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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Details
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Generation of gene editing mouse model and expression analysis
Protein expression analysis (heterozygous mice)
Species specific IL6 expression analysis in B-NDG hIL6 mice by ELISA. Serum from B-NDG and heterozygous B-NDG hIL6 (H/+) mice were analyzed ELISA with anti-IL6 antibodies. Mouse IL6 was detectable in B-NDG and heterozygous B-NDG hIL6 mice. Human IL6 were exclusively detectable in heterozygous B-NDG hIL6 but not B-NDG mice.
B-NDG hIL6 mice are well suited for human immune system engraftment
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL6 and B-NDG mice (female, 6 week-old, n=15). All mice were treated with 1.6gry-irradiation. Representative flow cytometric analysis of PBMCs from mice after engraftment with human CD34+ cells. B-NDG hIL6 show a higher percentage of human myeloid cells and monocyte compared with B-NDG.
B-NDG hIL6 mice are well suited for human immune system engraftment
Human CD34+ cells(0.15M) were intravenous implanted into homozygote B-NDG hIL6 and B-NDG mice (female, 6 week-old, n=14). All mice were treated with 1.6gry-irradiation. Representative flow cytometric analysis of PBMCs from mice after engraftment with human CD34+ cells. B-NDG hIL6 show a higher percentage of human leukocytes, myeloid cells and monocyte compared with B-NDG.
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Summary
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1.Species specific IL6 expression analysis in B-NDG hIL6 mice by ELISA. Human IL6 were exclusively detectable in serum of heterozygous B-NDG hIL6 mice
2.Human Immune System Engraftment in B-NDG hILl6 mice. The results showed that higher percentage of myeloid cells and monocyte in B-NDG hIL6 mice compared with B-NDG mice.
