Immunostimulatory CpG-oligodeoxynucleotides induce a factor that inhibits macrophage adhesion

Abstract

Selected phosphorothioate oligodeoxynucleotides containing CpG (CpG-ODN) activate immune responses, including immunoglobulin synthesis, B cell proliferation, and cytokine production by monocytes. We examined the effect of a CpG-ODN (#1760) on the adhesion of macrophages derived from human blood monocytes in vitro. CpG-ODN (6 microg/mL) completely inhibited the adherence of macrophages to plastic or glass during 7 or more days of culture. A non-CpG control ODN (#1814) was without effect. Two other CpG-ODNs (#1826 and #1842) also completely inhibited macrophage adherence. The specific inhibitor of CpG-ODN, quinacrine (0.1 micromol/L), blocked this action. CpG-ODN reduced the rate of senescence and cell death of monocytes in culture but did not influence their phagocytosis, procoagulant activity, or support of the mixed lymphocyte response. Four days of exposure of monocytes to CpG-ODN up-regulated the expression of the endotoxin receptor CD14 and down-regulated the mannose (scavenger) receptor, a result that is consistent with blocking the maturation of monocytes to macrophages. Incubation of peripheral blood monocytes (PBMCs) with CpG-ODN resulted in the generation of a heat labile factor that inhibited macrophage differentiation and accounts for the efficacy of the CpG-ODN. T cells selected from PBMCs by magnetic beads generated the majority of this factor. Cytokines (interleukin-3 (IL-3), IL-4, IL-6, IL-10, interferon-gamma, tumor necrosis factor-alpha, transforming growth factor-beta, granulocyte-macrophage colony-stimulating factor, monocyte chemotactic protein-1) did not inhibit macrophage adherence like CpG-ODN did. Antibodies to IL-6 or IL-10 did not block the activity of CpG-ODN. Dexamethasone inhibited macrophage adherence, and lipopolysaccharide had a minor effect. We conclude that immunostimulatory CpG-ODNs inhibit macrophage adherence by provoking the production of an unidentified heat-labile factor.