Antagonism of immunostimulatory CpG-oligodeoxynucleotides by quinacrine, chloroquine, and structurally related compounds.
Phosphorothioate oligodeoxynucleotides containing CpG (CpG-ODN) activate immune responses. We report that quinacrine, chloroquine, and structurally related compounds completely inhibit the antiapoptotic effect of CpG-ODN on WEHI 231 murine B lymphoma cells and inhibit CpG-ODN-induced secretion of IL-6 by WEHI 231. They also inhibit IL-6 synthesis and thymidine uptake by human unfractionated PBMC induced by CpG-ODN. The compounds did not inhibit LPS-induced responses. Half-maximal inhibition required 10 nM quinacrine or 100 nM chloroquine. Inhibition was noncompetitive with respect to CpG-ODN. Quinine, quinidine, and primaquine were much less powerful. Quinacrine was effective even when added after the CpG-ODN. Near-toxic concentrations of ammonia plus bafilomycin A1 (used to inhibit vesicular acidification) did not reduce the efficacy of the quinacrine, but the effects of both quinacrine and chloroquine were enhanced by inhibition of the multidrug resistance efflux pump by verapamil. Agents that bind to DNA, including propidium iodide, Hoechst dye 33258, and coralyne chloride did not inhibit CpG-ODN effect, nor did 4-bromophenacyl bromide, an inhibitor of phospholipase A2. Examination of the structure-activity relationship of seventy 4-aminoquinoline and 9-aminoacridine analogues reveals that increased activity was conferred by bulky hydrophobic substituents on positions 2 and 6 of the quinoline nucleus. No correlation was found between published antimalarial activity and ability to block CpG-ODN-induced effects. These results are discussed in the light of the ability of quinacrine and chloroquine to induce remission of rheumatoid arthritis and lupus erythematosus.
Macfarlane
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Department of Medicine, Veterans Affairs Medical Center, University of Iowa, Iowa City 52242, USA. donald-macfarlane@uiowa.edu
Manzel
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D000143:Acids / Q000037:antagonists & inhibitors / Q000378:metabolism; D000276:Adjuvants, Immunologic / Q000037:antagonists & inhibitors* / Q000502:physiology; D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000276:immunology; D001667:Binding, Competitive / Q000276:immunology; D001692:Biological Transport / Q000187:drug effects / Q000276:immunology; D002455:Cell Division / Q000187:drug effects / Q000276:immunology; D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology*; D018899:CpG Islands / Q000187:drug effects* / Q000276:immunology; D018432:Drug Resistance, Multiple / Q000276:immunology; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D007364:Intercalating Agents / Q000494:pharmacology; D015850:Interleukin-6 / Q000096:biosynthesis; D007700:Kinetics; D007963:Leukocytes, Mononuclear / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D051379:Mice; D009838:Oligodeoxyribonucleotides / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D011796:Quinacrine / Q000494:pharmacology*; D013329:Structure-Activity Relationship; D013873:Thionucleotides / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism
D000143:Acids; D000276:Adjuvants, Immunologic; D007166:Immunosuppressive Agents; D007364:Intercalating Agents; D015850:Interleukin-6; D008070:Lipopolysaccharides; D009838:Oligodeoxyribonucleotides; D013873:Thionucleotides; D002738:Chloroquine; D011796:Quinacrine