Aqueous C60 fullerene solution effects on cell viability

Publisher Copyright: © 2021 Proceedings of the Latvian Academy of Sciences. Section B. Natural, Exact, and Applied Sciences.. All right reserved.Fullerenes are carbon nanoparticles with the ability to quench reactive oxygen species. The biomedical potential of fullerenes is diminished by their low solubility in water, but many approaches have been developed to bypass this problem, like chemical modification of the carbon cage and the use of the solvent exchange method to transfer fullerenes from one solvent to the other. These two approaches were used in this study. Carboxylated fullerene aqueous solution was acquired using solvent exchange method transferring fullerene nanoparticles (C60) from toluene to water. Effects of varying concentration (0.5, 1, 1.5, 2, 2.5, 3, 5, 10 µM) of aqueous fullerene solution on cell viability and their antioxidative capabilities were evaluated on PC-3 and on monocytes isolated from a blood donor using Resazurin Cell Viability Assay. PC-3 cell viability was drastically affected by the 10 µM fullerene solution but remained relatively stable when treated with other concentrations even after longer periods of incubation with resazurin dye. Elevated cell viability was observed in monocytes treated with various fullerene concentrations, possibly indicative of fullerene protective activity against oxidative stress.publishersversionPeer reviewe

Investigation of the Involvement of HHV-6 Encoded Viral Chemokine Receptors in Autoimmune Thyroiditis Development

Funding Information: We have no competing interests to declare. This work was supported by grant No.1.1.1.2/VIAA/1/16/202, Agreement No. 9.-14.5/ 257 “Human herpesvirus-6 involvement in development of autoimmune thyroiditis.” Publisher Copyright: Copyright © 2022 Sultanova et al.The study of human herpesvirus-6 (HHV-6) involvement in autoimmunity development is very challenging, due to the complex nature of this virus. HHV-6 is a ubiquitous, lifelong persistent, and immunomodulating virus, which mainly spreads in solid tissues using cell-to-cell mechanics, and thus can escape from the host’s immune response.Human herpesvirus-6 (HHV-6) contains two genes (U12 and U51) that encode putative homologues of human G-protein-coupled receptors like CCR1, CCR3, and CCR5. It has been shown that these viral proteins can be expressed on the surface of epithelial and some peripheral blood mononuclear cells, suggesting that they could potentially induce autoimmunity. We aimed to investigate the possibility of HHV-6 encoded viral chemokine receptors (U12 and U51) involvement in autoimmune thyroiditis (AIT) development by detecting viral peptide specific antibodies in AIT patient samples. Seventy-nine AIT patients whose thyroid tissues were shown to be positive for HHV-6 and 32 blood donors were enrolled in this study. Twenty-eight synthetic peptides derived from HHV-6 U12 and U51 proteins’ amino acid sequences, as well as recombinant human CCR1, CCR3, and CCR5 proteins were used in suspension multiplex immunological assay to detect specific IgG and IgM antibodies. HHV-6 peptide specific IgG and IgM antibodies were found in patients’ samples. AIT patients' samples were found to be more frequently positive for peptide IgGs in comparison to control group’s samples. Even though peptide antibody cross-reactivity with human CCRs was not demonstrated, our results show a new immunogenic HHV-6 antigen—a possible new player in the HHV-6 induced autoimmunity exacerbation. IMPORTANCE The study of human herpesvirus-6 (HHV-6) involvement in autoimmunity development is very challenging, due to the complex nature of this virus. HHV-6 is a ubiquitous, lifelong persistent, and immunomodulating virus, which mainly spreads in solid tissues using cell-to-cell mechanics, and thus can escape from the host’s immune response. It has been implicated as an environmental factor in several autoimmune diseases. An association between HHV-6 and autoimmune thyroiditis has been demonstrated, yet clear mechanism of involvement remains to be elucidated, since the virus can be detected in nearly all autoimmune thyroiditis patient thyroid glands. Our results show new potentially immunogenic human herpesvirus-6 antigens—possible new players in the HHV-6 induced autoimmunity exacerbation, which could be subjects for further research. Together with previously published results, this study described possible mechanisms which may underlie the induction of autoimmune reactivities against thyroid tissues in AIT.publishersversionPeer reviewe

Thyrocytes as the Target Cells for Hhv-6 Infection in Patients with Autoimmune Thyroiditis

