B-hIL23A/hIL12B mice

Basic Information

Strain name
C57BL/6-Il23atm1(IL23A)Il12btm1(IL12B)/Bcgen
Common name
B-hIL23A/hIL12B mice
Background
C57BL/6
Catalog number
120553
Related genes
IL23A (interleukin 23 subunit alpha);  IL12B (interleukin 12B)

Description

IL23A encodes a subunit of the heterodimeric cytokine interleukin 23 (IL23). IL23 is composed of this protein and the p40 subunit of interleukin 12 (IL12B). The receptor of IL23 is formed by the beta 1 subunit of IL12 (IL12RB1) and an IL23 specific subunit, IL23R. Both IL23 and IL12 can activate the transcription activator STAT4, and stimulate the production of interferon-gamma (IFNG). In contrast to IL12, which acts mainly on naive CD4(+) T cells, IL23 preferentially acts on memory CD4(+) T cells.

IL12B encodes a subunit of interleukin 12, a cytokine that acts on T and natural killer cells, and has a broad array of biological activities. Interleukin 12 is a disulfide-linked heterodimer composed of the 40 kD cytokine receptor like subunit encoded by this gene, and a 35 kD subunit encoded by IL12A. This cytokine is expressed by activated macrophages that serve as an essential inducer of Th1 cells development. This cytokine has been found to be important for sustaining a sufficient number of memory/effector Th1 cells to mediate long-term protection to an intracellular pathogen. Overexpression of this gene was observed in the central nervous system of patients with multiple sclerosis (MS), suggesting a role of this cytokine in the pathogenesis of the disease. The promoter polymorphism of this gene has been reported to be associated with the severity of atopic and non-atopic asthma in children.

Details

Protein expression analysis

Strain specific IL12B expression analysis in heterozygous B-hIL23A/hIL12B mice by ELISA.

Serum were collected from WT and heterozygous B-hIL23A/hIL12b (H/+) mice stimulated with LPS in vivo, and analyzed by ELISA with species-specific IL12B ELISA kit. Mouse Il12b was detectable in WT (+/+) mice and heterozygous B-hIL23A/hIL12B (H/+) mice. Human IL12B was exclusively detectable in heterozygous B-hIL23A/hIL12b (H/+) mice but not WT (+/+) mice.

mRNA expression analysis

Strain specific analysis of IL23A and IL12B gene expression in B-hIL23A/hIL12B mice by RT-PCR. Mouse Il23a and Il12b mRNA was detectable in thymocyte of wild-type (+/+) mice. Human IL23A and IL12B mRNA was detectable only in homozygous B-hIL23A/hIL12B mice (H/H) but not in wild-type C57BL/6 mice.

Phenotypic analysis

Analysis of spleen leukocytes cell subpopulations in B-hIL23A/hIL12Bmice

Analysis of spleen leukocyte subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hIL23A/hIL12B 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 cells, B cells, NK cells, DCs, Granulocytes, Monocyte and macrophages in homozygous B-hIL23A/hIL12B mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL23A and hIL12B 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-hIL23A/hIL12B mice

Analysis of spleen T cell subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hIL23A/Hil12b 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+ T cells were gated for CD3+ T cell population and used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD4+ T cells, CD8+ T cells and Treg cells in homozygous B-hIL23A/hIL12B mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL23A and hIL12B 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 leukocyte subpopulations in B-hIL23A/hIL12B mice

Analysis of subpopulation of leukocytes in lymph node by FACS

Lymph node were isolated from female C57BL/6 and B-hIL23A/hIL12Bmice (n=3, 7-week-old). Flow cytometry analysis of the lymph node was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ T cells were used for further analysis as indicated here. B. Results of FACS analysis. Percent of T cells, B cells, and NK cells in homozygous B-hIL23A/hIL12B mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL23A and hIL12B in place of its mouse counterpart does not change the overall development, differentiation or distribution of these cell types in lymph node. Values are expressed as mean ± SEM.

Analysis of lymph node T cell subpopulations in B-hIL23A/hIL12B mice

Analysis of subpopulation of T cells in lymph node by FACS

Lymph node were isolated from female C57BL/6 and B-hIL23A/hIL12B mice (n=3, 7-week-old). Flow cytometry analysis of the lymph node was performed to assess leukocyte subpopulations. A. Representative FACS plots. Single live CD45+ T cells were used for further analysis as indicated here. B. Results of FACS analysis. Percent of CD8+ T cells, CD4+ T cells and Treg cells in homozygous B-hIL23A/hIL12B mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hIL23A and hIL12B in place of its mouse counterpart does not change the overall development, differentiation or distribution of these T cell subtypes in lymph node. Values are expressed as mean ± SEM.

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