Basic Information
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Targeting Strategy
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The exon 2 of mouse Pd-1 gene that encodes the extracellular domain was replaced by human PD-1 exon 2 in B-hPD-1/hPD-L1/hTNFR2 mice. The exon 3 of mouse Pd-l1 gene that encodes the extracellular domain was replaced by human PD-L1 exon 3 in B-hPD-1/hPD-L1/hTNFR2 mice. The exons 2~6 of mouse Tnfr2 gene that encode the extracellular domain was replaced by human TNFR2 exons 2~6 in B-hPD-1/hPD-L1/hTNFR2 mice.
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Details
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Protein expression analysis in Tregs
PD-1, PD-L1 and TNFR2 expression analysis in homozygous B-hPD-1/hPD-L1/hTNFR2 (H/H) mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hPD-1/hPD-L1/hTNFR2 (H/H) mice stimulated with anti-CD3ε in vivo, and analyzed by flow cytometry with species-specific anti-PD-1, anti-PD-L1 and anti-TNFR2 antibodies. Mouse PD-1, PD-L1 and TNFR2 were detectable in WT mice. Human PD-1, PD-L1 and TNFR2 were exclusively detected in homozygous B-hPD-1/hPD-L1/hTNFR2 but not WT mice.
Analysis of blood, spleen and lymph node leukocyte subpopulations in B-hPD-1/hPD-L1/hTNFR2 mice
Analysis of blood, spleen and lymph node leukocytes cell subpopulations by FACS
Blood, spleen and lymph node leukocytes cell were isolated from female mice in the panel(n=3, 6 week-old). Flow cytometry analysis was performed to assess leukocyte subpopulations. Percent of T, B, NK, Granulocytes, Monocyte, DC and macrophage cells in homozygous B-hPD-1/hPD-L1/hTNFR2 mice were similar to those in C57BL/6 mice, demonstrating that the humanized mouse does not change the overall development, differentiation or distribution of these cell types in blood, spleen and lymph node.
Analysis of blood, spleen and lymph node T cell subpopulations in B-hPD-1/hPD-L1/hTNFR2 mice
Analysis of blood, spleen and lymph node T cell subpopulations by FACS
Blood, spleen and lymph node leukocytes cell were isolated from female mice in the panel(n=3, 6 week-old). Flow cytometry analysis was performed to assess leukocyte subpopulations. Percentages of CD4+T, CD8+T and Foxp3+ Treg cells in homozygous B-hPD-1/hPD-L1/hTNFR2 mice were similar to those in C57BL/6 mice, demonstrating that the humanized mouse does not change the overall development, differentiation or distribution of these cell types in blood, spleen and lymph node.
