Eleonora Benedetti

An autochthonous case of Zika due to possible sexual transmission, Florence, Italy, 2014.

We report a case of Zika virus infection imported in Florence, Italy ex-Thailand, leading to a secondary autochthonous case, probably through sexual transmission. The two cases occurred in May 2014 but were retrospectively diagnosed in 2016 on the basis of serological tests (plaque reduction neutralisation) performed on stored serum samples. Our report provides further evidence that sexual transmission of Zika virus is possible.

Activation and repression of the 2-5A synthetase and p21 gene promoters by IRF-1 and IRF-2

The Interferon Regulatory Factors-1 and -2 (IRF-1 and IRF-2) were originally identified as transcriptional regulators of the interferon (IFN) and IFN-stimulated genes. These factors also modulate immune response and play a role in cell growth regulation. In this study we analysed the effect of the ectopic expression of IRF-1 and IRF-2 on the regulation of two potential IRF target genes involved in cell growth regulation, 2-5A synthetase and p21 (WAF/CP1), both of which contain consensus binding sites for IRF family members within their promoters. Following ectopic expression, IRF-1 transactivated 2-5A synthetase and p21 genes, an effect that was counterbalanced by concomitant ectopic expression of IRF-2. These effects were mediated by direct binding of IRF to the gene promoters. A construct expressing an IRF-2 antisense (FRI-2) was able to revert the inhibitory effect of IRF-2 on the IRF-1 transactivation. IRF-1 also induced expression of its homologous repressor IRF-2 as indicated by EMSA analysis using an IRF-E probe from the IRF-2 promoter; and by cotransfection of IRF-1 together with an IRF-2 promoter CAT construct. Therefore, the induction of IRF-1 by IFNs or other stimuli acts as a transactivator of genes involved in cell growth regulation, as well as of its own repressor IRF-2, thus providing autoinhibitory regulation of IRF-1 activated genes.

Circulation of West Nile virus lineage 1 and 2 during an outbreak in Italy.

In 2011, from 26 September to 16 October, a small outbreak of West Nile virus (WNV) disease occurred on the island of Sardinia (Italy). According to the national case definition, six cases with acute neurological disease were confirmed in hospitalized patients, and four of them died; one of these was only 34 years old. In two case, WNV RNA was detected in urine, suggesting renal involvement. Sequence analysis showed lineage 1 and 2 circulation.

Interferon regulatory factor-2 drives megakaryocytic differentiation.

IRFs [IFN (interferon) regulatory factors] constitute a family of transcription factors involved in IFN signalling and in the development and differentiation of the immune system. IRF-2 has generally been described as an antagonist of IRF-1-mediated transcription of IFN and IFN-inducible genes; however, it has been recently identified as a transcriptional activator of some genes, such as those encoding histone H4, VCAM-1 (vascular cell adhesion molecule-1) and Fas ligand. Biologically, IRF-2 plays an important role in cell growth regulation and has been shown to be a potential oncogene. Studies in knock-out mice have also implicated IRF-2 in the differentiation and functionality of haematopoietic cells. Here we show that IRF-2 expression in a myeloid progenitor cell line leads to reprogramming of these cells towards the megakaryocytic lineage and enables them to respond to thrombopoietin, as assessed by cell morphology and expression of specific differentiation markers. Up-regulation of transcription factors involved in the development of the megakaryocytic lineage, such as GATA-1, GATA-2, FOG-1 (friend of GATA-1) and NF-E2 (nuclear factor-erythroid-2), and transcriptional stimulation of the thrombopoietin receptor were also demonstrated. Our results provide evidence for a key role for IRF-2 in the induction of a programme of megakaryocytic differentiation, and reveal a remarkable functional diversity of this transcription factor in the regulation of cellular responses.

Detection of a chikungunya outbreak in Central Italy, August to September 2017

An autochthonous chikungunya outbreak is ongoing near Anzio, a coastal town in the province of Rome. The virus isolated from one patient and mosquitoes lacks the A226V mutation and belongs to an East Central South African strain. As of 20 September, 86 cases are laboratory-confirmed. The outbreak proximity to the capital, its late summer occurrence, and diagnostic delays, are favouring transmission. Vector control, enhanced surveillance and restricted blood donations are being implemented in affected areas.

Humoral immunity in natural infection by tick-borne encephalitis virus.

