MACERA LISA
@unipi.it
University of Pisa
Dept. Translational Research University of Pisa
EDUCATION
Biologist at the University of Pisa
PhD Researcher (2009-2011)
Post Doc Researcher (2011-now)
RESEARCH INTERESTS
Virology, Microbiology, Immunity, host virus relationship
Scopus Publications
Scopus Publications
Robertina Giacconi, Blanca Laffon, Solange Costa, Armanda Teixeira-Gomes, Fabrizio Maggi, Lisa Macera, Pietro Giorgio Spezia, Francesco Piacenza, Alexander Bürkle, María Moreno-Villanueva,et al.
S. Karger AG
<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.
Luigi Sansone, Antonio de Iure, Mario Cristina, Manuel Belli, Laura Vitiello, Federica Marcolongo, Alfredo Rosellini, Lisa Macera, Pietro Giorgio Spezia, Carlo Tomino,et al.
MDPI AG
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.
Denise Biagini, Maria Franzini, Paolo Oliveri, Tommaso Lomonaco, Silvia Ghimenti, Andrea Bonini, Federico Vivaldi, Lisa Macera, Laurence Balas, Thierry Durand,et al.
Elsevier BV
Lisa Macera, Pietro Giorgio Spezia, Chiara Medici, Eleonora Rofi, Marzia Del Re, Daniele Focosi, Paola Mazzetti, David Navarro, Guido Antonelli, Romano Danesi,et al.
Wiley
Torquetenovirus (TTV) viremia is emerging as a promising tool to assess functional immune competence, to predict posttransplant immune‐related complications, and eventually to customize immunosuppression.
Elisa Cardelli, Marco Calvigioni, Alessandra Vecchione, Lisa Macera, Diletta Mazzantini, Francesco Celandroni, Adelaide Panattoni, Mauro Pistello, Fabrizio Maggi, Emilia Ghelardi,et al.
Frontiers Media SA
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.
Daniele Focosi, Lisa Macera, Pietro Giorgio Spezia, Francesca Ceccarelli, Maria Lanza, and Fabrizio Maggi
Wiley
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.
Daniele Focosi, Alfredo Rosellini, Pietro Giorgio Spezia, Lisa Macera, Maria Lanza, Aldo Paolicchi, Denise Biagini, Andreina Baj, Mauro Pistello, and Fabrizio Maggi
Elsevier BV
Fabrizio Maggi, Alfredo Rosellini, Pietro Giorgio Spezia, Daniele Focosi, Lisa Macera, Michele Lai, Mauro Pistello, Antonio de Iure, Carlo Tomino, Stefano Bonassi,et al.
European Respiratory Society (ERS)
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.
Lorenzo Neri, Pietro Giorgio Spezia, Samuele Suraci, Lisa Macera, Stefano Scribano, Betti Giusti, Daniele Focosi, Fabrizio Maggi, and Simone Giannecchini
Elsevier BV
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.
Robertina Giacconi, Fabrizio Maggi, Lisa Macera, Pietro Giorgio Spezia, Mauro Pistello, Mauro Provinciali, Francesco Piacenza, Andrea Basso, Alexander Bürkle, María Moreno-Villanueva,et al.
Oxford University Press (OUP)
Abstract Torquetenovirus (TTV) viremia has been associated with increased mortality risk in the elderly population. This work aims to investigate TTV viremia as a potential biomarker of immunosenescence. We compared levels of circulating TTV in 1813 participants of the MARK-AGE project, including human models of delayed (offspring of centenarians [GO]) and premature (Down syndrome [DS]) immunosenescence. The TTV load was positively associated with age, cytomegalovirus (CMV) antibody levels, and the Cu/Zn ratio and negatively associated with platelets, total cholesterol, and total IgM. TTV viremia was highest in DS and lowest in GO, with intermediate levels in the SGO (spouses of GO) and RASIG (Randomly Recruited Age-Stratified Individuals From The General Population) populations. In the RASIG population, TTV DNA loads showed a slight negative association with CD3+T-cells and CD4+T-cells. Finally, males with ≥4log TTV copies/mL had a higher risk of having a CD4/CD8 ratio&lt;1 than those with lower viremia (odds ratio [OR] = 2.85, 95% confidence interval [CI]: 1.06–7.62), as well as reduced CD3+ and CD4+T-cells compared to males with lower replication rates (&lt;4log), even after adjusting for CMV infection. In summary, differences in immune system preservation are reflected in the models of delayed and premature immunosenescence, displaying the best and worst control over TTV replication, respectively. In the general population, TTV loads were negatively associated with CD4+ cell counts, with an increased predisposition for an inverted CD4/CD8 ratio for individuals with TTV loads ≥4log copies/mL, thus promoting an immune risk phenotype.
