Basic Information
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Targeting strategy
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Gene targeting strategy for B-hCTLA4/hCCR8 mice. The exon 2 of mouse Ctla4 gene that encode the extracellular domain was replaced by human CTLA4 exon 2 in B-hCTLA4/hCCR8 mice. The exon 2 that encoding the full coding sequence of mouse Ccr8 gene was replaced by exon 2 of human CCR8 gene in B-hCTLA4/hCCR8 mice.
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Protein expression analysis (spleen)
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CTLA4 (CD152) protein expression analysis in B-hCTLA4/hCCR8 mice by FACS. Spleen was collected from wild-type mice and homozygous B-hCTLA4/hCCR8 mice stimulated with anti-mouse CD3ε antibody in vivo, and analyzed by flow cytometry with species-specific anti-CTLA4 antibody. Mouse CTLA4 was detectable in wild-type mice. Human CTLA4 was exclusively detectable in homozygous B-hCTLA4/hCCR8 but not in wild-type mice.
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Protein expression analysis (spleen)
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CTLA4 (CD152) protein expression analysis in B-hCTLA4/hCCR8 mice by FACS. Spleen was collected from wild-type mice and homozygous B-hCTLA4/hCCR8 mice stimulated with anti-mouse CD3ε antibody, and analyzed by flow cytometry with species-specific anti-CTLA4 antibody. Human CTLA4 was exclusively detectable in homozygous B-hCTLA4/hCCR8 mice. Mouse CTLA4 was exclusively detectable in wild-type C57BL/6 mice.
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Protein expression analysis (lymph node)
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CTLA4 (CD152) protein expression analysis in B-hCTLA4/hCCR8 mice by FACS. Lymph node was collected from wild-type mice and homozygous B-hCTLA4/hCCR8 mice stimulated with anti-mouse CD3ε antibody, and analyzed by flow cytometry with species-specific anti-CTLA4 antibody. Mouse CTLA4 was detectable in wild-type mice. Human CTLA4 was exclusively detectable in homozygous B-hCTLA4/hCCR8 but not in wild-type mice.
CTLA4 (CD152) protein expression analysis in B-hCTLA4/hCCR8 mice by FACS. Lymph node was collected from wild-type mice and homozygous B-hCTLA4/hCCR8 mice stimulated with anti-mouse CD3ε antibody, and analyzed by flow cytometry with species-specific anti-CTLA4 antibody. Human CTLA4 was exclusively detectable in homozygous B-hCTLA4/hCCR8 mice. Mouse CTLA4 was exclusively detectable in wild-type C57BL/6 mice.
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Protein expression analysis (tumor-infiltrating cells)
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CTLA4 (CD152) protein expression analysis in B-hCTLA4/hCCR8 mice by FACS. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hCTLA4/hCCR8 mice (female, 7 week-old, n=3). Tumor were harvested at tumor volume approximately 400-700 mm3 and analyzed by flow cytometry. Human CTLA4 was detectable in T cells in tumors of homozygous B-hCTLA4/hCCR8 mice.
CCR8 (CD198) protein expression analysis in B-hCTLA4/hCCR8 mice by FACS. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hCTLA4/hCCR8 mice (female, 7 week-old, n=3). Tumor were harvested at tumor volume approximately 400-700 mm3 and analyzed by flow cytometry. Human CCR8 was both detectable in CD4+ T cells and Treg cells in tumors of homozygous B-hCTLA4/hCCR8 mice.
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Summary
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Protein expression analysis:
1.Human CTLA4 was exclusively detectable in T cells of homozygous B-hCTLA4/hCCR8 mice but not in wild-type mice stimulated with anti-mouse CD3ε antibody.
2.Human CTLA4 and CCR8 were detectable in tumors infiltrating T cells of homozygous B-hCTLA4/hCCR8 mice but not in wild-type mice.
