ADENOVIRUS E3 14.7-KILODALTON PROTEIN WHICH INHIBITS CYTOLYSIS BY TUMOR-NECROSIS-FACTOR INCREASES THE VIRULENCE OF VACCINIA VIRUS IN A MURINE PNEUMONIA MODEL

The 14.7-kilodalton protein (14.7K protein) encoded by the adenovirus (Ad) E3 region inhibits tumor necrosis factor alpha (TNF-alpha)-mediated lysis of cells in tissue culture experiments, but the relevance of this effect in vivo is incompletely understood. To examine the effect of the ability of the Ad 14.7K protein to block TNF lysis upon viral pathogenesis in a murine model, we cloned the 14.7K protein-encoding gene into vaccinia virus (W), permitting its study in isolation from other Ad E3 immunomodulatory proteins. The gene for murine TNF-alpha was inserted into the same W containing the 14.7K gene to ensure that each cell infected with the W recombinant would express both the agonist (TNF) and its antagonist (14.7K). W was utilized as the vector because it accommodates large and multiple inserts of foreign DNA with faithful, high-level expression of the protein products. In addition, infection of mice with W induces disease with quantifiable morbidity, mortality, and virus replication. The results of intranasal infections of BALB/c mice with these W recombinants indicate that the Ad 14.7K protein increases the virulence of W carrying the TNF-cr gene by reversing the attenuating effect of TNF-alpha on W pathogenicity. This was demonstrated by increased mortality, pulmonary pathology, and viral titers in lung tissue following infection with W coexpressing the 14.7K protein and TNF-alpha, compared with the control virus expressing TNF-alpha alone. These results suggest that the 14.7K protein, which is nonessential for Ad replication in tissue culture, is an immunoregulatory protein which functions in vivo to help counteract the antiviral effects of TNF-alpha.