In vitro evaluation of an enhanced human Serum Amyloid A (SAA2) promoter-regulated soluble TNF receptor fusion protein for anti-inflammatory gene therapy

M. Rygg, C. M. Uhlar, C. Thorn, L. E. Jensen, D. J. Gaughan, A. W. Varley, R. S. Munford, R. Göke, Y. Chen, A. S. Whitehead

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

Tumour necrosis factor (TNF)-α contributes to the pathogenesis of many inflammatory diseases. Recombinant soluble TNF receptor fusion proteins (sTNFR:Ig) are potent TNF antagonists, both in vitro and in vivo. The concentration of serum amyloid A (SAA) increases by up to 1000-fold during inflammation, largely owing to cytokine-driven transcriptional upregulation. A reporter plasmid, comprising the proximal 0.7 kb of the human SAA2 promoter fused to a luciferase gene, was used in transient transfection experiments in human HepG2 hepatoma cells to assess the quantitative and qualitative TNF antagonist properties of a construct in which sTNFR:Ig synthesis is under the control of a chimera of the SAA2 promoter and a tat/HIV element. The SAA2-tat/HIV-sTNFR:Ig construct retained the fine-tuned cytokine responsiveness of the SAA2 promoter, while exhibiting the quantitatively enhanced level of protein expression conferred by the tat/HIV element. It produced a biologically significant TNF inhibition that was at least as strong as that achieved using a CMV promoter-driven sTNFR:Ig construct. There was a dose- and time-dependent relationship between the pro-inflammatory cytokine used, and the generation of TNF antagonist activity by SAA2-tat/ HIV-sTNFR:Ig. Although sTNFR:Ig protein can be induced by either TNF-α or interleukin (IL)-lβ, its antagonist activity is limited to the former cytokine. The SAA2-tat/HIV-sTNFR:Ig construct, and derivatives thereof, may therefore be ideally suited to gene therapy applications that require the local production of potent and specific immune modifiers only when there is active pathology. It may consequently be of particular use in the future treatment of diseases such as rheumatoid arthritis.

Original languageEnglish (US)
Pages (from-to)588-595
Number of pages8
JournalScandinavian Journal of Immunology
Volume53
Issue number6
DOIs
StatePublished - 2001

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Serum Amyloid A Protein
Tumor Necrosis Factor Receptors
Genetic Therapy
Anti-Inflammatory Agents
Tumor Necrosis Factor-alpha
HIV
Cytokines
Proteins
Interleukins
Hep G2 Cells
Luciferases
Transfection
In Vitro Techniques
Hepatocellular Carcinoma
Rheumatoid Arthritis
Plasmids
Up-Regulation
Pathology
Inflammation
Genes

ASJC Scopus subject areas

  • Immunology

Cite this

In vitro evaluation of an enhanced human Serum Amyloid A (SAA2) promoter-regulated soluble TNF receptor fusion protein for anti-inflammatory gene therapy. / Rygg, M.; Uhlar, C. M.; Thorn, C.; Jensen, L. E.; Gaughan, D. J.; Varley, A. W.; Munford, R. S.; Göke, R.; Chen, Y.; Whitehead, A. S.

In: Scandinavian Journal of Immunology, Vol. 53, No. 6, 2001, p. 588-595.

Research output: Contribution to journalArticle

Rygg, M. ; Uhlar, C. M. ; Thorn, C. ; Jensen, L. E. ; Gaughan, D. J. ; Varley, A. W. ; Munford, R. S. ; Göke, R. ; Chen, Y. ; Whitehead, A. S. / In vitro evaluation of an enhanced human Serum Amyloid A (SAA2) promoter-regulated soluble TNF receptor fusion protein for anti-inflammatory gene therapy. In: Scandinavian Journal of Immunology. 2001 ; Vol. 53, No. 6. pp. 588-595.
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AU - Uhlar, C. M.

AU - Thorn, C.

AU - Jensen, L. E.

AU - Gaughan, D. J.

AU - Varley, A. W.

AU - Munford, R. S.

AU - Göke, R.

AU - Chen, Y.

AU - Whitehead, A. S.

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