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B-NDG hIL6 mice
Strain Name 


NOD.CB17-PrkdcscidIl2rgtm1Il6tm1(IL6)/Bcgen


Common Name 

B-NDG hIL6 mice

Background B-NDG Catalog number  110596
Related Genes 


Il6  (interleukin 6)


Description


IL6 (interleukin 6)this gene encodes a cytokine that functions in inflammation and the maturation of B cells. The encoded protein has been shown to be an endogenous pyrogen capable of inducing fever in people with autoimmune diseases or infections. Humanized mice are a powerful tool for the study of human hematopoiesis and immune function in vivo. However, the existing models cannot support robust adaptive immune responses, especially the generation of class-switched, antigen-specific antibody responses [1]. Interleukin 6 (IL-6) was initially identified as a B-cell differentiation factor both in vivo and in vitro. It is capable of inducing the final at uration of B cells into immunoglobulin-secreting plasma cells [2].In fact, IL-6 has been shown to stimulate the secretion of antibodies to such a degree that serum IgG1 levels can rise 120- to 400-fold [3].Because murine and human IL-6 show 65% sequence homology at the DNA level and 42% homology at the protein level [4],16 and murine IL-6 is not active in human cells,we reasoned that physiological expression of human IL-6 in the mouse may result in improved B-cell differentiation and human antibody production.

Biocytogen developed the B-NDG hIL6 mice, and the targeting strategy was that to replace the sequence encompassing the ORF of mouse Il6 with its human counterpart. This mice combines a B-NDG mouse background and expresses human IL6 cytokine, not only improves thymopoiesis and peripheral T-cell engraftment,but also significantly increases the level of total IgG and antigen-specific IgG, will become a suitable animal model to investigate human immune system.


Generation of gene editing mouse model and expression analysis  

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.


Protein expression analysis (heterozygous mice)


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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

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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.


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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.


Summary

  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 a higher percentage of myeloid cells and monocyte compared with B-NDG mice in B-NDG hIL6 mice.