Immunostimulatory oligodeoxynucleotide isolated from genome wide screening of Mycobacterium bovis chromosomal DNA.
Bacterial DNA has a variety of immunostimulatory activities, such as the activation of B cells and natural killer cells, the induction of interferon-gamma, and the induction of Th1-type immune responses. In contrast, mammalian DNA does not have these activities. To evaluate the genomic DNA sequences of Mycobacterium bovis that have immunostimulatory activity, we used a computer to analyze the M. bovis genome and we designed a series of synthetic, 20 base length, phosphodiester backbone oligodeoxynucleotides (ODNs) that contain CpG motifs (MB-ODNs). We screened the immunostimulatory MB-ODNs that induce the activation of the NF-kappaB-responsive IL-8 promoter in RAW 264.7 cells. Our experimental analyses demonstrate that the potent CpG DNA in the M. bovis genome has functional effects as a Th1-responsive adjuvant, and that it activates the transcription factor NF-kappaB. Moreover, we found that both the CpG motifs and the context of the sequence surrounding the CpG motif are important for the immunostimulatory activities. The identification of the potent immunostimulatory DNA sequence in a native bacterial genome may give insights to the optimal sequence for well-controlled immune responses.
Lee
Keun-Wook
KW
Research Institute for Bioscience & Biotechnology, College of Medicine, Hallym University, Gangwon 200-702, Republic of Korea.
Jung
Jinwon
J
Lee
Younghee
Y
Kim
Tae-Yoon
TY
Choi
Soo-Young
SY
Park
Jinseu
J
Kim
Doo-Sik
DS
Kwon
Hyung-Joo
HJ
D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology; D002460:Cell Line; D018899:CpG Islands / Q000276:immunology*; D004269:DNA, Bacterial / Q000276:immunology* / Q000494:pharmacology; D016680:Genome, Bacterial / Q000276:immunology*; D006801:Humans; D016209:Interleukin-8 / Q000276:immunology; D007694:Killer Cells, Natural / Q000276:immunology; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology; D051379:Mice; D009163:Mycobacterium bovis / Q000276:immunology*; D016328:NF-kappa B / Q000276:immunology; D009838:Oligodeoxyribonucleotides / Q000276:immunology* / Q000494:pharmacology; D011401:Promoter Regions, Genetic / Q000276:immunology; D018417:Th1 Cells / Q000276:immunology
D000276:Adjuvants, Immunologic; D004269:DNA, Bacterial; D016209:Interleukin-8; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides