Pisetsky D S, Reich C F
Medical Service, Durham VA Medical Center, NC 27705, USA.
Immunopharmacology. 1998 Nov;40(3):199-208. doi: 10.1016/s0162-3109(98)00044-7.
To assess the influence of base sequence on the immunostimulatory activities of DNA, cell binding and mitogenicity of a series of 30-mer phosphodiester oligonucleotides were tested using murine spleen cells. These compounds consisted of either a single base or a six base CpG motif in the context of 5' and 3' flanking sequences of each base. Among fluoresceinated oligonucleotides, (dG)30 had the highest binding of single base compounds tested while the presence of dG flanks increased binding of compounds with six base motifs, whether active on inactive. In assays of mitogenesis including incorporation of thymidine and uridine as well as expression of cell surface CD69, (dG)30 induced the highest responses among single base compounds. Among compounds with an active six base motif, the extent of proliferation varied with flanking sequence, with dG flanks producing the greatest stimulation in all assays tested. Together, these findings indicate that a variety of base sequences may affect the immunomodulatory properties of DNA, with the activity of dG sequences perhaps resulting from the formation of variant DNA structures.
为评估碱基序列对DNA免疫刺激活性的影响，使用小鼠脾细胞测试了一系列30聚体磷酸二酯寡核苷酸的细胞结合和促有丝分裂活性。这些化合物在每个碱基的5'和3'侧翼序列背景下，由单个碱基或六碱基CpG基序组成。在荧光标记的寡核苷酸中，(dG)30在测试的单个碱基化合物中具有最高的结合能力，而dG侧翼的存在增加了具有六碱基基序的化合物的结合能力，无论其是否具有活性。在包括胸苷和尿苷掺入以及细胞表面CD69表达的促有丝分裂测定中，(dG)30在单个碱基化合物中诱导了最高反应。在具有活性六碱基基序的化合物中，增殖程度随侧翼序列而变化，在所有测试的测定中，dG侧翼产生的刺激最大。总之，这些发现表明，多种碱基序列可能影响DNA的免疫调节特性，dG序列的活性可能源于变体DNA结构的形成。
