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The IL-1 family comprises 11 cytokines, consisting of 7 ligands with agonist activity (IL-1α, IL-1β, IL-18, IL-33, IL-36α, IL-36β, and IL-36γ) and 4 members with antagonist activity [IL-1 Receptor Antagonist (IL-1Ra), IL-36Ra, IL-37, and IL-38]. IL-1α, IL-1β, IL-18, IL-33, IL-36α, IL-36β, and IL-36γ trigger the activation of MAP kinases and NF-κB, thereby inducing inflammatory responses.

Similar to other cytokine members of the IL-1 family, IL-36 agonists (IL-36α, β, and γ) bind to a heterodimeric receptor complex composed of IL-36R and the IL-1 receptor accessory protein (IL-1RAcP). IL-36R is expressed by numerous cell types, including keratinocytes, pulmonary fibroblasts and epithelial cells, endothelial cells, and various immune cells. IL-36R is also highly expressed in human M0 and M2 macrophages, though it is not highly expressed in M1 macrophages. IL-36Ra and IL-38 exert negative regulatory effects on the IL-36 signaling pathway. IL-36Ra manifests its antagonistic function by competitively binding to IL-36R, thereby inhibiting the recognition of IL-36 agonists and the recruitment of IL-1RAcP; this action effectively blocks the receptor activation mediated by the agonist members of this family. In addition to blocking the IL-36R-mediated activation of the MAP kinase and NF-κB pathways, IL-36Ra can also suppress the expression of Th17 cytokines.

The Jurkat E6.1 Human IL36 Effector Reporter Cell Model—effectively simulates the signal transduction process of  IL36 *in vivo*. The underlying principle is illustrated in the figure below.

Figure 1. Jurkat E6.1 Human IL36 Effector Reporter Cell Model

Figure 2. Dose Response of Recombinant Human IL-36 alpha/IL-1F6 Protein in IL36 Effector Reporter Cell(Suspension, C6).

Figure 3. Inhibition of hIL36-induced Reporter Activity by Blocking Ab in IL36 Effector Reporter Cell(Suspension, C6).

Cell Passage Procedures

1.This cell line grows in suspension.
2.Upon receipt, cells should be thawed immediately or stored in liquid nitrogen until use.
3.Before thawing, pre-warm the water bath and culture medium to 37 °C, and prepare a small amount of dry ice.
4.Remove the cryovial from storage and transport it to the cell culture laboratory on dry ice.
5.Rapidly thaw the cells in a 37 °C water bath. Once the cells are completely thawed, spray the cryovial with 70% ethanol for disinfection and transfer it to a biosafety cabinet.
6.Add 10 mL of pre-warmed culture medium into a 15 mL centrifuge tube. Transfer the contents of the cryovial into the tube and centrifuge at 1000 rpm for 5 minutes.
7.Carefully discard the supernatant. Resuspend the cell pellet in 5 mL of pre-warmed culture medium by gentle pipetting. Immediately perform cell counting and adjust the cell density to 3–6 × 10⁵ cells/mL based on the counting results, then transfer the cells into a culture flask.
8.Count the cells every 1–2 days. When the cell density exceeds 1 × 10⁶ cells/mL, passage the cells promptly or add fresh culture medium. Maintain the cell density between 2 × 10⁵ and 1 × 10⁶ cells/mL.

Suspension Cell Cryopreservation Procedure:

1.Collect 8 × 10⁶ cells, centrifuge, and discard the supernatant.
2.Add 1 mL of cell freezing medium (90% FBS + 10% DMSO) and gently pipette to mix thoroughly. Transfer the suspension into a cryovial.
3.Immediately place the cryovial into a controlled-rate freezing container (Nalgene 5100-0001), fill with isopropanol up to the indicated level, and store at −80 °C.
4.After 24 hours, transfer the cryovial to liquid nitrogen for long-term storage.