Basic Information
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Gene Targeting Strategy
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Gene targeting strategy for B-hCD3E/h4-1BB/HLA-A2.1 mice. Exons 2-6 of mouse Cd3e gene that encode the extracellular domain were replaced by human CD3E exons 2-7 in B-hCD3E/h4-1BB/HLA-A2.1 mice. The exons 2-7 of mouse Tnfrsf9 gene that encode the extracellular domain were replaced by human TNFRSF9 exons 3-8 in B-hCD3E/h4-1BB/HLA-A2.1 mice. The B2M gene (Exon1 to Exon3) of mouse were replaced by the sequence encompassing the human B2M CDS and HLA-A*0201 gene that included leader sequence, α1 and α2 domains ligated to a fragment of the murine H-2Db gene containing the α3, transmembrane and cytoplasmic domains.
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Protein expression analysis
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Species-specific CD3E protein expression analysis in wild-type and humanized B-hCD3E/h4-1BB/HLA-A2.1 mice. Splenocytes were isolated from wild-type C57BL/6 and B-hCD3E/h4-1BB/HLA-A2.1 mice, and analyzed by flow cytometry using species-specific anti-CD3E antibodies. Mouse CD3E was detected in wild-type mice, while human CD3E was detected in B-hCD3E/h4-1BB/HLA-A2.1 mice (CD3EH/H; 4-1BBH/H; HLA-A2.1H/+) and B-hCD3E/h4-1BB/HLA-A2.1 mice (CD3EH/H; 4-1BBH/H; HLA-A2.1H/H).
Species-specific HLA protein expression analysis in wild-type and humanized B-hCD3E/h4-1BB/HLA-A2.1 mice. Splenocytes were isolated from wild-type C57BL/6 and B-hCD3E/h4-1BB/HLA-A2.1 mice and analyzed by flow cytometry. Mouse H-2Db was detected in wild-type and B-hCD3E/h4-1BB/HLA-A2.1 mice (CD3EH/H; 4-1BBH/H; HLA-A2.1H/+). Human HLA-A2.1 was detected in B-hCD3E/h4-1BB/HLA-A2.1 mice (CD3EH/H; 4-1BBH/H; HLA-A2.1H/+) and B-hCD3E/h4-1BB/HLA-A2.1 mice (CD3EH/H; 4-1BBH/H; HLA-A2.1H/H).
