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The Interleukin-12 (IL-12) cytokine family—comprising IL-12, IL-23, IL-27, IL-35, and IL-39—plays a pivotal role in the induction and regulation of both innate and adaptive immune responses. IL-27 is primarily produced by antigen-presenting cells (APCs), including dendritic cells (DCs), plasma cells, macrophages, inflammatory monocytes, microglia, B cells, and endothelial cells. IL-27 is typically involved in the differentiation and activation of various CD4 T cell subsets. Antigen-presenting cells stimulated by tumor antigens and CD40 can secrete IL-27; notably, IL-27 exerts a dual function—acting as both a pro-inflammatory and an anti-inflammatory agent—in regulating immune responses against cancer. Furthermore, IL-27 exhibits antiviral properties against a range of human viruses, including influenza, Herpes Simplex Virus (HSV), Hepatitis B Virus (HBV), Hepatitis C Virus (HCV), Human Immunodeficiency Virus (HIV), and others.

IL-27 is a heterodimeric cytokine composed of two subunits: EBi3 and p28. IL-27 signals through a heterodimeric receptor complex consisting of two subunits: IL-27Rα (also known as WSX-1 or TCCR) and gp130 (IL6ST). Upon binding, IL-27 activates the Janus kinase (JAK) signaling pathway, the Signal Transducer and Activator of Transcription (STAT1 and STAT3) pathway, and the Mitogen-Activated Protein Kinase (MAPK) signaling pathway.

The IL27 Effector Reporter Cell model serves as an excellent *in vitro* mimic of the IL-27 signal transduction process observed *in vivo*; the underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the IL27 Effector Reporter Cell Model

Figure 2. Recombinant IL27 Effector Reporter Cell constitutively expressing IL27Rα.

Figure 3.Dose Response of Recombinant Human IL27 in IL27 Effector Reporter Cell (C18).

Figure 3.Inhibition of rhIL27-Induced Reporter Activity by IL27 Neutralization Ab in IL27 Eeffector Reporter Cell(C18).

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 IL27 Effector Reporter 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.