Download Vaccinia virus (VACV) is primarily known as a vaccine against its

Vaccinia virus (VACV) is primarily known as a vaccine against its relative variola virus, the causative
agent of smallpox. In the seventies of the 20th century, the vaccination campaign with VACV led to
eradication of smallpox. Consequently, vaccination of the general population was stopped. Currently,
the vaccination was reintroduced, namely among army and healthcare professionals. However,
vaccination with VACV is accompanied with a high incidence of vaccination-related complications,
namely
among immunocompromised individuals. One of the complications is eczema vaccinatum, occuring in
patients with atopic dermatitis.
The laboratory of Dr. Melkova has focused on development of a model of eczema vaccinatum in mice
Nc/Nga and on studies of pathogenesis of this complication. The goal of my diploma thesis is to
contribute to characterization of imunopathogenesis of eczema vaccinatum in mice Nc/Nga infected
either with VACV strain Western Reserve (WR) or with a recombinant VACV with the integrated
cDNA for IRF-3 (Interferon Regulatory Factor 3; WR-IRF3). IRF-3 regulates the expression of
interferon type I in response to viral infection. This recombinant virus has been constructed in the
laboratory of Dr. Melková. The objective of my work was to verify the expression of the integrated
cDNA for IRF-3 and to characterize the growth of this recombinant virus.
The results were obtained by RT-PCR from the skin of mice Nc/Nga with specific primers for certain
cytokines (IL-1, IL-18 a IL-33). They suggest that infection with VACV strain WR modifies the
expression of IL-1 and IL-33, while the infection has not affected expression of IL-18. The detection of
cDNA for each cytokine was affected also by the way of sensitization and inoculation of VACV.
Further, the properties of the recombinant virus WR-IRF3 were characterized in vitro and in vivo. The
results of these experiments suggest that immune responses against infection with
WR-IRF3 are increased.