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Extended sequence preferences for oligodeoxyribonucleotide activity
Author(s) -
Lenert Petar,
Goeken Adam J.,
Ashman Robert F.
Publication year - 2006
Publication title -
immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.297
H-Index - 133
eISSN - 1365-2567
pISSN - 0019-2805
DOI - 10.1111/j.1365-2567.2006.02320.x
Subject(s) - tlr9 , receptor , toll like receptor 9 , transfection , biology , microbiology and biotechnology , function (biology) , immunology , cell culture , gene , biochemistry , genetics , gene expression , dna methylation
Summary 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‐κB roughly parallels that seen in the biological assays in mouse B cells.