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TREM1 is a member of the immunoglobulin superfamily—a group of cell-surface receptors characterized by related extracellular Ig-like domains. In addition to its extracellular Ig domain, it comprises a transmembrane domain containing a conserved lysine residue and a short cytoplasmic domain devoid of signaling motifs. Studies suggest that High Mobility Group Box 1 (HMGB1), Peptidoglycan Recognition Protein 1 (PGLYRP1), Heat Shock Protein 70 (HSP70), CD177, Actin, and extracellular Cold-Inducible RNA-Binding Protein (eCIRP) may serve as natural ligands for TREM-1; notably, all of these endogenous ligands are associated with inflammatory responses.

TREM1 signal transduction relies on its association with the immunoreceptor adaptor protein DNAX-activating protein of 12 kDa (DAP12). TREM-1 possesses a positively charged lysine residue within its transmembrane domain, which pairs with a negatively charged aspartate residue on DAP12. Upon receptor activation, the Immunoreceptor Tyrosine-based Activation Motif (ITAM) within DAP12 undergoes phosphorylation, thereby recruiting and activating Spleen Tyrosine Kinase (Syk). Syk subsequently activates a multitude of downstream signaling pathways—including the PI3K/AKT, Ras/ERK/MAPK, and NF-κB pathways—and triggers the phosphorylation of Phospholipase C, ultimately leading to increased intracellular calcium levels and the secretion of pro-inflammatory cytokines.

The TREM1 Effector Reporter model effectively recapitulates the in vivo signal transduction processes of TREM1; the underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the Jurkat E6.1 Human TREM1 Effector Reporter Cell Model

Figure 2. Recombinant TREM1 Effector Reporter Cell constitutively expressing TREM1.

Figure 3. Dose Response of TREM1 Agonist Ab in TREM1 Effector Reporter Cell(C33).

Figure 4. Blocking of PGLYRP1 (plus 1μg/ml PGN)induced TREM1 Effector Reporter Cell(C33) Activity by TREM1 Blokcing Ab with PGLYRP1 Target Cell .

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.