Pietro Giorgio Spezia, Lisa Macera, Paola Mazzetti, Michele Curcio, Chiara Biagini, Ilaria Sciandra, Ombretta Turriziani, Michele Lai, Guido Antonelli, Mauro Pistello,et al.
Elsevier BV
Abstract
Background
Redondovirus (ReDoV) is a recently discovered circular, Rep-encoding single-stranded DNA (CRESS-DNA) virus in humans. Its pathogenesis and clinical associations are still completely unknown.
Methods
The presence of ReDoV DNA was investigated in biological specimens of 543 Italian subjects by in-house developed PCR assays.
Results
The overall ReDoV prevalence was about 4% (23 of 543 samples). The virus was detected in 22 of 209 (11 %) respiratory samples. One stool sample was also ReDoV positive. Viral DNA was not found in blood samples from immunocompetent and immunosuppressed subjects and cerebrospinal fluids from patients with neurological diseases. Genomic nucleotide differences were detected among the ReDoV isolates by sequencing a 582-nucleotide fragment of the capsid gene of the viral genome.
Conclusions
The results demonstrate that ReDoV is mainly present in the respiratory tract of infected people. Further investigations are needed to reveal possible clinical implications of this new CRESS-DNA virus in humans.
Joshua S. Davis, Ginger Chu, Prabuddhua Pathinayake, Denise Jones, Phil Giffard, Lisa Macera, Peter Choi, and Nathan W. Bartlett
Wiley
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.
D. Focosi, P.G. Spezia, L. Macera, S. Salvadori, D. Navarro, M. Lanza, G. Antonelli, M. Pistello, and F. Maggi
Elsevier BV
OBJECTIVES
Torquetenovirus (TTV) is an emerging marker of functional immune competence with the potential to predict transplant-related adverse events. A large-scale epidemiological study was performed to understand how basal values vary in healthy subjects according to age and gender.
METHODS
We tested plasma from 1017 healthy blood donors aged 18-69 years. The presence and load of TTV were determined by a real-time PCR assay. A sub-cohort of 384 donors was tested for anti-cytomegalovirus IgG antibodies, and 100 subjects were also tested for TTV viremia on a paired whole blood sample.
RESULTS
The overall prevalence of TTV was 65% (657/1017) with a mean (±standard deviation, SD) growth of 5 ± 4% every 10 years of age increase, but stably higher in males (465/690, 67%) than in females (192/327, 59%). Mean (±SD) TTV load was 2.3 ± 0.7 Log copies/ml with no sex difference. TTV viremia showed modest increases along 10-years age intervals (mean ± SD: 0.3 ± 0.1). TTV viremia in donors sampled two years later remained stable (mean ± SD: 2.3 ± 0.8 versus 2.2 ± 0.7 Log copies between samples). Twenty-six percent (9/34) of blood donors with TTV-negative plasma scored positive when whole blood was tested, and the donors with positive plasma showed a mean (±SD) 1.4 ± 0.5 Log increase in copy numbers when whole blood was tested.
CONCLUSIONS
This study establishes the mean value of TTV viremia in plasma in healthy blood subjects and suggests that ageing causes only minimal increases in TTV viremia.
Sara Galimberti, Mario Petrini, Claudia Baratè, Federica Ricci, Serena Balducci, Susanna Grassi, Francesca Guerrini, Elena Ciabatti, Sandra Mechelli, Antonello Di Paolo,et al.
