Potent CpG oligonucleotides containing phosphodiester linkages: in vitro and in vivo immunostimulatory properties.
Bacterial and synthetic DNAs, containing CpG dinucleotides in specific sequence contexts, activate the vertebrate immune system. Unlike phosphorothioate (PS) CpG DNAs, phosphodiester (PO) CpG DNAs require either palindromic sequences and/or poly(dG) sequences at the 3(')-end for activity. Here, we report 'PO-immunomers' having two PO-CpG DNA molecules joined through their 3(')-ends. These PO-imunomers permitted us, for the first time, to assess immunostimulatory properties of PO-CpG DNAs in vitro and in vivo without the need for palindromic and/or poly(dG) sequences. In medium containing 10% fetal bovine serum, PO-immunomers were more resistant than PO-CpG DNAs to nucleases. Compared to PS-CpG DNA in BALB/c and C3H/HeJ mice spleen cell culture assays, PO-immunomers showed increased IL-12 secretion and minimal amounts of IL-6 secretion. PO-immunomers activated NF-kappa B and induced cytokine secretion in J774 cell cultures. In addition, PO-immunomers showed antitumor activity in nude mice bearing human breast (MCF-7) and prostate (DU145) cancer xenografts.
Yu
Dong
D
Hybridon, Inc., 345 Vassar Street, Cambridge, MA 02139, USA.
Zhu
Fu-Gang
FG
Bhagat
Lakshmi
L
Wang
Hui
H
Kandimalla
Ekambar R
ER
Zhang
Ruiwen
R
Agrawal
Sudhir
S
D000276:Adjuvants, Immunologic; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D018899:CpG Islands; D016207:Cytokines / Q000378:metabolism; D003851:Deoxyribonucleases / Q000378:metabolism; D004795:Enzyme Stability; D007111:Immunity, Cellular / Q000187:drug effects*; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D016328:NF-kappa B / Q000378:metabolism; D009841:Oligonucleotides / Q000737:chemistry / Q000494:pharmacology*; D013154:Spleen / Q000166:cytology; D014407:Tumor Cells, Cultured
D000276:Adjuvants, Immunologic; D000970:Antineoplastic Agents; D016207:Cytokines; D016328:NF-kappa B; D009841:Oligonucleotides; D003851:Deoxyribonucleases