Strain Name
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C57BL/6-Cd40tm1(CD40)Bcgen Cd40lgtm1(CD40LG)Bcgen/Bcgen
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Common Name
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B-hCD40/hCD40L mice
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Background
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C57BL/6
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Catalog number
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121335
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Related Genes
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CD40 (Bp50, CDW40, TNFRSF5, p50)
CD40LG (CD154, CD40L, HIGM1, IGM, IMD3, T-BAM, TNFSF5, TRAP, gp39, hCD40L)
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NCBI Gene ID
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958,21947
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mRNA expression analysis
Strain specific analysis of CD40/CD40L gene expression in wild-type C57BL/6 mice and B-hCD40/CD40L mice by RT-PCR. Mouse CD40/CD40L mRNA was detectable in splenocytes of wild-type C57BL/6 mice (+/+). Human CD40/CD40L mRNA was detectable only in homozygous B-hCD40/CD40L (H/H), but not in wild-type mice.
Protein expression analysis
Strain specific CD40 expression analysis in homozygous B-hCD40/hCD40L mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD40/hCD40L mice (H/H) stimulated with anti-CD3ε in vivo (7.5 μg/mice, stimulation for 24 hours, i.p.), and analyzed by flow cytometry with species-specific anti-CD40 antibody. Mouse CD40 was detectable in wild-type mice. Human CD40 was exclusively detectable in homozygous B-hCD40/hCD40L but not in wild-type mice.
Strain specific CD40L expression analysis in homozygous B-hCD40/hCD40L mice by flow cytometry. Thymocytes were collected from wild-type C57BL/6 mice (+/+) and homozygous B-hCD40/hCD40L mice (H/H) and stimulated with PMA & Ionomycin, then analyzed by flow cytometry with species-specific anti-CD40L antibody. Mouse CD40L was detectable in wild-type mice. Human CD40L was exclusively detectable in homozygous B-hCD40/hCD40L but not in wild-type mice.
In vivo efficacy of anti-CD40 and anti-CD40L antibodies
B-hCD40/hCD40L mice was used to establish The T-Dependent Antibody Response assay (TDAR assay) and evaluate the efficacy of anti-CD40 and anti-CD40L antibody. B-hCD40/hCD40L mice (n=5) were intraperitoneally immunized with 100 μg KLH on Day 1 and treated with anti-CD40L (provided by the client) or bleselumab (anti-CD40) (in house). Blood was collected on Day 7 and Day 14 and analyzed by ELISA with KLH specific IgG antibody and KLH specific IgM antibody. (A) Body weight of B-hCD40/CD40L mice increased steadily; (B, C) Concentration of mouse KLH specific IgG and IgM were significantly increased after immunization. But the concentration of KLH specific IgG and IgM in the groups treated with anti-CD40L antibody or bleselumab were significantly decreased when compared to that in the control group, demonstrating that the B-hCD40/hCD40L mice provide a powerful preclinical model for in vivo evaluation of anti-CD40 and anti-CD40L antibodies.