Document Detail


Optimal oligonucleotide sequences for TLR9 inhibitory activity in human cells: lack of correlation with TLR9 binding.
MedLine Citation:
PMID:  21393636     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Toll-like receptor (TLR)9 performs our innate response to bacterial DNA, warning us of the presence of infection. Inhibitory oligodeoxyribonucleotides (INH-ODN) have been developed that selectively block activation of mouse TLR9. Their inhibitory motif consisting of CCx(not-C)(not-C)xxGGG (x = any base) also reduces anti-DNA antibodies in lupus mice. The current study demonstrates that this motif also provides the sequences required to block TLR9 in human B cells and human embryonic kidney (HEK) cells transfected with human TLR9. However, extending the sequence by four to five bases at the 5' end enhanced activity and this enhancement was greater when a phosphorothioate (pS) backbone replaced the native phosphodiester (pO) backbone. A series of pO-backbone INH-ODN representing a 500-fold range of activity in biologic assays was shown to cover less than a 2.5-fold range of avidity for binding human TLR9-Ig fusion protein, eliminating TLR9 ectodomain binding as the explanation for sequence-specific differences in biologic activity. With few exceptions, the relative activity of INH-ODN in Namalwa cells and HEK/human TLR9 cells was similar to that seen in mouse B cells. INH-ODN activity in human peripheral blood B cells correlated significantly with the cell line data. These results favor the conclusion that although the backbone determines strength of TLR9 binding, critical recognition of the INH-ODN sequence necessary for biologic activity is performed by a molecule that is not TLR9. These studies also identify the strongest INH-ODN for human B cells, helping to guide the selection of INH-ODN sequences for therapeutics in any situation where inflammation is enhanced by TLR9.
Authors:
Robert F Ashman; J Adam Goeken; Eicke Latz; Petar Lenert
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural    
Journal Detail:
Title:  International immunology     Volume:  23     ISSN:  1460-2377     ISO Abbreviation:  Int. Immunol.     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2011-03-11     Completed Date:  2011-07-05     Revised Date:  2013-06-30    
Medline Journal Info:
Nlm Unique ID:  8916182     Medline TA:  Int Immunol     Country:  England    
Other Details:
Languages:  eng     Pagination:  203-14     Citation Subset:  IM    
Affiliation:
Carver College of Medicine, University of Iowa, Iowa City, IA 52242, USA.
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MeSH Terms
Descriptor/Qualifier:
Animals
Anti-Inflammatory Agents / pharmacology
B-Lymphocytes / immunology*
Base Sequence
Cell Line
DNA-Binding Proteins*
Flow Cytometry
Humans
Mice
Molecular Sequence Data
Oligodeoxyribonucleotides / chemistry,  genetics,  pharmacology*
Phosphorothioate Oligonucleotides / chemistry,  pharmacology
Protein Binding / drug effects
Toll-Like Receptor 9 / antagonists & inhibitors*,  immunology
Grant Support
ID/Acronym/Agency:
R01 AI083713/AI/NIAID NIH HHS; R01 HL093262/HL/NHLBI NIH HHS; R01AI0437374/AI/NIAID NIH HHS
Chemical
Reg. No./Substance:
0/Anti-Inflammatory Agents; 0/DNA-Binding Proteins; 0/Oligodeoxyribonucleotides; 0/Phosphorothioate Oligonucleotides; 0/Toll-Like Receptor 9
Comments/Corrections

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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