Basic Information

Common Name
B-hIL27/hIL27RA mice
Background
C57BL/6
Catalog number
121566
NCBI Gene ID

Gene Targeting Strategy

Gene targeting strategy for B-hIL27/hIL27RA mice. The murine Il27 gene was replaced by the full coding region of human IL27 in B-hIL27/hIL27RA mice. The murine extracellular Il27ra gene sequences were replaced with the human IL27RA counterpart in B-hIL27/hIL27RA mice.

Protein Expression Analysis

Species-specific IL27 protein expression analysis in humanized B-hIL27/hIL27RA mice. BMDC culture supernatant was collected from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA (H/H) mice, stimulated with LPS (1μg/ml), and analyzed by ELISA using species-specific IL27 ELISA kits. Murine IL27 protein was detected in wild-type mice, while human IL27 protein was exclusively detected in B-hIL27/hIL27RA mice.

Species-specific IL27RA protein expression analysis in humanized B-hIL27/hIL27RA mice. CD4+ T cells were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA (H/H) mice, stimulated with anti-CD3ε and anti-CD28 in vitro for 48h, and analyzed by flow cytometry using species-specific anti-IL27RA antibodies. Murine IL27RA protein was detected in wild-type mice, while human IL27RA protein was exclusively detected in B-hIL27/hIL27RA mice.

Intracellular staining of pSTAT1 induced by IL27

Intracellular pSTAT1 analysis in humanized B-hIL27/hIL27RA mice. Splenic T cells were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA (H/H) mice. CD4+ T cells were purified using magnetic beads, stimulated with mouse or human IL27 in vitro for 15min, and analyzed by flow cytometry using anti-p-STAT1 antibodies. Both mouse and human IL27 can induce STAT1, indicating humanization of IL27RA does not impact receptor function.

Analysis of Splenic Leukocyte Subpopulations in Humanized B-hIL27/hIL27RA Mice

Analysis of splenic leukocyte subpopulations in humanized B-hIL27/hIL27RA mice. Splenocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA mice (female, n=3, 6-week-old), and analyzed by flow cytometry to assess splenic leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45 and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, monocytes, dendritic cells, granulocytes and macrophages in homozygous B-hIL27/hIL27RA mice were similar to those in wild-type mice, demonstrating that hIL27/hIL27RA in place of their murine counterparts does not change the overall development, differentiation or distribution of splenic leukocytes. Values are expressed as mean ± SEM.

Analysis of Splenic T Cell Subpopulations in Humanized B-hIL27/hIL27RA Mice

Analysis of splenic T cell subpopulations in humanized B-hIL27/hIL27RA mice. Splenic lymphocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA mice (n=3, 6-week-old), and analyzed by flow cytometry to assess T cell subpopulations. There were no differences between wild-type and B-hIL27/hIL27RA mice, demonstrating that hIL27/hIL27RA in place of their murine counterparts does not change the overall development, differentiation or distribution of splenic T cell subtypes. Values are expressed as mean ± SEM.

Analysis of Lymph Node Leukocyte Subpopulations in Humanized B-hIL27/hIL27RA Mice

Analysis of lymph node leukocyte subpopulations in humanized B-hIL27/hIL27RA mice. Lymphocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA (H/H) mice (female, n=3, 6-week-old), and analyzed by flow cytometry to assess lymph node leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45 and used for further analysis as indicated. (B) Percent of T cells, B cells and NK cells in B-hIL27/hIL27RA mice were similar to those in wild-type mice, demonstrating that hIL27/hIL27RA in place of their murine counterparts does not change the overall development, differentiation or distribution of lymph node leukocytes. Values are expressed as mean ± SEM.

Analysis of Lymph Node T Cell Subpopulations in Humanized B-hIL27/hIL27RA Mice

Analysis of lymph node T cell subpopulations in humanized B-hIL27/hIL27RA mice. Lymphocytes were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA (H/H) mice (n=3, 6-week-old), and analyzed by flow cytometry to assess T cell subpopulations. There were no differences between wild-type and B-hIL27/hIL27RA mice, demonstrating that hIL27/hIL27RA in place of their murine counterparts does not change the overall development, differentiation or distribution of lymph node T cell subtypes. Values are expressed as mean ± SEM.

Analysis of Blood Leukocyte Subpopulations in Humanized B-hIL27/hIL27RA Mice

Analysis of blood leukocyte subpopulations in humanized B-hIL27/hIL27RA mice. Blood cells were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA (H/H) mice (female, n=3, 6-week-old), and analyzed by flow cytometry to assess blood leukocyte subpopulations. (A) Representative flow cytometry plots. Single live cells were gated on CD45 and used for further analysis as indicated. (B) Percent of T cells, B cells, NK cells, monocytes, dendritic cells, granulocytes and macrophages in homozygous B-hIL27/hIL27RA mice were similar to those in wild-type mice, demonstrating that hIL27/hIL27RA in place of their murine counterparts does not change the overall development, differentiation or distribution of blood leukocytes. Values are expressed as mean ± SEM.

Analysis of Blood T Cell Subpopulations in Humanized B-hIL27/hIL27RA Mice

Analysis of blood T cell subpopulations in humanized B-hIL27/hIL27RA mice. Blood cells were isolated from wild-type C57BL/6 (+/+) and homozygous B-hIL27/hIL27RA (H/H) mice (n=3, 6-week-old), and analyzed by flow cytometry to assess blood T cell subpopulations. There were no differences between wild-type and B-hIL27/hIL27RA mice, demonstrating that hIL27/hIL27RA in place of their murine counterparts does not change the overall development, differentiation or distribution of blood T cell subtypes. Values are expressed as mean ± SEM.