Aggregation and secondary loop structure of oligonucleotides do not determine their ability to inhibit TLR9

Robert F Ashman; J Adam Goeken; Petar S Lenert

doi:10.1016/j.intimp.2011.02.023

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Aggregation and secondary loop structure of oligonucleotides do not determine their ability to inhibit TLR9

Journal article

Peer reviewed

Aggregation and secondary loop structure of oligonucleotides do not determine their ability to inhibit TLR9

Robert F Ashman, J Adam Goeken and Petar S Lenert

International immunopharmacology, Vol.11(8), pp.1032-1037

08/2011

DOI: 10.1016/j.intimp.2011.02.023

PMCID: PMC3119753

PMID: 21376154

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Abstract

Toll-like receptor 9 (TLR9) is an endosomal DNA sensor that warns us of the presence of infectious danger and triggers a rapid pro-inflammatory response in dendritic cells, macrophages, and B cells. The consequences of uncontrolled TLR9 activation can be detrimental for the host, contributing to the pathogenesis of bacterial septic shock or autoimmune diseases, such as systemic lupus erythematosus. Therefore, we need to develop TLR9 antagonists. We and others have created inhibitory oligonucleotides (INH-ODN) that are capable of sequence-dependent inhibition of TLR9-induced activation in both human and mouse cells. However, it is not clear whether marked differences in INH-ODN activity related to base sequence derived from polymerization of INH-ODNs or their ability to complex with stimulatory CpG-oligonucleotides (ST-ODN). Furthermore, the 5' end of INH-ODNs may assume a particular loop configuration that may be needed for binding to a critical site on TLR9. Here, we show that 1) G-tetrads required for ODN stacking were compatible with INH-ODN activity but were not necessary; 2) there was no relationship between activity and self-association at endosomal pH; 3) there was no evidence for direct binding between ST-ODNs and INH-ODNs; 4) when a 3G sequence was disrupted, despite a preserved stem-loop formation, INH-ODN activity was abolished. These results support the conclusion that certain features of the primary linear sequence are critical for TLR9 inhibition, but changes in secondary structure or in ODN aggregation are irrelevant.

Cell Line

Oligonucleotides - chemistry

Toll-Like Receptor 9 - antagonists & inhibitors

Humans

Oligodeoxyribonucleotides - chemistry

B-Lymphocytes

Polymerization

Structure-Activity Relationship

Animals

Oligonucleotides - pharmacology

Base Sequence

Protein Binding

Mice

Oligodeoxyribonucleotides - pharmacology

Toll-Like Receptor 9 - metabolism

Binding Sites

Details

Title: Subtitle: Aggregation and secondary loop structure of oligonucleotides do not determine their ability to inhibit TLR9
Creators: Robert F Ashman - Department of Internal Medicine, Carver College of Medicine, The University of Iowa, Iowa City, IA 52242, United States
J Adam Goeken
Petar S Lenert
Resource Type: Journal article
Publication Details: International immunopharmacology, Vol.11(8), pp.1032-1037
DOI: 10.1016/j.intimp.2011.02.023
PMID: 21376154
PMCID: PMC3119753
NLM abbreviation: Int Immunopharmacol
ISSN: 1567-5769
eISSN: 1878-1705
Grant note: R01 AI047374 / NIAID NIH HHS AI047374 / NIAID NIH HHS R56 AI064736 / NIAID NIH HHS AI064736 / NIAID NIH HHS R01 AI047374-07 / NIAID NIH HHS R56 AI064736-01A2 / NIAID NIH HHS
Language: English
Date published: 08/2011
Academic Unit: Pathology; Immunology; Internal Medicine
Record Identifier: 9984094710302771

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