The in vitro and in vivo pharmacology of antisense oligonucleotides targeted to murine Stat6.
OBJECTIVE
This study was designed to establish whether phosphorothioate (PS) antisense oligonucleotides (AS-ODN) targeted to Stat6 were active in vivo in a mouse model of active sensitisation.


METHODS
Female Balb/c mice (6-8) per group were used for in vivo study.


METHODS
Mice were treated with active PS AS-ODNs determined in initial in vitro studies. Compounds were dosed daily (3-30mg/kg i.v.) over the course of sensitisation to ovalbumin.


METHODS
Stat6 mRNA and protein levels were determined in the spleen after treatment (quantitative northern and western analysis respectively), in addition to serum IgE (ELISA). ANOVA was used to determine any significant differences between groups.


RESULTS
Both of the AS-ODNs tested in vivo, down regulated Stat6 mRNA and protein levels in the spleen by 40-50% although there was no effect on serum IgE. These treatments also induced splenomegaly in vivo and caused splenocyte proliferation in vitro.


CONCLUSIONS
The AS-ODNs used can down regulate Stat6 mRNA and protein although not sufficiently to influence IgE-levels. These effects are likely to be complicated in vivo by the immune-stimulation evident as splenomegaly.
Danahay
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Novartis Horsham Research Centre, West Sussex, UK. henry.danahay@pharma.novartis.com
Hill
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Natt
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Owen
C E
CE
D000818:Animals; D015152:Blotting, Northern; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005347:Fibroblasts; D007073:Immunoglobulin E / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D050800:STAT6 Transcription Factor; D015398:Signal Transduction / Q000187:drug effects; D013154:Spleen / Q000166:cytology / Q000187:drug effects; D013163:Splenomegaly / Q000139:chemically induced / Q000473:pathology; D015534:Trans-Activators / Q000187:drug effects*
D016376:Oligonucleotides, Antisense; D050800:STAT6 Transcription Factor; C494100:Stat6 protein, mouse; D015534:Trans-Activators; D007073:Immunoglobulin E