Tick‐borne encephalitis (TBE) virus is one of the most important flaviviruses associated with neurological disease in Europe. Cross‐reactive antibodies elicited by different flaviviruses can make difficult the interpretation of ELISA and hemagglutination‐inhibition (HI) tests for the diagnosis of TBE. Neutralization tests, which are more specific, are not in common use because they are difficult to perform and standardize. A plaque reduction neutralization test (PRNT), optimized previously in vaccinated children, was evaluated in sera from acute cases of TBE, collected for diagnostic purposes, and from healthy human population and wild ruminants, collected for serosurvey purposes. The PRNT results were compared with the results of ELISA and HI tests. In acute TBE disease, most sera were positive for IgM antibodies by ELISA and with high HI antibody titers; neutralizing antibodies were detected in 71.4% of patients, at a very low titer (1:10 NT50) in almost all cases. Seroprevalences of 8% and 6.5% for anti‐TBE ELISA antibodies were found in healthy subjects and wild ruminants, respectively. Among anti‐TBE positive healthy subjects, a very low 1:10 NT50 titer was detected in 17.4% of cases, while NT80 titers ranging from 1:10 to 1:80 were detected in 65.2% of cases. Among wild ruminants, 90.9% of ELISA and HI positive samples showed a positive, ≥1:10 NT80 titer. In conclusion, neutralization assays can be useful for the diagnosis and serosurveys of TBE. J. Med. Virol. 81:665–671, 2009

Measles elimination in Italy: data from laboratory activity, 2011–2013

BACKGROUND The European Regional Office of the World Health Organization developed a strategic approach to halt the indigenous transmission of measles in its 53 Member States by 2015, World Health Organization [1]. Many European countries, including Italy began the implementation of national programs to reach this goal. OBJECTIVES To describe and discuss the results of laboratory activity in measles surveillance, performed from January 2011 to December 2013 by the Italian National Reference Laboratory for Measles and Rubella. STUDY DESIGN Samples of suspected measles cases were collected from different Italian regions to confirm clinical diagnosis. Anti-measles IgM antibodies detection by Enzyme-Linked Immunosorbent Assay and/or molecular detection by Reverse Transcriptase-Polymerase Chain Reaction assay were performed. Positive samples were sequenced for viral characterization. RESULTS AND CONCLUSIONS According to results from the National Reference Laboratorys activity urine and blood seem to be the best specimens for measles laboratory surveillance. Phylogenetic analysis revealed a co-circulation of the genotypes D4 and D8 during the reviewed period, a cluster of B3 and sporadic cases of D9 and H1.

Prevalence of antibodies to phleboviruses and flaviviruses in Peja, Kosovo

In order to investigate the current and past activity of phlebovirus and flavivirus in Kosovo, a seroprevalence study among 200 blood donors was performed. Positive results were obtained for the phleboviruses TOSV and SFNV, and for a flavivirus of the Japanese Encephalitis group. No positive results for TBEV were observed.

Imported arboviral infections in Italy, July 2014-October 2015: a National Reference Laboratory report

BackgroundImported cases of infections due to Dengue (DENV) and Chikungunya (CHIKV) viruses and, more recently, Zika virus (ZIKV) are commonly reported among travelers returning from endemic regions. In areas where potentially competent vectors are present, the risk of autochthonous transmission of these vector-borne pathogens is relatively high. Laboratory surveillance is crucial to rapidly detect imported cases in order to reduce the risk of transmission. This study describes the laboratory activity performed by the National Reference Laboratory for Arboviruses (NRLA) at the Italian National Institute of Health in the period from July 2014 to October 2015.MethodsSamples from 180 patients visited/hospitalized with a suspected DENV/CHIKV/ZIKV infection were sent to the NRLA from several Italian Hospitals and from Regional Reference Laboratories for Arboviruses, in agreement with the National Plan on human surveillance of vector-borne diseases. Both serological (ELISA IgM test and Plaque Reduction Neutralization Test—PRNT) and molecular assays (Real Time PCR tests, RT-PCR plus nested PCR and sequencing of positive samples) were performed.ResultsDENV infection was the most frequently diagnosed (80 confirmed/probable cases), and all four genotypes were detected. However, an increase in imported CHIKV cases (41 confirmed/probable cases) was observed, along with the detection of the first ZIKV cases (4 confirmed cases), as a consequence of the recent spread of both CHIKV and ZIKV in the Americas.ConclusionsMain diagnostic issues highlighted in our study are sensitivity limitations of molecular tests, and the importance of PRNT to confirm serological results for differential diagnosis of Arboviruses. The continuous evaluation of diagnostic strategy, and the implementation of laboratories networks involved in surveillance activities is essential to ensure correct diagnosis, and to improve the preparedness for a rapid and proper identification of viral threats.

Authors' reply: diagnostic challenges to be considered regarding Zika virus in the context of the presence of the vector Aedes albopictus in Europe.

G Venturi 1 , L Zammarchi 2 3 , C Fortuna 1 , ME Remoli 1 , E Benedetti 1 , C Fiorentini 1 , M Trotta 3 , C Rizzo 4 , A Mantella 2 , G Rezza 1 , A Bartoloni 2 3 1. Department of Infectious, Parasitic and Immune-Mediate Diseases, Istituto Superiore di Sanita, Rome, Italy 2. Clinica Malattie Infettive, Dipartimento di Medicina Sperimentale e Clinica, Universita Degli Studi di Firenze, Florence, Italy 3. SOD Malattie Infettive e Tropicali, Azienda Ospedaliero-Universitaria Caeggi, Florence, Italy 4. National Center for Epidemiology and Health Promotion, Istituto Superiore di Sanita, Rome, Italy.