Frontiers Media SA
SARS-CoV-2 is the viral agent responsible for the pandemic that in the first months of 2020 caused about 400,000 deaths. Among compounds proposed to fight the SARS-CoV-2-related disease (COVID-19), tyrosine kinase inhibitors (TKIs), already effective in Philadelphia-positive acute lymphoblastic leukemia (Ph+ ALL) and chronic myeloid leukemia (CML), have been proposed on the basis of their antiviral action already demonstrated against SARS-CoV-1. Very few cases of COVID-19 have been reported in Ph+ ALL and in CML Italian cohorts; authors suggested that this low rate of infections might depend on the use of TKIs, but the biological causes of this phenomenon remain unknown. In this study, the CML model was used to test if TKIs would sustain or not the viral replication and if they could damage patient immunity. Firstly, the infection and replication rate of torquetenovirus (TTV), whose load is inversely proportional to the host immunological control, have been measured in CML patients receiving nilotinib. A very low percentage of subjects were infected at baseline, and TTV did not replicate or at least showed a low replication rate during the follow-up, with a mean load comparable to the measured one in healthy subjects. Then, after gene expression profiling experiments, we found that several “antiviral” genes, such as CD28 and IFN gamma, were upregulated, while genes with “proviral” action, such as ARG-1, CEACAM1, and FUT4, were less expressed during treatment with imatinib, thus demonstrating that TKIs are not detrimental from the immunological point of view. To sum up, our data could offer some biological explanations to the low COVID-19 occurrence in Ph+ ALL and CML patients and sustain the use of TKIs in COVID-19, as already proposed by several international ongoing studies.
Paola Quaranta, Giulia Lottini, Giulia Chesi, Flavia Contrafatto, Roberta Russotto, Lisa Macera, Michele Lai, Pietro Giorgio Spezia, Annalaura Brai, Maurizio Botta,et al.
Elsevier BV
Picornaviridae are positive-sense single stranded RNA viruses with a similar genomic structure lacking a cap at the 5' end, but with a highly structured 5'-untranslated region (UTR) containing an internal ribosome entry site (IRES). IRES allows ribosomes to be recruited by the viral RNA and initiate translation in a cap-independent manner. Coxsackie virus type B (CV-B) belong to Picornaviridae and are widespread in human population. They usually cause subclinical infections but, occasionally, also severe diseases with various clinical manifestations. CV-B have no specific therapy. DEAD-box polypeptide 3 (DDX3) is a member of the Asp-Glu-Ala-Asp (DEAD)-box family with an ATP-dependent RNA unwinding helicase activity. Recently, several positive-sense single strand RNA viruses have been shown to need DDX3 for their translation. Here, we show that several DDX3 inhibitors reduced CV-B replication and production of viral protein, particularly when added within 12 h of infection. Based on in vitro and in silico data, we hypothesized that DDX3 inhibitors hamper interaction between DDX3 and viral IRES in a stereodynamic fashion. Accordingly, the DDX3 inhibitors tested have no activity against the Vesicular Stomatitis virus and Measles virus, which are negative-sense single stranded RNA viruses and use cap-dependent translation. This study suggests that DDX3 is required by RNA viruses lacking a cap and show that this enzyme is a valuable target to design antiviral molecules against CV-B. Thus, DDX3 is dispensable for cap-dependent translation, but required for translation of transcripts containing secondary structure in their UTRs.
Lisa Macera, Pietro Giorgio Spezia, Daniele Focosi, Paola Mazzetti, Guido Antonelli, Mauro Pistello, and Fabrizio Maggi
Wiley
Marseilleviridae is a family of viruses which have only been propagated in acanthamoeba. Marseillevirus sequences have been recently detected in different human matrices by viral metagenomics. Single‐center studies worldwide have estimated a low prevalence of marseillevirus both in symptomatic patients and in healthy donors but, to date, no informations are available on the prevalence of this giant virus in Italy. By a polymerase chain reaction targeting the ORF152 viral sequence, we tested sera from 197 immunosuppressed patients and 285 healthy donors, and 63 and 30 respiratory and cerebrospinal fluid samples, respectively, of patients with various clinical conditions and referring the Virology Division for diagnostic purposes. We observed no evidence of Marseillevirus DNA in all 575 samples tested. Marseillevirus probably does not cause infection in human.
Alessia Santoro, Carlo Tomino, Giulia Prinzi, Vittorio Cardaci, Massimo Fini, Lisa Macera, Patrizia Russo, and Fabrizio Maggi
Bentham Science Publishers Ltd.
