B-hTSLP/hTSLPR mice

Basic Information

Targeting strategy

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.

Protein expression analysis-TSLP

Strain specific TSLP expression analysis in heterozygous B-hTSLP mice by ELISA. Calcipotriol (MC903) was dissolved in ethanol and topically applied on ears of either wild type C57BL/6 mice (+/+) or heterozygous B-hTSLP mice (H/+) for 3 days. Ear grinding supernatant from the two mice was analyzed  by ELISA. Mouse TSLP was detectable in wild type C57BL/6 mice and heterozygous B-hTSLP mice. Human TSLP was exclusively detectable in heterozygous B-hTSLP mice but not in wild type mice.

mRNA expression analysis-TSLPR

 

Strain specific analysis of TSLPR gene expression in wild type C57BL/6 mice and heterozygous B-hTSLP/hTSLPR mice by RT-PCR. Mouse Tslpr mRNA was detectable in splenocytes of wild type C57BL/6 mice (+/+;+/+) and heterozygous B-hTSLP/hTSLPR mice (H/+;H/+). Human TSLPR mRNA was detectable only in heterozygous B-hTSLP/hTSLPR mice but not in wild type mice.

Protein expression analysis-TSLPR in macrophages

 

Strain specific TSLPR expression analysis in heterozygous B-hTSLP/hTSLPR mice by flow cytometry. Macrophages were collected from wild type C57BL/6 mice (+/+;+/+) and heterozygous B-hTSLP/hTSLPR mice (H/+;H/+), and analyzed by flow cytometry with species-specific anti-TSLPR antibody. Mouse TSLPR was detectable in wild type C57BL/6 mice and heterozygous B-hTSLP/hTSLPR mice. Human TSLPR was detectable in heterozygous B-hTSLP/hTSLPR mice but not in wild type mice.

Protein expression analysis-TSLPR in DCs

Strain specific TSLPR expression analysis in heterozygous B-hTSLP/hTSLPR mice by flow cytometry. Dendritic cells were induced from bone marrow of wild type C57BL/6 mice (+/+;+/+) and heterozygous B-hTSLP/hTSLPR mice (H/+;H/+), and stimulated with LPS. Protein expression was analyzed by flow cytometry with species-specific anti-TSLPR antibodies. Mouse TSLPR was detectable in wild type C57BL/6 mice and heterozygous B-hTSLP/hTSLPR mice. Human TSLPR was exclusively detectable in heterozygous B-hTSLP/hTSLPR mice but not in wild type mice.

Protein expression analysis-TSLPR

Human TSLPR expression analysis in homozygous B-hTSLP/hTSLPR mice by flow cytometry. Splenocytes were collected from wild type C57BL/6 mice and homozygous B-hTSLP/hTSLPR mice, and analyzed by flow cytometry with species-specific anti-TSLPR antibody. Human TSLPR was detectable on total spenocytes, CD11b+ cells (monocytes and neutrophils) and CD11c+ cells (DCs) of homozygous B-hTSLP/hTSLPR mice but not on those cells of wild type C57BL/6 mice.

Protein expression analysis-TSLPR

Human TSLPR expression analysis in homozygous B-hTSLP/hTSLPR mice by flow cytometry. Splenocytes were collected from wild type C57BL/6 mice and homozygous B-hTSLP/hTSLPR mice, and analyzed by flow cytometry with species-specific anti-TSLPR antibody. Human TSLPR was detectable on cDC2, mo-DC, cDC1 and pre-DC of homozygous B-hTSLP/hTSLPR mice but not on those cells of wild type C57BL/6 mice.

Summary

1.mRNA expression analysis:

• Mouse Tslpr mRNA was detectable in splenocytes of wild type C57BL/6 mice and heterozygous B-hTSLP/hTSLPR mice. Human TSLPR mRNA was detectable only in heterozygous B-hTSLP/hTSLPR mice but not in wild type mice.

2.Protein expression analysis:

• Mouse TSLP was detectable in wild type C57BL/6 mice and heterozygous B-hTSLP mice. Human TSLP was exclusively detectable in heterozygous B-hTSLP mice.
• Mouse TSLPR was detectable in wild type C57BL/6 mice and heterozygous B-hTSLP/hTSLPR mice. Human TSLPR was detectable on macrophages, total DCs, CD11b+ cells, cDC1, cDC2, mo-DC and pre-DC of B-hTSLP/hTSLPR mice.