Development of a SFTSV DNA vaccine that confers complete protection against lethal infection in ferrets.

Although the incidence of severe fever with thrombocytopenia syndrome virus (SFTSV) infection has increased from its discovery with a mortality rate of 10-20%, no effective vaccines are currently available. Here we describe the development of a SFTSV DNA vaccine, its immunogenicity, and its protective efficacy. Vaccine candidates induce both a neutralizing antibody response and multifunctional SFTSV-specific T cell response in mice and ferrets. When the vaccine efficacy is investigated in aged-ferrets that recapitulate fatal clinical symptoms, vaccinated ferrets are completely protected from lethal SFTSV challenge without developing any clinical signs. A serum transfer study reveals that anti-envelope antibodies play an important role in protective immunity. Our results suggest that Gn/Gc may be the most effective antigens for inducing protective immunity and non-envelope-specific T cell responses also can contribute to protection against SFTSV infection. This study provides important insights into the development of an effective vaccine, as well as corresponding immune parameters, to control SFTSV infection.

Community-based comprehensive measures to prevent severe fever with thrombocytopenia syndrome, China.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging disease caused by the SFTS virus (SFTSV) of the family Bunyaviridae. Since the virus was first isolated in 2009, it has become widespread in China, with an increasing number of cases year on year. Although the disease has been researched extensively in past years, there are still no effective measures to suppress the epidemic situation. This article reports a pilot study of comprehensive measures, including health education and risk communication, weed removal, livestock management, and tick control, to prevent this emerging disease in an endemic region of China. The density of ticks decreased dramatically month by month after acaricides were sprayed in the areas surrounding recreational and agricultural settings. The number of SFTS cases and villages involved declined in the years after the integrated measures were applied. Comprehensive measures, especially community-based tick control, may be a promising means of preventing SFTS in endemic regions.

Current status of Severe Fever with Thrombocytopenia Syndrome vaccine development.

Severe Fever with Thrombocytopenia Syndrome (SFTS) is a new emerging tick-borne disease caused by the phlebovirus, SFTS virus (SFTSV). The virus was discovered in central China in 2009 and has since been identified in both Japan and South Korea. Significant progress has been made on the molecular biology of the virus, and this has been used to develop diagnostic assays and reagents. Less progress has been made on the epidemiology, maintenance and transmission, clinical manifestations, immunological responses, and treatment regimens. A number of animal models have been investigated but, to date, none recapitulate all the clinical manifestations seen in humans. Vaccine development is at an early discovery phase.

[Recent topics in the research field of severe fever with thrombocytopenia syndrome (SFTS)].

Seven years have passed since the discovery of a novel infectious disease, severe fever with thrombocytopenia syndrome (SFTS) caused by a novel Phlebovirus, SFTS virus (SFTSV), in PR China. It was also confirmed that SFTS was endemic to Japan through an identification of a woman, who died of SFTSV infection in Yamaguchi prefecture in late 2012. Approximately 6 years have passed since the discovery of SFTS-endemicity in Japan. At present, SFTS is endemic to PR China, South Korea and western Japan. SFTSV is maintained between several species of ticks such as Haemaphysalis longicornis and wild and domestic animals in nature. Therefore, we cannot escape from the risk of being infected with SFTSV. Based on the similarity in the characteristics of the clinical symptoms including the high case fatality rate, mode of infection to humans, pathology and virology between SFTS and Crimean-Congo hemorrhagic fever (CCHF), SFTS should be classified as viral hemorrhagic fever. Although the time from the discovery of SFTS is still short, there have been many scientific reports on the epidemiological, clinical, and/or pathological, and virological studies on SFTS. Favipiravir was reported to show an efficacy in the prevention and treatment of SFTSV infections in an animal model. A clinical study to evaluate the efficacy of favipiravir in the treatment of SFTS patients has been initiated in Japan. Specific and effective treatment with antiviral drugs for and preventive measures of SFTS with vaccination shoued be developed through scientific, clinical, and basic research.

