C57BL/6-Pdcd1tm1(PDCD1)Bcgen Btlatm1(BTLA)Bcgen Tnfrsf14tm1(TNFRSF14)Bcgen/Bcgen • 113166
| Product name | B-hPD-1/hBTLA/hHVEM mice |
|---|---|
| Catalog number | 113166 |
| Strain name | C57BL/6-Pdcd1tm1(PDCD1)Bcgen Btlatm1(BTLA)Bcgen Tnfrsf14tm1(TNFRSF14)Bcgen/Bcgen |
| Strain background | C57BL/6 |
| NCBI gene ID | 5133,151888,8764 (Human) |
| Aliases | CD279, PD-1, PD1, SLEB2, hPD-1, hPD-l, hSLE1; BTLA1, CD272; ATAR, CD270, HVEA, HVEM, LIGHTR, TR2 |
Gene targeting strategy for B-hPD-1/hBTLA/hHVEM mice.
The exon 2 of mouse PD-1 gene that encodes the IgV domain was replaced by human PD-1 exon 2 in B-hPD -1/hBTLA/hHVEM mice.
The exon 2 of mouse Btla gene that encodes the extracellular domain was replaced by human BTLA exon 2 in B-hPD -1/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-hPD -1/hBTLA/hHVEM mice.
Strain specific BTLA expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hPD-1/hBTLA/hHVEM mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1/hBTLA/hHVEM mice (H/H;H/H;H/H). Protein expression was analyzed with anti-mouse BTLA antibody (Biolegend, 134809) and anti-human BTLA antibody (Biolegend, 344509) by flow cytometry. Mouse BTLA was only detectable in wild-type C57BL/6 mice. Human BTLA was exclusively detectable in homozygous B-hPD-1/hBTLA/hHVEM mice, but not in wild-type C57BL/6 mice.
Strain specific HVEM expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hPD-1/hBTLA/hHVEM mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1/hBTLA/hHVEM mice (H/H;H/H;H/H). Protein expression was analyzed with anti-mouse HVEM antibody (Biolegend, 136305) and anti-human HVEM antibody (Biolegend, 318806) by flow cytometry. Mouse HVEM was only detectable in wild-type C57BL/6 mice. Human HVEM was exclusively detectable in homozygous B-hPD-1/hBTLA/hHVEM mice, but not in wild-type C57BL/6 mice.
Strain specific PD-1 expression analysis in wild-type C57BL/6 mice and homozygous humanized B-hPD-1/hBTLA/hHVEM mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hPD-1/hBTLA/hHVEM mice (H/H;H/H;H/H) stimulated without or with anti-mouse CD3ε antibody (7.5 μg, i.p.) in vivo for 24 hrs. Protein expression was analyzed with anti-mouse PD-1 antibody (Biolegend, 109104) and anti-human PD-1 antibody (Biolegend, 329904) by flow cytometry. Mouse PD-1 was only detectable in wild-type C57BL/6 mice. Human PD-1 was exclusively detectable in homozygous B-hPD-1/hBTLA/hHVEM mice, but not in wild-type C57BL/6 mice.
Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6 mice and homozygous B-hPD-1/hBTLA/hHVEM mice (female, 7-week-old, n=3). A. Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. B. Frequencies of T cell subpopulations. Frequencies of T cells, B cells, NK cells, DCs, monocytes, macrophages, neutrophils, CD4+ T cells, CD8+ T cells and Tregs in B-hPD-1/hBTLA/hHVEM mice were similar to those in C57BL/6 mice. The frequency of leukocyte subpopulations in blood and lymph nodes of B-hPD-1/hBTLA/hHVEM mice were also comparable to wild-type C57BL/6 mice (Data not shown).Values are expressed as mean ± SEM.
Subcutaneous tumor growth of B-hHVEM MC38 cells. B-hHVEM MC38 cells (1x106, 2x106) and wild-type MC38 cells (5x105) were subcutaneously implanted into homozygous B-hPD-1/hBTLA/hHVEM mice (female, 6-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B) Body weight. Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. Results indicate that B-hHVEM MC38 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.
