Interaction of soluble CD163 with activated T lymphocytes involves its association with non-muscle myosin heavy chain type A.
CD163 is a monocyte/macrophage-specific scavenger receptor that undergoes ectodomain shedding upon an inflammatory stimulus. Soluble CD163 (sCD163) actively inhibits lymphocyte proliferation, but to date exactly how it interacts with these cells has remained elusive. We screened T lymphocytes and endothelial cells for proteins binding to sCD163. In both cell types a high affinity binding protein was detected. Partial sequencing of the protein revealed sequence identity to a non-muscle myosin heavy chain type A. Employing labelled sCD163 we found little specific binding of sCD163 to the extracellular domains of T lymphocytes and human umbilical vein endothelial cells (HUVEC). In activated T lymphocytes we demonstrated specific binding of sCD163 to intracellular structures as well as the presence of the native protein within the cell after co-incubation with purified sCD163. Furthermore, we developed a novel ELISA for highly specific detection of sCD163-myosin complexes. These complexes were present in activated T lymphocytes after incubation with shed sCD163. Co-localization of sCD163 and cellular myosin in T lymphocytes was further confirmed by fluorescence microscopy. Our results suggest that sCD163 associates with cellular myosin, thereby possibly modulating the cells' response to an inflammatory stimulus.
Timmermann
Meike
M
Institut für Pharmazie und Lebensmittelchemie, Bayerische Julius-Maximilians-Universität, Würzburg, Germany.
Buck
Friedrich
F
Sorg
Clemens
C
Högger
Petra
P
D000595:Amino Acid Sequence; D015703:Antigens, CD / Q000378:metabolism*; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism*; D004730:Endothelium, Vascular / Q000166:cytology; D005456:Fluorescent Dyes; D006801:Humans; D008213:Lymphocyte Activation; D020409:Molecular Motor Proteins / Q000378:metabolism*; D018995:Myosin Heavy Chains / Q000378:metabolism*; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism*; D017421:Sequence Analysis; D012995:Solubility; D013601:T-Lymphocytes / Q000737:chemistry / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate; D014471:Umbilical Veins / Q000166:cytology; D000098902:CD163 Antigen
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D000098902:CD163 Antigen; D005456:Fluorescent Dyes; C103814:MYH9 protein, human; D020409:Molecular Motor Proteins; D011956:Receptors, Cell Surface; D018995:Myosin Heavy Chains; D013755:Tetradecanoylphorbol Acetate