Browsing by Author "Kozireva, Svetlana"

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    Anemia as a complication of parvovirus B19 infection : In renal transplant recipients
    (2012) Čapenko, Svetlana; Kozireva, Svetlana; Folkmane, Inese; Bernarde, Kristina; Rozentals, Rafails; Murovska, Modra; Institute of Microbiology and Virology
    Background: The frequency of B19 infection in renal transplant donors and recipients was studied to determine the significance of active viral infection in the development of anemia. Material and Methods. Serum, plasma, and peripheral blood leukocyte samples of 47 renal transplant donors, 38 recipients with anemia (Group 1), and 25 without anemia (Group 2) after renal transplantation were evaluated for the presence of anti-B19 specific antibodies (ELISA) and B19 DNA (nPCR). Results: Active persistent B19 infection after renal transplantation was detected in 12 of the 38 in the Group 1 (10 had reactivation and 2 primary infection), and none of the recipients in the Group 2 had it. Of the 12 recipients in the Group 1, 10 were seropositive and 2 seronegative before renal transplantation; 10 received the transplants from the seropositive and 2 from seronegative donors. rHuEPO therapy-resistant severe anemia was detected only in the recipients with active B19 infection after renal transplantation in the Group 1 (7/12). The logistic regression analysis revealed a significant relationship between active B19 infection and severe anemia (OR, 0.039; 95% CI, 0.006-0.257; P=0.001). Conclusions: Active B19 infection was documented only in the anemic recipients and could be associated with the development of severe anemia after renal transplantation. This allows us to recommend concurrent screening for viral DNA in plasma and detection of anti-B19 IgM class antibodies. To find the association between B19 infection and the development of anemia, further investigations are necessary.
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    Chemokine Receptors CCR1 and CCR2 on Peripheral Blood Mononuclear Cells of Newly Diagnosed Patients with the CD38-Positive Chronic Lymphocytic Leukemia
    (2020-07-21) Kholodnyuk, Irina; Rivkina, Alla; Hippe, Laura; Svirskis, Simons; Kozireva, Svetlana; Ventina, Ildze; Spaka, Irina; Soloveichika, Marina; Pavlova, Jelena; Murovska, Modra; Lejniece, Sandra
    Chemokines and their receptors direct migration and infiltration of immune cells. CCR1 and CCR2 maintain sequence similarity and respond to a number of the same chemokines secreted in lymphoid organs. Expression of CD38 on leukemic cells has been associated with poor clinical outcomes in patients with chronic lymphocytic leukemia (CLL) and is considered as the negative predictor of progression. In our study of newly diagnosed CLL patients, which included 39 CD38-positive and 22 CD38-negative patients, CCR1 and/or CCR2 were always detected, using flow cytometry, on the peripheral blood (PB) CD19+CD5+ lymphocytes in patients with >30% of the CD38+ CD19+CD5+ lymphocytes (n = 16). Spearman’s rank correlation analysis determined correlations between the frequency of the CCR1- and CCR2-expressing PB CD19+CD5+ lymphocytes and the frequency of the CD38-positive CD19+CD5+ lymphocytes (rs = 0.50 and rs = 0.38, respectively). No significant correlations were observed between ZAP70 mRNA expression levels in PB mononuclear cells and the frequency of the circulating CCR1+ or CCR2+ CD19+CD5+ lymphocytes. Further association studies are needed to verify prognostic relevance of the CCR1/CCR2 expression on leukemic cells in CLL patients at diagnosis. We suggest that CCR1/CCR2 signaling pathways could represent attractive targets for development of CLL anti-progression therapeutics.
