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At least 19 recordsLinked to original sources

In vitro studies on cell-mediated immune response to tick-borne encephalitis virus: findings in convalescents and human subclinical infections.

Peripheral blood leukocytes from tick-borne encephalitis (TBE) convelescents (manifest and inapparent forms) working with TBE virus-containing material and from individuals without specific serum virus neutralizing antibodies (VNA) were studied in capillary tube leukocyte migration experiments. Partially purified TBE virus preparations were used as antigen. Under standardized conditions a strong inhibitory reaction was observed in convalescents with significantly higher values in persons recovered from an abortive form of infection only. These results differed markedly from values recorded in persons without specific VNA. Significance of correlation between humoral VNA titres and the intensity of the cell-mediated component of the immune response (CM IR), as indicated by leukocyte migration inhibition (LMI) values, was less than P equals 0.25.

Adolescent↗

Concentration and purification of tick-borne encephalitis virus grown in suspensions of chick embryo cells.

Tick-borne encephalitis (TBE) virus grown in suspensions of chick embryo cells was precipitated by various concentrations of polyethylene glycol (PEG) 6000. Quantitative recovery was obtained at PEG concentrations of 5% and higher. As precipitation of contaminating non-viral protein increases with increasing PEG concentrations, best results with respect to purity and recovery were obtained at 5% PEG 6000. Analysis of virus concentrates by rate zonal centrifugation revealed two peaks--rapidly sedimenting haemagglutinin (RHA) associated with infectivity and slowly sedimenting haemagglutinin (SHA)--characteristic of Flaviviruses. Purification factors after PEG precipitation followed by rate zonal centrifugation in sucrose density gradients ranged from 50 to 200 for HA activity and from 40 to 50 for infectivity.

Animals↗

Replication dynamics of tick-borne encephalitis virus in, and glycolytic activity of, human diploid cells.

Tick-borne encephalitis (TBE) virus (Western subtype) strains replicated very rapidly in human diploid embryonic lung (HDEL) cells, namely within the first 24 hours of infection. Greater amounts of both intra- and extracellular virus were produced after inoculation of a virulent than of an attenuated strain. With the virulent strain, more infectious virus was found in the medium than in the cells: the opposite was true for the attenuated strain. No cytopathic effect was observed during the period studied. The rates of glucose uptake and lactate production by cells were markedly influenced by the infection. Lactate production was retarded by both virus strains to a comparable degree. Glucose uptake was markedly increased, appreciably more so after infection by the virulent than the attenuated strain. A method for differentiating between attenuated and virulent strains based on the determination of glucose conthe medium was developed.

Cell Line↗

Phenothiazine-induced alterations of immune response in experimental tick-borne encephalitis: morphological model analysis of events.

The depressive effect of trifluoperazine (TFP), a phenothiazine derivative, on the morphology of the development of immune response (IR) (humoral and cell-mediated component) was studied in mice given tick-borne encephalitis (TBE) virus, sheep red blood cells (SRBC) or the BCG vaccine. This effect was manifested by a decrease in the mitotic activity of lymphocytes and in the number of blastic transformations after antigenic stimulation. In virus-infected and TFP-given mice, lowered levels of specific virus neutralizing antibody (VNA), together with a pronounced reduction of the inflammatory response in the brain were found. No signs of cytotoxicity following administration of the drug were observed. The mechanism of the immunodepressive action of TFP are discussed.

Animals↗

Non-infectious tick-borne encephalitis antigen.

After formalin treatment of sucrose-acetone antigens, prepared from different tick-borne encephalitis (TBE) virus strains, their haemagglutination (HA) activity was preserved or even higher than that of untreated antigens. Formalin-treated TBE antigen was non-infectious and thus may be recommended for routine work to avoid laboratory infections.

Animals↗

Interference stimulated by tick-borne encephalitis virus: influence of divalent cations.

The effects of divalent cations (barium, cobalt, calcium, copper, magnesium, zinc and iron) on the interference of tick-borne encephalitis (TE) virus with vaccinia virus in Detroit-6 cell cultures were investigated. An increase in interference was shown in the presence of cobalt, zinc, copper, and magnesium. Barium and iron displayed a marked inhibitory action. But an increase in interference was not parallel with an increased production of interferon.

Barium↗

Experimental live tick-borne encephalitis vaccine (Labgat E5 "14" virus clone): volunteers 1 and 2 years after single-dose immunization.

Groups of volunteers given intramuscularly 5 or 6.5 dex or perorally 6.5 dex newborn mouse icLD50 of the plaquesegregated "14" clone of the Langat E5 virus strain (tick-borne encephalitis comples) were followed for periods of 12 and 24-27 months. Circulating specific virus neutralizing antibodies persisted in them in the absence of apparent reaction as evidenced by clinical, electroencephalographic and cerebrospinal fluid findings.

Administration, Oral↗

Field-isolate recombinant tick-borne encephalitis viruses define reporter-stability guidelines for antiviral testing in flaviviruses.