Publisher Copyright: © 2016 by Alina Sultanova.Human herpesvirus-6 (HHV-6) is a ubiquitous betaherpesvirus with immunomodulating properties that have been suggested to play an important role in the development of several autoimmune disorders. Although the primary targets for HHV-6 replication, both in vitro and in vivo, are CD4+ and CD8+ T lymphocytes, some studies have reported the presence of HHV-6 sequences in different solid organs, including in the thyroid gland, showing possible involvement of this herpesvirus in development of autoimmune thyroid disease. The aim of this study was to determine loads of HHV-6 in thyroid gland tissue in comparison to those in peripheral blood of patients with autoimmune thyroiditis. Seven patients [women mean age 45 (28-65)] with histologically confirmed autoimmune thyroiditis were enrolled in this study. Fluorescence-activated cell sorting was used to distinguish and sort lymphocyte populations from peripheral blood mononuclear cells of patients. HHV-6 load was determined by real-time PCR for peripheral blood and thyroid gland tissue samples. Additionally, all results from molecular analyses were compared with histological results obtained by light microscopy. Viral load was detected only in one (46 viral copies/1×106cells) blood sample; others were under the detection limit of the used kit. However, in all HHV-6 positive tissue samples viral load was detected in the range of 132-1620 viral copies/106 cells. Substantial HHV-6 load in lymphocyte subpopulations was detected in two of seven patients. HHV-6 load was detected in NK and CD95+ cells of two patients. The obtained results show that thyroid gland cells (tyrocytes) act as target cells for HHV-6.publishersversionPeer reviewe

COVID-19: the third wave of coronavirus infection outbreak.

The novel coronavirus SARS-CoV-2 poses a great public health crisis. As of December 2019, it has spread all over the world with cases reported in more than 100 countries and the number of infected individuals surpassing 1,000,000. On March 11 World Health Organization officially characterized the COVID-19 as a pandemic. Although genetic analysis revealed some similarities between the novel coronavirus and the causative agent of the 2002 SARS epidemic, both viruses have significant differences. Research done on SARS-CoV has greatly aided the understanding of SARS-CoV-2, as it served as a knowledge base and helped to identify the cell entry receptor and some other features of the disease, but not all of them. Several critical questions remain unanswered and specific therapeutics and vaccine candidates are lacking

Beta-herpesvīrusu (HHV-6, HHV-7) infekcijas nozīme imūnā disbalansa apstākļos. Promocijas darbs

This work has been carried out at Rīga Stradiņš University (RSU) August Kirchenstein Institute of Microbiology and Virology in collaboration with Pauls Stradins Clinical University Hospital (Latvian Transplantation Center), Riga East Clinical University Hospital (Latvian Oncology Center and Clinic “Gaiļezers”) and Riga 1st Hospital. Defence: on 3rd of June 2014, at 15.00 during Rīga Stradiņš University Promotional Council of Medicine meeting in 16 Dzirciema Street, in the Lecture theatre Hippocrates.Human herpesvirus-6 (HHV-6) and human herpesvirus-7 (HHV-7) are ubiquitous beta-herpesviruses widely distributed in the general population. They primary infection usually occurs in the early years of life and remains latent in the host for the lifelong period. At present there are numbers of studies trying to find and evaluate the role of beta-herpesviruses infection in the development of various chronic diseases, but still there is no final answer to this question. It could be due to ther ubiquitous nature and different mechanisms of interference with the host organism that these viruses are using. The aim of the present study was to ascertain the involvement of beta-herpesviruses infection in the pathogenesis and clinical course of chronic diseases and development of post-transplant complications due to their ability to change host-pathogen interaction. Three groups of patients were enrolled in this study: renal transplant recipients with immunosuppressive therapy (after renal transplantation), patients with underlying disease (gastrointestinal cancer) and patients with autoimmune process (autoimmune thyroiditis). Practically healthy blood donors and thyroid tissue autopsy specimens were included as a control. Qualitative and quantitative polymerase chain reactions (PCR) were used to detect presence of viral genomic sequences, infection activity stage, HHV-6 strain and viral load. Viral specific antibodies were detected by enzyme-linked immunosorbent assay (ELISA) and chemiluminiscence test, cytokines’ expression level – by ELISA, immunocompetent cells’ subpopulations – by Becton Dickinson (USA) laser flow cyto-fluorimeter, sorted and further analysed. The results showed various frequency rates and activity stages of HHV-6 and HHV-7 infection in these three patient groups with different immune system dysfunctions, as well as differences in immune system responses and changes in immunocompetent cells’ populations. Comparison of three mentioned patients’ groups revealed that patients with anti-rejection medical treatment had the most severe immunosuppression and higher beta-herpesvirus activation risk leading to the development of different complications. The lowest beta-herpesviruses infection frequency and activation rate was detected in patients with gastrointestinal cancer, at the same time strong association of HHV-6 and HHV-7 active infection with lymphopenia was demonstrated and higher mortality rate was observed between the patients with lymphopenia than without it. In patients with autoimmune thyroiditis high HHV-6 and HHV-7 infection frequency was found, indicating that viral infection could be implicated in the disease development. Moreover, higher viral load in thyroid tissue as in whole blood of autoimmune thyroiditis patients’ was an additional evidence of HHV-6 involvement in the disease development and was as indicator that thyroid gland is one of the places of HHV-6 latency creating a precondition for autoimmune process development.Doctoral Thesis was supported by ESF project “Support for doctoral study programs and research degrees RSU” 2009/0147/1DP/1.1.2.1.2/09/IPIA/VIAA/009

