Association of Torquetenovirus Viremia with Physical Frailty and Cognitive Impairment in Three Independent European Cohorts Robertina Giacconi, Blanca Laffon, Solange Costa, Armanda Teixeira-Gomes, Fabrizio Maggi, Lisa Macera, Pietro Giorgio Spezia, Francesco Piacenza, Alexander Bürkle, María Moreno-Villanueva, Stefano Bonassi, Vanessa Valdiglesias, Joao Paulo Teixeira, Martijn E.T. Dollé, M. Liset Rietman, Eugène Jansen, Tilman Grune, Efstathios S. Gonos, Claudio Franceschi, Miriam Capri, Birgit Weinberger, Ewa Sikora, Wolfgang Stuetz, Olivier Toussaint, Florence Debacq-Chainiaux, Antti Hervonen, Mikko Hurme, P. Eline Slagboom, Christiane Schön, Jürgen Bernhardt, Nicolle Breusing, Eduardo Pásaro, Ana Maseda, Laura Lorenzo-López, José Carlos Millán-Calenti, Mauro Provinciali, Marco Malavolta Gerontology, 2023 <b><i>Introduction:</i></b> Immunosenescence and inflammaging have been implicated in the pathophysiology of frailty. Torquetenovirus (TTV), a single-stranded DNA anellovirus, the major component of the human blood virome, shows an increased replication rate with advancing age. An elevated TTV viremia has been associated with an impaired immune function and an increased risk of mortality in the older population. The objective of this study was to analyze the relation between TTV viremia, physical frailty, and cognitive impairment. <b><i>Methods:</i></b> TTV viremia was measured in 1,131 nonfrail, 45 physically frail, and 113 cognitively impaired older adults recruited in the MARK-AGE study (overall mean age 64.7 ± 5.9 years), and then the results were checked in two other independent cohorts from Spain and Portugal, including 126 frail, 252 prefrail, and 141 nonfrail individuals (overall mean age: 77.5 ± 8.3 years). <b><i>Results:</i></b> TTV viremia ≥4log was associated with physical frailty (OR: 4.69; 95% CI: 2.06–10.67, <i>p</i> &#x3c; 0.0001) and cognitive impairment (OR: 3.49, 95% CI: 2.14–5.69, <i>p</i> &#x3c; 0.0001) in the MARK-AGE population. The association between TTV DNA load and frailty status was confirmed in the Spanish cohort, while a slight association with cognitive impairment was observed (OR: 1.33; 95% CI: 1.000–1.773), only in the unadjusted model. No association between TTV load and frailty or cognitive impairment was found in the Portuguese sample, although a negative association between TTV viremia and MMSE score was observed in Spanish and Portuguese females. <b><i>Conclusions:</i></b> These findings demonstrate an association between TTV viremia and physical frailty, while the association with cognitive impairment was observed only in the younger population from the MARK-AGE study. Further research is necessary to clarify TTV’s clinical relevance in the onset and progression of frailty and cognitive decline in older individuals.
