Basic Information
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Targeting Strategy
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Gene targeting strategy for B-hCCR8/hCCL1 mice. The full coding region sequence of mouse Ccl1 was replaced by the corresponding human CCL1 gene sequences in B-hCCR8/hCCL1 mice. Exon 2 of mouse Ccr8 gene (encoding the full coding sequence) was replaced by exon 2 of human CCR8 in B-hCCR8/hCCL1 mice.
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CCL1 Protein Expression
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Strain specific CCL1 expression analysis in homozygous B-hCCR8/hCCL1 mice by ELISA. Serum were collected from wild-type mice and homozygous B-hCCR8/hCCL1 mice that stimulated with 7.5ug anti-mCD3ε antibody in vivo (n=2 or 3), and analyzed by ELISA with species-specific CCL1 ELISA kit. Mouse CCL1 was detectable in wild-type mice. Human CCL1 was exclusively detectable in homozygous B-hCCR8/hCCL1 mice but not in wild-type mice.
Detection of CCL1 in tumor microenvironment of C57BL/6 and B-hCCR8/hCCL1 mice bearing MC38 cells by ELISA. Murine colon cancer MC38 cells were subcutaneously implanted into C57BL/6 and B-hCCR8/hCCL1 mice (n=2 or 3). Tumor tissues were harvested when tumor volume ~ 600mm3 and analyzed by ELISA. Human CCL1 was detectable in homozygous B-hCCR8/hCCL1 mice, the content is about 150pg per mg total protein.
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CCR8 Protein Expression and TIL Assessment
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CCR8 protein expression analysis in B-hCCR8/hCCL1 mice by FACS. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hCCR8/hCCL1 mice. TILs were analysis in tumor volume at approximately 600 mm3. Human CCR8 were detectable in Th cells and Treg cells of homozygous B-hCCR8/hCCL1 mice. However, according to data, human CCR8 were also detectable in CTL cells of wild type C57BL/6 mice and humanized mice. It could be the antibodies have more unspecific binding with CTLs.
Tumor-infiltrating lymphocytes analysis in different mice. The percentage of Treg cells (% in Th cells) in tumor-bearing B-hCCR8/hCCL1 mice has no significant difference compared with the C57BL/6 mice and B-hCCR8 mice.
