Purification of a monocyte chemotactic factor secreted by nonhuman primate vascular cells in culture

Abstract

A protein chemotactic for peripheral blood monocytes (SMC-CF) of potential importance in their recruitment to the arterial intima in atherogenesis was purified from serum-free medium conditioned by cultured baboon aortic medial smooth muscle cells. The purification of SMC-CF was monitored by a filter assay using human peripheral blood mononuclear cells and was achieved by batch separation on a cation-exchange gel followed by gel permeation chromatography, ion-exchange high-performance liquid chromatography (HPLC), and reversed-phase HPLC. The overall recovery was approximately 10% of the initial activity and yielded 0.5-1 microgram of SMC-CF/L of conditioned medium. On analytical sodium dodecyl sulfate-polyacrylamide gel electrophoresis, SMC-CF migrated as a monomeric protein with an apparent molecular weight of 14,500. A dose-dependent relationship was observed between SMC-CF concentration and monocyte chemotactic activity, with maximal and half-maximal biologic activity being observed at approximately 5 and 0.1 nM, respectively. Cultured baboon aortic smooth muscle cells also express the genes for both the A and B polypeptide chains of platelet-derived growth factor, which has been reported to be chemotactic for blood monocytes and neutrophils [Deuel, T. F., Senior, R. M., Huang, J. S., & Griffin, G. L. (1982) J. Clin. Invest. 69, 1046-1049]. Amino acid composition analyses indicate that SMC-CF is not derived either from polypeptide chain of this growth factor or from certain potentially chemotactic connective tissue proteins.

Human monocyte chemoattractant protein‐1 (MCP‐1) Full‐length cDNA cloning, expression in mitogen‐stimulated blood mononuclear leukocytes, and sequence similarity to mouse competence gene JE

“…The amino acid composition of a monocyte chemoattractant produced by aortic smooth muscle cells of the baboon [27] is identical to that of MCP-1 [3]. Hybridization of the MCP-1 cDNA probe with baboon DNA ( fig.6) is added evidence for the relationship between MCP-1 and the smooth muscle product, and indicates that both lymphocytes and vascular smooth muscle cells can produce this attractant.…”

A Mechanism for the Impaired IFN-γ Production in C-C Chemokine Receptor 2 (CCR2) Knockout Mice: Role of CCR2 in Linking the Innate and Adaptive Immune Responses

“…Chemokine receptor 2 is the receptor for monocyte chemoattractant proteins (MCPs) 3 [1][2][3][4][5]. The MCPs are potent chemoattractants for monocytes (7)(8)(9), memory T cells (10), NK cells (11,12), and immature dendritic cells (DCs) (13). We and others have shown that deletion of CCR2 reduces the number of macrophages recruited into the peritoneum after thioglycolate administration (14 -16), consistent with the possibility that CCR2 is normally present on murine monocytes/macrophages.…”

Monocyte chemotactic protein-1 as a chemoattractant for human hepatic stellate cells

“…Measurement of lymphomonocyte chemotaxis was performed by using 5 µm filters and modified Boyden chambers, as described by Valente et al 23 The cell preparation (200 µL) was placed above the filters, and the chambers were incubated at 37°C for 90 minutes. After staining with Giemsa, cells migrating to the underside of the filters were quantitated as the mean number of cells in 10 high power fields.…”