Kaposi's sarcoma-associated herpesvirus vIRF2 protein utilizes an IFN-dependent pathway to regulate viral early gene expression.

Kaposi's sarcoma-associated herpesvirus (KSHV; human herpesvirus 8) belongs to the subfamily of Gammaherpesvirinae and is the etiological agent of Kaposi's sarcoma as well as of two lymphoproliferative diseases: primary effusion lymphoma and multicentric Castleman disease. The KSHV life cycle is divided into a latent and a lytic phase and is highly regulated by viral immunomodulatory proteins which control the host antiviral immune response. Among them is a group of proteins with homology to cellular interferon regulatory factors, the viral interferon regulatory factors 1-4. The KSHV vIRFs are known as inhibitors of cellular interferon signaling and are involved in different oncogenic pathways. Here we characterized the role of the second vIRF protein, vIRF2, during the KSHV life cycle. We found the vIRF2 protein to be expressed in different KSHV positive cells with early lytic kinetics. Importantly, we observed that vIRF2 suppresses the expression of viral early lytic genes in both newly infected and reactivated persistently infected endothelial cells. This vIRF2-dependent regulation of the KSHV life cycle might involve the increased expression of cellular interferon-induced genes such as the IFIT proteins 1, 2 and 3, which antagonize the expression of early KSHV lytic proteins. Our findings suggest a model in which the viral protein vIRF2 allows KSHV to harness an IFN-dependent pathway to regulate KSHV early gene expression.

The inflammatory kinase MAP4K4 promotes reactivation of Kaposi's sarcoma herpesvirus and enhances the invasiveness of infected endothelial cells.

Kaposi's sarcoma (KS) is a mesenchymal tumour, which is caused by Kaposi's sarcoma herpesvirus (KSHV) and develops under inflammatory conditions. KSHV-infected endothelial spindle cells, the neoplastic cells in KS, show increased invasiveness, attributed to the elevated expression of metalloproteinases (MMPs) and cyclooxygenase-2 (COX-2). The majority of these spindle cells harbour latent KSHV genomes, while a minority undergoes lytic reactivation with subsequent production of new virions and viral or cellular chemo- and cytokines, which may promote tumour invasion and dissemination. In order to better understand KSHV pathogenesis, we investigated cellular mechanisms underlying the lytic reactivation of KSHV. Using a combination of small molecule library screening and siRNA silencing we found a STE20 kinase family member, MAP4K4, to be involved in KSHV reactivation from latency and to contribute to the invasive phenotype of KSHV-infected endothelial cells by regulating COX-2, MMP-7, and MMP-13 expression. This kinase is also highly expressed in KS spindle cells in vivo. These findings suggest that MAP4K4, a known mediator of inflammation, is involved in KS aetiology by regulating KSHV lytic reactivation, expression of MMPs and COX-2, and, thereby modulating invasiveness of KSHV-infected endothelial cells.

Genetic Relationship Between Hard Ticks (Ixodidae) Infesting Cattle from Select Areas of a Wildlife-Livestock Interface Ecosystem at Mikumi National Park, Tanzania.

Background: There has recently been a substantial increase in the number of tick species and tick-borne infectious agents in Tanzania. Owing to their impact on human, livestock, and wild animal health, increased knowledge of ticks is needed. So far, no published data on the genetic relationship between hard tick (Ixodidae) sequences collected from cattle are available in Tanzania. Methods: Ticks from cattle in the wards, which lie at the border of Mikumi National Park, were collected in the dry season, November to December 2019. Morphological identification of ticks was initially performed at the genus level. To identify ticks at the species level, molecular analysis based on the 16S rRNA gene was performed. Evolutionary relationships and genetic distances between ticks were determined using MaximumLikelihood and Kimura 2-parameter methods, respectively. Results: Based on morphology, two genera (Rhipicephalus and Hyalomma) were identified in the 630 adult ticks collected from a total of 252 cattle. Six species (Rhipicephalus microplus, Rhipicephalus evertsi, Hyalomma marginatum, Hyalomma rufipes, Hyalomma truncatum, and Hyalomma turanicum) were confirmed by BLASTn and phylogenetic analyses. Considerable mean and pairwise genetic distances were observed for Rhipicephalus and Hyalomma genera. Conclusion: The presence of different phylogenetic clusters and considerable mean and pairwise genetic distances observed reflect possible biological diversity of hard ticks present in the study area. Considering the value of the cattle in the livelihoods and economies of people and the country, the outcomes of this study will be useful in planning integrated control strategies for ticks and tick-borne diseases in Tanzania.

Molecular Diversity of Hard Tick Species from Selected Areas of a Wildlife-Livestock Interface Ecosystem at Mikumi National Park, Morogoro Region, Tanzania.

Ticks are one of the most important arthropod vectors and reservoirs as they harbor a wide variety of viruses, bacteria, fungi, protozoa, and nematodes, which can cause diseases in human and livestock. Due to their impact on human, livestock, and wild animal health, increased knowledge of ticks is needed. So far, the published data on the molecular diversity between hard ticks species collected in Tanzania is scarce. The objective of this study was to determine the genetic diversity between hard tick species collected in the wildlife-livestock interface ecosystem at Mikumi National Park, Tanzania using the mitochondrion 16S rRNA gene sequences. Adult ticks were collected from cattle (632 ticks), goats (187 ticks), and environment (28 ticks) in the wards which lie at the border of Mikumi National Park. Morphological identification of ticks was performed to genus level. To identify ticks to species level, molecular analysis based on mitochondrion 16S rRNA gene was performed. Ticks representing the two genera (Hyalomma and Rhipicephalus) were identified using morphological characters. Six species were confirmed based on mitochondrion 16S rRNA gene, including Rhipicephalus microplus, Rhipicephalus evertsi, Hyalomma rufipes, Hyalomma truncatum, Hyalomma marginatum, and Hhyalomma turanicum. The presence of different clusters of tick species reflects the possible biological diversity of the hard ticks present in the study region. Further studies are however required to quantify species of hard ticks present in the study region and the country in general over a larger scale.

Application of Viral Metagenomics for Study of Emerging and Reemerging Tick-Borne Viruses.

Ticks are important vectors for different tick-borne viruses, some of which cause diseases and death in humans, livestock, and wild animals. Tick-borne encephalitis virus, Crimean-Congo hemorrhagic fever virus, Kyasanur forest disease virus, severe fever with thrombocytopenia syndrome virus, Heartland virus, African swine fever virus, Nairobi sheep disease virus, and Louping ill virus are just a few examples of important tick-borne viruses. The majority of tick-borne viruses have RNA genomes that routinely undergo rapid genetic modifications such as point mutations during their replication. These genomic changes can influence the spread of viruses to new habitats and hosts and lead to the emergence of novel viruses that can pose a threat to public health. Therefore, investigation of the viruses circulating in ticks is important to understand their diversity, host and vector range, and evolutionary history, as well as to predict new emerging pathogens. The choice of detection method is important, as most methods detect only those viruses that have been previously well described. On the other hand, viral metagenomics is a useful tool to simultaneously identify all the viruses present in a sample, including novel variants of already known viruses or completely new viruses. This review describes tick-borne viruses, their historical background of emergence, and their reemergence in nature, and the use of viral metagenomics for viral discovery and studies of viral evolution.