The “microbiome” is the operative term to refer to a collection of all taxa constituting microbial communities, such as bacteria, archaea, fungi and protists (originally microbiota). The microbiome consists of the indigenous microbial communities and of the host environment that they inhabit. Actually, it has been shown that there is a close relationship between the microbiome and human health and disease condition. Although, initially, the lung was considered sterile, actually, the existence of a healthy lung microbiome is usually accepted. Lung microbiome changes are reported in Chronic Obstructive Pulmonary Disease (COPD) and in its exacerbation. Viral and bacterial infections of the respiratory system are a major cause of COPD exacerbations (AECOPD) leading to increased local and systemic inflammation. Detection rates of virus in AECOPD are variable between 25-62% according to the detection method. The study of human airway and lung disease virome is quite recent and still very limited. The purpose of this review is to summarize recent findings on the lung microbiome composition with a special emphasis on virome in COPD and in AECOPD. Some drugs of natural origins active against resistant bacteria and virus are described.
Eliseo Albert, Carlos Solano, Estela Giménez, Daniele Focosi, Ariadna Pérez, Lisa Macera, José Luis Piñana, Eva María Mateo, Juan Carlos Hernández Boluda, Fabrizio Maggi,et al.
Springer Science and Business Media LLC
Torque teno virus (TTV) plasma DNA load has been consistently shown to be a surrogate biomarker of immunosuppression in solid organ transplant recipients. It is uncertain whether it may behave similarly in allogeneic hematopoietic stem cell transplant recipients (allo-HSCT). Here, we characterized the dynamics of TTV DNAemia in patients undergoing T-cell replete allo-SCT at late times after transplantation (> day + 100). This retrospective single-center observational study included 33 allo-HSCT patients. Plasma TTV DNA loads were quantified by real-time PCR before initiating the conditioning regimen and at different time points after transplant. Absolute lymphocyte counts (ALC) were measured by flow cytometry. Overall, TTV DNA load increased steadily after engraftment, reaching a peak by day + 90; afterwards, it remained relatively constant until day + 210. TTV DNA loads measured within days + 120 and + 210 correlated inversely with paired ALC, while both parameters did correlate directly within days + 20 and + 60. The median TTV DNA area under a curve between days + 90 and + 210 [(AUC)90–210] was significantly higher in patients who received corticosteroids within this time frame for treatment of graft versus host disease (either acute, chronic or both) than in controls (P = 0.025). In summary, TTV DNA load may mirror the degree of immunosuppression at late times after allo-HSCT.
Fabrizio Maggi, Daniele Focosi, Maura Statzu, Gabriele Bianco, Cristina Costa, Lisa Macera, Pietro Giorgio Spezia, Chiara Medici, Eliseo Albert, David Navarro,et al.
Springer Science and Business Media LLC
Monitoring the human virome has been recently suggested as a promising and novel area of research for identifying new biomarkers which would help physicians in the management of transplant patients. Imbalance of the immune system in transplant recipients has a significant impact on replication of Torquetenovirus (TTV), the most representative and abundant virus of human virome. TTV kinetic was studied by real-time PCR in 280 liver or kidney transplant recipients who underwent different drug regimens to maintain immunosuppression. During one-year post-transplant follow-up, TTV viremia fluctuated irrespective of transplanted organ type but consistent with the immunosuppression regimen. TTV kinetic in patients who manifested cytomegalovirus (CMV) reactivation within the first four months post-transplant differed from that observed in patients who did not experience CMV complications. Importantly, plasma TTV load measured between day 0 and 10 post-transplant was significantly higher in CMV DNA positive than in CMV DNA negative patients. TTV viremia above 3.45 log DNA copies/ml within the first 10 days post-transplant correlates with higher propensity to CMV reactivation following transplantation. This study provides further evidence for using early post-transplant TTV viremia to predict CMV reactivation in liver or kidney transplant recipients.
Robertina Giacconi, Fabrizio Maggi, Lisa Macera, Mauro Pistello, Mauro Provinciali, Simone Giannecchini, Francesco Martelli, Pietro Giorgio Spezia, Erminia Mariani, Roberta Galeazzi,et al.
