Basic Information

Strain name
C57BL/6-Cd3etm2(CD3E)Bcgen Tnfrsf9tm1(TNFRSF9)BcgenB2mtm2(B2M/HLA-A2.1/H2-D)Bcgen/Bcgen
Common Name
B-hCD3E/h4-1BB/HLA-A2.1 mice
Background
C57BL/6
Catalog Number
111960
Aliases
CD3E, CD3e molecule, IMD18, T3E, TCRE; TNFRSF9, TNF receptor superfamily member 9, 4-1BB, CD137, CDw137, ILA; Ly-m1, Ly-m11, beta2-, beta2-m, beta2m, IMD43, HLAA

Gene targeting strategy

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.

Protein expression analysis

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).

Species-specific 4-1BB 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 stimulated with anti-CD3ε antibody in vivo, and analyzed by flow cytometry using species-specific anti-4-1BB antibodies. Mouse 4-1BB was detected in wild-type mice, while human 4-1BB 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).

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