Nicotine in Combination with SARS-CoV-2 Affects Cells Viability, Inflammatory Response and Ultrastructural Integrity Luigi Sansone, Antonio de Iure, Mario Cristina, Manuel Belli, Laura Vitiello, Federica Marcolongo, Alfredo Rosellini, Lisa Macera, Pietro Giorgio Spezia, Carlo Tomino, Stefano Bonassi, Matteo A. Russo, Fabrizio Maggi, Patrizia Russo International Journal of Molecular Sciences, 2022 The aims of our study are to: (i) investigate the ability of nicotine to modulate the expression level of inflammatory cytokines in A549 cells infected with SARS-CoV-2; (ii) elucidate the ultrastructural features caused by the combination nicotine+SARS-CoV-2; and (iii) demonstrate the mechanism of action. In this study, A549 cells pretreated with nicotine were either exposed to LPS or poly(I:C), or infected with SARS-CoV-2. Treated and untreated cells were analyzed for cytokine production, cytotoxicity, and ultrastructural modifications. Vero E6 cells were used as a positive reference. Cells pretreated with nicotine showed a decrease of IL6 and TNFα in A549 cells induced by LPS or poly(I:C). In contrast, cells exposed to SARS-CoV-2 showed a high increase of IL6, IL8, IL10 and TNFα, high cytopathic effects that were dose- and time-dependent, and profound ultrastructural modifications. These modifications were characterized by membrane ruptures and fragmentation, the swelling of cytosol and mitochondria, the release of cytoplasmic content in extracellular spaces (including osmiophilic granules), the fragmentation of endoplasmic reticulum, and chromatin disorganization. Nicotine increased SARS-CoV-2 cytopathic effects, elevating the levels of inflammatory cytokines, and inducing severe cellular damage, with features resembling pyroptosis and necroptosis. The protective role of nicotine in COVID-19 is definitively ruled out.
MS-based targeted profiling of oxylipins in COVID-19: A new insight into inflammation regulation Denise Biagini, Maria Franzini, Paolo Oliveri, Tommaso Lomonaco, Silvia Ghimenti, Andrea Bonini, Federico Vivaldi, Lisa Macera, Laurence Balas, Thierry Durand, Camille Oger, Jean-Marie Galano, Fabrizio Maggi, Alessandro Celi, Aldo Paolicchi, Fabio Di Francesco Free Radical Biology and Medicine, 2022
Delivery Mode Shapes the Composition of the Lower Airways Microbiota in Newborns Elisa Cardelli, Marco Calvigioni, Alessandra Vecchione, Lisa Macera, Diletta Mazzantini, Francesco Celandroni, Adelaide Panattoni, Mauro Pistello, Fabrizio Maggi, Emilia Ghelardi, Paolo Mannella Frontiers in Cellular and Infection Microbiology, 2021 Radical alterations in the human microbiota composition are well-known to be associated with many pathological conditions. If these aberrations are established at the time of birth, the risk of developing correlated pathologies throughout life is significantly increased. For this reason, all newborns should begin their lives with a proper microbiota in each body district. The present study aimed at demonstrating a correlation between the mode of delivery and the development of a well-balanced microbiota in the lower airways of newborns. 44 pregnant women were enrolled in this study. Microbiological comparative analysis was carried out on tracheobronchial secretions of babies born through vaginal delivery (VD) or caesarean section (CS). All samples showed the presence of bacterial DNA, regardless of the mode of delivery. No viable cultivable bacteria were isolated from the CS samples. On the contrary, VD allowed colonization of the lower airways by alive cultivable bacteria. The identification of bacterial species revealed that Lactobacillus spp. and Bacteroides vulgatus were the most common microorganisms in the lower airways of vaginally-delivered newborns. Data obtained from quantitative PCRs showed a significantly higher total bacterial load, as well as Firmicutes and Lactobacillus spp. amount, in VD samples than CS ones, while no statistically significant difference was found in Torque Teno Virus (TTV) load between samples. Taken together, our findings confirm the hypothesis that passage through the maternal vaginal canal determines more beneficial colonization of the lower airways in newborns.
