B-hCCR4-luc EL4_Biocytogen Pharmaceuticals (Beijing) Co., Ltd._Biocytogen, drug development, antibody discovery

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B-hCCR4-luc EL4

Common name	B-hCCR4-luc EL4	Catalog number	321851
Aliases	CCR4, CC-CKR-4, CD194, CKR4, CMKBR4, ChemR13, HGCN:14099, K5-5	Disease	Lymphoma
Organism	Mouse	Strain	C57BL/6
Tissue types	Lymphoma	Tissue	T lymphocyte

Description

The mouse Ccr4 gene was replaced by human CCR4 coding sequence and the luciferase sequence in B-hCCR4-luc EL4 cells. Human CCR4 is highly expressed on the surface of B-hCCR4-luc EL4 cells.

Application

B-hCCR4-luc EL4 cells have the capability to establish tumors in vivo and can be used for efficacy studies.

Targeting strategy

Gene targeting strategy for B-hCCR4-luc EL4 cells. The exogenous promoter and human CCR4 coding sequence and the luciferase sequence were inserted to replace part of murine exon 2. The insertion disrupts the endogenous murine Ccr4 gene, resulting in a non-functional transcript.

Protein expression analysis

CCR4 expression analysis in B-hCCR4-luc EL4 cells by flow cytometry.Single cell suspensions from wild-type EL4 and B-hCCR4-luc EL4 cultures were stained with species-specific anti-CCR4 antibody. Human CCR4-luc was detected on the surface of B-hCCR4-luc EL4 cells but not wild-type EL4 cells. The clones 3-F10 and 3-F11 of B-hCCR4-luc EL4 cells were used for in vivo tumor growth assays.

The luminescence signal intensity analysis

The luminescence signal intensity of B-hCCR4-luc EL4 cells.The luminescence intensity was detected by Bright-Glo^TM luciferase Assay System (Promega, Cat E2610), and The clones 3-F10 and 3-F11 of B-hCCR4-luc EL4 cells has strong luminescence signal.

Tumor growth curve & Body weight changes

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Subcutaneous homograft tumor growth of B-hCCR4-luc EL4 cells. B-hCCR4-luc EL4 cells (2x10⁵) and wild-type EL4 cells (2x10⁵) were subcutaneously implanted into C57BL/6 mice (female, 7-week-old, n=5). Tumor volume and body weight were measured twice a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean± SEM). Volume was expressed in mm³ using the formula: V=0.5 X long diameter X short diameter². As shown in panel A, B-hCCR4-luc EL4 cells were able to form tumors in vivo and can be used for efficacy studies.

Protein expression analysis of tumor cells

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B-hCCR4-luc EL4 cells were subcutaneously transplanted into C57BL/6 mice (n=5). At the end of the experiment, tumor cells were harvested and assessed for human CCR4 expression by flow cytometry. As shown, human CCR4 was highly expressed on the surface of tumor cells. Therefore, B-hCCR4-luc EL4 cells can be used for in vivo efficacy studies of CCR4 therapeutics.

Quantitative imaging data & body weight changes

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Tumor growth and in vivo imaging of B-hCCR4-luc EL4 cells. B-hCCR4-luc EL4 cells (2x10⁵) were injected into the tail vein of wild-type C57BL/6 mice. Signal intensity and body weight were measured twice a week. (A) Imaging was performed twice a week. (B) Body weight (Mean ± SEM). B-hCCR4-luc EL4 cells can be used for in vivo efficacy evaluation.