Table III.

Effect of distance between motifs on ODN activitya

ODNCytokine-Producing Cells (% maximum)
IL-6IL-12IFN-γ
CpG ODNb100 ± 11100 ± 7100 ± 10
CpG ODNb + Cont ODN97 ± 1498 ± 9100 ± 17
CpG ODNb + Sup ODN16 ± 621 ± 618 ± 5
LPS100 ± 299 ± 2100 ± 2
LPS+ Sup ODN94 ± 794 ± 592 ± 7
[CpG-Sup] ODNb87 ± 12>100 ± 1492 ± 14
[CpG-5 bases-Sup] ODNb>100 ± 4>100 ± 21>100 ± 22
[CpG-10 bases-Sup] ODN38 ± 664 ± 1542 ± 7
[CpG-20 bases-Sup] ODN7 ± 448 ± 1324 ± 8
[CpG-Cont] ODNb94 ± 7>100 ± 1499 ± 11
[Sup-CpG] ODN0 ± 00 ± 00 ± 0
[Sup-20 bases-CpG] ODN8 ± 59 ± 32 ± 1
[CpG-Sup] ODNb + CpG ODNb>100 ± 16>100 ± 1598 ± 13
[CpG-5 bases-Sup] ODNb + CpG ODNb>100 ± 18>100 ± 1198 ± 20
[CpG-10 bases-Sup] ODN+ CpG ODNb58 ± 775 ± 966 ± 9
[CpG-20 bases-Sup] ODN+ CpG ODNb27 ± 526 ± 1030 ± 8
[Sup-CpG] ODN+ CpG ODNb9 ± 411 ± 48 ± 5
[Sup-20 bases-CpG] ODN+ CpG ODNb5 ± 19 ± 313 ± 2
  • a BALB/c spleen cells were stimulated in vitro with 1 μM of each ODN (or 5 μg/ml LPS), and the number of cells activated to secrete cytokine was determined 8 h later by ELISPOT. The percentage of cells activated to secrete cytokine was calculated by the formula: (number of cells activated by test ODN) − (background)/(number of cells activated by CpG ODN) − (background) × 100%. Multiple combinations of the CpG, suppressive and control 10-mer motifs described in Table II were used in these studies and gave similar results in these experiments. Results represent the average of two to four assays per data point. Table II shows typical numbers of cytokine-secreting cells per 106 cells.

  • b Stimulatory ODN, p < 0.05.