Molecular validation of pathogen-reduction technologies using rolling-circle amplification coupled with real-time PCR for torquetenovirus DNA quantification Daniele Focosi, Lisa Macera, Pietro Giorgio Spezia, Francesca Ceccarelli, Maria Lanza, Fabrizio Maggi Transfusion Medicine, 2021 BACKGROUND Pathogen reduction technologies (PRT) based on nucleic-acid damaging chemicals and/or irradiation are increasingly being used to increase safety of blood components against emerging pathogens, such as convalescent plasma in the ongoing COVID-19 pandemic. Current methods for PRT validation are limited by the resources available to the blood component manufacturer, and quality control rely over pathogen spiking and hence invariably require sacrifice of the tested blood units: quantitative real-time PCR is the current pathogen detection method but, due to the high likelihood of detecting nonviable fragments, requires downstream pathogen culture. We propose here a new molecular validation of PRT based on the highly prevalent human symbiont torquetenovirus (TTV) and rolling circle amplification (RCA). MATERIALS AND METHODS Serial apheresis plasma donations were tested for TTV before and after inactivation with Intercept® PRT using real-time quantitative PCR (conventional validation), RCA followed by real-time PCR (our validation), and reverse PCR (for cross-validation). RESULTS While only 20% of inactivated units showed significant decrease in TTV viral load using real-time qPCR, all donations tested with RCA followed by real-time PCR showed TTV reductions. As further validation, 2 units were additionally tested with reverse PCR, which confirmed absence of entire circular genomes. DISCUSSION We have described and validated a conservative and easy-to-setup protocol for molecular validation of PRT based on RCA and real-time PCR for TTV.
Lack of neutralizing activity in nonconvalescent sera, regardless of ABO blood group and anti-A isoagglutinin titer Daniele Focosi, Alfredo Rosellini, Pietro Giorgio Spezia, Lisa Macera, Maria Lanza, Aldo Paolicchi, Denise Biagini, Andreina Baj, Mauro Pistello, Fabrizio Maggi Journal of Clinical Virology Plus, 2021 Background: Several ABO blood groups have been associated with the likelihood of infection, severity, and/or outcome of COVID-19 in hospitalized cohorts, raising the hypothesis that anti-A isoagglutinins in non-A-group recipients could act as neutralizing antibodies against SARS-CoV-2. Materials and methods: We run live virus neutralization tests using sera from 58 SARS-CoV-2 seronegative blood donors (27 O-group and 31 A-group) negatives for SARS-CoV-2 IgG to investigate what degree of neutralizing activity could be detected in their sera and eventual correlation with anti-A isoagglutinin titers. Results: We could not find clinically relevant neutralizing activity in any blood group, regardless of anti-isoagglutinin titer. Discussion: Our findings suggest that mechanisms other than neutralization explain the differences in outcomes from COVID19 seen in different ABO blood groups.
Nicotine upregulates ace2 expression and increases competence for sars-cov-2 in human pneumocytes Fabrizio Maggi, Alfredo Rosellini, Pietro Giorgio Spezia, Daniele Focosi, Lisa Macera, Michele Lai, Mauro Pistello, Antonio de Iure, Carlo Tomino, Stefano Bonassi, Patrizia Russo Erj Open Research, 2021 The coronavirus disease 2019 (COVID-19) pandemic has a variable degree of severity according to underlying comorbidities and life-style. Several research groups have reported an association between cigarette smoking and increased severity of COVID-19. The exact mechanism of action is largely unclear.We exposed low angiotensin-converting enzyme 2 (ACE2)-expressing human pulmonary adenocarcinoma A549 epithelial cells to nicotine and assessed ACE2 expression at different times. We further used the nicotine-exposed cells in a virus neutralisation assay.Nicotine exposure induces rapid and long-lasting increases in gene and protein expression of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor ACE2, which in turn translates into increased competence for SARS-CoV-2 replication and cytopathic effect.These findings show that nicotine worsens SARS-CoV-2 pulmonary infection and have implications for public health policies.
