Sequence motifs in adenoviral DNA block immune activation by stimulatory CpG motifs.
Unmethylated CpG dinucleotides in particular base contexts (CpG-S motifs) are relatively common in bacterial DNA but are rare in vertebrate DNA. B cells and monocytes have the ability to detect such CpG-S motifs that trigger innate immune defenses with production of Th1-like cytokines. Despite comparable levels of unmethylated CpG dinucleotides, DNA from serotype 12 adenovirus is immune-stimulatory, but serotype 2 is nonstimulatory and can even inhibit activation by bacterial DNA. In type 12 genomes, the distribution of CpG-flanking bases is similar to that predicted by chance. However, in type 2 adenoviral DNA the immune stimulatory CpG-S motifs are outnumbered by a 15- to 30-fold excess of CpG dinucleotides in clusters of direct repeats or with a C on the 5' side or a G on the 3' side. Synthetic oligodeoxynucleotides containing these putative neutralizing (CpG-N) motifs block immune activation by CpG-S motifs in vitro and in vivo. Eliminating 52 of the 134 CpG-N motifs present in a DNA vaccine markedly enhanced its Th1-like function in vivo, which was increased further by the addition of CpG-S motifs. Thus, depending on the CpG motif, prokaryotic DNA can be either immune-stimulatory or neutralizing. These results have important implications for understanding microbial pathogenesis and molecular evolution and for the clinical development of DNA vaccines and gene therapy vectors.
Interdisciplinary Immunology Program and Department of Internal Medicine, University of Iowa College of Medicine, Iowa City, IA 52242, USA. arthur-krieg@uiowa.edu
A M
AM
Krieg
T
T
Wu
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R
Weeratna
S M
SM
Efler
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L
Love-Homan
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L
Yang
A K
AK
Yi
D
D
Short
H L
HL
Davis
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D001483:Base Sequence; D018899:CpG Islands / Q000502:physiology*; D016207:Cytokines / Q000096:biosynthesis; D004279:DNA, Viral / Q000235:genetics*; D005260:Female; D051379:Mice; D008807:Mice, Inbred BALB C; D018417:Th1 Cells / Q000276:immunology
D016207:Cytokines; D004279:DNA, Viral