Amidic derivatives of valproic acid, valpromide and valnoctamide, inhibit HSV-1 infection in oligodendrocytes

Herpes simplex virus type 1 (HSV-1) is a ubiquitous infectious agent that can establish latency in neurons, and in some cases, viral retrograde transport results in infection of the central nervous system (CNS). Several antivirals have been identified with the ability to inhibit HSV-1 replication in human cells to a greater or lesser degree, most of which are nucleoside analogues that unfortunately exhibit teratogenic potential, embryotoxicity, carcinogenic or antiproliferative activities and resistances in immunocompromised patients, specially. In the present study, we assessed two amidic derivatives of valproic acid (VPA) – valpromide (VPD) and valnoctamide (VCD) – which are already used in clinic treatments, as feasible HSV-1 antivirals in glial cells. Both VPD and VCD have exhibited increased efficacy in bipolar disorders and as anticonvulsant drugs compared to VPA, while being less teratogenic and hepatotoxic. Cytotoxicity assays carried out in our laboratory showed that VPD and VCD were not toxic in a human oligodendroglioma cell line (HOG), at least at the concentrations established for human treatments. Infectivity assays showed a significant inhibition of HSV-1 infection in HOG cells after VPD and VCD treatment, being more pronounced in VPD-treated cells, comparable to the effects obtained with acyclovir. Furthermore, the same antiherpetic effects of VPD were observed in other oligodendrocytic cell lines and rat primary oligodendrocytes (OPCs), confirming the results obtained in HOG cells. Altogether, our results allow us to propose VPD as a potential antiherpetic drug that is able to act directly on oligodendrocytes of the CNSFinancial support for the study was provided by the Fundación Severo Ochoa-Aeromédica Canaria and Ministerio de Ciencia, Investigación e Innovación, Spain (SAF2016-77575-R and RD16-0060- 0019

Potent virucidal activity in vitro of photodynamic therapy with Hpericum extract as photosensitizer and white light against human coronavirus HCoV-229E

The emergent human coronavirus SARS-CoV-2 and its high infectivity rate has highlighted the strong need for new virucidal treatments. In this sense, the use of photodynamic therapy (PDT) with white light, to take advantage of the sunlight, is a potent strategy for decreasing the virulence and pathogenicity of the virus. Here, we report the virucidal effect of PDT based on Hypericum extract (HE) in combination with white light, which exhibits an inhibitory activity of the human coronavirus HCoV-229E on hepatocarcinoma Huh-7 cells. Moreover, despite continuous exposure to white light, HE has long durability, being able to maintain the prevention of viral infection. Given its potent in vitro virucidal capacity, we propose HE in combination with white light as a promising candidate to fight against SARS-CoV-2 as a virucidal compoundThis research was funded by Fundación Universidad Autónoma de Madrid, grant number PI21/00315 and by Instituto de Salud Carlos III, grant number PI21/00953. Institutional Review Board Statement: Not applicabl

Examining the immune signatures of SARS-CoV-2 infection in pregnancy and the impact on neurodevelopment: Protocol of the SIGNATURE longitudinal study.

The COVID-19 pandemic represents a valuable opportunity to carry out cohort studies that allow us to advance our knowledge on pathophysiological mechanisms of neuropsychiatric diseases. One of these opportunities is the study of the relationships between inflammation, brain development and an increased risk of suffering neuropsychiatric disorders. Based on the hypothesis that neuroinflammation during early stages of life is associated with neurodevelopmental disorders and confers a greater risk of developing neuropsychiatric disorders, we propose a cohort study of SARS-CoV-2-infected pregnant women and their newborns. The main objective of SIGNATURE project is to explore how the presence of prenatal SARS-CoV-2 infection and other non-infectious stressors generates an abnormal inflammatory activity in the newborn. The cohort of women during the COVID-19 pandemic will be psychological and biological monitored during their pregnancy, delivery, childbirth and postpartum. The biological information of the umbilical cord (foetus blood) and peripheral blood from the mother will be obtained after childbirth. These samples and the clinical characterisation of the cohort of mothers and newborns, are tremendously valuable at this time. This is a protocol report and no analyses have been conducted yet, being currently at, our study is in the recruitment process step. At the time of this publication, we have identified 1,060 SARS-CoV-2 infected mothers and all have already given birth. From the total of identified mothers, we have recruited 537 SARS-COV-2 infected women and all of them have completed the mental health assessment during pregnancy. We have collected biological samples from 119 mothers and babies. Additionally, we have recruited 390 non-infected pregnant women

Examining the immune signatures of SARS-CoV-2 infection in pregnancy and the impact on neurodevelopment: Protocol of the SIGNATURE longitudinal study

