Basic Information
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Gene Targeting Strategy
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Gene targeting strategy for B-hTNFA/hTNFR2 mice. The targeted mouse Tnf whole genomic sequence was replaced by human TNF whole genomic sequence in B-hTNFA/hTNFR2 mice. The exons 2~6 of mouse Tnfr2 gene that encode the extracellular domain was replaced by human TNFR2 exons 2~6 in B-hTNFA/hTNFR2 mice.
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mRNA Expression Analysis
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Strain specific analysis of TNFA and TNFR2 gene expression in WT and B-hTNFA/TNFR2 mice by RT-PCR. Mouse Tnfα and Tnfr2 mRNA were detectable only in splenocytes of wild-type (+/+) mice. Human TNFA and TNFR2 mRNA were detectable only in H/H, but not in +/+ mice.
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Protein Expression Analysis
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Strain specific TNFα expression analysis in homozygous B-hTNFA/hTNFR2 mice by ELISA. Serum was collected from WT and homozygous B-hTNFA/hTNFR2 (H/H) mice stimulated with LPS in vivo, and analyzed by ELISA with species-specific TNFα ELISA kit. Mouse TNFα was detectable in WT mice. Human TNFα was exclusively detectable in homozygous B-hTNFA/hTNFR2 but not WT mice.
Strain specific TNFR2 expression analysis in homozygous B-hTNFA/hTNFR2 mice by flow cytometry. Splenocytes were collected from WT and homozygous B-hTNFA/hTNFR2 (H/H) mice, and analyzed by flow cytometry with species-specific anti-TNFR2 antibody. Mouse TNFR2 was detectable in WT mice. Human TNFR2 was exclusively detectable in homozygous B-hTNFA/hTNFR2 but not WT mice.
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In Vivo Efficacy of Anti-Human TNFR2 Antibody
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Antitumor activity of anti-human TNFR2 antibody in B-hTNFA/hTNFR2 mice. (A) Anti-human TNFR2 antibody get from cooperation company inhibited MC38 tumor growth in B-hTNFA/hTNFR2 mice. Murine colon cancer MC38 cells were subcutaneously implanted into homozygous B-hTNFA/hTNFR2 mice (female, 7-week-old). Mice were grouped when tumor volume reached approximately 100 mm3, at which time they were treated with anti-human TNFR2 antibody. (B) Body weight changes during treatment. As shown in panel A, anti-human TNFR2 antibody was efficacious in controlling tumor growth in B-hTNFA/hTNFR2 mice. B-hTNFA/hTNFR2 mice provide a powerful preclinical model for in vivo evaluation of anti-human TNFR2 antibody. Values are expressed as mean ± SEM.
