• The IL-10/IL-10R signaling pathway is a crucial anti-inflammatory and immunoregulatory pathway in the immune system. Acting on both innate and adaptive immune cells, it exhibits broad immunomodulatory activity by inhibiting the proliferation, cytokine secretion, and co-stimulatory molecule expression of pro-inflammatory immune cells. It plays a key role in regulating intestinal mucosal homeostasis.
• The genome of the mouse Il10 gene encoding the full-length protein was replaced with human IL10 counterpart in B-hIL10/hIL10RA/hIL10RB mice. The genome of the mouse Il10ra gene encoding the extracellular domain was replaced with human IL10RA counterpart in B-hIL10/hIL10RA/hIL10RB mice. The genome of the mouse Il10rb gene encoding the extracellular domain was replaced with human IL10RB counterpart in B-hIL10/hIL10RA/hIL10RB mice.
• Human IL10, IL10RA, IL10RB protein were exclusively detectable in homozygous B-hIL10/hIL10RA/hIL10RB mice. The hIL10 could reduce the production of downstream cytokines in homozygous B-hIL10/hIL10RA/hIL10RB mice.
• This product is used for the evaluation of the pharmacodynamics and safety of anti-human IL10/IL10R antibodies in autoimmune diseases such as inflammatory bowel disease.

Gene targeting strategy for B-hIL10/hIL10RA/hIL10RB mice.

The exons 1-5 of mouse Il10 gene that encode the full-length protein were replaced by human IL10 exons 1-5 in B-hIL10/hIL10RA/hIL10RB mice.

The exons 1-6 of mouse Il10ra gene that encode the extracellular protein were replaced by human IL10RA exons 1-6 in B-hIL10/hIL10RA/hIL10RB mice.

The exons 2-6 of mouse Il10rb gene that encode the extracellular protein were replaced by human IL10RB exons 2-6 in B-hIL10/hIL10RA/hIL10RB mice.

Strain specific IL10 expression analysis in homozygous B-hIL10/hIL10RA/hIL10RB mice by ELISA. Serum were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hIL10/hIL10RA/hIL10RB mice (H/H;H/H;H/H) stimulated with LPS in vivo for 2 hrs, and analyzed by ELISA with species-specific mouse IL10 ELISA kit (Biolegend, 431414) and human IL10 ELISA kit (Biolegend, 430604). Mouse IL10 was only detectable in wild-type C57BL/6JNifdc mice. Human IL10 was only detectable in homozygous B-hIL10/hIL10RA/hIL10RB mice but not in wild-type mice. Values are expressed as mean ± SEM. ND: not detectable.

Strain specific IL10RA expression analysis in homozygous B-hIL10/hIL10RA/hIL10RB mice by flow cytometry. Peritoneal washes were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hIL10/hIL10RA/hIL10RB mice (H/H;H/H;H/H), protein expression was analyzed by flow cytometry with species-specific anti-mouse IL10RA antibody (Biolegend, 112705) and anti-human IL10RA antibody (Biolegend, 308811). Mouse IL10RA was detectable on macrophages of wild-type C57BL/6JNifdc mice. Human IL10RA was detectable on macrophages of homozygous B-hIL10/hIL10RA/hIL10RB mice.

Strain specific IL10RB expression analysis in homozygous B-hIL10/hIL10RA/hIL10RB mice by flow cytometry. Peritoneal washes were collected from wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hIL10/hIL10RA/hIL10RB mice (H/H;H/H;H/H), protein expression was analyzed by flow cytometry with anti-mouse IL10RB antibody (Miltenyi, 130-114-689) and anti-human IL10RA antibody (Biolegend, 396803). Mouse IL10RB was detectable on macrophages of wild-type C57BL/6JNifdc mice and homozygous B-hIL10/hIL10RA/hIL10RB mice, as the anti-mouse IL10RB antibody was cross-reactive between mouse and human. Human IL10RB was detectable on macrophages of homozygous B-hIL10/hIL10RA/hIL10RB mice.

Mouse TNF-α expression analysis in wild-type C57BL/6JNifdc and homozygous B-hIL10/hIL10RA/hIL10RB mice by ELISA. Bone marrow-derived macrophages were isolated from the wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hIL10/hIL10RA/hIL10RB mice (H/H;H/H), and stimulated with 100 ng/mL LPS  in vitro for 6 h. The protein expression of mouse TNF-α in cell culture supernatant was measured using the mouse TNF-α ELISA kit (Biolegend, 430904). The mIL10 and hIL10 could reduce the production of mouse TNF-α in wild-type C57BL/6JNifdc mice. The hIL10 could reduce the production of mouse TNF-α in homozygous B-hIL10/IL10RA/hIL10RB mice. The results suggested that the mouse IL10 receptor could recognize both mouse IL10 and human IL10, while the human IL10 receptor only recognizes human IL10. Significance was determined by two-way ANOVA test.  *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.

Mouse IL6 expression analysis in wild-type C57BL/6JNifdc and homozygous B-hIL10/hIL10RA/hIL10RB mice by ELISA. Bone marrow-derived macrophages were isolated from the wild-type C57BL/6JNifdc mice (+/+) and homozygous B-hIL10/hIL10RA/hIL10RB mice (H/H;H/H), and stimulated with 100 ng/mL LPS  in vitro for 6 h. The protein expression of mouse IL6 in cell culture supernatant was measured using the mouse IL6 ELISA kit (Biolegend, 431304). The mIL10 and hIL10 could reduce the production of mouse IL6 in wild-type C57BL/6JNifdc mice. The hIL10 could reduce the production of mouse IL6 in homozygous B-hIL10/hIL10RA/hIL10RB mice. The results suggested that the mouse IL10 receptor could recognize both mouse IL10 and human IL10, while the human IL10 receptor only recognizes human IL10. Significance was determined by two-way ANOVA test.  *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001.