The antiviral activity of RNA-dye combinations

The results of our previous studies (Jamison et al. 1988, 1989, 1990 a, b, c, d, e) have shown that the ability of intercalative dyes to modulate the antiviral activity of poly r(A-U) is related to the groove through which the dyes intercalate into the poly r(A-U). When poly r(A-U) is combined with the minor groove intercalating dyes or the minor/major groove intercalating dyes, optimum enhancement of antiviral activity is observed at the dye/ribonucleotide ratio predicted by the neighbor exclusion model (usually 1/4 or 1/6). No enhancement is observed when poly r(A-U) is combined with major groove intercalating dyes. When poly r(A-U) is combined with additional intercalative dyes to produce a dye/ribonucleotide ratio of 1/4 and a ribonucleotide concentration of 200 microM, the antiviral activity of poly r(A-U) is enhanced 8- to 20-fold, while 50% effective doses of the poly r(A-U) and the dyes decreases 18- to 347-fold. Interferon neutralization assays demonstrate that the interferon-inducing capability of the dye/poly r(A-U) combinations approximates the sum of the interferon-inducing capabilities of the poly r(A-U) and the dyes employed and suggests that the dyes potentiate the antiviral activity of poly r(A-U) without affecting the amount of interferon induced. Direct viral inactivation studies demonstrate that the dyes, poly r(A-U), and the dye/poly r(A-U) combinations do not inactivate VSV at concentrations near the 50% viral inhibitory dose. Assessment of cytotoxicity by microscope examination of HSF cell morphology and trypan blue exclusion indicates that the dye/poly r(A-U) combinations exhibit antiviral activity at concentrations well below those that induce cyto-toxicity. Several of the dyes and the dye/poly r(A-U) combinations exhibit anti-HIV-1 activity, suggesting that the enhancement phenomenon is not virus-specific nor host cell-specific. The enhancement phenomenon is sensitive to the base sequence of the polynucleotide with dye/poly r(A-U) and dye/poly r(G-C) combinations displaying enhanced antiviral activity, while dye/poly (rI).poly (rC) and dye/poly d(A-T) combinations do not. These results suggest that while intercalation of the dye and interferon induction are necessary for enhanced antiviral activity, neither intercalation nor interferon induction alone is sufficient to potentiate the antiviral activity of polyribonucleotides.