Basic Information
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Targeting strategy
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Gene targeting strategy for B-hBTLA/hHVEM mice.
The exon 2 of mouse Btla gene that encodes the extracellular domain was replaced by human BTLA exon 2 in B-hBTLA/hHVEM mice. The exons 1~6 of mouse Tnfrsf14 gene that encode the extracellular domain were replaced by human TNFRSF14 exons 1~6 in B-hBTLA/hHVEM mice.
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mRNA expression analysis
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Strain specific analysis of BTLA and HVEM gene expression in wild type (WT) mice and B-hBTLA/hHVEM mice by RT-PCR.
Mouse Btla and Hvem mRNA was detectable only in splenocytes of WT mice (+/+). Human BTLA and HVEM mRNA was detectable only in homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+).
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Protein expression analysis in B cells
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Strain specific BTLA expression analysis in homozygous B-hBTLA/hHVEM mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hBTLA/hHVEM mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-BTLA antibody. Mouse BTLA was detectable in WT mice (+/+). Human BTLA was exclusively detectable in homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+).
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Protein expression analysis in T cells
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Strain specific HVEM expression analysis in homozygous B-hBTLA/hHVEM mice by flow cytometry.
Splenocytes were collected from wild type (WT) mice (+/+) and homozygous B-hBTLA/hHVEM mice (H/H;H/H), and analyzed by flow cytometry with species-specific anti-HVEM antibody. Mouse HVEM was detectable in WT mice (+/+). Human HVEM was exclusively detectable in homozygous B-hBTLA/hHVEM mice (H/H;H/H) but not in WT mice (+/+).
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Tumor growth curve & Body weight changes
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Subcutaneous homograft tumor growth of B-hHVEM MC38 cells.
B-hHVEM MC38 cells (1×106) and wild-type MC38 cells (5×105) were subcutaneously implanted into homozygous B-hBTLA/hHVEM mice (female, 6-week-old, n=7). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean ± SEM). Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel A, B-hHVEM MC38 cells were able to form tumors in vivo and can be used for efficacy studies.