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    Effect of Human Herpesviruses 6 and 7 Infection on the Clinical Course of Rheumatoid Arthritis
    (2016-08-01) Kadiša, Anda; Nora-Krūkle, Zaiga; Kozireva, Svetlana; Svirskis, Simons; Studers, Peteris; Groma, Valerija; Lejnieks, Aivars; Murovska, Modra; Department of Internal Diseases; Institute of Microbiology and Virology; Joint Laboratory of Traumatology and Orthopaedics; Institute of Anatomy and Anthropology
    Rheumatoid arthritis (RA) is a chronic systemic autoimmune inflammatory disease affecting joints and causing symmetrical chronic progressive aseptic synovitis and erosive-destructive changes. Viruses and viral infections are considered to be the main risk factors for autoimmune disease development (especially for individuals with genetic predisposition). The goal of this study was to evaluate the frequency of HHV-6 and HHV-7 persistent infection and its activity phase in RA and osteoarthritis (OA) patients, and healthy persons. We examined also the influence of HHV-6 and-7 infections on RA activity, aggressiveness, radiographical stage, and frequency of complications as well as the presence of HHV-6 infection markers in synovial fluid and synovial tissues of RA joints of affected patients. Despite the lack of significant correlation between frequency of persistent single HHV-6, single HHV-7, and concurrent HHV-6 and HHV-7 infection and RA clinical course, we found that both active and latent HHV-6 and/or HHV-7 infection increased RA activity and progression in several clinical and laboratory parameters. Regarding the severity of the course of RA, we observed also a high prevalence of RA complications in the patient group with active single HHV-6 infection and also a more severe radiographical stage in RA patients with active concurrent HHV-6 and HHV-7 infection. Moreover, viral infection markers were found in synovial fluid and synovial tissues of affected joints of RA patients. This suggests that HHV-6 and/or HHV-7 infection has effect on the disease clinical course, but virus reactivation may be a consequence of immunosuppressive treatment.
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    Influence of the Time-Phase Varied Magnetic Field on the Nucleic Acids Delivery Into the Cancer Cells. Summary of the Doctoral Thesis
    (Rīga Stradiņš University, 2016) Avotiņa, Dace; Bariševs, Mihails; Kozireva, Svetlana
    Gene delivery is one of the critical steps in gene therapy, including both efficient delivery of the therapeutic gene to the target cells and attachment to the cell membrane with following internalization and transfer to the nucleus. Magnetofection is one of the most effective physical gene delivery methods based on the increased delivery and concentration of nucleic acids coupling superparamagnetic iron oxide particles (SPION) onto the cell surface by the influence of an external magnetic field. In the static magnetic field generated by permanent magnets the movement of SPION in solution is axial towards the magnet. The time-phase varied magnetic field which is generated with the magnetofection device DynaFECTOR is based on the orbital rotation of the permanent magnets plate in the parallel plane of the cell culture plate. In the result of computer modelling it was shown that the orbital rotation of magnets causes the axial-lateral movement of SPION in parallel and perpendicular plane against the magnet. This led to the more uniform distribution of SPION onto the surface and could be a cause of the enhanced SPION-nucleic acids complexes internalization. In this study the influence of the time-phase varied magnetic field on the delivery efficiency of SPION-nucleic acids-liposomal component complexes into the cancer cells was experimentally investigated. Obtained results showed that in the time-phase varied magnetic field the SPION sedimentation onto the surface is more uniform and the amount of internalized SPION increases when compared to static magnetic field. Under the influence of the time-phase varied magnetic field, the delivery of SPION-nucleic acids-liposomal component complexes also increases, as evidenced by the significant increase in both the number of transfected cells and the total amount of an expressed protein in transfected cells in comparison with other gene delivery methods. While SPION-nucleic acids-liposomal component complexes delivery into the cells with less cytotoxic effect in provided by more uniform sedimentation onto the surface. In this study enhanced effective gene delivery method – the liposomal magnetofection in the time-phase varied magnetic field is reported. The method could be used for the replacement of mutated genes and for the investigation of the inhibition effect of novel therapeutic siRNAs in vitro cell lines. The method could potentially used also for the therapeutic gene delivery into the cancer cells in vivo.