Beta-herpesvīrusu (HHV-6, HHV-7) infekcijas nozīme imūnā disbalansa apstākļos. Promocijas darba kopsavilkums

This work has been carried out at Rīga Stradiņš University (RSU) August Kirchenstein Institute of Microbiology and Virology in collaboration with Pauls Stradins Clinical University Hospital (Latvian Transplantation Center), Riga East Clinical University Hospital (Latvian Oncology Center and Clinic “Gaiļezers”) and Riga 1st Hospital. Defence: on 3rd of June 2014, at 15.00 during Rīga Stradiņš University Promotional Council of Medicine meeting in 16 Dzirciema Street, in the Lecture theatre Hippocrates.Human herpesvirus-6 (HHV-6) and human herpesvirus-7 (HHV-7) are ubiquitous beta-herpesviruses widely distributed in the general population. They primary infection usually occurs in the early years of life and remains latent in the host for the lifelong period. At present there are numbers of studies trying to find and evaluate the role of beta-herpesviruses infection in the development of various chronic diseases, but still there is no final answer to this question. It could be due to ther ubiquitous nature and different mechanisms of interference with the host organism that these viruses are using. The aim of the present study was to ascertain the involvement of beta-herpesviruses infection in the pathogenesis and clinical course of chronic diseases and development of post-transplant complications due to their ability to change host-pathogen interaction. Three groups of patients were enrolled in this study: renal transplant recipients with immunosuppressive therapy (after renal transplantation), patients with underlying disease (gastrointestinal cancer) and patients with autoimmune process (autoimmune thyroiditis). Practically healthy blood donors and thyroid tissue autopsy specimens were included as a control. Qualitative and quantitative polymerase chain reactions (PCR) were used to detect presence of viral genomic sequences, infection activity stage, HHV-6 strain and viral load. Viral specific antibodies were detected by enzyme-linked immunosorbent assay (ELISA) and chemiluminiscence test, cytokines’ expression level – by ELISA, immunocompetent cells’ subpopulations – by Becton Dickinson (USA) laser flow cyto-fluorimeter, sorted and further analysed. The results showed various frequency rates and activity stages of HHV-6 and HHV-7 infection in these three patient groups with different immune system dysfunctions, as well as differences in immune system responses and changes in immunocompetent cells’ populations. Comparison of three mentioned patients’ groups revealed that patients with anti-rejection medical treatment had the most severe immunosuppression and higher beta-herpesvirus activation risk leading to the development of different complications. The lowest beta-herpesviruses infection frequency and activation rate was detected in patients with gastrointestinal cancer, at the same time strong association of HHV-6 and HHV-7 active infection with lymphopenia was demonstrated and higher mortality rate was observed between the patients with lymphopenia than without it. In patients with autoimmune thyroiditis high HHV-6 and HHV-7 infection frequency was found, indicating that viral infection could be implicated in the disease development. Moreover, higher viral load in thyroid tissue as in whole blood of autoimmune thyroiditis patients’ was an additional evidence of HHV-6 involvement in the disease development and was as indicator that thyroid gland is one of the places of HHV-6 latency creating a precondition for autoimmune process development.Doctoral Thesis was supported by ESF project “Support for doctoral study programs and research degrees RSU” 2009/0147/1DP/1.1.2.1.2/09/IPIA/VIAA/009

Significance of Beta-Herpesviruses (HHV-6, HHV-7) Infection under the Conditions of Immune Disorders. Summary of the Doctoral Thesis

