Strategies to increase longevity of adenoviral-mediated transgene expression in muscle

P. Prellop, D. E. Good, Arthur W. Zieske, P. Ye, P. O. Schwarzenberger, J. E. Shellito, J. K. Kolls

Research output: Contribution to journalArticle

Abstract

Adenoviral vectors are reasonably efficient at transducing a wide variety of tissues including lung, liver, and muscle. However, transgene expression is limited by a combination of innate immune responses, apoptosis, and cell-mediated immunity against vector-encoded proteins including the transgene. Recently, tumor necrosis factor (TNF) has been implicated as a critical cytokine in anti-adenoviral immune responses based on the fact that adenoviral-mediated transgene expression in TNF knockout mice. However, TNF knockout mice lack germinal centers in their lymph nodes and thus, it remains unclear whether these findings could be generalized to a therapeutic model. To investigate this point, we pre-treated mice with a single dose of either a polyclonal goat-anti rat TNF antibody or non-immune IgG prior to the administration of 109 pfu of AdCMVLacZ into the tibialis anterior (TA) muscle. A subgroup of mice were treated with 109 pfu of AdCMVLacZ and AdTNF-R, which encodes a TNF inhibitor consisting of a TNF-R:Fc fusion protein. Mice were sacrificed at 4, 14, or 28 days. At sacrifice, the TA was harvested, incubated in a PBS sucrose gradient, frozen in OCT and then sectioned. Sections were stained with X-gal solution and then counter-stained with safranin, or H and E. A blinded pathologist scored stained sections for gene transfer and inflammation. Mice receiving the anti-TNF had significantly less muscle injury as measured by the number of central nuclei. Moreover, there was significantly less inflammation at both 4 and 14 days. All TA muscles from the anti-TNF group demonstrated gene expression at 28 days whereas all control mice were negative beyond 14 days. These data demonstrate that TNF is a critical cytokine to anti-adenoviral immune responses in muscle. We speculate that transient blockade of TNF bioactivity may be used therapeutically to prolong transgene expression in vivo.

Original languageEnglish (US)
JournalJournal of Investigative Medicine
Volume47
Issue number2
StatePublished - Jan 1 1999

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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