B-hCD180 mice

Basic Information

Targeting strategy

Gene targeting strategy for B-hCD180 mice.

The exons 2-3 of mouse Cd180 gene were replaced by human CD180 exons 2-3 in B-hCD180 mice.

Protein expression analysis

Protein expression analysis in T cells

Species-specific CD180 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD180 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD180 antibody. CD180 was not detectable in T cells.

Protein expression analysis in B cells

Species-specific CD180 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD180 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD180 antibody. Mouse CD180 was only detectable in B cells in wild-type C57BL/6 mice, while hCD180 was exclusively detectable in B cells in homozygous mice.

Protein expression analysis in NK cells

Species-specific CD180 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD180 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD180 antibody. CD180 was not detectable in NK cells.

Protein expression analysis in DCs

Species-specific CD180 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD180 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD180 antibody. Mouse CD180 was only detectable in DCs in wild-type C57BL/6 mice, while hCD180 was exclusively detectable in DCs in homozygous mice.

Protein expression analysis in neutrophils

Species-specific CD180 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD180 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD180 antibody. CD180 was not detectable in neutrophils.

Protein expression analysis in monocytes

Species-specific CD180 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD180 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD180 antibody. Mouse CD180 was only detectable in monocytes in wild-type C57BL/6 mice, while hCD180 was exclusively detectable in monocytes in homozygous mice.

Protein expression analysis in macrophages

Species-specific CD180 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD180 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD180 antibody. Mouse CD180 was only detectable in macrophages in wild-type C57BL/6 mice, while hCD180 was exclusively detectable in macrophages in homozygous mice.

Protein expression analysis

Species-specific CD180 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD180 mice (H/H), and analyzed by flow cytometry with species-specific anti-CD180 antibody. CD180 was not detectable in T cells, NK cells, and neutrophils. Mouse CD180 was only detectable in B cells, DCs, monocytes, and macrophages in wild-type C57BL/6 mice, while hCD180 was exclusively detectable in B cells, DCs, monocytes, and macrophages in homozygous mice.

Functional assay: B cell activation

CD86 expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) (n=2, female) and homozygous B-hCD180 mice (H/H) (n=2, 1 female and 1 male), and cultured in 96-well plate with or without anti-human CD180 antibody stimulation (1, 5 μg/mL) for 24 h. Then cells were harvested and analyzed by flow cytometry. After anti-human CD180 antibody stimulation, CD86 expression was up-regulated in B cells from homozygous B-hCD180 mice (A) but not in B cells from wild-type mice (B). (C) Frequency of CD86 expression level in both wild-type and homozygous B-hCD180 mice.

MHCII expression analysis in homozygous B-hCD180 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) (n=2, female) and homozygous B-hCD180 mice (H/H) (n=2, 1 female and 1 male), and cultured in 96-well plate with or without anti-human CD180 antibody stimulation (1, 5 μg/mL) for 24 h. Then cells were harvested and analyzed by flow cytometry. After anti-human CD180 antibody stimulation, MHCII expression was up-regulated in B cells from homozygous B-hCD180 mice (A) but not in B cells from wild-type mice (B). (C) Frequency of CD86 expression level in both wild-type and homozygous B-hCD180 mice.

Summary

Protein expression analysis:

• CD180 was not detectable in T cells, NK cells, and neutrophils.
• Mouse CD180 was only detectable in B cells, DCs, monocytes, and macrophages in wild-type C57BL/6 mice, while hCD180 was exclusively detectable in B cells, DCs, monocytes, and macrophages in homozygous B-hCD180 mice.

Functional analysis:

• Anti-human CD180 antibodies can only activate the B cells activation of B-hCD180 mice, as CD86 and MHC II are up-regulated.