Effect of chemical modifications of cytosine and guanine in a CpG-Motif of oligonucleotides: Structure-immunostimulatory activity relationships

被引:70
作者
Kandimalla, ER [1 ]
Yu, D [1 ]
Zhao, QY [1 ]
Agrawal, S [1 ]
机构
[1] Hybridon Inc, Cambridge, MA 02139 USA
关键词
D O I
10.1016/S0968-0896(00)00316-3
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Oligodeoxynucleotides containing unmethylated CpG-motifs stimulate the innate immune system, including inducing B-cell proliferation and cytokine production. However, the mechanism of immunostimulation by CpG-oligonucleotides and the precise structural requirements and specific functional groups of cytosine and guanine necessary for recognition of and interaction with protein/receptor factors that are responsible for immune stimulation have not been elucidated. We sought to understand the critical role of each functional group of the cytosine and guanine moieties in a CpG-motif in inducing immunostimulatory activity. To this end, we examined structure-immunostimulatory activity relationships of phosphorothio ate oligodeoxynucleotides (PS-oligos) containing YpG- and CpR-motifs (Y and R stand for pyrimidine and purine analogues, respectively). The PS-oligos containing a YpG- motif in which the natural deoxycytidine was replaced with deoxy-5-hydroxycytidine or deoxy-N4-ethylcytidine showed immunostimulatory activity. Substitution of deoxycytidine with a deoxy-5-methylisocytidine, deoxyuridine, or deoxy-P-base-nucleoside in the YpG-motif completely abolished the immune stimulatory activity, similar to the results observed with deoxy-5-methylcytidine. In the case of PS-oligos containing a CpR-motif, 7-deazaguanine substitution for natural guanine showed immunostimulatory activity similar to that of a parent PS-oligo. These studies suggest that the 2-keto, 3-imino and 4-amino groups of cytosine, and the 1-imino, 2-amino and 6-keto groups of guanine in a CpG-motif are important for the immunostimulatory activity of CpG-PS-oligos. The absence of N7 on guanine of the CpG-motif does not affect immunostimulatory activity significantly. These studies suggest that it is possible to develop YpG- and CpR-motifs as an alternative to CpG-motifs in PS-oligos for immunostimulatory studies. (C) 2001 Published by Elsevier Science Ltd. All rights reserved.
引用
收藏
页码:807 / 813
页数:7
相关论文
共 23 条
[1]   Antisense therapeutics: is it as simple as complementary base recognition? [J].
Agrawal, S ;
Kandimalla, ER .
MOLECULAR MEDICINE TODAY, 2000, 6 (02) :72-81
[2]  
Carpentier AF, 2000, CLIN CANCER RES, V6, P2469
[3]   CpG oligodeoxynucleotides act as adjuvants that switch on T helper 1 (Th1) immunity [J].
Chu, RS ;
Targoni, OS ;
Krieg, AM ;
Lehmann, PV ;
Harding, CV .
JOURNAL OF EXPERIMENTAL MEDICINE, 1997, 186 (10) :1623-1631
[4]   Repeated administration of cytosine-phosphorothiolated guanine-containing oligonucleotides together with peptide/protein immunization results in enhanced CTL responses with anti-tumor activity [J].
Davila, E ;
Celis, E .
JOURNAL OF IMMUNOLOGY, 2000, 165 (01) :539-547
[5]  
DUNFORD PJ, 1997, ANTISENSE 97 TARGETI, P40
[6]   CpG oligodeoxynucleotides and interleukin-12 improve the efficacy of Mycobacterium bovis BCG vaccination in mice challenged with M-tuberculosis [J].
Freidag, BL ;
Melton, GB ;
Collins, F ;
Klinman, DM ;
Cheever, A ;
Stobie, L ;
Suen, W ;
Seder, RA .
INFECTION AND IMMUNITY, 2000, 68 (05) :2948-2953
[7]  
Häcker H, 2000, CURR TOP MICROBIOL, V247, P77
[8]   CpG motifs present in bacterial DNA rapidly induce lymphocytes to secrete interleukin 6, interleukin 12, and interferon gamma [J].
Klinman, DM ;
Yi, AK ;
Beaucage, SL ;
Conover, J ;
Krieg, AM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1996, 93 (07) :2879-2883
[9]   Mechanisms and therapeutic applications of immune stimulatory CpG DNA [J].
Krieg, AM ;
Yi, AK ;
Hartmann, G .
PHARMACOLOGY & THERAPEUTICS, 1999, 84 (02) :113-120
[10]   CPG MOTIFS IN BACTERIAL-DNA TRIGGER DIRECT B-CELL ACTIVATION [J].
KRIEG, AM ;
YI, AK ;
MATSON, S ;
WALDSCHMIDT, TJ ;
BISHOP, GA ;
TEASDALE, R ;
KORETZKY, GA ;
KLINMAN, DM .
NATURE, 1995, 374 (6522) :546-549