Assessing putative interplay between human herpesvirus-6 infection and alcohol abuse in substantia nigra

The previous studies have demonstrated that the central nervous system (CNS) is particularly vulnerable to alcohol induced changes, for example, alcohol increases the risk of Parkinson’s disease by affecting Substantia nigra (SN). Also called the “Black Substance”, it is the dopaminergic neurons rich part of the basal ganglia located in the midbrain. Human herpesvirus-6 (HHV-6) is a linear double stranded DNA virus; infection is ubiquitous and can induce various neurological diseases. HHV-6 replicates most efficiently in activated primary T cells, however, studies show that the virus can also replicate in a wide array of host cells, for example in monocytes, macrophages, astrocytes, oligodendrocytes and neurons. The aim of the study is to detect the presence of HHV-6 in SN region of chronic alcoholics and healthy individuals. Post mortem tissue samples of SN grey and white matter from 42 individuals (control group, age matched alcoholics and non-age matched alcoholics) were analysed in this study. DNA was extracted using black PREP FFPE DNA kit. To detect viral genomic sequences and variant, we were using nPCR technique. Viral loads were detected using HHV-6 Real-TM Quant kit. Fluorescent immunehistochemical staining and confocal microscopy were applied. The presence of HHV-6 DNA was detected in 19, 05% (8/42) of the SN region. All positive HHV-6 FFPE tissue samples were from alcoholic individuals. In white matter HHV-6 was detected in 62, 5% (5/8), and in grey matter – 87,5% (7/8) out of HHV-6 positive cases. All control individuals were HHV-6 negative. HHV-6B variant was detected in all positive individuals. Viral load was detected in the one alcoholic individuals’ white matter – 101207, 97 copies/1x106 cells.HHV-6 immunopositivity was detected in both grey and white matter. These findings provide evidence that HHV-6 can integrate and replicate in the SN region. In addition, the evidence from this study shows then the potential role of HHV-6 and alcohol use may affect brain homeostasis.publishersversionPeer reviewe

SARS-CoV-2 infection in COVID-19 patients in Latvia

Šī darba ietvaros tika pētīta SARS-CoV-2 infekcija COVID-19 pacientiem Latvijā, raksturojot vīrusa genoma klātbūtni un slodzi vairākos klīniskajos paraugos un antivielas dinamikā. Pētījumā iesaistīti 134 pacienti, kuriem bija detektēts SARS-CoV-2 RNS nazofaringeālājās iztriepēs ar RT-PĶR metodi, pirms hospitalizācijas. Tika ievākti COVID-19 pacientu nazofaringeālās iztriepes, fēces, urīns, asins plazma, PMBC SARS-CoV-2 genoma klātbūtnes un vīrusa slodzes noteikšanai, izmantojot RT-kPĶŖ metodi. Tika ievākti pacientu plazmas paraugi antivielu detektēšanai un raksturošanai dinamikā, izmantojot ELISA metodi. SARS-CoV-2 genoma sekvence visbiežāk tika detektēta nazofaringeālajās iztriepēs (100/134, 74,6%) un fēcēs (76/104, 73,1%). Visaugstākā mediānā slodze tika konstatēta fēču paraugos (14529 [IQR 3224 – 93704] kopijas/ml). IgM antivielu titrs sāk samazināties pēc 12 dienām, un IgG titrs augstā līmenī spēj saglabāties līdz 119 dienām.The aim of this study was to evaluate SARS-CoV-2 infection in COVID-19 patients in Latvia, exploring the presence of SARS-CoV-2 and viral load in various clinical specimens and evaluating antibody response to SARS-CoV-2 in dynamics. Nasopharyngeal swabs, faeces, urine, blood plasma and PBMC samples from COVID-19 patients were collected to detect SARS-CoV-2 genome and viral load, using the RT-qPCR method. Patient plasma samples were collected for antibody detection and analysis in dynamics, using ELISA method. SARS-CoV-2 was detected most frequently in nasopharyngeal swab (100/134, 74,6%) and fecal (76/104, 73,1%) samples. The highest median load was observed in faecal samples (14529 [IQR 3224 - 93704] copies/ml). IgM class antibody titers decreased after 12 days, and IgG titer maintained at high level for up to 119 days

