Multiplex analysis of serum cytokines in humans with hantavirus pulmonary syndrome

© 2015 Morzunov, Khaiboullina, St. Jeor, Rizvanov and Lombardi. Hantavirus pulmonary syndrome (HPS) is an acute zoonotic disease transmitted primarily through inhalation of virus-contaminated aerosols. Hantavirus infection of endothelial cells leads to increased vascular permeability without a visible cytopathic effect. For this reason, it has been suggested that the pathogenesis of HPS is indirect with immune responses, such as cytokine production, playing a dominant role. In order to investigate their potential contribution to HPS pathogenesis, we analyzed the serum of hantavirus-infected subjects and healthy controls for 68 different cytokines, chemokines, angiogenic, and growth factors. Our analysis identified differential expression of cytokines that promote tissue migration of mononuclear cells including T lymphocytes, natural killer cells, and dendritic cells. Additionally, we observed a significant upregulation of cytokines known to regulate leukocyte migration and subsequent repair of lung tissue, as well as cytokines known to increase endothelial monolayer permeability and facilitate leukocyte transendothelial migration. Conversely, we observed a downregulation of cytokines associated with platelet numbers and function, consistent with the thrombocytopenia observed in subjects with HPS. This study corroborates clinical findings and extends our current knowledge regarding immunological and laboratory findings in subjects with HPS

Hantaviral proteins: Structure, functions, and role in hantavirus infection

© 2015 Muyangwa, Martynova, Khaiboullina, Morzunov and Rizvanov. Hantaviruses are the members of the family Bunyaviridae that are naturally maintained in the populations of small mammals, mostly rodents. Most of these viruses can easily infect humans through contact with aerosols or dust generated by contaminated animal waste products. Depending on the particular Hantavirus involved, human infection could result in either hemorrhagic fever with renal syndrome or in Hantavirus cardiopulmonary syndrome. In the past few years, clinical cases of the Hantavirus caused diseases have been on the rise. Understanding structure of the Hantavirus genome and the functions of the key viral proteins are critical for the therapeutic agents' research. This paper gives a brief overview of the current knowledge on the structure and properties of the Hantavirus nucleoprotein and the glycoproteins

Hantavirus infection suppresses thrombospondin-1 expression in cultured endothelial cells in a strain-specific manner

© 2016 Khaiboullina, Morzunov, St. Jeor, Rizvanov and Lombardi.Hantavirus infection is associated with two frequently fatal diseases in humans: Hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). The pathogenesis of hantavirus infection is complex and not fully understood; however, it is believed to involve virus-induced hyperinflammatory immune responses. Thrombospondin-1 (THBS1) is a large homotrimeric protein that plays a putative role in regulating blood homeostasis. Hyperresponsiveness to inflammatory stimuli has also been associated with defects in the THBS1 gene. Our data suggest that hantavirus infection of human umbilical cord vein endothelial cells (HUVEC) suppress the accumulation of THBS1 in the extracellular matrix. Additionally, this suppression is dependent on virus replication, implying a direct mechanism of action. Our data also imply that the pathogenic Andes and Hantaan strains inhibit THBS1 expression while the non-pathogenic Prospect Hill strain showed little inhibition. These observations suggest that a dysregulation of THBS1 may contribute to the pathogenesis of hantavirus infection

Death-domain associated protein-6 (DAXX) mediated apoptosis in hantavirus infection is counter-balanced by activation of interferon-stimulated nuclear transcription factors

Hantaviruses are negative strand RNA species that replicate predominantly in the cytoplasm. They also activate numerous cellular responses, but their involvement in nuclear processes is yet to be established. Using human umbilical vein endothelial cells (HUVECs), this study investigates the molecular finger-print of nuclear transcription factors during hantavirus infection. The viral-replication-dependent activation of pro-myelocytic leukemia protein (PML) was followed by subsequent localization in nuclear bodies (NBs). PML was also found in close proximity to activated Sp100 nuclear antigen and interferon-stimulated gene 20. kDa protein (ISG-20), but co-localization with death-domain associated protein-6 (DAXX) was not observed. These data demonstrate that hantavirus triggers PML activation and localization in NBs in the absence of DAXX-PLM-NB co-localization. The results suggest that viral infection interferes with DAXX-mediated apoptosis, and expression of interferon-activated Sp100 and ISG-20 proteins may indicate intracellular intrinsic antiviral attempts. © 2013

