B-hIL17A Mice

Basic Information

Strain Name
C57BL/6-Il17atm1(IL17A)/Bcgen
Common Name
B-hIL17A mice
Background
C57BL/6
Catalog number
110053
Related Genes
IL17A (interleukin 17A)

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 and IL17RC, 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.

Targeting strategy

Details

Protein Expression Analysis

Serum from wild type (WT) C57BL/6 and homozygous B-hIL17A mice were analyzed by ELISA.

Mouse IL17A was detectable in the WT mice, while human IL17A was detectable in the homozygous B-hIL17A mice.

 

Schematic diagram-Spleen

Analysis of leukocytes cell subpopulation in B-hIL17A mice

Analysis of leukocytes subpopulation in spleen.Splenocytes were isolated from C57BL/6 and B-hIL17A mice (n=3) and profiled by flow cytometry. As shown in the panels above, leukocytes subpopulations in homozygous B-hIL17A mice are similar to those in the C57BL/6 mice, indicating that development of T, NK, Monocyte, DC and macrophage cells are not affected by the knock-in of hIL17A.

 

Analysis of T cell subpopulation in B-hIL17A mice

Analysis of T cell subpopulation in spleen.Lymphocytes were isolated from spleen in C57BL/6 and B-hIL17A mice (n=3). T cells subpopulations were profiled by flow cytometry. As shown above, the T cell subpopulations in homozygous B-hIL17A mice are similar to those in the C57BL/6 mice, indicating that T cell development is not affected by the introduction of hIL17A.

 

Analysis of leukocytes cell subpopulation in B-hIL17A mice

Analysis of leukocytes subpopulation in blood

Blood were isolated from C57BL/6 and B-hIL17A mice (n=3) and profiled by flow cytometry. As shown in the panels above, leukocytes subpopulations in homozygous B-hIL17Amice are similar to those in the C57BL/6 mice, indicating that development of T, NK, Monocyte, DC and macrophage cells are not affected by the knock-in of hIL17A.

 

Analysis of T cell subpopulation in B-hIL17A mice

Analysis of T cell subpopulation in blood Lymphocytes were isolated from blood in C57BL/6 and B-hIL17A mice (n=3). T cells subpopulations were profiled by flow cytometry. As shown above, the T cell subpopulations in homozygous B-hIL17A mice are similar to those in the C57BL/6 mice, indicating that T cell development is not affected by the introduction of hIL17A.

 

ResultsBlood routine test

Complete blood counts (CBC). Blood from the C57BL/6 and B-hIL17Amice (n=3) was collected and analyzed. As shown above, there  are no significant differences in terms of CBC between C57BL/6 and B-hIL17A mice.

 

Results-Blood Biochemical test

Blood biochemistry test. Serum ALT/AST levels of C57BL/6 and B-hIL17A mice were tested (n=3). Results indicate that there are no significant differences in ALT/AST levels between C57BL/6 and B-hIL17A mice.

 

MS/EAE model

Schematic of EAE model

Experimental Autoimmune Encephalomyelitis (EAE) is an induced demyelinating disease model that closely resembles the progression and symptoms of the human neurological disease Multiple Sclerosis (MS).

Clinical score of EAE model

EAE induction by MOG35–55/CFA Emulsion PTX in 10-week-old B-hIL17A mice. Data are expressed as mean ± SEM from a typical experiment (n = 5 except n = 4 for B-hIL17A-F PBS). MOG: myelin-oligodendrocyte glycoprotein; PTX: pertussis toxin. F: female; M: male.

H&E staining and IHC staining in mouse EAE model

Local inflammation of the CNS in B-hIL17A mice (female,n=5) during EAE. On day 45 after MOG/CFA and PTx immunization, spinal cords were removed. The tissue sections were stained with H&E(A,B) and IHC (C,D)(Green, MBP; Blue, DAPI). The sections at the lumbar level are shown. The results showed that the infiltration of inflammatory cells in the MOG group was significantly increased, and the myelin protein was greatly reduced.

IL17A+ cells in lymph node of EAE model

IL-17 was primarily produced by CD4 Th17 cells during the development of EAE. To detect IL-17 production,  cells from lymph node of B-hIL17A mice(female,n=5) immunized with MOG/CFA were stimulated for 6 hours by PMA and ionomycin in the presence of brefeldin A. IL-17-producing cells were analyzed by FACS, along with IFNg. The percentage of IL-17+CD3+CD4+ T cells in CD3+CD4+ T cells was increased in response to MOG immunization in B-hIL17A mice (left panel).  So was percentage of IFNg+ T cells.

IL17A+ cells in CNS of EAE model

IL-17 was primarily produced by CD4 Th17 cells during the development of EAE. To detect IL-17 production,  cells from CNS(Brain cell) of B-hIL17A mice(female,n=5) immunized with MOG/CFA were stimulated for 6 hours by PMA and ionomycin in the presence of brefeldin A. IL-17-producing cells were analyzed by FACS, along with IFNg. The percentage of IL-17+CD3+CD4+ T cells in CD3+CD4+ T cells was increased in response to MOG immunization in B-hIL17A mice (left panel).  So was percentage of IFNg+ T cells.

hIL17A Ab efficacy evaluationB-hIL17A mice

hIL17A Ab efficacy evaluationB-hIL17A mice

EAE induction by MOG35–55/CFA Emulsion PTX in B-hIL17A mice. The clinical score and body weight was collected after the treatment of anti-hIL17A antibody. After treatment of anti-hIL17A antibody, the clinical score level was much lower than the control in homozygous B-hIL17A mice.Data are expressed as mean ± SEM from a typical experiment (n = 6). MOG: myelin-oligodendrocyte glycoprotein; PTX: pertussis toxin.

 

Imiquimod-Induced Psoriasis Model

IMQ-induced psoriasis model

IMQ cream was applied topically on left ear and back, QD x 8

Ear thickness and clinical score in psoriasis model in B-hIL17A mice

Animals(n=5) was scored daily for clinical signs(B) of skin inflammation following treatment with either control Vaseline and imiquimod cream (IMQ) and measured Skin lesion(A) on day 8. The results showed that IMQ significantly increase clinical signs of skin inflammation.

 H&E Staining and Skin Thickness in Psoriasis-like Model

A. H&E stained sections of Ear & Back skin of mice treated with vehicle Vaseline and imiquimod cream. The skin thickness of 5 points was randomly measured. The results showed that IMQ significantly increase epidermal thickness in Ear and Back skin.

Clinical score and ear thickness measurement in psoriasis model in C57BL/6 mice

Ear thickness and clinical score in psoriasis model in C57BL/6 mice

IMQ increased thickness of sections of ear and back skin of mice treated with imiquimod cream (by H&E). B. IMQ increased skin thickness measurement. Five random points were selected and measured. The results showed that IMQ significantly increased epidermal thickness in ear and back skin. Scale bar: 100 um.

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