR mRNA was only detectable in homozygous B-hTSLP/hTSLPR mice but not in wild type mice.
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Protein expression analysis-TSLP
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Strain specific TSLP expression analysis in homozygous B-hTSLP/hTSLPR mice by ELISA. Calcipotriol (MC903) was dissolved in ethanol and topically applied on ears of either wild type C57BL/6 mice (+/+;+/+) or homozygous B-hTSLP/hTSLPR mice (H/H;H/H) for 7 days. n=3. Ear grinding supernatant from the two mice was analyzed by ELISA. Mouse TSLP was detectable in wild type C57BL/6 mice. Human TSLP was exclusively detectable in homozygous B-hTSLP/hTSLPR mice but not in wild type mice. ND: not detectable.
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TSLPR mRNA Expression Analysis
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Species-specific TSLPR gene expression analysis in wild type and heterozygous B-hTSLP/hTSLPR mice. Murine Tslpr mRNA was detected in splenocytes isolated from wild-type C57BL/6 (+/+) and heterozygous B-hTSLP/hTSLPR (H/+;H/+) mice. Human TSLPR mRNA was exclusively detected in humanized B-hTSLP/hTSLPR mice.
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TSLPR protein expression analysis in T cells and ILC2 of blood
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Human TSLPR expression analysis on T cells and ILC2 of homozygous B-hTSLP/hTSLPR mice by flow cytometry. Blood cells were collected from wild-type C57BL/6 mice (+/+;+/+) and homozygous B-hTSLP/hTSLPR mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-TSLPR antibody. Mouse TSLPR was detectable on T cells and ILC2 of wild-type C57BL/6 mice but not in homozygous B-hTSLP/hTSLPR mice. Human TSLPR was exclusively detectable on the two cells of homozygous B-hTSLP/hTSLPR mice.
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In vivo efficacy of anti-human TSLP antibody in OXA induced AD-like mouse model
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Efficacy of anti-human TSLP antibody in B-hTSLP/hTSLPR mice. Mice in each group were treated with anti-hTSLP antibody tezepelumab (in house). (A) Statistical analysis of ear thickness in each group. Epidermis of ear began to desquamate from day 18. So the ear thicknesses were decreased from day 18 as shown in figure. (B) Body weight changes during the treatment. (C) Total IgE levels in serum. Serum was collected on day 26 and total IgE levels were measured by ELISA. (n = 8).
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Poster
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Functional Analysis of Anti-TSLP Antibodies Using Humanized B-hTSLP/hTSLPR Mice
