@article {10757, title = {Antibodies reacting to mimotopes of Simian virus 40 large T antigen, the viral oncoprotein, in sera from children.}, journal = {J Cell Physiol}, volume = {234}, year = {2019}, month = {2019 Apr}, pages = {3170-3179}, abstract = {

Recent data indicate that the Simian virus 40 (SV40) infection appears to be transmitted in humans independently from early SV40-contaminated antipolio vaccines. Serum antibodies against SV40 large T antigen (Tag) were analyzed in children/adolescents and young adults. To investigate antibodies reacting to SV40 Tag antigens, serum samples ( n = 812) from children and young adults were analyzed by indirect ELISAs using specific SV40 Tag mimotopes. Mimotopes were synthetic peptides corresponding to SV40 Tag epitopes. In sera ( n = 412) from healthy children up to 17 years old, IgG antibodies against SV40 Tag mimotopes reached an overall prevalence of 15\%. IgM antibodies against SV40 Tag were detected in sera of children 6-8 months old confirming and extending the knowledge that SV40 seroconversion occurs early in life. In children/adolescents affected by different diseases ( n = 180) SV40 Tag had a prevalence of 18\%, being the difference no significant compared to healthy subjects ( n = 220; 16\%) of the same age. Our immunological data indicate that SV40 circulates in children and young adults, both in healthy conditions and affected by distinct diseases. The IgM detection in sera from healthy children suggests that the SV40 infection/seroconversion occurs early in life (>6 months). Our immunological data support the hypothesis that SV40, or a closely related still unknown polyomavirus, infects humans. The SV40 seroprevalence is lower than common polyomaviruses, such as BKPyV and JCPyV, and other new human polyomaviruses. In addition, our immunological surveillance indicates a lack of association between different diseases, considered herein, and SV40.

}, issn = {1097-4652}, doi = {10.1002/jcp.27490}, author = {Mazzoni, Elisa and Frontini, Francesca and Rotondo, John Charles and Zanotta, Nunzia and Fioravanti, Arianna and Minelli, Francesca and Torreggiani, Elena and Campisciano, Giuseppina and Marcuzzi, Annalisa and Guerra, Giovanni and Tommasini, Alberto and Touz{\'e}, Antoine and Martini, Fernanda and Tognon, Mauro and Comar, Manola} } @article {10763, title = {Cervico-vaginal secretion cytokine profile: A non-invasive approach to study the endometrial receptivity in IVF cycles.}, journal = {Am J Reprod Immunol}, volume = {81}, year = {2019}, month = {2019 Jan}, pages = {e13064}, abstract = {

PROBLEM: Cytokines have a significant role in the process of embryo implantation, trophoblast growth, and differentiation by modulating the immune and endocrine system. The aim of this study was to investigate the profile of a large set of cytokines in the cervico-vaginal washing of women undergoing IVF, to explore the association of these proteins with a good receptive endometrium.

METHOD OF STUDY: A cohort of 155 women scheduled for IVF cycle was recruited. All patients were asymptomatic for genitourinary infections and had been screened for chlamydia, mycoplasma, and other bacterial infections. All IVF subjects were treated according to standard clinical and laboratory protocols. A panel of 48 immune factors was analyzed on cervico-vaginal washing, using magnetic bead-based multiplex immunoassays (Bio-Plex, BIO-RAD Laboratories, Milano, Italy).

RESULTS: A total of 99 patients reached embryo transfer, of which 31 had a clinical pregnancy. A pattern of four pro-inflammatory immune molecules, IL-12p40, IFN-a, MIF, and MCP3 (P~<~0.001), was found significantly up-regulated in the cervico-vaginal fluid of women with clinical pregnancy. A significantly increased expression of IL-9, Gro , and SDF-1 (P~<~0.05) was observed in the presence of endometriosis, while high levels of IL-13 and L-15 were associated with ovulatory infertility factor (P~<~0.05).

CONCLUSION: In this pilot study, we demonstrated that the expression of specific cytokines in the cervico-vaginal washing on the day of oocyte retrieval might have a positive correlation with the potential clinical pregnancy. Therefore, cervico-vaginal secretion cytokine profiling might be a new, non-invasive approach to study the endometrial receptivity in IVF management.

}, issn = {1600-0897}, doi = {10.1111/aji.13064}, author = {Zanotta, Nunzia and Monasta, Lorenzo and Skerk, Kristina and Luppi, Stefania and Martinelli, Monica and Ricci, Giuseppe and Comar, Manola} } @article {10781, title = {Four-miRNA Signature to Identify Asbestos-Related Lung Malignancies.}, journal = {Cancer Epidemiol Biomarkers Prev}, volume = {28}, year = {2019}, month = {2019 Jan}, pages = {119-126}, abstract = {

BACKGROUND: Altered miRNA expression is an early event upon exposure to occupational/environmental carcinogens; thus, identification of a novel asbestos-related profile of miRNAs able to distinguish asbestos-induced cancer from cancer with different etiology can be useful for diagnosis. We therefore performed a study to identify miRNAs associated with asbestos-induced malignancies.

METHODS: Four groups of patients were included in the study, including patients with asbestos-related (NSCLC) and asbestos-unrelated non-small cell lung cancer (NSCLC) or with malignant pleural mesothelioma (MPM), and disease-free subjects (CTRL). The selected miRNAs were evaluated in asbestos-exposed population.

RESULTS: Four serum miRNAs, that is miR-126, miR-205, miR-222, and miR-520g, were found to be implicated in asbestos-related malignant diseases. Notably, increased expression of miR-126 and miR-222 were found in asbestos-exposed subjects, and both miRNAs are involved in major pathways linked to cancer development. Epigenetic changes and cancer-stroma cross-talk could induce repression of miR-126 to facilitate tumor formation, angiogenesis, and invasion.

