Extended sequence preferences for oligodeoxyribonucleotide activity.
Synthetic type B phosphorothioate oligodeoxyribonucleotides (ODN) activate mouse B cells via Toll-like receptor 9 (TLR9). Starting with closely related 15-mer prototype ODN, the sequence requirements for stimulatory (ST-) and inhibitory (IN-) activity were contrasted, by measuring apoptosis protection, G(1) entry and interleukin-6 secretion. ST-ODN and IN-ODN differ in that (1) ST-ODN require a 5' T, (2) the central CG is obligatory, (3) CG must be flanked 3' specifically by TT at the position where IN-ODN have GG, and (4) IN-ODN tolerate truncation of the 3' end better than ST-ODN. Features shared by ST-ODN and IN-ODN include (1) requiring CC adjacent to the 5' end, and (2) avoiding CC immediately 5' to the CG. This pattern is used to create a model of how ST-ODN binding might function to aggregate TLR9 so as to initiate the signal, and how the 5' ends of ST-ODN and IN-ODN compete for binding. Further justification for considering TLR9 to be the ODN receptor was provided by a demonstration that in HEK293 cells transfected with TLR9, the potency of a panel of ODN for activating NF-kappaB roughly parallels that seen in the biological assays in mouse B cells.
Division of Rheumatology, Department of Internal Medicine, Carver College of Medicine, The University of Iowa, Iowa City, IA 52242, USA. petar-lenert@uiowa.edu
Petar
P
Lenert
Adam J
AJ
Goeken
Robert F
RF
Ashman
D000818:Animals; D017209:Apoptosis / Q000276:immunology*; D001402:B-Lymphocytes / Q000276:immunology; D001483:Base Sequence; D002478:Cells, Cultured; D018899:CpG Islands / Q000276:immunology*; D004268:DNA-Binding Proteins / Q000276:immunology; D004306:Dose-Response Relationship, Immunologic; D005260:Female; D016193:G1 Phase / Q000276:immunology; D015850:Interleukin-6 / Q000378:metabolism; D051379:Mice; D008815:Mice, Inbred Strains; D016328:NF-kappa B / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000235:genetics / Q000276:immunology*; D013329:Structure-Activity Relationship; D051217:Toll-Like Receptor 9 / Q000276:immunology
C408982:CPG-oligonucleotide; D004268:DNA-Binding Proteins; D015850:Interleukin-6; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D051217:Toll-Like Receptor 9