Basic Information
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Targeting strategy
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Gene targeting strategy for B-NDG hB2M/HLA-A11.1 mice plus. The promoter and signal peptide sequences of HLA-A2.1, the coding sequences of human B2M gene and the genomic sequences of HLA-A11.1 gene (from the first coding base on exon1 to the last base of the 3‘ UTR) were inserted into mouse Hipp11 locus of B-NDG mice. Thus, a B-NDG background mouse strain with human B2M/HLA genes was obtained. The exons 2-8 of mouse H2-K1 and H2-D1 genes were knocked out in B-NDG H2-K1/H2-D1 KO mice. B-NDG hB2M/HLA-A11.1 mice plus was developed by cross-mating the B2M/HLA humanized mice and B-NDG H2-K1/H2-D1 KO mice together.
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Protein expression analysis
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Strain specific B2M expression analysis in wild-type B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus by flow cytometry. Splenocytes, blood cells and bone marrow were collected from wild-type B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus, and analyzed by flow cytometry with species-specific anti-B2M antibody. Mouse B2M was detectable in wild-type mice but not in homozygous B-NDG hB2M/HLA-A11.1 mice plus. Human B2M was exclusively detectable in homozygous B-NDG hB2M/HLA-A11.1 mice plus but not in wild-type mice.
Strain specific HLA-A expression analysis in wild-type B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus by flow cytometry. Splenocytes, blood cells and bone marrow were collected from wild-type B-NDG mice and homozygous B-NDG hB2M/HLA-A11.1 mice plus, and analyzed by flow cytometry with species-specific anti-HLA-ABC antibody and anti-H2-K1/H2-D1 antibody. Mouse H2-K1/H2-D1 was detectable in wild-type mice but not in homozygous B-NDG hB2M/HLA-A11.1 mice plus. Human HLA-A was exclusively detectable in homozygous B-NDG hB2M/HLA-A11.1 mice plus but not in wild-type mice.
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Summary
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Protein expression analysis:
Mouse B2M, H2-K1/H2-D1 were detectable in B-NDG mice but not in B-NDG hB2M/HLA-A11.1 mice. Human B2M and HLA-A were exclusively detectable in homozygous B-NDG hB2M/HLA-A11.1 mice plus but not in B-NDG mice.
