Diminution of Phagocytosed Micro/Nanoparticles by Tethering with Immunoregulatory CD200 Protein

Abstract

CD200 is known as an anti-inflammatory transmembrane glycoprotein in the immunoglobulin superfamily. CD200 interacts with its receptor CD200R which is highly expressed on myeloid cells such as macrophages and neutrophils. CD200-CD200R interaction has known to reduce macrophage activation and chronic inflammation. To harness the immunomodulatory property of CD200 for surface modification, CD200-streptavidin fusion protein was expressed from bacteria transformed with pET20b plasmid encoded with CD200 extracellular domain and core streptavidin. The purified CD200-SA protein was bound to biotin-coated fluorescent polystyrene particles of various sizes ranging from 0.15 to 2 µm. THP-1 macrophages were cultivated with CD200-modified micro/nanoparticles in comparison with controls. Our results showed that both nano- and micro-sized particles decorated with CD200 decreased phagocytosis activities of THP-1 macrophages. Such diminution of phagocytosis was examined to be associated with downregulation of Toll-like receptor 4 (TLR4) expression on the surface of macrophages. Moreover, THP-1 macrophages treated with CD200-coated particles decreased the secretion of tumor necrosis factor-α (TNF-α).