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This work describes the role of membrane bridges in the spread of ectromelia virus (ECTV) Moscow strain (ECTV-MOS) in vitro in BALB/3T3 and Vero cells. In this research, immunofluorescence techniques were used: cells were fixed, stained for the presence of viral antigens, F-actin, and DNA, and viewed under a fluorescence microscope. In both cell lines, ECTV-MOS infection induces production of two types of actin-rich protrusions: tiny actin tails or long filopodia-like structures. It appears that both types contribute to the spread of ECTV-MOS particles and may be classified as membrane bridges.
Ectromelia virus (ECTV) is a member of the Orthopoxvirus genus of the Poxviridae family. It is a causative agent of mouse pox in genetically sensitive mice strains of H-2ᵈ (e.g. BALB/c) and H-2ᵃ (eg.A, A/J) haplotypes. Mouse pox virus is a well recognized model for studying generalized viral infections in natural hosts. The aim of this study was to determine the role of heat shock proteins (namely hsp90, hsp70 and hsp2) during the replication of the Moscow strain of ECTV (ECTV-MOS) in BALB/c mice. The internal organs of mice are important sites for primary virus replication whereas ECTV penetration into the brain may seriously influence mechanisms supervising immune and nervous systems cooperation. Based on studies carried out in BALB/c mice infected with ECTV-MOS, it was found that the hsp (hsp90, hsp70 and hsp27) expression in brain cells reach peak values during the incubation period and clinical manifestation of mouse pox but the relative numbers of hsp⁺ cells in the brains decreased during the recrudescence of the infection to values observed in the control mice. The high expression of hsp (hsp90, hsp70 and hsp27) on oligodendrocytes in BALB/c mice during the incubation and clinical periods may reflect the protective action of heat shock proteins within the brain.
Apoptosis is the process involved in embryonic development and homeostasis maintenance of developed organisms. Furthermore, apoptosis is the mechanism of NK and T cells (CTL) cytotoxicity. The family of bcl-2 proteins plays a crucial role in regulation of cell death and consists both of proteins with pro-apoptotic (Bax, Bad, Bid, Bik), and anti-apoptotic (Bcl-2, Bcl-xL) activity. The present work was aimed at elucidating the role of Bax and Bcl-2 expression in pathogenesis of mousepox caused by virulent Moscow strain of Ectromelia virus (ECTV-MOS) in sensitive animals - BALB/c mice (H-2ᵈ). BALB/c mice were infected via footpad with ECTV-MOS and at 5, 10 and 15 day of infection, lymph nodes, livers and spleens were isolated and subjected to further research. The tests included measurement of: apoptosis using TUNEK assay, caspase-9 activity and bcl-2 and Bax expression. The results showed: (a) influence of ECTV-MOS upon expression of Bcl-2/Bax and subsequent transduction of mitochondrial apoptotic pathway and (b) the role of Bcl-2 protein in suppression of apoptosis in Mø depending on specific immunological environment of infected organs.
The present work describes the involvement of Fas/FasL receptors in apoptosis during the pathogenesis of ectromelia virus (ECTV) infection in BALB/c mice. The involvement of Fas/FasL in apoptosis observed during ECTV infection is related to: (a) the influence of ECTV upon the expression of Fas/FasL and subsequent transduction pathways and (b) the specific immunological environment of infected organs and its influence on the functioning of immunocompetent cells.
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