As arthropod-borne viruses continue to threaten populations globally, there is a pressing need for experimental systems that enable rapid antiviral discovery. Reverse-genetics platforms producing recombinant reporter orthoflaviviruses have been developed to address this gap. Here, we present two new recombinant tick-borne encephalitis viruses (TBEVrec) generated on a European-subtype Haselmühl Tiho1 isolate backbone. A reporter gene, either eGFP or Nluc, was inserted in the capsid-coding region of the genome downstream of the capsid RNA regulatory signal and separated from the complete viral polyprotein by a 2A self-cleaving peptide. TBEVrec was better rescued using the circular polymerase extension reaction (CPER) than with the infectious subgenomic amplicon (ISA) method. TBEVrec replicated efficiently in relevant human cell lines, with comparable replication to wild-type TBEV in a neuronal cell line and moderately reduced titers and RNA levels in immune-derived cell lines. Using either eGFP or Nluc, we illustrate how TBEVrec enabled high-content RNAi screening, highlighting Nucleolin and PRKD1 as potential TBEV host factors, and drug testing on a benchtop plate reader. Nanopore sequencing of the eGFP insert revealed that the reporter is excised without affecting flanking regions. Comparative analysis of eGFP and Nluc further shows that this instability is time- and cell type-dependent, and that Nluc is comparatively more stable. From these observations, we outline safeguards and design principles that are broadly applicable both to the rescue of existing constructs and to the design of future recombinant reporter virus platforms.

CPER↗

Widespread circulation of Alongshan virus in Austria and serological evidence for infection in humans: a nationwide molecular and serological observational study.

BACKGROUND: Alongshan virus, a segmented RNA virus in the Jingmenvirus group of flaviviruses, was first identified in 2017 in China in patients with tick-borne encephalitis-like illness. Alongshan virus has since been detected in ticks and mammals in several European countries. In this study, we aimed to characterise the temporal and geographical distribution of Alongshan virus in Austria through nationwide tick surveillance combined with serological and molecular screening in individuals with suspected tick-borne encephalitis or tick exposure. METHODS: In this nationwide molecular and serological observational study, we conducted a PCR-based screening of ticks collected across Austria in 2024, using flagging, animal hosts, and citizen submissions. We also collated genomic data from stored nucleic acid extracts from ticks collected in Austria in 2005 and 2013 in previous surveillance studies and stored extracts of paired tick-human samples collected between 2015 and 2018. Alongshan virus-positive tick samples were subjected to whole-genome sequencing and phylogenetic analysis. In addition to tick samples, blood samples from Austrian patients with reported tick bite or suspected tick-borne encephalitis, submitted to the National Reference Center for Human Arbovirus Infections, Austria, were screened for Alongshan virus infection and subjected to serological and molecular testing. FINDINGS: 2952 ticks were collected between Feb 1, 2024, and Dec 6, 2024, from 29 (83%) of 35 NUTS-3 regions in Austria; the median detection rate for Alongshan virus was 1·2% (IQR 0·4-3·5). In addition, 1816 archived tick samples were analysed, with three testing positive for Alongshan virus. For the virus-positive samples, phylogenetic analysis showed that sequences from Austria grouped within the European clade, with Austrian sequences from the same region showing high sequence similarity. 1361 human serum samples collected between March 1, 2023, and May 16, 2025, were assessed for anti-Alongshan virus IgG antibodies. Two individuals had high antibody titres against Alongshan virus proteins VP1a and VP2. INTERPRETATION: Our study shows detection of Alongshan virus in archived tick samples dating back to 2005, representing the earliest documented occurrence of the virus to date and suggesting that Alongshan virus has been circulating in Austria for at least two decades. The detection of Alongshan virus-specific antibody titres in two individuals suggests past infection and previously unrecognised exposure. These results highlight the need for continued molecular surveillance of tick populations and serological monitoring in humans to define the epidemiology and public health relevance of Alongshan virus in Europe. FUNDING: One Health surveillance and Vector monitoring for cross-border pathogens (OH SURVector) and UNITED4Surveillance.

Adolescent↗

The opposite temperature-sensitivity character (ts) in two attenuated flaviviruses, used for human immunization: 17D yellow fever and E5"14" (Langat) viruses. A reappraisal of thoughts.

For the reproduction of the man-attenuated E5"14" clone of the Langat virus (tick-borne encephalitis complex) in pig kidney epithelial cells, the temperature of 39 degrees C was shown as restrictive, whereas it was permissive for the 17D strain of yellow fever virus and three virulent strains of tick-borne encephalitis (western subtype) virus. The temperature of 36 degrees C permitted the reproduction of all viruses studied. The implication of genetic marker studies in the assessment of human neuropathogenicity of flaviviruses is discussed.

Arboviruses↗

Buoyant density of some togaviruses in sucrose density gradient and capacity of their haemagglutinin fractions to interact with antibody.

