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By J. Thomas August (Eds.)

ISBN-10: 0120329417

ISBN-13: 9780120329410

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Internal modifications of base, sugar, and backbone have also been reported to enhance nuclease stability at or near the modified nucleoside (Manoharan, 1993). , 1995). The demonstration that modifications may induce nuclease stability sufficient to enhance activity in cells in tissue culture and in animals has proved to be much more complicated because of the presence of 5'exonucleases and endonucleases. , 1991). In fact, even a 5-nucleotide-long phosphodiester gap in the middle of a phosphoro- 34 Stanley T.

1992). In the latter study, the oligonucleotides that displayed natural killer (NK) cell-stimulating activity contained specific palindromic sequences and tended to be guanosine rich. Collectively, these observations indicate that nucleic acids may have broad immunostimulatory activity. It has been shown that phosphorothioate oligonucleotides stimulate B lymphocyte proliferation in a mouse splenocyte preparation (analogous t o bacterial DNA) (Psietsky and Reich, 1993), and the response may underlie the observations of lymphoid hyperplasia in the spleen and lymph nodes of rodents caused by repeated administration of these compounds (see Section V,F,3).

In this study, both human keratinocytes and an in vitro model of human skin released interleukin la (IL-la) when treated with 250 mM to 1 mM of phosphorothioate oligonucleotides. The effects seemed to be dependent on the phosphorothioate backbone and independent of sequence Pharmacology of Antisense Oligonucleotides 27 or 2' modification. This has been extrapolated to suggest that the CpG motif may be required for immune stimulation of oligonucleotide analogs such as phosphorothioates. This clearly is not the case with regard to release of I L - l a from keratinocytes (S.

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Advances in Pharmacology by J. Thomas August (Eds.)

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