Basic Information
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Targeting strategy
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Gene targeting strategy for B-hTRBC1/hTRBC2 mice.
The exon1~2, partial exon 3 and intron 1~2 of mouse Trbc1 and Trbc2 were replaced by human The exon1~2, partial exon 3 and intron 1~2 of human TRBC1 and TRBC2 in B-hTRBC1/hTRBC2 mice.
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mRNA expression analysis in humanized B-hTRBC1/hTRBC2 mice
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Species specific analysis of TRBC1 and TRBC2 gene expression in wild-type C57BL/6 mice and homozygous humanized B-hTRBC1/hTRBC2 mice by RT-PCR. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTRBC1/hTRBC2 mice (H/H). Mouse TRBC1 and TRBC2 mRNA were detectable only in wild-type C57BL/6 mice. Human TRBC1 and TRBC2 mRNA were detectable only in homozygous B-hTRBC1/hTRBC2 mice, but not in wild-type C57BL/6 mice.
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Protein expression analysis in spleen T cells
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Strain specific TRBC1 expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hTRBC1/hTRBC2 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTRBC1/hTRBC2 mice (H/H), and analyzed by flow cytometry. Mouse TRBC1 was detectable in NK cells of wild-type C57BL/6 mice. Human TRBC1 was detectable in NK cells of homozygous B-hTRBC1/hTRBC2 mice. Anti-TRBC1 antibody is crossly reactive with TRBC1 in human and mice.
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Protein expression analysis in spleen B cells
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Strain specific TRBC1 expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hTRBC1/hTRBC2 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hTRBC1/hTRBC2 mice (H/H), and analyzed by flow cytometry. Mouse TRBC1 was detectable in B cells of wild-type C57BL/6 mice. Human TRBC1 was detectable in B cells of homozygous B-hTRBC1/hTRBC2 mice. Anti-TRBC1 antibody is crossly reactive with TRBC1 in human and mice.
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Frequency of leukocyte subpopulations in spleen
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Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice and homozygous B-hTRBC1/hTRBC2 mice (n=3, 7-week-old). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequency of T cell subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, granulocytes, monocytes, macrophages, CD4+ T cells, CD8+ T cells and Tregs in B-hTRBC1/hTRBC2 mice were similar to those in C57BL/6 mice, demonstrating that humanization of TRBC1 and TRBC2 do not change the frequency or distribution of these cell types in spleen. The frequency of leukocyte subpopulations in blood and lymph node of B-hTRBC1/hTRBC2 mice were also comparable to wild-type C57BL/6 mice (Data not shown). Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test. *P < 0.05, **P < 0.01, ***p < 0.001.
