Immunostimulatory nucleic acids
An immune stimulation and nucleic acid technology, applied in biocides, organic chemistry, animal husbandry, etc., can solve the problem of weak activation ability
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Embodiment 1
[0463] The levels of interferon-α (IFN-α), IFN-γ, IL-10, IL-6, and TNF-α secreted by human PBMC after exposure to the CpG oligonucleotides described herein are shown in the accompanying drawings 1-5. The detection oligonucleotides are indicated by ▲ in the figure. Oligonucleotides serving as positive control oligonucleotides are indicated by -. Detection oligonucleotides shown in Figures 1A, 2A, 3A, 4A, and 5A include SEQ ID NO:322, SEQ ID NO:323, and SEQ ID NO:324. Detection oligonucleotides shown in Figures 1B, 2B, 3B, 4B, and 5B include SEQ ID NO:325, SEQ ID NO:326, SEQ ID NO:327, and SEQ ID NO:328. The concentration of oligonucleotide used to generate each data point is indicated along the X-axis (μΜ). Below the graphs are listed the levels of cytokines secreted by cells treated with a negative control (medium), and in some cases LPS was also used in each experiment.
[0464] As demonstrated in Figures 1-5, the oligonucleotides tested in the assay were able to elicit s...
Embodiment 2
[0466] The level of CD69 expression (MFI) of NK cells in response to treatment with the detection oligonucleotide relative to the control oligonucleotide was examined. CD69 expression is a hallmark of T cell and NK cell activation. Cells exposed to detection oligonucleotides are indicated by ▲ in FIG. 6 . Oligonucleotides serving as positive control oligonucleotides are indicated by -. Detection oligonucleotides shown in Figure 6A include SEQ ID NO:322, SEQ ID NO:323, and SEQ ID NO:324. Detection oligonucleotides shown in Figure 6B include SEQ ID NO:325, SEQ ID NO:326, SEQ ID NO:327, and SEQ ID NO:328. The positive control oligonucleotide used in these studies was SEQ ID NO:329. The levels of CD69 expression in T cells and NK cells treated with negative control (medium) and LPS for each experiment are listed below the graph.
[0467] As demonstrated in Figure 6, the oligonucleotides tested in the assay were able to induce the expression of CD69 at levels comparable to or h...
Embodiment 3
[0469] The levels of interferon-α (IFN-α) and IL-10 secreted by human PBMC following exposure of these cells to the CpG oligonucleotides described herein are shown in Figures 7-12 and 17 . The detected nucleotides are indicated by ■ in the figure. The oligonucleotide of SEQ ID NO: 242 as the positive control oligonucleotide is indicated by ●. The oligonucleotide of SEQ ID NO: 330 as a negative control oligonucleotide is indicated by ♦. Figure 7A and 7B The detection oligonucleotide shown is SEQ ID NO:313. Figure 8A and Figure 8BThe detection oligonucleotide shown is SEQ ID NO:314. Figure 9A and 9B The detection oligonucleotide shown is SEQ ID NO:319. Figure 10A and Figure 10B The detection oligonucleotide shown is SEQ ID NO:316. Figure 11A and 11B The detection oligonucleotide shown is SEQ ID NO:317. Figure 12A and 12B The detection oligonucleotide shown is SEQ ID NO:320. Figure 17A and 17B The detection oligonucleotide shown is SEQ ID NO:321. The concen...
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Abstract
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