Elsevier BV
ABSTRACT An age‐related dysregulation of immune response, known as immunosenescence, contributes to increased susceptibility to infections, frailty and high risk of mortality in the elderly. Torquetenovirus (TTV), a circular, single‐stranded DNA virus, is highly prevalent in the general population and it may persist in the organism, also in association with other viruses such as cytomegalovirus (CMV), causing chronic viremia. The relationship that TTV establishes with the immune system of infected hosts is not clear. It is known that TTV encodes microRNAs (miRNAs) that might contribute to immune evasion and that the highest viral loads are found in peripheral blood cells. Moreover, it is suspected that TTV infection lead to increased production of inflammatory mediators, thus playing a role in immunosenescence. We investigated the association of TTV load and miRNAs expression with inflammatory and immune markers and the influence of TTV load on mortality within a cohort of 379 elderly subjects who were followed up for 3years. TTV DNA load in polymorphonuclear leukocytes was slightly positively correlated with age and negatively associated with serum albumin levels and NK cell activity. A marginal positive correlation between TTV DNA load, monocytes and IL‐8 plasma levels was found in females and males respectively. TTV DNA copies ≥4.0 log represented a strong predictor of mortality (Hazard ratio=4.78, 95% CI: 1.70–13.44, after adjusting for age, sex and the main predictors of mortality rate) and this association remained significant even after the CMV IgG antibody titer was included in the model (HR=9.83; 95% CI: 2.48–38.97; N=343 subjects). Moreover, multiple linear regression model showed that TTV miRNA‐t3b of genogroup 3 was inversely associated with triglycerides, monocytes and C‐reactive protein, and directly associated with IL6. Overall these findings suggest a role of TTV in immunesenescence and in the prediction of all‐cause mortality risk in Italian elderly subjects. Further studies are needed to fully understand the pathogenic mechanisms of TTV infection during aging. HIGHLIGHTSTTV DNA load in polymorphonuclear leukocytes positively correlated with age.TTV DNA load negatively associated with serum albumin levels and NK cell activity.TTV DNA copies ≥4.0 log associated with three year mortality in elderly people.TTV and its miRNAs may play a role in immunesenescence.
Francesco Martelli, Lisa Macera, Pietro Giorgio Spezia, Chiara Medici, Mauro Pistello, Daniele Guasti, Paolo Romagnoli, Fabrizio Maggi, and Simone Giannecchini
Springer Science and Business Media LLC
BackgroundTorquetenovirus (TTV) belongs to Anelloviridae family, infects nearly all people indefinitely without causing overt disease establishing a fine and successful interaction with the host. Increasing evidence have shown some human viruses exploit extracellular vesicles thereby helping viral persistence in the host. Here, the presence of TTV in extracellular vesicles circulating in human plasma was investigated.MethodsTTV DNA was quantified in plasma-derived exosomes from 122 samples collected from 97 diseased patients and 25 healthy donors. Exosomes enriched vesicles (EEVs) were extracted from plasma and characterized by Nanoparticle tracking analysis, by western blot for presence of tetraspanin CD63, CD81 and annexin II protein and, finally, by electron microscopy (EM). Presence and quantitation of TTV DNA were assessed with an universal single step real-time TaqMan PCR assay.ResultsPreliminary investigation showed that the human plasma extracted extracellular vesicles exhibited a main size of 70 nm, had concentration of 2.5 × 109/ml, and scored positive for tetraspanin CD63, CD81 and annexin II, typical characteristic of the exosomes vesicles. EEVs extracted from pooled plasma with TTV DNA viremia of 9.7 × 104 copies/ml showed to contain 6.3 × 102 TTV copies/ml, corresponding to 0.65% of total viral load. Important, TTV yield changed significantly following freezing/thawing, detergents and DNAse treatment of plasma before EEVs extraction. EEVs purified by sucrose-density gradient centrifugation and analysis of gradient fraction positive for exosomes marker CD63 harbored 102 TTV copies/ml. Moreover, EM evidenced the presence of TTV-like particles in EEVs. Successive investigation of plasma EEVs from 122 subjects (37 HIV-positive, 20 HCV infected, 20 HBV infected, 20 kidney transplant recipients, and 25 healthy) reported TTV DNA detection in 42 (34%) of the viremic samples (37 were from diseased patients and 5 from healthy people) at a mean level of 4.8 × 103 copies/ml. The examination of EEVs selected samples reported the presence of TTV genogroup 1, 3, 4 and 5, with genogroup 3 highly observed.ConclusionsCollectively, although these observations should be confirmed by further studies, circulation of TTV particles in EEVs opens new avenues and mechanistic insights on the molecular strategies adopted by anelloviruses to persist in the host.
Eliseo Albert, Ignacio Torres, Alberto Talaya, Estela Giménez, José Luis Piñana, Juan Carlos Hernández-Boluda, Daniele Focosi, Lisa Macera, Fabrizio Maggi, Carlos Solano,et al.
