Severe Fever with Thrombocytopenia Syndrome (SFTS) is a newly emerging tick-borne disease caused by the SFTS virus (SFTSV), which belongs to the phlebovirus in the Bunyaviridae family. SFTSV is enveloped with a tripartite ambisense RNA genome. The L segment encodes the viral RNA-dependent RNA polymerase, the M segment encodes the two glycoproteins, Gc and Gn, and the S segment encodes the nucleoprotein (NP) and the nonstructural protein (NSs). NP participates in ribonucleoprotein (RNP) packaging and commonly detected early after infection, suggesting that the N protein is possible to be used as a target antigen for early diagnosis of SFTSV infection. In this study, we analyzed a highly immunogenic multi-epitope using GnGc and NP genes from a consensus sequence of SFTSV strain isolated from infected patients in Korea. The selected genes are constructed to the expression vector plasmid pJHL65 and the recombinant plasmid vector was transformed into the Δasd Δlon ΔcpxR Salmonella Typhimurium attenuated strain JOL912 and the expression of these antigens was verified by immunoblotting assay. We observed the significant levels of systemic IgG and mucosal IgM responses against the JOL912-derived antigen in the immunized BALB/c mice. The level of CD3+CD4+, CD3+CD8+ T lymphocyte subpopulation and TNF-α were also highly regulated in splenic T cells re-stimulated in vitro with NP and Gn/Gc multi-epitope selected antigens. Therefore, immunized mice with NP and Gn/GC multi-epitope recombinant proteins of attenuated Salmonella delivery system elicited T cell-related immune response, inducing an effective immune response. In conclusion, the attenuated Salmonella expressing NP-GnGc multi-epitopes could be a novel vaccine candidate against the SFTS virus.

Ixodidae에 속하는 일부 참진드기들은 중증열성혈소판감소증후군(severe fever with thrombocytopenia syndrome; SFTS)을 비롯한 질병을 매개하는 공중보건학적으로 중요한 해충이다. 우리나라에서는 2013년 이후로 SFTS 환자 발생수가 지속적으로 증가하는 경향을 보이며, 경상북도는 전국적으로 2번째로 많은 SFTS 환자가 발생하는 지역이다. 본 연구에서는 2019년 경북 상주 지역의 진드기 분포를 조사하기 위해 시민의 생활 반경 주변을 도심 녹지지역, 관리 취약지역 및 농촌지역으로 구별하여 flagging 방법을 통해 진드기를 채집하였고, 채집 진드기내 SFTS 바이러스 보균 여부를 조사하였다. 채집된 진드기 수를 Collection Index (CI = tick number / 1h / 2 people)로 산출한 결과, 상주시 내 총 26개 지점에서 작은소피참진드기와 개피참진드기, 일본참진드기를 포함하는 총 CI 143의 진드기가 채집되었고, 그 중 작은소피참진드기가 96.5%(CI 138)로 우점종으로 확인되었다. 채집된 진드기의 약 92%(131 CI)는 인적이 드물고 시청이나 관계 당국의 관리가 이루어지지 않는 관리 취약지역에서 채집되었으나, 도심의 녹지지역와 농촌 지역에서는 8.4%(CI 12)의 진드기만 관찰되었다. 총 CI 143의 진드기를 26개 pooling 한 후 SFTS 바이러스 존재 여부를 조사하였으나, 모두 음성으로 확인되었다. 본 연구의 결과는 지역주민들로 하여금 진드기 매개 질병으로부터 안전한 생활을 하기 위한 권고 자료로 활용할 수 있을 것으로 기대된다.

Heamaphysalis longicornis is a major vector for Severe Fever with Thrombocytopenia Syndrome (SFTS) virus and the density of the vector has been increasing because of the climate change. The incidence of fatalities due to SFTS is increasing every year. In this study, to evaluate the SFTS transmission by ticks, the density of ticks mediating SFTS was monitored. Tick was collected every month from four different sites (Grass land, Mountain path, Grave, Copse) in Andong with the traps containing dry ice as CO2 attractants. Among 2,572 ticks of 3 species; H. longicornis, H. flava, and Ixodes nipponensis were most abundantly collected from April to August. H. longicornis is the richest species (92.8%), whereas Ixodes nipponensis was the least species (0.8%). The 54.5% of the sample were nymph stage and female/male ratio was 64.3%. According to the pathogene analysis, SFTS virus was detected from H. longicornis adult and larvae stages collected in July and August. In July samples, SFTS virus was detected only from grassland site, but the virus was observed in the sample from all four sites in August. For effective prevention of SFTS, the tick density should be continuously monitored based on the onset time of SFTS with the consideration of habitate, habit & life history of ticks.

The Ixodid ticks has been suspected to be the vector of severe fever with thrombocytopenia syndrome virus (SFTSV) in Korea. In order to confirm transovarial transmission of Ixodid ticks, a total of blood fed 284 ticks were collected from six host animals. They were identified as 3 genera and 4 species, Haemaphysalis longicornis (97.2%), H. flava (0.7%), Ixodes nipponensis (1.8%) and Amblyomma testudinarium (0.4%). Among them, 192 female adults hatched their eggs successfully. Our results showed a high prevalence of SFTSV among collected ticks (3.1%, 6/192 ticks), and we detected SFTSV from their larvae. This result suggests that the parent generation of H. longicornis can transmit SFTSV transovarially into their F1 progeny and H. longicornis might be one of major vectors of SFTSV in Korea.

Severe fever with thrombocytopenia syndrome (SFTS) is an emerging tick-borne zoonosis and SFTS virus is transmitted to humans by tick bites. The aim of this study was to monitor tick population and its SFTS virus infection. During March 2014 to October 2015, ticks were monthly sampled using dry-ice bait trap and flag method from 4 collecting points in Boeun-gun. A total of 16,500 ticks, including 14,646 Haemaphysalis longicornis (88.7%), 1,825 H. flava (11.1%), and 29 Ixodes nipponensis (0.2%), were collected. Ticks were pooled (5,156 ticks in 456 pools) and tested by RT-PCR and nested PCR and the minimum infection rate of ticks was 0.27% (14 pools) consisting of 0.13% H. longicornis (6 pools), 1.19% H. flava (8 pools). This annual surveillance study will be needed for long-term monitoring of ixodid ticks in Korea.

Ixodid ticks are notorious as the vector of severe fever with thrombocytopenia syndrome virus (SFTSV) in Korea. To confirm the transovarial transmission of SFTSV, we collected blood-fed adult ticks from animals in SFTS case reported area of Jeju island in 2015. A total of 476 ticks were collected from four host animal species, horse (81.3%), dog (9.0%), roe deer (7.1%) and wild boar (2.5%). They were identified as 1 genus and 2 species, Haemaphysalis longicornis (87.8%) and H. flava (12.2%). Among them, 131 adults hatched their eggs successfully. The body weight of females was highly correlated with the number of egg masses laid (P < 0.05). We confirmed that one adult H. flava was infected with SFTSV and its infection rate was 0.77%. The eggs of the SFTSV-infected adults also were infected with SFTSV and its transovarial transmission was confirmed. This result suggests that the parent generation of H. flava could transmit SFTSV transovarially into their F1 progeny and H. flava might be one of major vectors of SFTSV in Korea.