Andes-virus-induced cytokine storm is partially suppressed by ribavirin

Background: Microbe-induced over-activation of cytokines, especially tumour necrosis factor (TNF)-α, is key to the pathogenesis of hantavirus infection leading to severe inflammation with high mortality rate. Although ribavirin showed promise in inhibiting viral replication in vitro, its clinical efficacy remains controversial. Methods: Various concentrations of ribavirin were used to determine its effect on cytokine activation in our infectious model system. Results: Ribavirin decreased the virus load and dosedependently inhibited the accumulation of RANTES messenger RNA in Andes-virus (ANDV)-infected human endothelial cells, but failed to suppress TNF-α-induced activation of RANTES and interleukin-6 in ANDV-inoculated cultures. This report also shows, for the first time, that the deleterious over-stimulation by TNF-α is mediated by nuclear factor-kB, and describes the effect of ribavirin on cytokine production following ANDV infection. Conclusions: Although highly effective in preventing ANDV replication and suppressing activation of select inflammatory mediators, the therapeutic efficacy of ribavirin is limited due to its inability to fully inhibit cytokine outburst triggered by hantavirus infection. © 2013 International Medical Press

Human dendritic cells transfected with a human papilloma virus-18 construct display decreased mobility and upregulated cytokine production

The marked depletion of dendritic cells (DCs) in skin cancers, as well as preneoplastic and neoplastic cervical epithelium, suggests a central role for DCs in productive human papillomavirus (HPV) infection and cancer promotion. It has been suggested that HPV may facilitate tumor development by reducing DC density, contributing to a decrease in local immune surveillance. In this study, we have examined the response of human DCs transfected with a construct containing the HPV18 genome and their subsequent expression of papilloma virus proteins. Transfected cells expressed the L1 major capsid protein and upregulated E6 and E7 oncoprotein transcripts as detected by RT-PCR. Transfection of DCs also resulted in a significant increase in cytokine production. Finally, we observed that HPV18 transfection decreased the migratory activity of DCs. Our data indicate that HPV transfection of DCs leads to changes in migratory activity and cytokine production, which potentially can suppress or delay immune responses to viral antigens. Additionally, changes in cytokine production by HPV-transformed human fibroblasts and human cervical epithelial cells revealed that the migratory and antigen-presenting functions of DCs may be impaired by the suppressive effects of cytokines produced by HPV-infected epithelial and stromal cells

Serum cytokine profiles differentiating hemorrhagic fever with renal syndrome and hantavirus pulmonary syndrome