Wiley
Plasma torque teno virus (TTV) DNA load directly correlates with the degree of T‐cell immune reconstitution early after allogeneic hematopoietic stem cell transplantation (allo‐HSCT). Here, the kinetics of oral TTV DNA shedding was examined to assess whether quantitation of TTV DNA load in saliva may either replace or complement that in plasma for predicting lymphocyte (ALC) reconstitution after engraftment. This prospective observational study enrolled 38 nonconsecutive allo‐HSCT recipients. Saliva and plasma specimens were collected at baseline (pretransplant) and at around days +30, +50, and +90 after allo‐HSCT. TTV DNA was quantitated in both specimen types by real‐time PCR. ALCs were measured by cytometry. A total of 104 paired saliva and plasma specimens were available for TTV PCR analyses. TTV DNA was detected more frequently in saliva than in plasma specimens at all time points (overall, 94.2% vs 86.5%). Increasing levels of TTV DNA were seen in both specimen types from day +30 to day +90 after transplantation. Overall, TTV DNA loads were significantly higher in saliva than in plasma specimens (P = .0002) and correlated significantly (P ≤ .0001). A direct correlation between TTV DNA loads in saliva and plasma and ALCs was observed after engraftment (P = .034 and P = .002, respectively). Future studies should be aimed at determining whether monitoring of oral TTV DNA shedding may be of any utility for inference of immune reconstitution after allo‐HSCT.
Lorenzo Iovino, Francesco Mazziotta, Giovanni Carulli, Francesca Guerrini, Riccardo Morganti, Valentina Mazzotti, Fabrizio Maggi, Lisa Macera, Enrico Orciuolo, Gabriele Buda,et al.
Elsevier BV
INTRODUCTION
Zinc plays an important role in thymic function and immune homeostasis. We performed a prospective clinical trial using a high-dose zinc oral supplementation to improve the immune reconstitution after hematopoietic stem cell transplant (HSCT).
PATIENTS AND METHODS
We enrolled 18 patients undergoing autologous HSCT for multiple myeloma. Nine patients were randomized to receive only a standard antimicrobial prophylaxis; whereas, nine patients received in addition 150 mg/day of zinc from day +5 to day +100 after transplant.
RESULTS
CD4+ naïve lymphocytes and TRECs showed a significant increase from day +30 until day +100 only in the zinc-treated group. Moreover, the load of Torquetenovirus, a harmless virus that replicates in course of immunedepression, increased at day +100 only in the control group. No severe adverse events were reported during the zinc consumption.
CONCLUSION
First data from the ZENITH trial suggest that high-dose zinc supplementation is safe and may enhance the thymic reconstitution after HSCT. Registered: http://Clinicaltrials.gov (NCT03159845); and EUDRACT: 2014-28 004499-47.
E Albert, C Solano, E Giménez, D Focosi, A Pérez, L Macera, J L Piñana, J C H Boluda, F Maggi, and D Navarro
Springer Science and Business Media LLC
Monitoring Torque teno virus (TTV) DNA load helps to estimate the risk of opportunistic infections in solid organ transplant recipients. We investigated whether the early kinetic pattern of plasma TTV DNA load after allogeneic hematopoietic stem cell transplantation (allo-HSCT) associates with subsequent CMV and EBV DNAemia. This study included 71 allo-HSCT patients. We found that the area under the curve (AUC) for log10 TTV DNA loads quantified by days 20 and 30 after transplantation (TTV DNA load AUC20-30), was significantly lower (P=0.036) in patients who subsequently developed CMV DNAemia requiring preemptive antiviral therapy (n=17) than in those who did not (n=8) or had no CMV DNAemia (n=19). Patients displaying TTV DNA load AUC20-30⩽2.8 copies × days × mL−1 were more likely to have high-level CMV DNAemia. A trend towards a direct correlation between TTV DNA AUC20-30 and CMV-specific interferon-γ CD8+ T-cell counts by day +30 was noted (P=0.095). However, this dynamic parameter was not useful for anticipating the occurrence of either CMV recurrences (n=12) or EBV DNAemia (n=34). In summary, it may be possible to identify a subset of allo-HSCT patients at a high risk of developing high-level CMV DNAemia by analyzing the kinetics of plasma TTV DNA load early after engraftment.