Journal of Clinical Virology - Volume 82 - Supplement 1 - September 2016

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S138 Abstracts / Journal of Clinical Virology 82S (2016) S1–S142
presence of viruses at birth in children with ASD and in healthy
controls.
Materials and methods: The genomes of Cytomegalovirus
(CMV), Epstein Barr virus, Herpes simplex virus, Herpesvirus 6-8
and Varicella Zoster virus were investigated in dried blood spots
collected at birth from 82 children, 38 with ASD and 44 healthy
controls. Extracted DNA was amplified using specific nested-PCR
for each herpetic virus.
Results: Cytomegaloviruswas the only virus foundwith a preva-
lence of congenital CMV infection of 5.3% (2/38) in ASD cases and
0% (0/44) in controls.
Conclusions: The rate of congenital CMV infection was about
10-fold higher inASDpatients than in the Italian general population
and this result underlines the importance of the retrospective diag-
nosis by using DBS-test. Only investigating a wider ASD population
it will be possible to determine the real burden of Cytomegalovirus
in the evolution of neurodevelopmental disorders.
Abstract no: 13
Presentation at ESCV 2016: Poster 238
Aberrant expression of miR-21, miR-376c
miR-145 and their target host genes in Merkel
cell polyomavirus-positive non-small cell lung
cancer
Ismini Lasithiotaki ∗ liza Tsitoura
Anastasios Koutsopoulos , Eleni Lagoudaki ,
Chara Koutoulaki orge Pitsidianakis
Demetrios A. Spandidos , Nikolaos M. Siafakas ,
Katerina M. Antoniou , George Sourvinos
1 University of Crete, Heraklion, Crete 71110, Greece
2 Laboratory of Cellular and Molecular
Pneumonology, Medical School, University of Crete,
Heraklion, Crete 71110, Greece
3 Laboratory of Clinical Virology, Medical School,
University of Crete, Heraklion, Crete 71110, Greece
Merkel Cell Polyoma Virus (MCPyV) has already been associ-
ated with non-small cell lung cancer (NSCLC). In this study, we
evaluated expression profiles of miR-21, miR-145, miR-146a, miR-
155, miR-302c, miR-367 and miR-376c in MCPyV +ve and MCPyV
−ve paraffin-embedded NSCLC tissue samples as well as in MCPyV
+ve and MCPyV −ve samples from “healthy” lung tissue. Signifi-
cant differences were found only in the levels of miR-21, miR-376c
and miR-145 in the MCPyV +ve samples compared to the MCPyV
−ve tumour samples. Overall, miR-21 and miR-376c expression
was higher in tumour compared to healthy tissue samples; how-
ever, miR-21 and miR-376c expression was higher in MCPyV +ve
compared to MCPyV −ve tumour samples. The expression of tar-
get genes of miR-21(Pten, Bcl-2, Daxx, Pkr, Timp3), miR-376c (Grb2,
Alk7, Mmp9) andmiR-145 (Oct-4, Sox2, Fascin1) and their associated
pathways (Braf, Akt-1, Akt-2, Bax, Hif1a, p53) was altered between
MCPyV +ve tumor samples and their corresponding controls. miR-
145 was downregulated in MCPyV +ve compared to MCPyV −ve
tumour samples and the corresponding controls. These results
show a novel association between miR-21, miR-376c and miR-145
and their host genes with the presence of MCPyV, suggesting a
mechanism of virus-specific microRNA signature in NSCLC.
Abstract no: 134
Presentation at ESCV 2016: Poster 239
Establishment and characterization of an
in vitro model of human Polyomavirus BK
(BKV) infected prostate normal cells
S. Villani ∗, N. Gagliano , P. Procacci , M. Dolci ,
L. Signorini , F. Elia , R. Ticozzi , P. Ferrante ,
S. Delbue
1 Biomedical, Surgical and Dental Sciences,
University of Milano, Milano, Italy
2 Biomedical Sciences for Health, University of
Milano, Milano, Italy
3 School of Medicine and Surgery, University of
Bicocca, Milano, Italy
Introduction: Prostate cancer (PCa) is one of the most common
male neoplasm in the Western word, being the most commonly
diagnosed non-skin cancer and the second leading cause of cancer
death. Various potential risk factors exist for the initial triggering
events, including exposure to infectious agents, such as the human
Polyomavirus BK (BKV). BKV is a good candidate as risk factor of
PCa because it naturally infects the human reno-urinary tract, it
establishes latency, and encodes oncoproteins that interfere with
tumor suppressors pathways, thus altering the normal progression
of cell cycle. The aim of the study is to establish an in vitro model
of infection and to investigate the possible co-factorial role of BKV
in PCa onset and progression.
Materials and methods: To investigate the potential rela-
tionship between BKV infection and PCa, an vitro model
was established using the normal epithelial prostate cell line
RWPE-1. The titration of the viral load was performed by
means of BKV specific-quantitative real time PCR (qPCR) and
droplet digital PCR (ddPCR). 3-(4,5-dimethylthiazol-2-yl)-2,5-
diphenyltetrazolium bromide (MTT) assay was performed at
different time points to evaluate the effect of BKV infection on
the growth of RWPE-1 cells. The expression profiles of a panel of
48 cytokines/chemokines were analyzed to identify differences in
their kinetics in infected and uninfected cells by multiplex assay.
To assess whether BKV infection was able to modify the cells
morphology, ultrastructural analysis and analysis of epithelial-
mesenchymal transition (EMT)markers in BKV infected, uninfected
and cleared cells were conducted.
Preliminary results: RWPE-1 cell line was found to be both
susceptible and permissive to BKV infection, reaching a peak of
infection after 3 days (3.9 ×106 copies/mL) and the infection lasted
for 14 days. The infected cells showed an higher rate of prolifer-
ation than the uninfected cells ranging from +37% to +18%. The
expression of IL-6, -9 -18 and TNF- was higher in the infected
cells than in the uninfected. Regarding the EMTmarkers, E-cadherin
was expressed in some uninfected RWPE-1 at cell boundaries and
upon BKV infection, E-cadherin expression was mainly located in
the cytoplasm and in the perinuclear region. Moreover, after infec-
tion some big multinucleated cells could be detected. This pattern
was maintained also after 30 days post infection.
Conclusions: The RWPE-1 cell line could be used as a model of
BKV infection. The viral infection induces molecular and morpho-
logical changes in the cells, but the possible cancer progression due
to the virus needs to be still elucidated.