Sandfly fever in Afghanistan-a sometimes overlooked disease of military importance: a case series and review of the literature.

Sandfly fever, sometimes known as pappataci fever or Phlebotomus fever, is a vector transmitted viral illness with a history of affecting naïve military formations that travel through or fight in areas in which the infection is endemic. We present a series of 4 hospitalized cases of sandfly fever (2 presumptive, 2 laboratory confirmed) that were admitted to a Role 3 hospital in Afghanistan for evaluation and treatment following medical evacuation from a forward area for marked fevers and malaise. Laboratory evaluation of these cases was significant for leukopenia and thrombocytopenia, consistent with historical descriptions of sandfly fever. In the correct geographic and clinical setting, the finding of mild leukopenia among a cluster of febrile patients should prompt the clinician to at least consider a diagnosis of sandfly fever. A cluster investigation conducted by preventive medicine personnel identified numerous other presumed cases of sandfly fever in this forward special operations camp. Response efforts emphasized enforcement of standard vector-borne disease control measures by operational leadership in order to limit effect on tactical operations. We review historical instances of sandfly fever affecting military operations, and present a review of clinical presentation, transmission, management, and prevention.

Immunization with recombinant SFTSV/NSs protein does not promote virus clearance in SFTSV-infected C57BL/6J mice.

The severe fever with thrombocytopenia syndrome (SFTS), caused by a novel Phlebovirus in the Bunyaviridae family named SFTS virus (SFTSV), is an emerging hemorrhagic fever with a wide distribution and high case-fatality rate. Neither effective treatment nor vaccines are available to treat and prevent this disease to date. It was recently reported that SFTSV nonstructural protein in S segment (SFTSV/NSs) functioned as the interferon (IFN) antagonist targeting for suppressing host's innate immunity. This study was designed to investigate the potential of recombinant SFTSV (rSFTSV)/NSs protein for inducing anti-NSs antibodies by pre-exposure vaccination to block SFTSV/NSs in the SFTSV-infected C57BL/6J mice. All mice in the rSFTSV/NSs-vaccinated group, negative control group, and blank control group survived with no visible clinical abnormities throughout the experiment, except for their sacrifice for sampling at each observation point. However, unexpectedly, a negative effect on the bodyweight of rSFTSV/NSs-vaccinated mice was observed after 21 days postinoculation. Pre-exposure vaccination with rSFTSV/NSs did not accelerate virus removal in mice though high titer of anti-NSs antibodies and elevated IFN-γ were detected in sera. Before virus challenge, the rSFTSV/NSs-vaccinated mice and negative control mice had a larger amount of platelets (PLT) than the blank control mice, which indicated that Freund's adjuvants could stimulate PLT production. In the aspect of cytokines, the rSFTSV/NSs-vaccinated mice had a 5- to 10-fold increase in interleukin (IL)-2, IL-5, IL-6, IFN-γ, and tumor necrosis factor-α, which probably just had a negative effect on the bodyweight of mice. In general, therefore, previous vaccination with rSFTSV/NSs did not accelerate virus clearance in the SFTSV-infected mice.

Sandfly-borne phleboviruses of Eurasia and Africa: epidemiology, genetic diversity, geographic range, control measures.

Sandfly-borne phleboviruses may cause a transient febrile illness (sandfly fever) or more severe neuroinvasive disease. In the Old World, they are vectored by phlebotomine flies, which are widely distributed in the Mediterranean basin, North Africa, the Indian subcontinent, the Middle East and central Asia. High seroprevalence rates have been recorded in humans and domestic animals in areas where sandflies are present. Most published studies have focused on phlebovirus infections of travelers and of soldiers stationed in endemic areas, but the health impact on local populations should not be underestimated, as seroprevalence studies indicate massive circulation of these viruses, even if disease is seldom documented. Except for Toscana virus, which shows a marked neurotropism and is a leading cause of aseptic meningitis in endemic regions, phlebovirus infections are inadequately considered by physicians and are generally underestimated. However, several properties of these viruses suggest that they will extend their geographic range. First, changes in the areas occupied by sandflies as a result of climate change have a direct impact on the epidemiology of associated human and animal diseases. Second, phleboviruses exhibit a high mutation rate, and their tri-segmented genome is prone to reassortment and recombination. Third, distinct virus strains can be transmitted by the same arthropod species. Recent studies have documented the distribution of sandfly-borne phleboviruses in Western Europe, but data for Eastern Europe, the Middle East and Africa are very limited. With the goal of filling knowledge gaps and planning new research programs, we have examined available information and present it as a comprehensive review, with a specific focus on understudied regions. We also discuss the need to conduct studies aimed at developing new antiviral drugs and vaccines.

