Description

• IL22RA1 is primarily expressed on non-hematopoietic cells, such as epithelial cells, keratinocytes, and gastrointestinal epithelial cells. The expression of IL10RB is more widespread, not only in non hematopoietic cells such as epithelial cells and endothelial cells, but also in certain hematopoietic cells such as macrophages, T cells, and B cells.  After IL22 binds to IL22RA1, it forms a heterodimeric receptor complex with IL10RB, activating the downstream JAK-STAT signaling pathway, which participates in processes like cell proliferation, differentiation, survival, and inflammatory responses.
• The genome of the extracellular domain of mouse Il22ra1 and Il10rb gene was replaced by corresponding human IL22RA1 and IL10RB genome in B-hIL22RA1/hIL10RB mice.
• Human IL22 recombinant protein induces IL22 downstream related gene changes in B-hIL22RA1/hIL10RB mice through the receptor composed of hIL2RA1 and hIL0RB. .
• This product is used for the pharmacological and safety evaluation of autoimmune diseases such as inflammatory bowel disease and psoriasis.

Targeting strategy

Gene targeting strategy for B-hIL22RA1/hIL10RB mice.

The exons 2-5 of mouse Il22ra1 gene that encode extracellular domain and were replaced by human counterparts in B-hIL22RA1/hIL10RB mice. The genomic region of mouse Il22ra1 gene that encodes signal peptide, transmembrane domain and cytoplasmic portion was retained. The promoter, 5’UTR and 3’UTR region of the mouse gene were also retained. The chimeric IL22RA1 expression was driven by endogenous mouse Il22ra1 promoter, while mouse Il22ra1 gene transcription and translation will be disrupted.

The exons 2-6 of mouse Il10rb gene that encode extracellular domain and were replaced by human counterparts in B-hIL22RA1/hIL10RB mice. The genomic region of mouse Il10rb gene that encodes signal peptide, transmembrane domain and cytoplasmic portion was retained. The promoter, 5’UTR and 3’UTR region of the mouse gene were also retained. The chimeric IL10RB expression was driven by endogenous mouse Il10rb promoter, while mouse Il10rb gene transcription and translation will be disrupted.

Functional analysis in colon

Relative expression of IL-22 target genes from wild-type C57BL/6JNifdc mice and homozygous B-hIL22RA1/hIL10RB mice was analyzed by RT-qPCR. Colon was collected from wild-type C57BL/6JNifdc and homozygous B-hIL22RA1/hIL10RB mice (female, n=3, 7-week-old) stimulated with PBS or human IL22 (hIL22) recombinant protein (provided by the clent, 200 μg/mice, i.p.) in vivo for 6 hours. Human IL22RA1 and IL10RB RNA were only detected in homozygous B-hIL22RA1/hIL10RB mice (A). Relative expression of IL-22 target genes analyzed by RT-qPCR (B). The mRNA expression of Reg3b, Reg3g, Muc1, Il1b, Il6, Tnfsf10, and Isg15 in homozygous B-hIL22RA1/hIL10RB mice treated with hIL22 recombinant protein showed no significant changes. The mRNA expression of Cxcl1 and Tnfrsf1b was significantly increased in homozygous B-hIL22RA1/hIL10RB mice after the treatment of hIL22 recombinant protein. The expression of target genes in the skin did not change, and the data was not displayed. This indicates that hIL22 recombinant protein induces IL22 downstream related gene changes in B-hIL22RA1/hIL10RB mice through the receptor composed of hIL2RA1 and hIL0RB. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA. ns: non-significant, *P < 0.05, **P < 0.01, ***P < 0.001.

Functional analysis in liver

Relative expression of IL-22 target genes from wild-type C57BL/6JNifdc mice and homozygous B-hIL22RA1/hIL10RB mice was analyzed by RT-qPCR. Liver was collected from wild-type C57BL/6JNifdc and homozygous B-hIL22RA1/hIL10RB mice (female, n=3, 7-week-old) stimulated with PBS or human IL22 (hIL22) recombinant protein (provided by the clent, 200 μg/mice, i.p.) in vivo for 6 hours. Human IL22RA1 and IL10RB RNA were only detected in homozygous B-hIL22RA1/hIL10RB mice (A). Relative expression of IL-22 target genes analyzed by RT-qPCR (B). The mRNA expression of Tnfrsf1b, Muc1, Il1b, Il6, Tnfsf10, and Isg15 in homozygous B-hIL22RA1/hIL10RB mice treated with hIL22 recombinant protein showed no significant changes. The mRNA expression of Cxcl1 was significantly increased in homozygous B-hIL22RA1/hIL10RB mice after the treatment of hIL22 recombinant protein. The expression of target genes in the skin did not change, and the data was not displayed. This indicates that hIL22 recombinant protein induces IL22 downstream related gene changes in B-hIL22RA1/hIL10RB mice through the receptor composed of hIL2RA1 and hIL0RB. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA. ns: non-significant, *P < 0.05, **P < 0.01, ***P < 0.001.