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Franceschi, V.

Publications and source records attributed to Franceschi, V..

3 recordsLinked to original sources

Generation, characterization and exploiting caprine herpesvirus 1 secreted glycoprotein D

Caprine herpesvirus 1 (CpHV-1), a member of the Herpesvirales order, Herpesviridae family, Alphaherpesvirinae subfamily, and Simplexvirus genus, is classically associated to two distinct clinical syndromes. In kids, CpHV-1 induces severe systemic disease with high morbidity and mortality, in adult goats, the infection leads to genital lesions such as vulvovaginitis or balanoposthitis, with abortions occurring mainly in the second half of gestation. CpHV-1 shares several biological characteristics with human herpesvirus 2 (HSV-2), including molecular features, tropism for vaginal epithelium, genital lesion nature and latency in the sacral ganglia. These features make CpHV-1-infected goats a reliable animal model for studying human herpesvirus-induced genital disease, employable for pathogenic research, as well as the development of new vaccines and antiviral agents. Recent full sequencing of CpHV-1 genome has identified at least ten genes encoding glycoproteins. Among these, glycoprotein D (gD) has been characterized but not yet exploited for immunogenic or diagnostic purposes. In this study, the structural features of CpHV-1 gD were predicted using in silico analysis. A truncated version of gD lacking the transmembrane domain (Sec-gD) was subsequently generated and expressed in mammalian cells, enabling its secretion into the culture medium. Despite the structural modifications, Sec-gD retained a conserved glycosylation pattern, as confirmed by PNGase F treatment. Furthermore, the antigenic properties of Sec-gD were preserved, as demonstrated by reverse serum neutralization assays. Notably, the culture supernatant containing Sec-gD was directly usable in diagnostic enzyme-linked immunosorbent assays, supporting its potential as a valuable tool for both diagnostic and immunization strategies. ImportanceCpHV-1-infected goats represent a large animal model for studying human herpesvirus-induced genital disease, and could be utilized for pathogenic research, as well as for the development of new vaccines and antiviral agents. CpHV-1 gD can be efficiently produced and rescued from the supernatant of transfected mammalian cells, retaining its immunogenic properties and could be employed for immunogenic and diagnostic purposes.

microbiology↗

Vaccination of cattle with a virus-vectored vaccine against a major membrane protein of Mycobacterium a. subsp. paratuberculosis elicits CD8 cytotoxic T cells that kill intracellular bacteria

Infection of cattle with Mycobacterium a. subsp. Paratuberculosis (Map), the causative agent of paratuberculosis, induces an immune response directed toward a 35 kD major membrane protein (MMP) of Map. CD8 cytotoxic T cells (CTL) are elicited when peripheral blood mononuclear cells from healthy cattle are incubated ex vivo with antigen-presenting cells (APC) primed with bacterial recombinant MMP. Ex vivo development of CTL was MHC-restricted and required the presence of both CD4 and CD8 T cells. The gene MAP2121c, encoding MMP, was modified to express a modified form of MMP (p35NN) in a mammalian cell line, also capable of eliciting an ex vivo CTL response. In the present study, the modified gene for p35NN was placed into a BoHV4 vector to determine the potential use of BoHV-4A{Delta}TK-p35NN as a peptide-based vaccine. Subcutaneous vaccination of healthy cattle with BoHV-4A{Delta}TK-p35NN elicited a CTL recall response, as detected ex vivo. Further studies are warranted to conduct a challenge study to determine if CD8 CTL elicited by vaccination with BoHV-4A{Delta}TK-p35NN prevents the establishment of a persistent infection by Map.

immunology↗

Characterization of BoHV-4 ORF45

Bovine herpesvirus 4 (BoHV-4) is a Gammaherpesvirus of the genus Rhadinovirus, his natural host is the bovine whereas the African Buffalo the natural reservoir. Anyhow, BoHV-4 infection is not associated to a specific disease. Genome structure and genes are well conserved in Gammaherpesvirus, and orf45 gene and its product, ORF45, is one of those. BoHV-4 ORF45 has been suggested to be a tegument protein, however, BoHV-4 ORF45 structure and function have not yet been experimentally characterized. In the present study, it is shown that BoHV-4 ORF45, despite its poor homology with other characterized Rhadinovirus ORF45s, is structurally related to Kaposis sarcoma-associated herpesvirus (KSHV), is a phosphoprotein and localizes in the host cell nuclei. Through the generation of an ORF45-null mutant BoHV-4 and its pararevertant, it was possible to demonstrate that ORF45 is essential for BoHV-4 lytic replication and is associated to the viral particles, as for the other characterized Rhadinovirus ORF45s. Finally, the impact of BoHV-4 ORF45 on cellular transcriptome was investigated, an aspect poorly explored or not at all for other Gammaherpesvirus. Many cellular transcriptional pathways were found to be alterate, mainly those involving p90 ribosomal S6 kinase (RSK) and signal-regulated kinase (ERK) complex (RSK/ERK), thus highlighting the authentic character of BoHV-4 ORF45 and paving the way to further investigations.

microbiology↗