Basic Information
Description
Biocytogen developed the highly immunodeficient B-NDG hIL6 mouse model, in which the targeting strategy was to replace the sequence encompassing the ORF of mouse Il6 with its human counterpart. This immunodeficient B-NDG model expresses human IL6 cytokine to improve engraftment and survival of human leukocytes, myeloid cells, and monocytes. B-NDG hIL6 mice will become a suitable animal model to investigate the role of the human immune system in xenograft studies.
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Targeting strategy
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Gene targeting strategy for B-NDG hIL6 mice. The Il6 sequence encompassing the ORF of mouse were replaced by human counterpart (a 4.8 kb human IL6 sequence extending from ATG in exon 1 to exon 5 with 16 nucleotides of 3’ downstream sequence), maintain the promoter and other regulatory elements of mouse origin in B-NDG hIL6 mice.
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Protein expression analysis
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Strain specific analysis of IL6 expression in B-NDG mice and B-NDG hIL6 mice by ELISA. Serum was collected from B-NDG mice (+/+) and homozygous B-NDG hIL6 mice (H/H) stimulated with LPS, and analyzed by ELISA with anti-IL6 antibodies. Mouse IL6 was detectable in B-NDG mice. Human IL6 was exclusively detectable in homozygous B-NDG hIL6 mice but not in B-NDG mice.
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Human CD34+ HSCs immune system engraftment
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B-NDG hIL6 mice are well suited for human monocytes reconstitution. Human CD34+ HSCs (0.15 M) were intravenous implanted into homozygous B-NDG hIL6 and B-NDG mice (female, 6 week-old, n=15). All mice were treated with 1.6 Gy-irradiation. Flow cytometry analysis were performed with PBMCs from mice engrafted with human CD34+ HSCs. B-NDG hIL6 mice show higher percentages of human myeloid cells and monocytes compared with B-NDG mice.
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Summary
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- Protein expression analysis:
Mouse IL6 was detectable in B-NDG mice. Human IL6 was exclusively detectable in homozygous B-NDG hIL6 mice
- Human CD34+ HSCs immune system engraftment:
B-NDG hIL6 mice show higher percentages of human myeloid cells and monocytes compared with B-NDG mice.