Torque teno virus microRNA detection in cerebrospinal fluids of patients with neurological pathologies Lorenzo Neri, Pietro Giorgio Spezia, Samuele Suraci, Lisa Macera, Stefano Scribano, Betti Giusti, Daniele Focosi, Fabrizio Maggi, Simone Giannecchini Journal of Clinical Virology, 2020 BACKGROUND Torque teno virus (TTV) is a widespread anellovirus that establishes persistent infections in humans and represents the most abundant component of the human virome. TTV encodes microRNAs (miRNA) which are found both in viremic and not viremic subjects being potentially ideal tools for the virus to evade the immune system response and to maintain chronic infection in the host. OBJECTIVE To investigate TTV-DNA loads and TTV-miRNAs expression in cerebrospinal fluids (CSF) from subjects under analysis for the assessment of neurological diseases. STUDY DESIGN Detection of TTV-DNA and TTV-miRNAs (e. g. miRNA t1a, t3b, and tth8) were carried out from CSF samples of 93 subjects with neurological diseases by using universal real-time PCR, real-time RT-PCR, and next-generation sequencing (NGS) analyses. RESULTS TTV-DNA was detected in 11 of 93 (12 %) CSFs with a mean TTV load of 155 copies/mL. Conversely, 29 CSF samples (31 %) were positive for at least one TTV-miRNA, while 15 (16 %) CSFs contained all the TTV-miRNAs examined. Overall, TTV-miRNA tth8 was detected in 62 % of samples, followed by TTV miRNA t3b (56 %), and t1a (29 %). Interestingly, TTV-miRNAs were found in CSF samples that were negative for the presence of TTV-DNA. Next-generation sequencing analysis carried out from 4 TTV-DNA negative CSF samples detected reads mapped in TTV-miRNA sequences region. CONCLUSIONS These results shed novel light on the relationship between TTV and the central nervous system and make compelling furthered studies for investigating the potential role of TTV-miRNAs in neurological disorders.
Seroprevalence of Torque Teno Virus in hemodialysis and renal transplant patients in Australia: A cross-sectional study Joshua S. Davis, Ginger Chu, Prabuddhua Pathinayake, Denise Jones, Phil Giffard, Lisa Macera, Peter Choi, Nathan W. Bartlett Transplant Infectious Disease, 2020 Torque teno virus (TTV) is a non‐pathogenic anellovirus commonly found in the blood of human beings. Emerging data suggest that TTV viral load is proportional to the degree of immunosuppression, but its seroprevalence is unknown in Australia. We aimed to determine the seroprevalence of TTV in an Australian population of renal patients.
Redondovirus DNA in human respiratory samples Pietro Giorgio Spezia, Lisa Macera, Paola Mazzetti, Michele Curcio, Chiara Biagini, Ilaria Sciandra, Ombretta Turriziani, Michele Lai, Guido Antonelli, Mauro Pistello, Fabrizio Maggi Journal of Clinical Virology, 2020
Prevalence and loads of torquetenovirus in the European mark-age study population Robertina Giacconi, Fabrizio Maggi, Lisa Macera, Pietro Giorgio Spezia, Mauro Pistello, Mauro Provinciali, Francesco Piacenza, Andrea Basso, Alexander Bürkle, María Moreno-Villanueva, Martijn E T Dollé, Eugène Jansen, Tilman Grune, Wolfgang Stuetz, Efstathios S Gonos, Christiane Schön, Jürgen Bernhardt, Beatrix Grubeck-Loebenstein, Ewa Sikora, Magdalena Dudkowska, Dorota Janiszewska, Olivier Toussaint, Florence Debacq-Chainiaux, Claudio Franceschi, Miriam Capri, Antti Hervonen, Mikko Hurme, Eline Slagboom, Nicolle Breusing, Eugenio Mocchegiani, Marco Malavolta Journals of Gerontology Series A Biological Sciences and Medical Sciences, 2020
Torquetenovirus (TTV) load is associated with mortality in Italian elderly subjects Robertina Giacconi, Fabrizio Maggi, Lisa Macera, Mauro Pistello, Mauro Provinciali, Simone Giannecchini, Francesco Martelli, Pietro Giorgio Spezia, Erminia Mariani, Roberta Galeazzi, Laura Costarelli, Lorenzo Iovino, Sara Galimberti, Lorenzo Nisi, Francesco Piacenza, Marco Malavolta Experimental Gerontology, 2018
