B-hILl7A/hIL17F mice

Basic Information

Strain Name
C57BL/6-Il17atm1(IL17A) Il17ftm1(IL17F)/Bcgen
Stock Number
120554
Common Name
B-hIL17A/hIL17F mice
Source/Investigator
Bcgen (Beijing Biocytogen Co., Ltd)
Related Genes
IL17A (interleukin 17A); IL17F (interleukin 17F)
Species
C57BL/6
Appearance
Black
Genotypes
Homozygous

Description

IL17A encodes the pro-inflammatory cytokine IL17A that is a member of the interleukin-17 family. IL17A is produced by activated T helper (Th17) cells and certain cell types of innate immune system (Cua and Tato, Nat Rev Immunol. 2010, 10(7):479-89). IL17A functions as either a homodimer or a heterodimer with other IL17 family members, e.g. IL17F and signals through the IL17 receptor (IL17R), which is composed of IL17RA ,IL17RC and IL17RD, to induce chemokine and cytokine production. High level of IL17A is associated with several autoimmune diseases, including psoriasis, multiple sclerosis, and rheumatoid arthritis. IL17A also plays a central role in host defense against bacterial and fungal pathogens.  The protein encoded by this IL17F is a cytokine that shares sequence similarity with IL17. This cytokine is expressed by activated T cells, and has been shown to stimulate the production of several other cytokines, including IL6, IL8, and CSF2/GM-CSF. This cytokine is also found to inhibit the angiogenesis of endothelial cells and induce endothelial cells to produce IL2, TGFB1/TGFB, and monocyte chemoattractant protein-1. The heterodimer formed by IL17A and IL17F is a ligand for the heterodimeric complex formed by IL17RA and IL17RC.

Details

Protein expression analysis

Strain specific IL17A expression analysis in heterozygous B-hIL17A/hIL17F mice by ELISA. Serum were collected from wild type (WT) and heterozygous B-hIL17A/hIL17F (H/+) mice stimulated with anti-mCD3 and anti-mCD28 antibody in vivo, and analyzed by ELISA with species-specific IL17A ELISA kit. Mouse IL17A was detectable in WT mice and heterozygous B-hIL17A/hIL17F mice. Human IL17A was exclusively detectable in heterozygous B-hIL17A/hIL17F but not WT mice. ND: Not detected.

Strain specific IL17F expression analysis in heterozygous B-hIL17A/hIL17F mice by ELISA. CD4+ T cells were sorted from splenocytes of wild type (WT) and heterozygous B-hIL17A/hIL17F (H/+) mice, and induced into Th17 cells. Th17 cells were stimulated by PMA and lonomycin. The Th17 cells culture supernatants were collected and analyzed by ELISA with species-specific IL17F ELISA kit. Mouse IL17F was detectable in WT mice and heterozygous B-hIL17A/hIL17F mice. Human IL17F was exclusively detectable in heterozygous B-hIL17A/hIL17F mice but not WT mice. ND: Not detected.

Analysis of spleen leukocytes cell subpopulations in B-hIL17A/hIL17F mice

Analysis of spleen leukocyte subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hIL17A/hIL17F mice (n=3, 7 week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for CD45 population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T, B, NK, Monocyte, DC and macrophage cells in homozygous B-hIL17A/hIL17F mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL17A/hIL17F in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in spleen.

Analysis of spleen T cell subpopulations in B-hIL17A/hIL17F mice

Analysis of spleen T cell subpopulations by FACS. Splenocytes were isolated from female C57BL/6 and B-hIL17A/hIL17F mice (n=3, 7 week-old). Flow cytometry analysis of the splenocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3 T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8, CD4, and Treg cells in homozygous B-hIL17A/hIL17F mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL17A/hIL17F in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in spleen. Values are expressed as mean ± SEM.