INVOLVEMENT OF HHV-6 INFECTION AND PROINFLAMMATORY CYTOKINES IN AUTOIMMUNE THYROIDITIS DEVELOPMENT

Šī darba ietvaros tika pētīta HHV-6, RANTES hemokīna un IL-6, INF-γ, TNF-α iekaisuma citokīnu iesaistes loma autoimūnā tireoidīta attīstībā. Pētījumā iesaistīti 63 pacienti ar AIT, kuriem tika veikta tireoīdektomija (Thyreoidectomia totalis) un 27 asins donori kā kontroles grupa. HHV-6 genoma sekvence tika detektēta 58 (92%) AIT pacientu audu paraugos, 10 (16%) asins paraugos un 3 (11%) kontroles grupas asins paraugos. Aktīva infekcija tika noteikta 25/58 (43%) AIT pacientu vairogdziedzeru audu paraugos. RANTES hemokīna un iekaisuma citokīnu līmeņa noteikšanai tika izmantota Luminex tehnoloģija, kas ir balstīta uz imūnfermentatīvo metodi. AIT pacientu asins paraugos tika detektēts augsts iekaisuma citokīnu līmenis. RANTES hemokīna līmenis asins donoriem (mediāna 1346 [IQR 645 – 2503] pg/ml) bija ievērojami paaugstināts nekā AIT pacientiem (mediāna 418,2 [IQR 113,1 – 803,1] pg/ml). Darba saturu veido 7 nodaļas ar apakšnodaļām. Kopumā darba izstrādāšanai izmantoti 42 literatūras avoti, 12 attēli. Darba apjoms sastāv no 48 lapaspusēm. Bakalaura darbs izstrādāts Rīgas Stradiņu universitātes mikrobioloģijas un virusoloģijas institūtā.The aim of this study was to evaluate the potential role of HHV-6 (human herpesvirus 6), RANTES chemokine and IL-6, INF-γ, TNF-α proinflammatory cytokines involvement in AIT development. This study included 63 AIT patients and 27 blood donors as control. HHV-6 genome sequence was detected in 58 (92%) AIT patients’ thyroid gland tissue, 10 (16%) blood samples and 3 (11%) in control groups’ blood samples. Active infection was detected in 25/58 (43%) AIT patients’ thyroid gland tissue. Immunfermentative method based on Luminex technology was used for RANTES chemokine and proinflammatory cytokines detection. Cytokines level in AIT patients’ blood samples was higher compared to blood donors. AIT patients’ RANTES level was lower (median 418,2 [IQR 113,1 – 803,1] pg/ml) compared to control group (median 1346 [IQR 645 – 2503] pg/ml). This study contains 7 chapter with subdivisions. The totals amount of 42 references, 12 figures were used in this thesis which has 47 pages. This work was done in Rigas Stradins University Institute of Microbiology and Virology

