Vaccinia virus (VACV) is an enveloped DNA virus from the Orthopoxvirus family, various strains of which were used in the successful eradication campaign against smallpox. Both original and newer VACV-based replicating vaccines reveal a risk of serious complications in atopic individuals. VACV encodes various factors interfering with host immune responses at multiple levels. In atopic skin, the production of type I interferon is compromised, while VACV specifically inhibits the phosphorylation of the Interferon Regulatory Factor 3 (IRF-3) and expression of interferons. To overcome this block, we generated a recombinant VACV-expressing murine IRF-3 (WR-IRF3) and characterized its effects on virus growth, cytokine expression and apoptosis in tissue cultures and in spontaneously atopic Nc/Nga and control Balb/c mice. Further, we explored the induction of protective immune responses against a lethal dose of wild-type WR, the surrogate of smallpox. We demonstrate that the overexpression of IRF-3 by WR-IRF3 increases the expression of type I interferon, modulates the expression of several cytokines and induces superior protective immune responses against a lethal poxvirus challenge in both Nc/Nga and Balb/c mice. Additionally, the results may be informative for design of other virus-based vaccines or for therapy of different viral infections.

• Klíčová slova
• IRF-3, Nc/Nga mice, atopic dermatitis, cytokines, eczema vaccinatum, immunization, interferon beta, interleukin-1 beta, smallpox, vaccinia virus,
• MeSH
• exprese genu genetika   MeSH
• imunita imunologie   MeSH
• infekce vyvolané poxviry imunologie   prevence a kontrola   MeSH
• interferon typ I metabolismus   MeSH
• interferonový regulační faktor 3 genetika   imunologie   MeSH
• interleukin-1beta imunologie   MeSH
• kůže imunologie   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• Poxviridae patogenita   MeSH
• regulace exprese virových genů genetika   MeSH
• replikace viru imunologie   MeSH
• vakcínie virologie   MeSH
• virové vakcíny imunologie   MeSH
• virus vakcinie genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• interferon typ I MeSH
• interferonový regulační faktor 3 MeSH
• interleukin-1beta MeSH
• virové vakcíny MeSH

Smallpox was declared eradicated in 1980. However recently, the need of agents effective against poxvirus infection has emerged again. In this paper, we report an original finding that two redox-modulating agents, the ethacrynic and alpha-lipoic acids (EA, LA), inhibit growth of vaccinia virus (VACV) in vitro. The effect of EA and LA was compared with those of beta-mercaptoethanol, DTT and ascorbic acid, but these agents increased VACV growth in HeLa G cells. The inhibitory effects of EA and LA on the growth of VACV were further confirmed in several cell lines of different embryonic origin, in epithelial cells, fibroblasts, macrophages and T-lymphocytes. Finally, we have analyzed the mechanism of action of the two agents. They both decreased expression of VACV late genes, as demonstrated by western blot analysis and activity of luciferase expressed under control of different VACV promoters. In contrast, they did not inhibit virus entry into the cell, expression of VACV early genes or VACV DNA synthesis. The results suggest new directions in development of drugs effective against poxvirus infection.

• MeSH
• antivirové látky farmakologie   MeSH
• exprese genu účinky léků   MeSH
• HeLa buňky MeSH
• kultivované buňky MeSH
• kyselina ethakrynová farmakologie   MeSH
• kyselina lipoová farmakologie   MeSH
• lidé MeSH
• luciferasy metabolismus   MeSH
• plakové testy MeSH
• replikace viru účinky léků   MeSH
• reportérové geny MeSH
• virus vakcinie účinky léků   růst a vývoj   MeSH
• western blotting MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH
• Názvy látek
• antivirové látky MeSH
• kyselina ethakrynová MeSH
• kyselina lipoová MeSH
• luciferasy MeSH