The yield of temporary exclusion of blood donors, exposed to emerging infections abroad.

BACKGROUND AND OBJECTIVES: Emerging infections abroad pose a threat to the safety of blood, donated by travelling blood donors. In this study, the yield of donor deferral after travelling was evaluated, by comparing the estimated numbers of infected donors returning from various affected areas. METHODS: A deterministic model was applied to calculate the number of infected donors, returning from six areas affected by outbreaks: Greece - Macedonia (West Nile fever), Italy - Emilia Romagna (West Nile fever), Thailand (chikungunya), Latvia (hepatitis A), central Turkey (Sicilian sandfly fever) and Italy - Tuscany (Toscana sandfly fever). RESULTS: The estimated number of infections among returning blood donors was surprisingly low, ranging from 0·32 West Nile virus-infected donors per year returning from Macedonia (Greece) to approximately 0·005 infected donors per year returning respectively from Tuscany (sandfly fever), Latvia (hepatitis A) and central Turkey (sandfly fever). CONCLUSION: The yield of the temporary exclusion of blood donors travelling to a specific, affected area is low, but the continuous monitoring of emerging infections and the timely assessment of new threats are laborious and imperfect. Safety measures may be instituted after the greatest threat of a new outbreak has passed. A general deferral of travelling donors may be more appropriate than targeted measures. It can be argued that all donors who stayed outside their country or continent of residency should be deferred for 4 weeks.

Prophylaxis with cationic liposome-DNA complexes protects hamsters from phleboviral disease: importance of liposomal delivery and CpG motifs.

Cationic liposome-DNA complexes (CLDC) are cationic/neutral lipid carriers complexed with plasmid DNA that when administered systemically results in a robust T(H)1 cytokine response. CLDC have been shown to be effective in prophylaxis and therapeutic treatment of animal models of viral disease. To determine the contribution of liposomal delivery and CpG content of the plasmid DNA to the efficacy of CLDC; plasmid, CpG-free plasmid DNA, or CpG-containing oligodeoxynucleotides (ODN) with and without liposomes, as well as poly(I:C(12)U), were evaluated for their ability to elicit protection against lethal Punta Toro virus (PTV, Bunyaviridae, phlebovirus) challenge in hamsters. CLDC-containing plasmid significantly improved survival, decreased systemic and liver viral loads, and reduced liver damage due to progression of viral infection. Mouse-reactive ODNs complexed with liposomes failed to protect hamsters, whereas ODNs known to cross-react with human and mouse (CpG 2006) or non-liposomal poly(I:C(12)U) showed survival benefit but did not limit liver injury. Liposomes complexed with a non-CpG motif-containing plasmid reduced liver viral load and tissue damage, but did not protect hamsters from death. To evaluate the mechanisms of the enhanced activity of CLDC, microarray experiments examined differences in the gene expression profile. The results suggest a broad T(H)1 response elicited by liposomal delivery of a diverse sequence containing CpG and non-CpG elements may be a more effective antiviral treatment than other nucleic acid based immunotherapeutics.

Immunization with Toscana virus N-Gc proteins protects mice against virus challenge.

