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CD200 (also known as OX-2) is a member of the Immunoglobulin Superfamily (IgSF), a family of proteins structurally similar to B7 family proteins. It contains two extracellular immunoglobulin domains and a small intracellular domain of 19 amino acids, lacking any known signal motifs. CD200 is expressed in various normal tissues, including immune cells such as B lymphocytes and activated T lymphocytes. Recent studies have shown that CD200 is also overexpressed in multiple human cancer cells, including human melanoma, ovarian cancer, brain gliomas, myeloid leukemias, as well as some B-cell malignancies and most endocrine malignancies (e.g., small cell lung cancer).

  
CD200R, as the homologous ligand for CD200, is also an IgSF protein. In both mice and humans, the expression pattern of CD200R is similar, with high expression in macrophages, neutrophils, and mast cells. Unlike most IgSF receptors, CD200R lacks an ITIM domain. However, its cytoplasmic tail, composed of 67 amino acids, contains three tyrosine residues. The third tyrosine residue is located within an NPXY motif, which is phosphorylated upon CD200R ligand binding. This phosphorylation does not directly recruit protein tyrosine phosphatases such as SHP1 and SHP2 or the phosphorylated inositol phosphatase SHIP, but instead recruits downstream tyrosine kinase adaptor proteins Dok-2 and Dok-1, which then associate with RasGAP and SHIP. In macrophages and mast cells, this cascade has been shown to inhibit the phosphorylation of ERK, P38, and JNK, and suppress myeloid cell activation. Although CD200R expression is primarily found in macrophages and neutrophils, further studies have indicated its presence in dendritic cells (DCs) and some T-cell subsets, suggesting that CD200R signaling may also have regulatory functions in these cell types.

The CD200R Effector Reporter Cell reporter gene drug target model effectively simulates the signal transduction process of CD200 and CD200R in vivo, with the principle shown in the figure below.

Figure 1. Schematic of the CD200R Effector Reporter Cell Model.

Figure 2. Recombinant CD200R Effector Reporter cell constitutively expressing CD200R.

Figure 3. Dose Response of CD200R Agonist Ab Induced Agonism Activity in CD200R Effector Reporter Cell( C34).

Figure 4. Dose Response of CD200R Blocking Ab in CD200R Effector Reporter Cell(C34) with CD200 Target cell(C18).

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.