B-hTNFA/hIL17A mice

Basic Information

Strain Name
C57BL/6-Tnftm1(TNF)Il17atm1(IL17A)/Bcgen
Common Name
B-hTNFA/hIL17A mice
Background
C57BL/6
Catalog Number
120548
Related Gene
TNF (tumor necrosis factor); IL17A (interleukin 17A)

Description

TNF gene encodes a multifunctional pro-inflammatory cytokine that belongs to the tumor necrosis factor (TNF) superfamily. The tumor necrosis factor (TNF) superfamily is a protein superfamily of type II transmembrane proteins containing TNF homology domain and forming trimers. Members of this superfamily can be released from the cell membrane by extracellular proteolytic cleavage and function as a cytokine. These proteins are expressed predominantly by immune cells and they regulate diverse cell functions, including immune response and inflammation, but also proliferation, differentiation, apoptosis and embryogenesis. TNFα Regulation of immune cells, induction of fever, cachexia, inflammation and apoptosis, inhibition of tumorigenesis and viral replication and response to sepsis.

IL17A, the protein encoded by this gene is a pro-inflammatory cytokine produced by activated T cells. This cytokine regulates the activities of NF-κB and mitogen-activated protein kinases. This cytokine can stimulate the expression of IL6 and cyclooxygenase-2 (PTGS2/COX-2), as well as enhance the production of nitric oxide (NO). High levels of this cytokine are associated with several chronic inflammatory diseases including rheumatoid arthritis, psoriasis and multiple sclerosis.

Details

Protein expression analysis

Strain specific TNFα expression analysis in homozygous B-hTNFA/hIL17A mice by ELISA.

Serum were collected from WT and homozygous B-hTNFA/hIL17A (H/H) mice stimulated with LPS in vivo, and analyzed by ELISA with species-specific TNFα ELISA kit. Mouse TNFα was detectable in WT mice. Human TNFα was exclusively detectable in homozygous B-hTNFA/hIL17A but not WT mice.

 

Strain specific IL17A expression analysis in homozygous B-hTNFA/hIL17A mice by ELISA.

Serum were collected from WT and homozygous B-hTNFA/hIL17A (H/H) mice stimulated with LPS in vivo, and analyzed by ELISA with species-specific IL17A ELISA kit. Mouse IL17A was detectable in WT mice. Human IL17A was exclusively detectable in homozygous B-hTNFA/hIL17A but not WT mice.

 

Phenotypic analysis 

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

 

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

Analysis of spleen T cell subpopulations by FACS

Splenocytes were isolated from female C57BL/6 and B-hTNFA/hIL17A mice (n=3, 6 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-hTNFA/hIL17A mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTNFA/hIL17A 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-hTNFA/hIL17A mice

Analysis of lymph node leukocyte subpopulations by FACS

Leukocytes were isolated from female C57BL/6 and B-hTNFA/hIL17A mice (n=3, 6 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-hTNFA/hIL17A mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTNFA/hIL17A 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-hTNFA/hIL17A mice

Analysis of lymph node T cell subpopulations by FACS

Leukocytes were isolated from female C57BL/6 and B-hTNFA/hIL17A mice (n=3, 6 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-hTNFA/hIL17A mice were similar to those in the C57BL/6 mice, demonstrating that introduction of hTNFA/hIL17A 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.

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