The spectrum of ixodid ticks that bite humans in Western Siberia has significantly changed over the past two decades. In this study, we determined tick species attacking people in the vicinity of Novosibirsk and the range of bacterial agents they were infected with. This study included 301 ticks taken from people and 46% were Ixodes pavlovskyi, followed by Ixodes persulcatus (19.6%), I. persulcatus/I. pavlovskyi interspecies hybrids (19.6%), Dermacentor reticulatus (12.8%), and single Dermacentor marginatus and Dermacentor nuttalli/Dermacentor silvarum. Human DNA was determined in ticks, first demonstrating that all Ixodes spp., including hybrids, can effectively feed on humans. The DNA of Borrelia spp., Rickettsia spp., and Anaplasmataceae bacteria was detected in different tick species. Borrelia garinii prevailed in Ixodes species, being found in 8.8% of ticks, whereas B. afzelii and B. bavariensis were found in single ticks. Borrelia miyamotoi was revealed in 3.7% of ticks. “Candidatus Rickettsia tarasevichiae” and R. raoultii were identified mainly in I. persulcatus and D. reticulatus (44.8% and 26.3%, respectively), while Rickettsia helvetica was found only in 2.2% I. pavlovskyi. The prevalence of Anaplasma phagocytophilum, Ehrlichia muris, and Neoehrlichia mikurensis did not exceed 2%. The obtained results indicate a high risk for humans to be infected with agents of Lyme borreliosis, primarily B. garinii.

Omsk hemorrhagic fever virus (OHFV) is a member of the tick-borne encephalitis virus (TBEV) complex of the Flaviviridae family. Currently, there are no data on the cross-reactivity of antibodies to the NS1 proteins of OHFV and TBEV. Such data are of major interest for monitoring viral encephalitis of unknown etiology due to the increasing geographical distribution of OHFV. In this study, a recombinant OHFV NS1 protein was produced using the Escherichia coli expression system and purified. The recombinant OHFV NS1 protein was recognized by specific mice immune ascetic fluids to the native OHFV NS1 protein. A Western blot analysis and ELISA of the recombinant NS1 proteins of OHFV and TBEV were used to study the cross-reactivity of antibodies from immune ascites fluid obtained from OHFV-infected mice and mAbs against TBEV NS1. Anti-TBEV NS1 mouse monoclonal antibodies (mAbs) have been shown to not be cross-reactive to the OHFV NS1 protein. Sera from patients with confirmed tick-borne encephalitis (TBE) were examined by ELISA using recombinant OHFV NS1 and TBEV NS1 proteins as antigens. It was shown for the first time that cross-reactive antibodies to the OHFV NS1 protein were not detected in the sera of TBE patients, whereas the sera contained antibodies to the TBEV NS1 protein.

Omsk hemorrhagic fever virus (OHFV) is a member of the tick-borne encephalitis virus (TBEV) complex of the Flaviviridae family. Currently, there is no data on cross-reactivity of antibodies to the NS1 proteins of OHFV and TBEV. Such data are of major interest for monitoring viral encephalitis of unknown etiology due to the increasing geographical distribution of OHFV. In this study, recombinant OHFV NS1 protein was produced using Escherichia сoli expression system and purified. The recombinant OHFV NS1 protein was recognized by specific mice immune ascetic fluids to the native OHFV NS1 protein. Western blot analysis and ELISA of recombinant NS1 proteins of OHFV and TBEV were used to study the cross-reactivity of antibodies from immune ascites fluid obtained from OHFV-infected mice and mAbs against TBEV NS1. Anti-TBEV NS1 mouse monoclonal antibodies (mAbs) have been shown not to be cross-reactive to OHFV NS1 protein. Sera from patients with confirmed tick-borne encephalitis (TBE) were examined by ELISA using recombinant OHFV NS1 and TBEV NS1 proteins as antigens. It was shown for the first time that cross-reactive antibodies to OHFV NS1 protein were not detected in the sera of TBE patients whereas the sera contained antibodies to TBEV NS1 protein.

