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CCR4 (also known as CD194 or CMKBR4) is a protein-coding gene for CC chemokine receptor 4, a key member of the G protein-coupled receptor family. This gene is primarily expressed in specific T-cell subsets, such as Th2 cells and regulatory T cells (Tregs), and to a lesser extent in B cells, NK cells, and macrophages; notably, it is highly expressed in various tumor cells, including those associated with adult T-cell leukemia/lymphoma (ATLL). Upon binding to ligands such as CCL17 and CCL22, the encoded receptor activates downstream pathways—including ERK, MAPK, and PI3K-AKT—thereby regulating the directional migration, chemotaxis, and activation of immune cells and participating in physiological processes such as the maintenance of immune homeostasis and inflammatory responses.

Aberrations in this gene are closely linked to the pathogenesis of allergic inflammation, autoimmune diseases (such as asthma and rheumatoid arthritis), and various malignancies. It serves as a significant drug target; notably, the targeted monoclonal antibody mogamulizumab has already been approved for the treatment of related lymphomas.

The CHO-K1 Human CCR4 Gα15 Cell Line model effectively simulates the in vivo CCR4 signal transduction process, the principle is illustrated in the figure below.

Figure 1. Schematic diagram of the CHO-K1 Human CCR4 Gα15 Cell Line model

Figure 1. Dose Response of Recombinant Human CCL22 in CCR4/Gα15 CHO-K1 cells(C11).

Cell Resuscitation
1)Rapidly thaw the frozen cells in a 37 °C water bath for approximately 60 seconds. Once thawed (which may take slightly less or more than 60 seconds), immediately transfer the cell suspension from the cryovial into a 15 mL centrifuge tube containing 10 mL of pre-warmed CHO-K1 Human CCR4 Gα15 Cell Line complete culture medium.
2)Centrifuge cells at 1000 rpm for 5 min to remove medium, then resuspend cells in 5 mL of pre-warmed complete medium.
3)Transfer the cell suspension into a T25 culture flask and incubate at 37 °C with 5% CO₂.
4)After approximately 24–36 hours, replace the medium or passage the cells to remove non-adherent dead cells.

Subculturing procedure
1)When the cell density reaches the appropriate confluency for passaging, wash the cells with PBS, then add 1 mL trypsin to detach the cells. When more than 80% of the cells detach upon gently tapping the culture flask, add complete culture medium to terminate digestion. Gently pipette to obtain a single-cell suspension, transfer to a 15 mL centrifuge tube, and centrifuge at 1000 rpm for 5 minutes.

2)Discard supernatant after centrifugation. Resuspend cells in fresh medium to a single-cell suspension and transfer to a new culture flask for continued growth.

Cell Freezing
After trypsinization and centrifugation of cells from each T75 flask or 10 cm culture dish, discard the supernatant. Add 2 mL of cryopreservation medium (90% FBS + 10% DMSO), gently resuspend thoroughly, and aliquot into two cryovials. Immediately place the cryovials into a controlled-rate freezing container (e.g., Nalgene 5100-0001), fill with isopropanol to the indicated level, and store at −80 °C. After 24 hours, transfer the cryovials to liquid nitrogen for long-term storage.