Toscana virus (TOSV) is an emerging virus, circulating in the Mediterranean area, that is responsible for aseptic meningitis, meningoencephalitis, and encephalitis. The development of a vaccine that could provide complete protection from TOSV infection is needed. In this study we investigated the capacity of TOSV structural proteins, nucleocapsid protein N and the two Gc and Gn glycoproteins, produced as recombinant proteins, in an animal model. In particular, we investigated their role in inducing specific and protective immune responses against virus infection. Mice were immunized intraperitoneally using TOSV antigens singly or in combination. The results show that only the N-Gc combination was able to protect 100% of animals from a lethal challenge with a neurovirulent strain of TOSV. This potential vaccine induces high serum antibody titres with neutralizing activity and it is safe for animals. Moreover, immunization induces a virus specific cell-mediated immune response, in particular a CD8+ T cell response associated with a marked expression of interferon gamma. These results indicate that the N+Gc viral antigen combination could be useful for future development of a vaccine controlling the spread of this emerging virus that could pose a new threat for humans.

[Sandflies in Italy: observations on their distribution and methods for control]. / I flebotomi in Italia: osservazioni sulla distribuzione e sui metodi di campionamento.

The paper analyzes published and unpublished data of sandfly collections carried out in Italy by one of the authors (M.M.) over a period of 18 years (1975-1993). These data are discussed in relation to the collection methods and the habitats. The leishmaniasis foci surveyed are localized in 11 regions of Central and Southern Italy and in the two main islands, Sicily and Sardinia. Five collection methods were used: i) sticky traps, ii) light traps, iii) window exit traps, iv) emergence traps, and v) hand catches. Captures were performed in domestic and sylvatic habitats in rural, urban and periurban areas. A total of 81,915 sandflies has been collected. Specimens were identified as belonging to 6 species: Phlebotomus perniciosus (46.1%), P. perfiliewi (43.8%), P. major (0.2%), P. mascittii (0.1%), P. papatasi (0.3%), and Sergentomyia minuta (9.5%). P. perniciosus, P. perfiliewi and S. minuta have been reported in almost all the regions samples, P. major only in 3 regions of South Italy (Apulia, Calabria and Sicily), P. mascitti in 4 regions of Central Italy (Tuscany, Latium, Abruzzo, Molise) and in Campania, P. papatasi in 6 regions without any definite geographical distribution. From the analysis of sticky trap captures P. perniciosus seems to be the prevalent species in the domestic habitat (65.6%) even if present in the sylvatic one (21.8%). The report of P. perniciosus in both situations might indicate the ability of the species to colonize a large range of habitats. P. perfiliewi resulted as the predominant species (88.1%) in the domestic area from collections with CDC light traps. S. minuta was the most abundant species in the sylvatic conditions (76.6%) while P. papatasi showed a clear endophilic behaviour reaching high percentages only in bedrooms and stables. Moreover, taxonomic characters of spermathecae utilized in species identification are reported.

[Data on a study of a natural focus of sandfly fever in Sevastopol during 1945-1988]. / Materialy po izucheniiu prirodnogo ochaga moskitnoi likhoradki v Sevastopole za 1945--1988 gg.

Sandfly [correction of Mosquito-borne] fever in Sevastopol was of a marked season character (June-September), with maximum cases registered in August. The morbidity curve corresponded to the sandfly [correction of mosquito] number curve, lagging behind it because of the incubation period. Since 1947 DDT and hexachlorane, having a lasting effect (40-50 days), have been widely used for sandfly [correction of mosquito] control. This was accompanied by intensive improvement of the living conditions in the city, regular sanitation and deratization. As a result of the above measures sandfly [correction of mosquito-borne] fever has not been registered in the city since 1954. The number of sandflies [correction of mosquitoes] reduced drastically, their species composition has changed: in 1959 sandflies [correction of mosquitoes] were found in 5.4% of the houses examined and in 1988, in 0.16% of houses. Since 1958 only 2 (Phlebotomus papatasii Scop and Ph. major Ann.) out of the former 7 species have been caught. Each species strives for self-preservation. Therefore, epidemiological survey of the former foci and sandfly-control [correction of antimosquito] measures are to be performed annually not to permit recovery of sandfly [correction of mosquito] population and repeated epidemics of sandfly [correction of mosquito-borne] fever.