Efficacy of adenoviral TNFα antisense is enhanced by a macrophage specific promoter

P. Sidiropoulos, H. Liu, S. Mungre, L. Anderson, B. Thimmapaya, R. M. Pope*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

4 Scopus citations

Abstract

Macrophage-derived TNFα is a critical mediator of inflammation and destruction in diseases such as rheumatoid arthritis and Crohn's disease. These studies were undertaken to develop an effective adenovirus-based strategy to specifically suppress and TNFα in primary human macrophages. A variety of promoters and LTRs were evaluated for effective expression in the macrophage cell line RAW 264.7. The CMV promoter and the Visna LTR were the most strongly expressed and were therefore used to drive the expression of TNFα antisense fragments. In transient transfection assays, the antisense fragment terminating at the 3′ end of the first exon (216 bp) was superior to the others (70 and 750 bp), when expressed under the control of either the CMV promoter or the Visna LTR. Adenoviral vectors expressing the 216 bp TNFα antisense fragment, controlled by the CMV promoter or the Visna LTR, were both effective at suppressing LPS-induced TNFα secretion by primary human macrophages. However, the Visna LTR was more effective not only at suppressing LPS-induced TNFα secretion, but also IL-6, which is highly sensitive to TNFα secretion. These results demonstrate that effective, specific, suppression of TNFα in macrophages is possible, employing a directed antisense approach and a promoter system that is highly efficient in human macrophages.

Original languageEnglish (US)
Pages (from-to)223-231
Number of pages9
JournalGene therapy
Volume8
Issue number3
DOIs
StatePublished - 2001

Keywords

  • Antisense
  • Macrophages
  • TNFα

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Genetics

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