Basic Information
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Targeting strategy
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Gene targeting strategy for B-hPD-1/hPD-L1/hCD3E mice.
The exon 2 of mouse Pdcd1 gene that encode the IgV domain was replaced by human PDCD1 exon 2 in B-hPD-1/hPD-L1/hCD3E mice. The exon 3 of mouse Cd274 gene that encode the IgV domain was replaced by human CD274 exon 3 in B-hPD-1/hPD-L1/hCD3E mice. The exons 2-6 of mouse Cd3e gene that encode the extracellular domain were replaced by human CD3E exons 2-7 in B-hPD-1/hPD-L1/hCD3E mice.
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Protein expression analysis in T cells
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Strain specific PD-1 expression analysis in homozygous B-hPD-1/hPD-L1/hCD3E mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) stimulated with anti-CD3ε in vivo (7.5 μg/mice, stimulation for 24 hours, i.p.), and analyzed by flow cytometry with species-specific anti-PD-1 antibody. Mouse PD-1 was detectable in WT mice (+/+). Human PD-1 was exclusively detectable in homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+).
Strain specific CD3E expression analysis in homozygous B-hPD-1/hPD-L1/hCD3E mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H), and analyzed by flow cytometry with species-specific anti-CD3E antibody. Mouse CD3E was detectable in WT mice (+/+). Human CD3E was exclusively detectable in homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+).
Strain specific PD-L1 expression analysis in homozygous B-hPD-1/hPD-L1/hCD3E mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) stimulated with anti-CD3ε in vivo (7.5 μg/mice, stimulation for 24 hours, i.p.), and analyzed by flow cytometry with species-specific anti-PD-L1 antibody. Mouse PD-L1 was detectable in WT mice (+/+). Human PD-L1 was exclusively detectable in homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+).
Strain specific PD-L1 expression analysis in homozygous B-hPD-1/hPD-L1/hCD3E mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) stimulated with anti-CD3ε in vivo (7.5 μg/mice, stimulation for 24 hours, i.p.), and analyzed by flow cytometry with species-specific anti-PD-L1 antibody. Mouse PD-L1 was detectable in WT mice (+/+). Human PD-L1 was exclusively detectable in homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+).
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Protein expression analysis in dendritic cells
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Strain specific PD-L1 expression analysis in homozygous B-hPD-1/hPD-L1/hCD3E mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) stimulated with anti-CD3ε in vivo (7.5 μg/mice, stimulation for 24 hours, i.p.), and analyzed by flow cytometry with species-specific anti-PD-L1 antibody. Mouse PD-L1 was detectable in WT mice (+/+). Human PD-L1 was exclusively detectable in homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+).
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Summary
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- Protein expression analysis:
Human PD-1 was exclusively detectable on T cells of homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+), and mouse PD-1 was detectable only in WT mice (+/+).
Human PD-L1 was exclusively detectable on T cells and dendritic cells of homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+), and mouse PD-L1 was detectable only in WT mice (+/+).
Human CD3E was exclusively detectable on T cells of homozygous B-hPD-1/hPD-L1/hCD3E mice (H/H;H/H;H/H) but not in WT mice (+/+), and mouse CD3E was detectable only in WT mice (+/+).
