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Platelet-Dendritic Cell Antigen 2 (BDCA2) is a C-type lectin expressed exclusively on the surface of human pDCs. BDCA2 consists of an extracellular domain that recognizes carbohydrates, a transmembrane region, and a short intracellular segment lacking any signal transduction domains. BDCA-2 forms an adaptor with the FcRγ chain in the transmembrane region to transmit intracellular signals, thereby inducing a signaling cascade similar to that of the B cell receptor (BCR).

  
Antibody-mediated binding to BDCA2 leads to the recruitment of SYK to the immunoreceptor tyrosine-based activation motif (ITAM) of phosphorylated FcεRIγ. Activation of SYK results in the activation of BTK and PLCγ2, leading to calcium ion mobilization. Binding to the BDCA2 receptor has been shown to inhibit IFN I and other pro-inflammatory mediators induced by TLR7 or TLR9.

The BDCA2 Effector Reporter gene-target model effectively simulates the signal transduction process of BDCA2 in vivo, as illustrated in the figure below.

Figure 1. Schematic of the BDCA2 Effector Reporter Cell Model.

Figure 2. Recombinant BDCA2 Effector Reporter Cell stably expressing BDCA2.

Figure 3. Dose Response of Litifilimab in BDCA2 Effector Reporter Cell (C12).

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.