Basic Information
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Targeting strategy
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Gene targeting strategy for B-hIL4/hIL4RA/hIL13/hIL13RA1 mice.
The exons 1-4 of mouse Il4 gene that encode the full length coding sequence were replaced by human IL4 exons 1-4 in B-hIL4/hIL4RA/hIL13/hIL13RA1 mice. The exons 4-7 of mouse Il4ra gene that encode the extracellular region coding sequences were replaced by human IL4RA exons 4-7 in B-hIL4/hIL4RA/hIL13/hIL13RA1 mice. The exons 1-4 of mouse Il13 gene that encode the full length coding sequence were replaced by human IL13 exons 1-4 in B-hIL4/hIL4RA/hIL13/hIL13RA1 mice. The exons 2-9 of mouse Il13ra1 gene that encode the extracellular protein were replaced by human IL13RA1 exons 2-9 in B-hIL4/hIL4RA/hIL13/hIL13RA1 mice. B-hIL4/hIL4RA/hIL13/hIL13RA1 mice were derived from crossing B-hIL4/hIL4RA/hIL13 mice and B-hIL13RA1 mice together.
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IL4 Protein expression analysis in serum
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Strain specific IL4 expression analysis in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice by ELISA. Serum were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1mice (H/H; H/H; H/H; H/H) stimulated with anti-mCD3Ɛ antibody in vivo, and analyzed by ELISA with species-specific IL4 ELISA kit. Mouse IL4 was detectable in wild-type C57BL/6 mice. Human IL4 was exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice, but not in wild-type mice.
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IL4RA Protein expression analysis in spleen
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Strain specific IL4RA expression analysis in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice (H/H; H/H; H/H; H/H), and analyzed by flow cytometry with species-specific anti-IL4RA antibody. Mouse IL4RA was detectable in wild-type mice, Human IL4RA was exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice, but not in wild-type mice.
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IL13 Protein expression analysis
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Strain specific IL13 expression analysis in homozygous B-hIL4//hIL4RA/hIL13/hIL13RA1 mice by ELISA. CD4+ T cells were collected from wild-type mice (+/+) and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice (H/H; H/H; H/H; H/H)，and then induction of CD4 + T cells differentiation into Th2 cells, supernatants from Th2 cells culture was analyzed by ELISA with species-specific IL13 ELISA kit. Mouse IL13 was detectable in wild-type mice. Human IL13 was exclusively detectable in homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice, but not in wild-type mice.
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Functional analysis
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MHCⅡ expression analysis in wild-type and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice by flow cytometry. Splenocytes from wild-type C57BL/6 mice and homozygous B-hIL4/hIL4RA/hIL13/hIL13RA1 mice were stimulated with mouse IL13 or human IL13 in vitro, resulting in increased MHCⅡ expression, respectively. Values are expressed as mean ± SD.