Analysis of lymph node leukocytes cell subpopulations in B-hIL17A/hIL17F mice

Analysis of lymph node leukocyte subpopulations by FACS. Leukocytes were isolated from female C57BL/6 and B-hIL17A/hIL17F mice (n=3, 7 week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live cells were gated for CD45 population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of T, B and NK cells in homozygous B-hIL17A/hIL17F mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL17A/hIL17F in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in lymph node.

Analysis of lymph node T cell subpopulations in B-hIL17A/hIL17F mice

Analysis of lymph node T cell subpopulations by FACS. Leukocytes were isolated from female C57BL/6 and B-hIL17A/hIL17F mice (n=3, 7 week-old). Flow cytometry analysis of the leukocytes was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ cells were gated for CD3 T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8, CD4, and Treg cells in homozygous B-hIL17A/hIL17F mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL17A/hIL17F in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell sub types in lymph node. Values are expressed as mean ± SEM.

Blood routine test in B-hIL17A/hIL17F mice

Complete blood count (CBC). Blood from female C57BL/6 and B-hIL17A/hILl7F mice (n=5, 8 week-old) was analyzed for CBC. There was no differences among any measurement between C57BL/6 and B-hIL17A/hILl7F mice, indicating that introduction of hIL17A and hIL17F in place of its mouse counterpart does not change blood cell composition and morphology. Values are expressed as mean ± SEM.

Blood chemistry of B-hIL17A/hIL17F mice

Blood chemistry tests of B-hIL17A/hIL17F mice. Serum from the C57BL/6 and B-hIL17A/hILl7F mice (n=3, 8 week-old) was analyzed for levels of ALT (alanine aminotransferase) and AST (aspartate aminotransferase). There was no differences on either measurement between C57BL/6 and B-hIL17A/hILl7F mice, indicating that introduction of hIL17A and hIL17F in place of its mouse counterpart does not change ALT and AST levels or health of liver. Values are expressed as mean ± SEM.

Experimental schedule for induction of psoriasis-like skin lesions and in vivo efficacy of anti-human IL17A and IL17F antibody

Experimental schedule for induction of psoriasis-like skin lesions in B-hIL17A/hIL17F mice. Mice at 10-12 week-old of age received a daily topical of commercially available IMQ cream on the shaved back for 7 consecutive days to induce psoriasis-like skin lesions. Control mice were treated similarly with vaseline cream. Severity of skin inflammation was daily scored and back skin was collected at the endpoint. IMQ: imiquimod.

In vivo efficacy of anti-human IL17A and IL17F antibody with psoriasis model induced in B-hIL17A/hIL17F mice

IMQ-induced skin inflammation in B-hIL17A/hIL17F mice phenotypically resembles psoriasis. Mice (female, 10 week-old, n=5) were scored daily for up to 6 days for body weight and clinical signs of skin inflammation following treatment with imiquimod (IMQ) cream. Mice in each group were treated with different dose of bimekizumab produced in house. Doses are shown in legend. (A) Phenotypical presentation of mouse back skin after 6 days of treatment. (B) Body weight changes during treatment. (C-D) Erythema and scaling score of the back was scored daily on a scale from 0 to 4. Additionally, the cumulative score (erythema plus scaling) is depicted. Values are expressed as mean ± SEM.

In vivo efficacy of anti-human IL17A and IL17F antibody with psoriasis model induced in B-hIL17A/hIL17F mice

Dose dependent effects of antibody on keratinocyte proliferation and inflammatory cell infiltration in IMQ induced psoriasis-like skin lesions in B-hIL17A/hIL17F mice. Back skin was collected at the endpoint and stained with Hematoxylin and eosin (H&E). (A) H&E staining of the back skin of mice. (B) Epidermal thickness of the mice. (C) Histological changes were scored on a scale from 0 to 11. Results indicated that bimekizumab (in house) significantly reduced psoriasis-like skin lesions in B-hIL17A/hIL17F mice, confirming that B-hIL17A/hIL17F mice is a powerful model for in vivo evaluation of anti-human IL17A and IL17F antibody.

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