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    Laika-fāzes variējoša magnētiska lauka ietekme uz nukleīnskābju piegādes efektivitāti vēža šūnās. Promocijas darba kopsavilkums
    (Rīgas Stradiņa universitāte, 2016) Avotiņa, Dace; Bariševs, Mihails; Kozireva, Svetlana
    Gēnu piegāde ir viens no problemātiskākajiem posmiem gēnu terapijā, jo ietver ne tikai terapeitiskā gēna aizvadīšanu līdz šūnām, bet arī efektīvu piesaisti šūnas membrānai ar sekojošu ieslēgšanu šūnā un migrāciju šūnas iekšējā vidē līdz kodolam. Magnetofekcija ir viena no efektīvākajām fizikālajām gēnu piegādes metodēm, kuras pamatā ir ar superparamagnētiskajām nanodaļiņām (SPION) saistītu nukleīnskābju paātrināta koncentrēšana uz šūnu virsmas un piegāde šūnās, izmantojot spēcīgu ārējo magnētisko lauku. Permanentu magnētu radītā statiskajā magnētiskajā laukā SPION kustība šķīdumā virzienā pret magnētu ir aksiāla. Laika-fāzes variējoša magnētiskā lauka, kas tiek ģenerēts ar magnetofekcijas iekārtas DynaFECTOR palīdzību, pamatā ir permanentu magnētu plates orbitāla rotācija šūnu kultūras platei paralēlā plaknē. Datormodelēšanas rezultātā tika noskaidrots, ka laika-fāzes variējoša magnētiskā lauka ietekmē notiek SPION aksiāli laterāla kustība magnētam paralēlā un perpendikulārā plaknē. Tas veicina SPION vienmērīgāku sedimentāciju uz virsmas un varētu būt par iemeslu paaugstinātai SPION-nukleīnskābju kompleksu ieslēgšanai šūnās. Šajā pētījumā eksperimentāli tika pārbaudīta laika-fāzes variējoša magnētiskā lauka ietekme uz SPION-nukleīnskābju-liposomālās komponentes kompleksu piegādes efektivitāti vēža šūnās. Iegūtie rezultāti parādīja, ka laika-fāzes variējošā magnētiskajā laukā, salīdzinot ar statisko magnētisko lauku notiek vienmērīgāka SPION sedimentācija uz virsmas un paaugstinās šūnās ieslēgto SPION daudzums. Laika-fāzes variējoša magnētiskā lauka ietekmē paaugstinās arī SPION-nukleīnskābju-liposomālās komponentes kompleksu piegāde šūnās, par ko liecina būtiski paaugstināts gan transficēto šūnu skaits, gan arī kopējais ekspresētā proteīna līmenis transficētajās šūnās, salīdzinot ar citām gēnu piegādes metodēm. Savukārt vienmērīgāka sedimentācija uz virsmas laika-fāzes variējoša magnētiskā lauka ietekmē nodrošina SPION-nukleīnskābju-liposomālās komponentes kompleksu piegādi šūnās ar samazinātu citotoksisko efektu. Šajā pētījumā aprakstīta uzlabota efektīva gēnu piegādes metode – liposomālā magnetofekcija laika-fāzes variējošā magnētiskajā laukā, kuru varētu izmantot mutēto gēnu aizstāšanai un jaunu terapeitisku siRNS inhibējošā efekta pētījumiem in vitro šūnu līnijās. Metode perspektīvā varētu tikt pielietota arī terapeitisko gēnu pārnesē vēža šūnās in vivo.
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    Laika-fāzes variējoša magnētiska lauka ietekme uz nukleīnskābju piegādes efektivitāti vēža šūnās. Promocijas darbs
    (Rīgas Stradiņa universitāte, 2016) Avotiņa, Dace; Bariševs, Mihails; Kozireva, Svetlana
    Gēnu piegāde ir viens no problemātiskākajiem posmiem gēnu terapijā, jo ietver ne tikai terapeitiskā gēna aizvadīšanu līdz šūnām, bet arī efektīvu piesaisti šūnas membrānai ar sekojošu ieslēgšanu šūnā un migrāciju šūnas iekšējā vidē līdz kodolam. Magnetofekcija ir viena no efektīvākajām fizikālajām gēnu piegādes metodēm, kuras pamatā ir ar superparamagnētiskajām nanodaļiņām (SPION) saistītu nukleīnskābju paātrināta koncentrēšana uz šūnu virsmas un piegāde šūnās, izmantojot spēcīgu ārējo magnētisko lauku. Permanentu magnētu radītā statiskajā magnētiskajā laukā SPION kustība šķīdumā virzienā pret magnētu ir aksiāla. Laika-fāzes variējoša magnētiskā lauka, kas tiek ģenerēts ar magnetofekcijas iekārtas DynaFECTOR palīdzību, pamatā ir permanentu magnētu plates orbitāla rotācija šūnu kultūras platei