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The IL-17 family comprises six cytokines: IL-17A, IL-17B, IL-17C, IL-17D, IL-17E (also known as IL-25), and IL-17F. Among them, IL-17A is a key driver of pro-inflammatory signaling; secreted by T-helper 17 (Th17) cells, it plays a crucial role in autoimmune diseases. These cytokines function in immune responses and inflammation, mediating signal transduction through the activation of various pathways. The IL-17 receptor family consists of five members (IL-17RA through IL-17RE). IL-17RA serves as a common subunit that forms heterodimeric complexes with ligand-specific subunits (IL-17RB binds IL-25; IL-17RC binds IL-17A/F; IL-17RE binds IL-17C).

IL-25 (also known as IL-17E) plays a complex, dual role within the immune system. As a core driver of type 2 immunity, it is primarily secreted by mucosal epithelial cells. By activating ILC2s and Th2 cells and inducing the release of IL-4, IL-5, and IL-13, it mediates anti-helminth immunity, allergic reactions (such as eosinophilic infiltration in asthma), and tissue repair (including goblet cell hyperplasia). Its overactivation is a central pathological mechanism in allergic rhinitis and atopic dermatitis.

The IL25 Effector Reporter Cell (adherent) model effectively simulates the IL25 signal transduction process *in vivo*; the underlying mechanism is illustrated in the figure below.

Figure 1. Schematic Diagram of the HEK293 Human IL25 Effector Reporter Cell(Adherent) Model

Figure 2.  Recombinant IL25 Effector Reporter Cell(Adherent) stably expressing IL17RA & IL17RB.

Figure 3. Dose Response of Recombinant Human IL25 in IL25 Effector Reporter Cell(Adherent, C19).

Figure 4.  Inhibition of Human IL25-induced Reporter Activity by IL25 Neutralization Ab or IL17RA&RB Blocking Abs in IL25 Effector Reporter Cell(Adherent, C19).

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 HEK293 Human IL25 Effector Reporter Cell(Adherent) Cell 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.