Basic Information
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Gene targeting strategy
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Gene targeting strategy for B-hNKP46 mice. The exons 1~7 of mouse NKp46 gene that encode the extracellular domain were replaced by human NKP46 exons 1~7 in B-hNKP46 mice.
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Protein expression analysis
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Species-specific NKP46 expression analysis in wild-type and humanized B-hNKP46 mice. Splenocytes were collected from wild-type C57BL/6 (+/+) and heterozygous B-hNKP46 (H/+) mice and analyzed by flow cytometry using species-specific NKP46 antibodies. Mouse NKP46 was detectable in wild-type and heterozygous B-hNKP46 mice, while human NKP46 protein was exclusively detected in heterozygous B-hNKP46 mice.
Species-specific NKP46 expression analysis in wild-type and humanized B-hNKP46 mice. Blood was collected from wild-type C57BL/6 (+/+) and heterozygous B-hNKP46 (H/+) mice and analyzed by flow cytometry using species-specific NKP46 antibodies. Mouse NKP46 was detectable in wild-type and heterozygous B-hNKP46 mice, while human NKP46 protein was exclusively detected in heterozygous B-hNKP46 mice.
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Analysis of spleen leukocytes
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Analysis of spleen leukocyte subpopulations. Splenocytes were isolated from female C57BL/6 and homozygous B-hNKP46 mice (n=3, 6-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45+ cells and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes and macrophages in homozygous B-hNKP46 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hNKP46 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these spleen cell types. Values are expressed as mean ± SEM.
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Analysis of spleen T cells
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Analysis of spleen T cells. Splenocytes were isolated from female C57BL/6 and homozygous B-hNKP46 mice (n=3, 6-week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. (A) Representative flow cytometry plots. Single live CD45+ cells were gated on CD3+ T cells and used for further analysis as indicated here. (B) Percent of CD8+ T cells, CD4+ T cells, and Tregs in homozygous B-hNKP46 mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hNKP46 in place of its mouse counterpart does not change the overall development, differentiation or distribution of these spleen T cell subtypes. Values are expressed as mean ± SEM.
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In vivo efficacy of an anti-human NKP46-based antibody
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Antitumor activity of anti-human NKP46-based Ab in B-hNKP46 mice. (A) Anti-human NKP46-based Ab inhibited MC38 tumor growth in B-hNKP46 mice. Murine colon cancer MC38 cells (5E5) were subcutaneously implanted into B-hNKP46 mice (female, 7-8 week-old, n=7). Mice were grouped when tumor volume reached approximately 100 mm3, at which time they were treated with anti-human NKP46-based Ab with doses and schedules indicated in panel A. (B) Body weight changes during treatment. As shown in panel A, anti-human NKP46-based Ab was efficacious in controlling tumor growth in B-hNKP46 mice. Values are expressed as mean ± SEM. (All antibodies were provided by the clients).