paralēlā plaknē. Datormodelēšanas rezultātā tika noskaidrots, ka laika-fāzes variējoša magnētiskā lauka ietekmē notiek SPION aksiāli laterāla kustība magnētam paralēlā un perpendikulārā plaknē. Tas veicina SPION vienmērīgāku sedimentāciju uz virsmas un varētu būt par iemeslu paaugstinātai SPION-nukleīnskābju kompleksu ieslēgšanai šūnās. Šajā pētījumā eksperimentāli tika pārbaudīta laika-fāzes variējoša magnētiskā lauka ietekme uz SPION-nukleīnskābju-liposomālās komponentes kompleksu piegādes efektivitāti vēža šūnās. Iegūtie rezultāti parādīja, ka laika-fāzes variējošā magnētiskajā laukā, salīdzinot ar statisko magnētisko lauku notiek vienmērīgāka SPION sedimentācija uz virsmas un paaugstinās šūnās ieslēgto SPION daudzums. Laika-fāzes variējoša magnētiskā lauka ietekmē paaugstinās arī SPION-nukleīnskābju-liposomālās komponentes kompleksu piegāde šūnās, par ko liecina būtiski paaugstināts gan transficēto šūnu skaits, gan arī kopējais ekspresētā proteīna līmenis transficētajās šūnās, salīdzinot ar citām gēnu piegādes metodēm. Savukārt vienmērīgāka sedimentācija uz virsmas laika-fāzes variējoša magnētiskā lauka ietekmē nodrošina SPION-nukleīnskābju-liposomālās komponentes kompleksu piegādi šūnās ar samazinātu citotoksisko efektu. Šajā pētījumā aprakstīta uzlabota efektīva gēnu piegādes metode – liposomālā magnetofekcija laika-fāzes variējošā magnētiskajā laukā, kuru varētu izmantot mutēto gēnu aizstāšanai un jaunu terapeitisku siRNS inhibējošā efekta pētījumiem in vitro šūnu līnijās. Metode perspektīvā varētu tikt pielietota arī terapeitisko gēnu pārnesē vēža šūnās in vivo.
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    A novel approach for nucleic acid delivery into cancer cells
    (2012) Vainauska, Dace; Kozireva, Svetlana; Karpovs, Andrejs; Čistjakovs, Maksims; Bariševs, Mihails; Institute of Microbiology and Virology
    Background. Liposomal magnetofection is based on the use of superparamagnetic particles and cationic lipids and shows better transfection efficiency than other common nonviral gene delivery methods; however, the distribution of aggregate complexes over the cell surface may be ununiform. The use of a dynamic gradient magnetic field could overcome this limitation. A newly developed device for magnetofection under a dynamic magnetic field was used to compare the transfection efficiency of prostate carcinoma cell line PC3 with that obtained by lipofection and magnetofection. Material and Methods. Reporter plasmid pcDNA3.1LacZ DNA was used in combination with Lipofectamine2000 reagent and superparamagnetic nanoparticles CombiMag. The effects of incubation time under a dynamic magnetic field and a rotation frequency of magnets on transfection efficiency for PC3 cell line were determined. Alternatively, lipofection and liposomal magnetofection were carried out. Transfection efficiency of delivery methods was estimated by β-galactosidase staining; cell viability, by acridine orange/ethidium bromide staining. Results. Liposomal magnetofection under a dynamic gradient magnetic field demonstrated the highest transfection efficiency: it was greater by almost 21% and 42% in comparison with liposomal magnetofection and lipofection, respectively. The optimal incubation time under dynamic magnetic field and the optimal magnet rotation frequency were 5 minutes and 5 rpm, respectively. Liposomal magnetofection under a dynamic gradient magnetic field was less cytotoxic (7%) than that under a permanent magnetic field (17%) and lipofection (11%). Conclusions. Our new approach, based on the use of a dynamic gradient magnetic field, enhanced the transfection efficiency and had a less cytotoxic effect on prostate cancer cells in comparison with the standard magnetofection and lipofection.