The buoyant densities of Western equine encephalomyelitis virus (an Alphavirus) and tick-borne encephalitis (TBE) virus (a Flavivirus) antigens prepared by different methods were studied. Sucrose density centrifugation revealed a heterogeneity in the density of the virions. The sedimentation pattern and height of peaks of the haemagglutinating activities and infectivity, other conditions being equal, depended both on the virus species and properties of its strains and on the mode of preparation and treatment of the virus-containing material. Different haemagglutinating antigen fractions differed in their capacity to interact with specific antibody. It was suggested that the kinetics of a serological reaction and its result depend on the functional activity of the antigen preparation and, in particular, on the proportion in the reaction mixture of virus particles with a dissimilar antigenic structure.

Antibody Specificity↗

Eyach--an arthropod-borne virus related to Colorado tick fever virus in the Federal Republic of Germany.

From Ixodes ricinus ticks collected in a tick-borne encephalitis focus in Baden-Württemberg, an agent pathogenic exclusively for suckling mice was isolated. Obviously a virus, it passed membrane filters with a pore width of 200 nm. It was resistant to ether and sodium deoxycholate, but not to chloroform. In the complement fixation test it showed a close, and in the neutralisation test a more one-sided relationship to Colorado tick fever virus.

Animals↗

A series of patients infected with the emerging tick-borne Yezo virus in China: an active surveillance and genomic analysis.

BACKGROUND: Yezo virus (YEZV) is an emerging tick-borne pathogen, which was initially reported in Japan in 2021. Only one patient had been reported in China so far. We aimed to describe the epidemiological, clinical, and laboratory findings of a series of patients, and to characterise the viral genomes of YEZV. METHODS: In this active surveillance and genomic analysis, we conducted active surveillance at Mudanjiang Forestry Central Hospital, Heilongjiang Province of northeast China. Participants were eligible for inclusion if they sought medical care for a recent tick bite between May 1 and July 31, in 2022 and 2023, and between May 1 and July 10, in 2024. We collected sera from participants to detect YEZV infection by meta-transcriptomic sequencing, real-time RT-PCR, and indirect immunofluorescence assay. We isolated YEZV by cell culture and characterised the pathogen by morphological and phylogenetic analyses. FINDINGS: A series of 18 patients with YEZV infection (12 male and six female; median age 53 years, IQR 45-60) were identified among 988 participants. The patients presented with fever (18 patients, 100%), headache (ten patients, 56%), dizziness (nine patients, 50%), malaise (three patients, 17%), lumbago (three patients, 17%), and cough (three patients, 17%). Nine (50%) patients had rash around the tick bite site and four (22%) had lymphadenopathy. Nine (50%) patients had gastrointestinal symptoms, and five (28%) had neurological symptoms. We observed leukopenia in ten (63%) and thrombocytopenia in five (31%) of 16 assessed patients. Elevated hepatic transaminase concentrations were identified in 13 (72%) of all 18 patients, lactate dehydrogenase or α-hydroxybutyric dehydrogenase in nine (50%), serum amyloid protein A in 13 (72%), and hypersensitive C-reactive protein in ten (56%). Eight (7%) of 119 Ixodes persulcatus ticks removed from participants were positive for YEZV. Three YEZV strains were isolated from the sera of patients. Ten viral genomes were obtained from five patients, a blood-sucking I persulcatus removed from a participant, and four host-questing tick samples collected in the areas where patients were identified or in the adjacent region. Phylogenetic analyses revealed that YEZVs in either patients or ticks were divided into two clades, each with distinct mutations. INTERPRETATION: Awareness of YEZV infection is important and clinicians should consider the virus when diagnosing patients with suitable symptoms. FUNDING: National Key Research and Development Program of China. TRANSLATION: For the Chinese translation of the abstract see Supplementary Materials section.

Humans↗

Establishment of reverse genetics systems for Colorado tick fever virus.

The Colorado tick fever virus (CTFV), which has 12-segmented double-stranded RNA genomes, is a pathogenic arbovirus that causes severe diseases in humans. However, little progress has been made in the analysis of replication mechanisms and pathogenicity. This virological constraint is due to the absence of a reverse genetics system for CTFV; therefore, we aimed to establish the system. Initially, the efficacy of CTFV replication was investigated in various cell lines. CTFV was found to grow in many cell types derived from different hosts and organs. Subsequently, BHK-T7 cells stably expressing T7 RNA polymerase were transfected with plasmids encoding each of the 12 CTFV gene segments, expression plasmids encoding all CTFV proteins, and a vaccinia virus RNA-capping enzyme. Following transfection, the cells were co-cultured with Vero or HeLa cells. Using this system, we rescued monoreassortants and recombinant viruses harboring peptide-tagged viral proteins. Furthermore, an improved system using Expi293F cells expressing T7 RNA polymerase was established, which enabled the generation of recombinant reporter CTFVs. In conclusion, these reverse genetics systems for CTFV will greatly contribute to the understanding of viral replication mechanisms, pathogenesis, and transmission, ultimately facilitating the development of rational treatments and candidate vaccines.

Animals↗