Promocijas darbs izstrādāts Rīgas Stradiņa universitātes (RSU) Augusta Kirhenšteina Mikrobioloģijas un virusoloģijas institūtā sadarbībā ar Paula Stradiņa Klīniskās universitātes slimnīcas Latvijas Transplantācijas Centru un Rīgas Austrumu klīniskās universitātes slimnīcas Latvijas Onkoloģijas centru un Rīgas 1. slimnīcu. Aizstāvēšana: 2014. gada 3. jūnijā plkst. 15.00 Rīgas Stradiņa universitātes Medicīnas promocijas padomes atklātā sēdē Rīgā, Dzirciema ielā 16, Hipokrāta auditorijā.Cilvēka herpesvīruss-6 (HHV-6) un herpesvīruss-7 (HHV-7) ir plaši izplatīti limfotropi herpesvīrusi, ar kuriem inficējas agrā bērnībā, un kas saglabājas persistenti visā dzīves periodā. Pēdējā laikā tiek pievērsta lielāka uzmanība beta-herpevīrusu infekcijas lomai vairāku hronisku slimību attīstībā, taču vienotas atbildes aizvien nav. Tas varētu būt saistīts ar šo vīrusu izplatību un dažādiem darbības mehānismiem. Darba mērķis bija noskaidrot beta-herpesvīrusu infekcijas iesaisti hronisku slimību patoģenēzē, slimības klīniskajā gaitā un pēctransplantācijas komplikāciju attīstībā. Šajā pētījumā tika iekļautas trīs pacientu grupas: pacienti ar imūnsupresīvo terapiju (pēc nieru transplantācijas), pacienti ar pamatslimības izsauktu imūnsupresiju (kuņģa-zarnu trakta vēzis) un pacienti ar imūnsistēmas izmaiņām autoimūna procesa rezultātā (autoimūnais tireoidīts). Praktiski veselu asins donoru grupa un vairog-dziedzera audu autopsijas paraugi bez patoloģiskām makro vai mikro izmaiņam tika iekļauti pētījumā kā kontroles. Lai detektēt vīrusu genomu secību klātbūtni, infekcijas latento vai aktīvo fāzi un vīrusu slodzi, tika izmantota kvalitatīvā un kvantitatīvā polimerāzes ķēdes reakcija. Vīrusu specifisko antivielu klātbūtne plazmā tika noteikta lietojot ELISA un IFA. Citokīnu ekspresijas līmeni plazmā noteica ar ELISA. Lietojot Becton-Dickinson plūsmas citofluorometru, tika atdalītas un analizētas imūnkompetento šūnu sub-populācijas. Rezultāti parādīja, ka HHV-6 un HHV-7 infekcijas biežums un aktivitātes fāze trīs pacientu grupās ar dažādu iemeslu radītiem imūnsistēmas traucējumiem ir atšķirīgi, konstatētas arī atšķirības imūnsistēmas atbildēs un izmaiņas imūnkompentento šūnu populācijās. Salīdzinot trīs pētījumā iekļautās pacientu grupas, parādīts, ka nieres transplantāta recipientiem, kas saņēmuši pretatgrūšanas medikamentozo terapiju, bija smagāka imūnsupresija un lielāks beta-herpesvīrusu aktivācijas risks, novedot pie dažādu komplikāciju attīstības. Lai gan pacientiem ar kuņģa-zarnu trakta vēzi vīrusu infekcijas sastopamība un aktivitāte bija zemāka, taču konstatēta izteikta asociācija starp HHV-6 un HHV-7 aktīvu 5 infekciju un limfopēniju, pie tam pacientiem ar limfopēniju tika novērota lielāka mirstība salīdzinot ar pacientiem bez limfopēnijas. Pacientiem ar autoimūno tireoidītu bieži atrasta HHV-6 un HHV-7 genoma secību klātbūtne gan perifēro asiņu, gan vairogdziedzera audu DNS paraugos. Turklāt vairogdziedzera audu DNS paraugos konstatēta lielāka HHV-6 slodze nekā asins DNS paraugos, tādējādi norādot uz vairogdziedzeri kā vienu no iespējamām HHV-6 latences vietām, kas savukārt var veicināt autoimūnā procesa attīstību.Darbs izpildīts ar ESF projekta “Atbalsts doktorantiem studiju programmas apguvei un zinātniskā grāda ieguvei Rīgas Stradiņa universitātē” Nr. 2009/ 0147/ 1DP/ 1.1.2.1.2/ 09/ IPIA/ VIAA/ 009 atbalstu