CD83, Fc Chimera, Recombinant, Human (B-cell activation protein, BL11, Cell Surface Protein HB15)

Human CD83 is a 40-50kD member of the Siglec (or sialic-acid-binding immunoglobulin-like lectin) family of transmembrane proteins (1, 2, 3). CD83 is synthesized as a type I transmembrane glycoprotein that contains a 125aa extracellular region, a 22 aa transmembrane segment, and 39aa cytoplasmic domain . It contains one V type Ig-like domain in the extracellular region with no inhibitory cytoplasmic motif(s). Although in vitro studies suggest CD83 may form membrane-bound covalent homodimers, in vivo this does not appear to be the case (1, 4). In the extracellular region, mouse and human CD83 are 66% aa identical (1, 2, 4, 5). Relative to human, mouse CD83 is 11 aa shorter in its extracellular domain and is expressed as a 30-35kD protein (1, 4, 5). Human CD83 is active in the mouse system (4). One alternate splice form has been reported. This leads to a small monomeric soluble form of 74aa that includes aa'20-52 and 164-205 (6, 7). In human, proteolytic cleavage and solubilization of CD83 has also been suggested, and this could lead to dimeric circulating CD83 (4, 6). CD83 is a primary marker for dendritic cells (3, 6, 8). It is also found on B cells (6, 9), neutrophils (10), monocytes and macrophages (11). Except for dendritic cells, CD83 expression is often transient. CD83 binds to sialic acids on target cells (12). The function of CD83 is only now becoming clear. Membrane CD83 appears to promote T cell proliferation, particularly of CD8+ cytotoxic T cells (13, 14). Soluble CD83, however, appears to be immunosuppressive and blocks T cell activation (15, 16). On monocytes, CD83 is suggested to drive monocytes into a fibrocyte phenotype (13). A lack of membrane-expressed CD83 leads to an unusual IL-4/IL-10 producing CD4+ T cell phenotype (17). Source: Human CD83 (Met 1-Ala 143) DIEGRMD Human IgG 1 (Pro 100-Lys 330). A DNA sequence encoding the extracellular domain of human CD83 (Met 1-Ala 143, Accession # Q01151) (Zhou, J.L. et. al., 1992, J. Immunol.149(2): 735-742) was fused with the Fc region of human IgG1 via a linker peptide. The chimeric protein was expressed in insect cells, Sf21. Molecular Mass: The recombinant human CD83/Fc chimera, generated by proteolytic removal of the signal peptide, is a disulfide-linked homodimer. Based on N-terminal amino acid sequencing, the recombinant human CD83/Fc starts at Thr 20. The predicted molecular mass of the monomer is ~40.7kD. As a result of glycosylation, the recombinant protein migrates as an ~51-54kD protein in SDS-PAGE under reducing conditions. Activity: Measured by the ability of immobilized protein to support the adhesion of human monocyte derived dendritic cells. When 5x10e4 cells/well are added to human CD83/Fc coated plates (5ug/ml, 100ul/well), approximately 50-75% will adhere after 30 minutes at 37° C , Storage and Stability: Lyophilized powder may be stored at -20°C. Stable for 12 months at -20°C. Reconstitute with sterile PBS. Aliquot to avoid repeated freezing and thawing. Store at -20°C. Reconstituted product is stable for 6 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.