Previously, we described the mechanisms of development of autoimmune encephalomyelitis (EAE) in 3-month-old C57BL/6, Th, and 2D2 mice. The faster and more profound spontaneous development of EAE with the achievement of deeper pathology occurs in hybrid 2D2/Th mice. Here, the cellular and immunological analysis of EAE development in 2D2/Th mice was carried out. In Th, 2D2, and 2D2/Th mice, the development of EAE is associated with a change in the differentiation profile of hemopoietic bone marrow stem cells, which, in 2D2/Th, differs significantly from 2D2 and Th mice. Hybrid 2D2/Th mice demonstrate a significant difference in these changes in all strains of mice, leading to the production of antibodies with catalytic activities, known as abzymes, against self-antigens: myelin oligodendrocyte glycoprotein (MOG), DNA, myelin basic protein (MBP), and five histones (H1–H4) hydrolyze these antigens. There is also the proliferation of B and T lymphocytes in different organs (blood, bone marrow, thymus, spleen, lymph nodes). The patterns of changes in the concentration of antibodies and the relative activity of abzymes during the spontaneous development of EAE in the hydrolysis of these immunogens are significantly or radically different for the three lines of mice: Th, 2D2, and 2D2/Th. Several factors may play an essential role in the acceleration of EAE in 2D2/Th mice. The treatment of mice with MOG accelerates the development of EAE pathology. In the initial period of EAE development, the concentration of anti-MOG antibodies in 2D2/Th is significantly higher than in Th (29.1-fold) and 2D2 (11.7-fold). As shown earlier, antibodies with DNase activity penetrate cellular and nuclear membranes and activate cell apoptosis, stimulating autoimmune processes. In the initial period of EAE development, the concentration of anti-DNA antibodies in 2D2/Th hybrids is higher than in Th (4.6-fold) and 2D2 (25.7-fold); only 2D2/Th mice exhibited a very strong 10.6-fold increase in the DNase activity of IgGs during the development of EAE. Free histones in the blood are cytotoxic and stimulate the development of autoimmune diseases. Only in 2D2/Th mice, during different periods of EAE development, was a sharp increase in the anti-antibody activity in the hydrolysis of some histones observed.

With the use of the ELISA method to detect an antigen, reverse transcription with quantitative real-time PCR with subtype-specific fluorescent probes, phylogenetic analysis of E and NS1 gene nucleotide sequences, bioassays with suckling mice, hemagglutination and neuroinvasiveness tests there was made a comparison of the tick-borne encephalitis virus (TBEV) infection of ixodid ticks Ixodes persulcatus P.Schulze and Ixodes pavlovskyi Pomerantsev 1946 in the area of sympatria of their natural habitats in the Novosibirsk region during growth period of their populations with the replacement ofprevailing species of monodominant type of the ixodid population structure. The ratio of 2 tick species didn’t depend on biotopes ofpine or birch forest but rather on the distance from the Novosibirsk Scientific Center: the lower anthropogenic pressure the smaller I.pavlovskyi proportion. The TBEV rate (including both pathogenic and apathogenic for laboratory mice virus), spectra of the TBEV3 main genetic types, neurovirulence and hemagglutination activity were similarfor both I.persulcatus and I.pavlovskyi. However, the proportion ofpathogenic for laboratory mice virus and the TBEV Far Eastern subtype, as well as viral loads of Siberian and European types for the TBEV from I.pavlovskyi were significantly higher than those from I.persulcatus.

Spectrum of ixodid ticks that bite people in Western Siberia significantly changed over the past two decades. In this study, we determined tick species attacking people in surroundings of Novosibirsk and the range of bacterial agents they infected with. This study included 301 ticks taken from people and 46% were Ixodes pavlovsky, followed by Ixodes persulcatus (19.6%), I. persulcatus / I. pavlovskyi interspecies hybrids (19.6%), and Dermacentor reticulatus (12.8%). Human DNA was determined in ticks, first demonstrating that all these tick species, including hybrids, were able effectively feed on humans. DNA of Borrelia spp., Rickettsia spp. and Anaplasmatacreae bacteria was detected in different tick species. Borrelia garinii prevailed in Ixodes species, being found in 8.8% ticks, whereas B. afzelii and B. bavariensis were found in single ticks. Borrelia miyamotoi was revealed in 3.7% ticks. Among Rickettsia spp., “Candidatus Rickettsia tarasevichiae” and R. raoultii were identified mainly in I. persulcatus and D reticulatus (44.8% and 26.3%, respectively), whereas Rickettsia helvetica was found only in 2.2% I. pavlovsky. The prevalence of Anaplasma phagocytophilum, Ehrlichia muris and Neoehrlichia mikurensis did not exceed 2%. The obtained results indicate a high risk for humans to be infected with agents of Lyme borreliosis, primarily B. garinii.