Basic Information

Strain Name
C57BL/6-Il10tm1(IL10)/Bcgen
Common Name
B-hIL10 mice
Background
C57BL/6
Catalog number
110060
Related Genes
Il10 (interleukin 10)

Description

The protein encoded by this gene is a cytokine produced primarily by monocytes and to a lesser extent by lymphocytes. This cytokine has pleiotropic effects in immunoregulation and inflammation. It down-regulates the expression of Th1 cytokines, MHC class II Ags, and costimulatory molecules on macrophages. It also enhances B cell survival, proliferation, and antibody production. This cytokine can block NF-kappa B activity, and is involved in the regulation of the JAK-STAT signaling pathway. Knockout studies in mice suggested the function of this cytokine as an essential immunoregulator in the intestinal tract. Mutations in this gene are associated with an increased susceptibility to HIV-1 infection and rheumatoid arthritis.

Details

1.Generation of gene editing mouse model and expression analysis

RT-PCR analysis

RT-PCR analysis of B-hIL10 mice

The hIL10 mRNA, but not mIl10 mRNA was detectable in splenocytes of the homozygous B-hIL10 mice .

Protein expression analysis

Serum from both wild type (WT) C57BL/6 and homozygous B-hIL10 mice were analyzed by ELISA.

When treated with LPS, mouse IL10 was only detectable in WT C57BL/6 mice, while human IL10 was only detectable in homozygous B-hIL10 mice. n=3.

2.Functional analysis

Analysis of leukocytes cell subpopulation in B-hIL10 mice

Analysis of leukocytes subpopulation in spleen

Splenocytes were isolated from C57BL/6 and B-hIL10 mice (n=3). The proportion of leukocytes subpopulation was tested by flow cytometry. As a result, the expression profile of leukocytes subpopulation in homozygous B-hIL10 mice is similar to that in the C57BL/6 mice, indicating that differentiation of T, B, NK, Monocyte, DC and macrophage cells are not affected by the humanization of hIL10.

Analysis of T cell subpopulation in B-hIL10 mice

Analysis of T cell subpopulation in spleen

The lymphocytes were isolated from spleen in C57BL/6 and B-hIL10 mice (n=3). The proportion of T cells subpopulation was tested by flow cytometry. As a result, the expression profile of lymphocyte subpopulation in homozygous B-hIL10 mice is similar to that in the C57BL/6 mice, indicating that T cell differentiation is not affected by the humanization of hIL10.

Analysis of leukocytes cell subpopulation in B-hIL10 mice

Analysis of leukocytes subpopulation in spleen

Blood were isolated from C57BL/6 and B-hIL10 mice (n=3). The proportion of leukocytes subpopulation was tested by flow cytometry. As a result, the expression profile of leukocytes subpopulation in homozygous B-hIL10 mice is similar to that in the C57BL/6 mice, indicating that differentiation of T, B, NK, Monocyte, DC and macrophage cells are not affected by the humanization of hIL10.

Analysis of T cell subpopulation in B-hIL10 mice

Analysis of T cell subpopulation in spleen

The lymphocytes were isolated from Blood in C57BL/6 and B-hIL10 mice (n=3). The proportion of T cells subpopulation was tested by flow cytometry. As a result, the expression profile of lymphocyte subpopulation in homozygous B-hIL10 mice is similar to that in the C57BL/6 mice, indicating that T cell differentiation is not affected by the humanization of hIL10.

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