Inhibition of respiratory syncytial virus replication by antisense oligodeoxyribonucleotides

• Published in: Antiviral research 1997-02, Vol.33 (3), p.201-213
• Main Authors: Jairath, S, Brown Vargas, P, Hamlin, H.A, Field, A.K, Kilkuskie, R.E
• Format: Article
• Language: English
• Subjects: Antibiotics. Antiinfectious agents. Antiparasitic agents; Antigens, Viral - biosynthesis; Antisense inhibition; Antiviral agents; Antiviral Agents - pharmacology; Antiviral Agents - toxicity; Base Sequence; Biological and medical sciences; Cell Division - drug effects; DNA, Viral; Genome, Viral; HN Protein; Humans; Medical sciences; Molecular Sequence Data; Oligonucleotides, Antisense - pharmacology; Oligonucleotides, Antisense - toxicity; Pharmacology. Drug treatments; Phosphorothioate oligonucleotides; Respiratory syncytial virus; Respiratory Syncytial Viruses - drug effects; Respiratory Syncytial Viruses - genetics; Respiratory Syncytial Viruses - growth & development; Respiratory Syncytial Viruses - physiology; RNA, Viral; Tumor Cells, Cultured; Viral Envelope Proteins; Viral Proteins - biosynthesis; Virus Replication - drug effects
• ISSN: 0166-3542; 1872-9096
• DOI: 10.1016/S0166-3542(96)01015-7

Oligodeoxyribonucleotides targeted against respiratory syncytial virus (RSV) genomic RNA inhibited RSV replication in cell culture by an apparent antisense mechanism. HEp-2 cells were infected with RSV strain A2 and incubated in the presence of oligonucleotides. Virus replication was measured by enzyme-linked immunosorbent assay (ELISA), virus yield assay, or production of specific RSV mRNAs. Using ELISA, 50% effective concentration (EC 50) values were about 0.5–1 μM for an antisense oligonucleotide targeted to the start of the NS2 gene. All oligonucleotides inhibited virus antigen production as measured by ELISA. In all assays, this antisense oligonucleotide was more potent than: (1) a control oligonucleotide containing the reverse sequence; (2) oligonucleotides targeted at RSV mRNA; (3) a random sequence oligonucleotide; and (4) ribavirin. Reverse transcriptase polymerase chain reaction (RT-PCR) showed sequence-specific depletion of the genomic RNA target following treatment of cells with the antisense oligonucleotide. Specific cleavage of the genomic target RNA has been detected at the antisense oligonucleotide binding site, suggesting that cellular RNase H participates in the reaction. These results indicate that antisense oligonucleotides targeted against RSV genomic RNA can effectively inhibit RSV replication and may have therapeutic value.