Basic Information

Strain Name
C57BL/6-IL4tm1(hIL4)IL4rtm1(hIL4R)/Bcgen
Stock Number
B-DM-025
Common Name
B-hIL4/hIL4RA mice
Source/Investigator
Bcgen (Beijing Biocytogen Co., Ltd)
Related Genes
IL4R (interleukin 4 receptor)
Species
C57BL/6
Appearance
Black
Genotypes
Homozygous

Details

Phenotype

Protein Expression Analysis

Anti-mCD3

Serum from both wild type (WT) C57BL/6 and homozygous B-hIL4/hIL4RA mice were analyzed by ELISA. Mouse IL-4 was detected in the WT mice, while human IL-4 was detected in the homozygous B-hIL4/hIL4RA mice, albeit being at a lower level. ND not detected.

b-Hil4il4r-Splenocytes

Splenocytes from both wild type (WT) C57BL/6 and homozygous B-hIL4/hIL4RA were analyzed by flow cytometry. Mouse IL4RA was detectable in the WT mice, while human hIL4RA was detectable (BioLegend and BD anti-hIL4RA antibody) in the homozygous B-hIL4/hIL4RA mice.

b-Hil4il4r-Splenocytes-wild

Splenocytes from both wild type (WT) C57BL/6 and homozygous B-hIL4/hIL4RA were analyzed by flow cytometry after mCD3 antibody activation. Mouse IL4RA was detectable in the WT mice, while human hIL4RA was detectable (BioLegend and BD anti-hIL4RA antibodies) in the homozygous B-hIL4/hIL4RA mice.

Antibody Binding Assay

Antibody-Binding-Assay-B-hIL4

Splenocytes from homozygous B-hIL4/hIL4RA were analyzed by flow cytometry.

B cells in homozygous B-hIL4/hIL4RA mice bind well to anti-hIL4RA antibody dupilumab.

Functional Analysis

Analysis of Leukocytes Subpopulation in B-hIL4/hIL4RA Mice

Leukocytes-Subpopulation-B-hIL4

b-Hil4il4r-Splenocytes-isolated

Splenocytes were isolated from C57BL/6 and B-hIL4/hIL4RA mice (n=3). Proportions of leukocyte subpopulations were determined by flow cytometry. As shown in panels here,the expression profile of leukocytes subpopulation in homozygous B-hIL4/hIL4RA 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 hIL4/hIL4RA.

Analysis of T Cell Subpopulation in B-hIL4/hIL4RA Mice

b-Hil4il4r-Lymphocyte

Lymphocytes were isolated from spleen in C57BL/6 and B-hIL4/hIL4RA mice (n=3). Proportion of T cells subpopulations was determined by flow cytometry. As shown here, the expression profile of lymphocyte subpopulation in homozygous B-hIL4/hIL4RA mice is similar to that in the C57BL/6 mice, indicating that T cell differentiation is not affected by the humanization of hIL4/hIL4RA.

Blood Routine Test in B-hIL4/hIL4RA Mice

b-Hil4il4r-C57BL6

Blood from the C57BL/6 and B-hIL4/hIL4RA mice (n=3) was collected and analyzed by CBC. There is no significant difference between the C57BL/6 and B-hL4/hIL4RA mice.

Blood Biochemistry Test in B-hIL4/hIL4RA Mice

b-Hil4il4r-Blood-Biochemistry

Serum AST/ALT levels of C57BL/6 and B-hIL4/hIL4RA mice were tested (n=3). Results indicate that there is no significant difference between the C57BL/6 and B-hIL4/hIL4RA mice.

IL-4 Induced IgE Secretion in B-hIL4/hIL4RA Mice

b-Hil4il4r-Splenic-B-Cells

Splenic B cells from C57BL/6 and B-hIL4/hIL4RA mice were cultured with LPS alone or together with 50 ng/mL mIL-4/hIL-4. Culture supplements  were harvested on day 6 for quantification of IgE  by ELISA. The result showed that B cells from B-hIL4/hIL4RA mice responded well to LPS and IL-4 with IgE production , as well as C57BL/6 mice.

Dupilumab Blockade Experiment

b-Hil4il4r-Dupilumab-Blockage

Splenic B cells from C57BL/6 and B-hIL4/hIL4RA mice were incubated with increasing doses of dupilumab for half of an hour before adding LPS and hIL-4 (50 ng/mL). Culture supernatant was  harvested on day 6 for quantification of IgE  by ELISA. The result showed that IL-4 induction of mIgE production in vitro in mouse B cells was blocked by dupilumab even at 0.01 ng/mL, the lowest concentration tested.

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