Table I.

Rules governing K ODN-induced immune activationa

Assay Subject No.Stimulation Index
IL-6ProliferationIgM
121212
A. Multiple CpGs induce more stimulation
1ATCGACTCTCGAGCGTTCTC5057133013871
2TCGAGCGTTCTC354015371979
3TCGAGGCTTCTC281225822
4TGCAGGCTTCTC105754
B. Minimum size of stimulatory K ODN
1TCGACTCTCGAGCGTTCTC (20)20233772>100100
2ACTCTCGAGCGTTCTC (16)20183046>100100
3TCTCGAGCGTTCTC (14)16402841>100100
4TCGAGCGTTCTC (12)23151325>10080
5CTCGAGCGTTCT (12)211416219592
6TCGAGCGTTCT (11)10415354578
7TCGAGCGTTC (10)6517403582
8TCGAGCGTT (9)17573225
9TCGAGCGT (8)151132512
10TCGAGCG (7)111594
C. CpG motifs located at the 5′ end of the ODN are most stimulatory
1TCGAGCGTTCTC1240525980>100
2TCGAGGCTTCTC6125161>100>100
3TGCTTCGAGCTC4312162060
4GCGAGGCTTCTC5121814>100>100
5TGCAGCGAGCTC5244116
6CGAGCGTTCTC<1<111<12
D. Optimization of the 5′CpG flanking region
1TCGATGCTTCTC51260671009
2TCGAGGCTTCTC61251611605
3ACGAGGCTTCTC33182311011
4GCGAGGCTTCTC311814726
5CCGAGGCTTCTC411525257
6TGCTTCGAGCTC3112166040
7TGCAGCGAGCTC2144168
E. Optimization of the 3′ CpG flanking region
1TCGTTTGTTCTC882831>100>100
2TCGTATGTACTC882632233
3TCGGATGAGCTC689202841
4TCGAATGCTCTC351422614
5TTGTTCGTTCTC3415141926
6TTGTTCGTACTC2414131572
7TTGTTCGAGCTC2464616
8TTGTTCGAACTC2211511
  • a Over 200 novel ODN were synthesized, and their ability to activate PBMC from multiple donors was examined. PBMC were stimulated for 72 h in the presence of ODN (1 μM added at time 0). Cytokine and Ab secretion in the supernatants were assessed by ELISA, and proliferation was determined by [3H] uptake. Examples of general findings are presented in this table. Results are expressed as fold increase over unstimulated cells.