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    Peripheral Blood Mononuclear Cells' Proliferative Response to Human Parvovirus B19 Antigens in Patients with Rheumatoid Arthritis
    (2016-08-01) Bratslavska, Olga; Kozireva, Svetlana; Kadisa, Anda; Svirskis, Simons; Pavlova, Jeļena; Lejnieks, Aivars; Murovska, Modra; Institute of Microbiology and Virology; Department of Internal Diseases
    This study aimed to determine peripheral blood mononuclear cells' (PBMC) proliferative response to parvovirus B19 (B19) antigens in rheumatoid arthritis (RA) patients and possible changes in proliferative response due to chemotherapy. Serum and blood samples of 52 RA patients and 25 sex and age matched healthy individuals were examined for the presence of anti-B19 IgG and IgM class antibodies and virus specific DNA sequence by the recomLine B19 test and nested polymerase chain reaction, respectively. The PBMC proliferative activity was estimated on the 3rd and 6th day of PBMC cultivation in the presence of virus and B19 VP1/VP2 peptide, using thymidine incorporation assay. On the 3rd day, PBMC response to B19 antigens was detected in 74.1% RA patients with active, in 44.8%-with remote and in 40%-with latent stage of persistent B19 infection, while in the control group the response was observed only in two individuals with active viral infection. On the 6th day, the response was found in 50% RA patients with active, 68.9%-with remote and in 80%-with latent stage of latent persistent infection as well as in 41.1% remotely infected control individuals. The highest PBMC mean stimulation indices were detected in the RA patients and control persons with active infection as well as in RA patients with latent stage of persistent viral infection. On the 3rd and 6th day, strong proliferative response was significantly more frequently observed in RA patients not receiving methotrexate treatment, compared to the patients receiving methotrexate treatment in different combinations with other drugs. RA patients had more frequent and faster response to B19 antigens than apparently healthy persons.
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    Upregulation of the chemokine receptor CCR2B in Epstein-Barr Virus-positive Burkitt lymphoma cell lines with the latency III program
    (2018-05-03) Kozireva, Svetlana; Rudevica, Zhanna; Baryshev, Mikhail; Leonciks, Ainars; Kashuba, Elena; Kholodnyuk, Irina; Institute of Microbiology and Virology
    CCR2 is the cognate receptor to the chemokine CCL2. CCR2–CCL2 signaling mediates cancer progression and metastasis dissemination. However, the role of CCR2–CCL2 signaling in pathogenesis of B-cell malignancies is not clear. Previously, we showed that CCR2B was upregulated in ex vivo peripheral blood B cells upon Epstein-Barr virus (EBV) infection and in established lymphoblastoid cell lines with the EBV latency III program. EBV latency III is associated with B-cell lymphomas in immunosuppressed patients. The majority of EBV-positive Burkitt lymphoma (BL) tumors are characterized by latency I, but the BL cell lines drift towards latency III during in vitro culture. In this study, the CCR2A and CCR2B expression was assessed in the isogenic EBV-positive BL cell lines with latency I and III using RT-PCR, immunoblotting, and immunostaining analyses. We found that CCR2B is upregulated in the EBV-positive BL cells with latency III. Consequently, we detected the migration of latency III cells toward CCL2. Notably, the G190A mutation, corresponding to SNP CCR2-V64I, was found in one latency III cell line with a reduced migratory response to CCL2. The upregulation of CCR2B may contribute to the enhanced migration of malignant B cells into CCL2-rich compartments.
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    Use of exploratory factor analysis to ascertain the correlation between the activities of rheumatoid arthritis and infection by human parvovirus B19
    (2015) Kakurina, Natalja; Kadisa, Anda; Lejnieks, Aivars; Mikazane, Helena; Kozireva, Svetlana; Murovska, Modra; Department of Internal Diseases; Institute of Microbiology and Virology
    Background and objective: We evaluated a possible correlation between the clinical activities of rheumatoid arthritis (RA) and human parvovirus B19 (B19) infection using exploratory factor analysis (EFA). Materials and methods: RA patients were organized into two groups: 100 patients in the main group and 97 in the RA(DAS28) group. Four subgroups were defined from the main group according to the presence or absence of certain infection-specific markers: group I comprised 43 patients who had IgG antibodies against B19; group II, 25 patients with active B19 infection (B19-specific IgM antibodies and/or plasma viremia); group III, 19 patients with latent/persistent B19 infection (virus-specific sequences in peripheral blood leukocytes' DNA with or without B19-specific IgG antibodies), and group IV, 13 patients without infection markers. The RA(DAS28) group was divided into four subgroups similarly to the main group: group I, 35; group II, 31; group III, 19; and group IV, 12 patients. Disease-specific clinical values in both groups were analyzed employing EFA, and the RA(DAS28) group was additionally assessed using Disease Activity Score (DAS)28. Results: RA activity was higher in patients who had markers of B19 infection. The highest activity of RA in both study groups was in patients with latent/persistent infection. In the RA(DAS28) group, according to DAS28, the highest activity of RA was in patients with active B19 infection. Conclusions: Using EFA and DAS28, a correlation between the clinical activity of RA and B19 infection was confirmed. These data suggest that EFA is applicable for medico-biological studies.