A sense phosphorothioate oligonucleotide directed to the initiation codon of transcription factor NF-kappa B p65 causes sequence-specific immune stimulation.
Antisense oligonucleotides have proved effective in achieving targeted inhibition of gene expression. In such experiments, sense oligonucleotides have frequently been used as a control for nonspecific effects, but the results have been variable, raising questions about the reliability of sense oligomers as a control. It is possible that some of the effects of sense oligonucleotides may be specific. We have shown that phosphorothioate antisense oligonucleotides to the p65 subunit of NF-kappa B, a transcription factor, cause a block in cell adhesion. In our efforts to test the efficacy of NF-kappa B p65 oligonucleotides in vivo, we unexpectedly observed that the control p65-sense, but not the p65-antisense, oligonucleotides caused massive splenomegaly in mice. In the current study we demonstrate a sequence-specific stimulation of splenic cell proliferation, both in vivo and in vitro, by treatment with p65-sense oligonucleotides. Cells expanded by this treatment are primarily B-220+, sIg+ B cells. The secretion of immunoglobulins by the p65-sense oligonucleotide-treated splenocytes is also enhanced. In addition, the p65-sense-treated splenocytes, but not several other cell lines, showed an upregulation of NF-kappa B-like activity in the nuclear extracts, an effect not dependent on new protein or RNA synthesis. These results demonstrate that phosphorothioate oligonucleotides can exert sequence-specific effects in vivo, irrespective of sense or antisense orientation.
McIntyre
K W
KW
Department of Inflammation/Autoimmune Diseases, Roche Research Center, Hoffmann-La Roche, Inc., Nutley, New Jersey 07110.
Lombard-Gillooly
K
K
Perez
J R
JR
Kunsch
C
C
Sarmiento
U M
UM
Larigan
J D
JD
Landreth
K T
KT
Narayanan
R
R
D000818:Animals; D001402:B-Lymphocytes / Q000166:cytology / Q000187:drug effects*; D001483:Base Sequence; D001665:Binding Sites; D002448:Cell Adhesion / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D008213:Lymphocyte Activation / Q000187:drug effects*; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000235:genetics* / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D013154:Spleen / Q000166:cytology / Q000187:drug effects*; D013163:Splenomegaly / Q000139:chemically induced; D013873:Thionucleotides / Q000494:pharmacology*; D015854:Up-Regulation
D007074:Immunoglobulin G; D007075:Immunoglobulin M; D016328:NF-kappa B; D009838:Oligodeoxyribonucleotides; D013873:Thionucleotides