The COVID-19 pandemic represents a valuable opportunity to carry out cohort studies that allow us to advance our knowledge on pathophysiological mechanisms of neuropsychiatric diseases. One of these opportunities is the study of the relationships between inflammation, brain development and an increased risk of suffering neuropsychiatric disorders. Based on the hypothesis that neuroinflammation during early stages of life is associated with neurodevelopmental disorders and confers a greater risk of developing neuropsychiatric disorders, we propose a cohort study of SARS-CoV-2-infected pregnant women and their newborns. The main objective of SIGNATURE project is to explore how the presence of prenatal SARS-CoV-2 infection and other non-infectious stressors generates an abnormal inflammatory activity in the newborn. The cohort of women during the COVID-19 pandemic will be psychological and biological monitored during their pregnancy, delivery, childbirth and postpartum. The biological information of the umbilical cord (foetus blood) and peripheral blood from the mother will be obtained after childbirth. These samples and the clinical characterisation of the cohort of mothers and newborns, are tremendously valuable at this time. This is a protocol report and no analyses have been conducted yet, being currently at, our study is in the recruitment process step. At the time of this publication, we have identified 1,060 SARS-CoV-2 infected mothers and all have already given birth. From the total of identified mothers, we have recruited 537 SARS-COV-2 infected women and all of them have completed the mental health assessment during pregnancy. We have collected biological samples from 119 mothers and babies. Additionally, we have recruited 390 non-infected pregnant women.This work has received support from the Fundación Alicia Koplowitz to realize the epigenetic wide association study and to the clinical assessment to the children. This work has also received public support from the Consejería de Salud y Familias para la financiación de la investigación, desarrollo e innovación (i + d + i) biomédica y en ciencias de la salud en Andalucía (CSyF 2021 - FEDER). Grant Grant number PECOVID- 0195-2020. Convocatoria financiada con Fondo Europeo de Desarrollo Regional (FEDER) al 80% dentro del Programa Operativo de Andalucía FEDER 2014-2020. Andalucía se mueve con Europa. NG-T received payment under Rio Hortega contract CM20-00015 with the Carlos III Health Institute.Peer reviewe

Estudio de la infección del virus Herpes simplex tipo 1 en oligodendrocitos humanos: entrada viral y antivirales

Tesis doctoral inédita leída en la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 02-07-2020Esta tesis tiene embargado el acceso al texto completo hasta el 02-01-202

Effect of PLP overexpression on HSV-1 infection.

<p>HOG and HOG-PLP cells were infected with HSV-1. PLP-transfected HOG cells showed higher susceptibility to HSV-1 than mock-transfected cells (A and B). A. Plaque assay showed an increase in the number of plaque forming units (p.f.u.) per ml in PLP-transfected cells compared to mock-transfected control cells. Two representative wells are also shown. The average plaque size of infected HOG-PLP cells is slightly larger than that of plaques in HOG cells. B. Cells were infected at a m.o.i. of 0.1 with HSV-1, and viral titers were determined 20 h p.i. by TCID₅₀. Virus yield was significantly increased in PLP-transfected cells. The increment on viral yield correlated with the presence of PLP-EGFP, as shown by immunoblotting (C) with an anti-GFP antibody. C. HOG and HOG-PLP cells were infected with HSV-1 at an m.o.i. of 0,1. After 24 h p.i., equal number of cells were subjected to SDS–PAGE and analyzed by immunoblotting with a rabbit polyclonal anti-GFP antibody.</p

Viral entry assays.

<p>A. HOG and HOG-PLP cells were infected with R120vGF at a m.o.i. of 1. After 24 h p.i., equal number of cells were subjected to SDS–PAGE and analyzed by immunoblotting with a rabbit polyclonal anti-HSV-1 antibody to detect immediate early proteins. In HOG-PLP cells, an increase in viral signal was observed. A positive control of HOG cells infected with HSV-1 was also included. The histogram corresponds to the quantification of the immunoblot signals expressed in arbitrary scanning units. B. Confluent monolayers of cells plated in 96-well tissue culture dishes were infected with a recombinant HSV-1 (KOS) gL86 at a m.o.i. of 10. After 6 h p.i., the β-galactosidase activity at 410 nm was analyzed in a microplate reader. Optical density (OD) was increased in HOG-PLP cells compared to HOG cells. C. To perform an antibody blocking assay, HOG cells blocked with an anti-PLP antibody were infected at a m.o.i. of 1 with K26GFP and processed for flow cytometry, analyzing fluorescence of GFP. Percentage (%) of max designates the number of cells relative to the maximum fraction. For each fluorescence intensity within positive cells, the percentage of cells incubated with blocking mouse anti-PLP antibody is considerably lower than control cells incubated without blocking antibodies (red plot) or cells incubated with a control mouse serum (yellow plot). Data are representative of 3 independent experiments.</p

Expression of PLP-EGFP in HOG cells.

<p>Cells stably transfected (A) or mock-transfected (B) with PLP-EGFP were fixed and processed for confocal indirect immunofluorescence analysis with a polyclonal anti-PLP antibody. As the images show, the localization pattern of exogenous PLP (green) is similar to that of endogenous PLP (red). (DIC: Differential Interference Contrast).</p

Colocalization between PLP and HSV-1.

<p>HOG cells incubated at 4°C for 1 h at a m.o.i. of 100 with HSV-1 were fixed and processed for confocal double-label indirect immunofluorescence analysis with an anti-gD LP2 antibody. Images correspond to confocal slices of 0.6 μm. Images show partial colocalization (yellow) between PLP-EGFP (green) and virions (red). (DIC: Differential Interference Contrast).</p

EM immunocolocalization of PLP-EGFP and HSV-1.

<p>HOG-PLP cells were mock-infected or infected with HSV-1 at a m.o.i. of 100 and incubated for 1 h at 4°C. Then, cells were fixed and processed for observation by electron microscopy. Panels A, B and C show the presence of virions (arrows) attached to the plasma membrane localized next to colloidal gold particles (arrowheads) corresponding to PLP.</p