INVOLVEMENT OF HHV-6 INFECTION AND PROINFLAMMATORY CYTOKINES IN AUTOIMMUNE THYROIDITIS DEVELOPMENT

Šī darba ietvaros tika pētīta HHV-6, RANTES hemokīna un IL-6, INF-γ, TNF-α iekaisuma citokīnu iesaistes loma autoimūnā tireoidīta attīstībā. Pētījumā iesaistīti 63 pacienti ar AIT, kuriem tika veikta tireoīdektomija (Thyreoidectomia totalis) un 27 asins donori kā kontroles grupa. HHV-6 genoma sekvence tika detektēta 58 (92%) AIT pacientu audu paraugos, 10 (16%) asins paraugos un 3 (11%) kontroles grupas asins paraugos. Aktīva infekcija tika noteikta 25/58 (43%) AIT pacientu vairogdziedzeru audu paraugos. RANTES hemokīna un iekaisuma citokīnu līmeņa noteikšanai tika izmantota Luminex tehnoloģija, kas ir balstīta uz imūnfermentatīvo metodi. AIT pacientu asins paraugos tika detektēts augsts iekaisuma citokīnu līmenis. RANTES hemokīna līmenis asins donoriem (mediāna 1346 [IQR 645 – 2503] pg/ml) bija ievērojami paaugstināts nekā AIT pacientiem (mediāna 418,2 [IQR 113,1 – 803,1] pg/ml). Darba saturu veido 7 nodaļas ar apakšnodaļām. Kopumā darba izstrādāšanai izmantoti 42 literatūras avoti, 12 attēli. Darba apjoms sastāv no 48 lapaspusēm. Bakalaura darbs izstrādāts Rīgas Stradiņu universitātes mikrobioloģijas un virusoloģijas institūtā.The aim of this study was to evaluate the potential role of HHV-6 (human herpesvirus 6), RANTES chemokine and IL-6, INF-γ, TNF-α proinflammatory cytokines involvement in AIT development. This study included 63 AIT patients and 27 blood donors as control. HHV-6 genome sequence was detected in 58 (92%) AIT patients’ thyroid gland tissue, 10 (16%) blood samples and 3 (11%) in control groups’ blood samples. Active infection was detected in 25/58 (43%) AIT patients’ thyroid gland tissue. Immunfermentative method based on Luminex technology was used for RANTES chemokine and proinflammatory cytokines detection. Cytokines level in AIT patients’ blood samples was higher compared to blood donors. AIT patients’ RANTES level was lower (median 418,2 [IQR 113,1 – 803,1] pg/ml) compared to control group (median 1346 [IQR 645 – 2503] pg/ml). This study contains 7 chapter with subdivisions. The totals amount of 42 references, 12 figures were used in this thesis which has 47 pages. This work was done in Rigas Stradins University Institute of Microbiology and Virology

Chronic Alcoholism and HHV-6 Infection Synergistically Promote Neuroinflammatory Microglial Phenotypes in the Substantia Nigra of the Adult Human Brain

Funding Information: Funding: The present study was funded by Fundamental & Applied Research Projects (FLPP), Latvian Council of Science wide no. lzp-2020/2-0069 (The role of human herpesvirus-6 infection and alcohol abuse in the development of neuroinflammation). Publisher Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland.Both chronic alcoholism and human herpesvirus-6 (HHV-6) infection have been identified as promoters of neuroinflammation and known to cause movement-related disorders. Substantia Nigra (SN), the dopaminergic neuron-rich region of the basal ganglia, is involved in regulating motor function and the reward system. Hence, we hypothesize the presence of possible synergism between alcoholism and HHV-6 infection in the SN region and report a comprehensive quantification and characterization of microglial functions and morphology in postmortem brain tissue from 44 healthy, age-matched alcoholics and chronic alcoholics. A decrease in the perivascular CD68+ microglia in alcoholics was noted in both the gray and white matter. Additionally, the CD68+/Iba1- microglial subpopulation was found to be the dominant type in the controls. Conversely, in alcoholics, dystrophic changes in microglia were seen with a significant increase in Iba1 expression and perivascular to diffuse migration. An increase in CD11b expression was noted in alcoholics, with the Iba1+/CD11b- subtype promoting inflammation. All the controls were found to be negative for HHV-6 whilst the alcoholics demonstrated HHV-6 positivity in both gray and white matter. Amongst HHV-6 positive alcoholics, all the above-mentioned changes were found to be heightened when compared with HHV-6 negative alcoholics, thereby highlighting the compounding relationship between alcoholism and HHV-6 infection that promotes microglia-mediated neuroinflammation.publishersversionPeer reviewe