© 2017 Khaiboullina, Levis, Morzunov, Martynova, Anokhin, Gusev, St Jeor, Lombardi and Rizvanov.Hantavirus infection is an acute zoonosis that clinically manifests in two primary forms, hemorrhagic fever with renal syndrome (HFRS) and hantavirus pulmonary syndrome (HPS). HFRS is endemic in Europe and Russia, where the mild form of the disease is prevalent in the Tatarstan region. HPS is endemic in Argentina, as well as other countries of North and South American. HFRS and HPS are usually acquired via the upper respiratory tract by inhalation of virus-contaminated aerosol. Although the pathogenesis of HFRS and HPS remains largely unknown, postmortem tissue studies have identified endothelial cells as the primary target of infection. Importantly, cell damage due to virus replication, or subsequent tissue repair, has not been documented. Since no single factor has been identified that explains the complexity of HFRS or HPS pathogenesis, it has been suggested that a cytokine storm may play a crucial role in the manifestation of both diseases. In order to identify potential serological markers that distinguish HFRS and HPS, serum samples collected during early and late phases of the disease were analyzed for 48 analytes using multiplex magnetic bead-based assays. Overall, serum cytokine profiles associated with HPS revealed a more pro-inflammatory milieu as compared to HFRS. Furthermore, HPS was strictly characterized by the upregulation of cytokine levels, in contrast to HFRS where cases were distinguished by a dichotomy in serum cytokine levels. The severe form of hantavirus zoonosis, HPS, was characterized by the upregulation of a higher number of cytokines than HFRS (40 vs 21). In general, our analysis indicates that, although HPS and HFRS share many characteristic features, there are distinct cytokine profiles for these diseases. These profiles suggest a strong activation of an innate immune and inflammatory responses are associated with HPS, relative to HFRS, as well as a robust activation of Th1-type immune responses. Finally, the results of our analysis suggest that serum cytokines profiles of HPS and HFRS cases are consistent with the presence of extracellular matrix degradation, increased mononuclear leukocyte proliferation, and transendothelial migration

Genetic evidence of Dobrava virus in Apodemus agrarius in Hungary.

Using nested polymerase chain reaction, we sequenced Dobrava virus (DOB) from the rodent Apodemus agrarius in Hungary. The samples we isolated group with DOB samples previously isolated from A. flavicollis. This grouping may indicate host switching

Genetic Characterization of Small (s)-Segment Genome Puumala Virus Strain Kazan

© 2016, Springer Science+Business Media New York.The Republic of Tatarstan is one of the most active endemic regions for nephropathia epidemica (NE) in the Russian Federation. Annually, over 1000 cases of NE are registered, with an average mortality rate of 0.43 %. NE is a zoonosis where human become infected by inhaling a hantavirus-contaminated aerosol. Puumala virus is commonly detected in NE cases. Although NE cases have been registered in the region since 1958, little is known about the genetic variability of Puumala virus circulating in Tatarstan. We conducted phylogenetic analysis of the full length (1828 nt) small (S) segment genome sequence of Puumala virus RNA isolated from four bank voles (Myodes glareolus) captured during the 2014 outbreak. These virus sequences were compared to known sequences of Puumala viruses from nearby regions such as Samara and the Republic of Bashkortostan, as well as to European isolates. We found that there was over 89 % nucleotide identity between Puumala virus sequences isolated from the bank voles captured in Tatarstan. Sequence identity between Puumala viruses from the Tatarstan bank voles and the sequences from Republic of Bashkortostan and the Samara region were 90–95 %. Less similarity was found between Tatarstan sequences and Puumala strains circulating in Europe (79.7–87.7 %). Taken together, these data suggest that Puumala viruses circulating within the bank vole population in the Republic of Tatarstan are phylogenetically closer to the viruses circulating in neighboring regions of Russia

Detection of Antibodies Recognizing Puumala Virus Nucleocapsid and Glycoprotein Peptides in NE Serum

© 2016, Springer Science+Business Media New York.Nephropatia epidemica (NE), a mild form of hemorrhagic fever with renal syndrome (HFRS), is an endemic zoonosis in the Republic of Tatarstan, Russia. Humans become infected by inhaling an aerosol contaminated with Puumala virus, a member of genus Hantavirus. NE diagnosis is based on detection of anti-hantavirus antibodies using ELISA. Antibodies to hantavirus nucleocapsid (N) protein are detected early in the course of infection, suggesting that this viral protein is the most immunogenic. Several epitopes were previously identified on N protein as well as glycoproteins of Puumala viruses endemic in Europe. However, there is limited knowledge about Puumala virus N protein antigenic epitopes in NE patients in the Republic of Tatarstan. The aim of the present study is to identify N protein and glycoprotein epitopes which induce a humoral immune response in NE cases. Analysis of NE serum using an array of overlapping N protein and glycoprotein peptides identified the most immunogenic epitopes, which can then be used for developing Puumala virus-specific vaccine