Humanized IL-4 and IL-4Rα animals
Non-human animals comprising a human or humanized IL-4 and/or IL-4Rα nucleic acid sequence are provided. Non-human animals that comprise a replacement of the endogenous IL-4 gene and/or IL-4Rα gene with a human IL-4 gene and/or IL-4Rα gene in whole or in part, and methods for making and using the non-human animals, are described. Non-human animals comprising a human or humanized IL-4 gene under control of non-human IL-4 regulatory elements is also provided, including non-human animals that have a replacement of non-human IL-4-encoding sequence with human IL-4-encoding sequence at an endogenous non-human IL-4 locus. Non-human animals comprising a human or humanized IL-4Rα gene under control of non-human IL-4Rα regulatory elements is also provided, including non-human animals that have a replacement of non-human IL-4Rα-encoding sequence with human or humanized IL-4Rα-encoding sequence at an endogenous non-human C IL-4Rα locus. Non-human animals comprising human or humanized IL-4 gene and/or IL-4Rα sequences, wherein the non-human animals are rodents, e.g., mice or rats, are provided.
1. A rodent, comprising a replacement of a genomic DNA of a rodent IL-4 gene at an endogenous rodent IL-4 locus with a genomic DNA of a human IL-4 gene to form a modified IL-4 gene, wherein the modified IL-4 gene encodes a human IL-4 protein and comprises exon 1 starting from the ATG initiation codon through exon 4 of the human IL-4 gene, wherein expression of the modified IL-4 gene is under control of rodent regulatory elements at the endogenous rodent IL-4 locus, and wherein the rodent is a mouse or a rat.
2. The rodent of claim 1 , wherein the rodent is a mouse that is incapable of expressing a mouse IL-4 protein.
3. The rodent of claim 1 , wherein the rodent is a mouse that expresses a mouse IL-4Rα protein encoded by an endogenous mouse IL-4Rα gene.
4. The rodent of claim 1 , wherein the rodent is a mouse that expresses a human or humanized IL-4Rα protein.
5. The rodent of claim 4 , wherein the humanized IL-4Rα protein comprises the ectodomain of a human IL-4Rα protein.
6. The rodent of claim 5 , wherein the humanized IL-4Rα protein comprises the transmembrane domain and cytoplasmic domain of the endogenous mouse IL-4Rα protein.
7. The rodent of claim 4 , wherein the mouse comprises a replacement at an endogenous mouse IL-4Rα locus of a mouse nucleic acid comprising an exon of a mouse IL-4Rα gene with a nucleic acid sequence encoding at least one exon of a human IL-4Rα gene to form a modified IL-4Rα gene, wherein expression of the modified IL-4Rα gene is under control of mouse regulatory elements at the endogenous mouse IL-4Rα locus.
8. The rodent of claim 1 , wherein the rodent is a mouse, and wherein said replacement is a replacement of a genomic DNA comprising exon 1 starting from the ATG initiation codon through exon 4 of a mouse IL-4 gene with a genomic DNA comprising exon 1 starting from the ATG initiation codon through exon 4 of the human IL-4 gene.
9. A rodent, comprising a humanized IL-4Rα gene and a humanized IL-4 gene,
wherein the humanized IL-4Rα gene results from a replacement of a rodent IL-4Rα ectodomain-coding sequence at an endogenous rodent IL-4Rα locus with a human IL-4Rα ectodomain-coding sequence, and the humanized IL-4Rα gene comprises a rodent IL-4Rα transmembrane domain coding sequence and a rodent IL-4Rα cytoplasmic domain coding sequence, and wherein the humanized IL-4Rα gene is under control of endogenous rodent IL-4Rα regulatory elements at the endogenous rodent IL-4Rα locus;
wherein the humanized IL-4 gene encodes a human IL-4 protein, and wherein the humanized IL-4 gene is under control of endogenous rodent IL-4 regulatory elements at an endogenous rodent IL-4 locus; and
wherein the rodent is a mouse or a rat.
10. The rodent of claim 9 , wherein the rodent is a mouse, and wherein the humanized IL-4 gene results from a replacement of a genomic DNA comprising exon 1 starting from the ATG initiation codon through exon 4 of a mouse IL-4 gene at an endogenous mouse IL-4 locus with a genomic DNA comprising exon 1 starting from the ATG initiation codon through exon 4 of a human IL-4 gene.
11. The rodent of claim 9 , wherein the rodent is a mouse that is incapable of expressing a mouse IL-4 protein and incapable of expressing a mouse IL-4Rα protein.
12. A method of screening for human-specific IL-4 or IL-4Rα antagonist, comprising:
providing a mouse according to claim 10 ,
inducing pulmonary inflammation in said mouse,
administering an agent to said mouse,
determining whether pulmonary inflammation is reduced by the agent, and
identifying the agent as a human specific IL-4 or IL-4Rα antagonist based on its ability to reduce pulmonary inflammation.
13. The method of claim 12 , wherein the extent of pulmonary inflammation is determined by measuring mucus accumulation in the airway, eosinophilic infiltrating cells in bronchoalveolar lavage fluid, and/or total circulating IgE.
14. A method of screening for human-specific IL-4 or IL-4Rα antagonist, comprising:
providing a mouse according to claim 10 ,
inducing skin inflammation in said mouse,
administering an agent to said mouse,
determining whether skin inflammation is reduced by the agent, and
identifying the agent as a human specific IL-4 or IL-4Rα antagonist based on its ability to reduce skin inflammation.