CONCLUSIONS: This study indicates that miRNAs are potentially involved in asbestos-related malignancies, and their expression outlines mechanism(s) whereby miRNAs may be involved in an asbestos-induced pathogenesis.

IMPACT: The discovery of a miRNA panel for asbestos-related malignancies would impact on occupational compensation and may be utilized for screening asbestos-exposed populations.

}, issn = {1538-7755}, doi = {10.1158/1055-9965.EPI-18-0453}, author = {Santarelli, Lory and Gaetani, Simona and Monaco, Federica and Bracci, Massimo and Valentino, Matteo and Amati, Monica and Rubini, Corrado and Sabbatini, Armando and Pasquini, Ernesto and Zanotta, Nunzia and Comar, Manola and Neuzil, Jiri and Tomasetti, Marco and Bovenzi, Massimo} } @article {10425, title = {Antibody response to polyomavirus primary infection: high seroprevalence of Merkel cell polyomavirus and lymphoid tissue involvement.}, journal = {J Neurovirol}, volume = {24}, year = {2018}, month = {2018 Jun}, pages = {314-322}, abstract = {

Human polyomaviruses (HPyVs) asymptomatically infect the human population establishing latency in the host, and their seroprevalence can reach 90\% in healthy adults. Few studies have focused on the pediatric population, and there are no reports regarding the seroprevalence of all the newly isolated HPyVs among Italian children. Therefore, we investigated the frequency of serum antibodies against 12 PyVs in 182 immunocompetent children from Northeast Italy, by means of a multiplex antibody detection system. Additionally, secondary lymphoid tissues were collected to analyze the presence of HPyV DNA sequences using a specific real-time PCRs or PCRs. Almost 100\% of subjects were seropositive for at least one PyV. Seropositivity ranged from 3\% for antibodies against simian virus 40 (SV40) in children from 0 to 3~years, to 91\% for antibodies against WU polyomavirus (WUPyV) and HPyV10 in children from 8 to 17~years. The mean number of PyV for which children were seropositive increased with the increasing of age: 4 standard deviations (SD) 1.8 in the 0-3-year group, 5 (SD 1.9) in the 4-7-year group, and 6 (SD 2.2) in the 8-17-year group. JC polyomavirus (JCPyV) DNA was detected in 1\% of the adenoids, WUPyV in 12\% of the tonsils, and 28\% of the adenoids, and Merkel cell polyomavirus (MCPyV) was present in 6 and 2\% of the tonsils and adenoids, respectively. Our study gives new insights on the serological evidence of exposure to PyVs during childhood, and on their possible respiratory route of transmission.

}, issn = {1538-2443}, doi = {10.1007/s13365-017-0612-2}, author = {Cason, Carolina and Monasta, Lorenzo and Zanotta, Nunzia and Campisciano, Giuseppina and Maestri, Iva and Tommasino, Massimo and Pawlita, Michael and Villani, Sonia and Comar, Manola and Delbue, Serena} } @article {10418, title = {Cerebrospinal Fluid Cytokine Expression Profile in Multiple Sclerosis and Chronic Inflammatory Demyelinating Polyneuropathy.}, journal = {Immunol Invest}, volume = {47}, year = {2018}, month = {2018 Feb}, pages = {135-145}, abstract = {

BACKGROUND: Cerebrospinal fluid (CSF) analysis in patients with particular neurologic disorders is a powerful tool to evaluate specific central nervous system inflammatory markers for diagnostic needs, because CSF represents the specific immune micro-environment to the central nervous system.

METHODS: CSF samples from 49 patients with multiple sclerosis (MS), chronic inflammatory demyelinating polyneuropathy (CIDP), and non-inflammatory neurologic disorders (NIND) as controls were submitted to protein expression profiles of 47 inflammatory biomarkers by multiplex Luminex bead assay to investigate possible differences in the inflammatory process for MS and CIDP.

RESULTS: Our results showed differences in CSF cytokine levels in MS and CIDP; in particular, IL12 (p40) was significantly highly expressed in MS in comparison with CIDP and NIND, while SDF-1α and SCGF-β were significantly highly expressed in CIDP cohort when compared to MS and NIND. IL-9, IL-13, and IL-17 had higher expression levels in NIND if compared with the other groups.

CONCLUSIONS: Our study showed that, despite some common pathogenic mechanisms, central and peripheral nervous system demyelinating diseases, such as MS and CIDP, differ in some specific inflammatory soluble proteins in CSF, underlining differences in the immune response involved in those autoimmune diseases.

}, keywords = {Adult, Aged, Biomarkers, Central Nervous System, Diagnosis, Differential, Female, Hematopoietic Cell Growth Factors, Humans, Interleukin-12, Lectins, C-Type, Male, Middle Aged, Multiple Sclerosis, Peripheral Nervous System, Polyradiculoneuropathy, Chronic Inflammatory Demyelinating, Proteins}, issn = {1532-4311}, doi = {10.1080/08820139.2017.1405978}, author = {Bonin, Serena and Zanotta, Nunzia and Sartori, Arianna and Bratina, Alessio and Manganotti, Paolo and Trevisan, Giusto and Comar, Manola} } @article {10445, title = {Cytokine profiles of women with vulvodynia: Identification of a panel of pro-inflammatory molecular targets.}, journal = {Eur J Obstet Gynecol Reprod Biol}, volume = {226}, year = {2018}, month = {2018 Jul}, pages = {66-70}, abstract = {

OBJECTIVE: The vulvar pain syndrome (VPS) is a multifactorial disease severely influencing the lifestyle of affected women. Among possible etiological factors, local injury, peripheral and/or central sensitization of the nervous system, and a chronic inflammatory status have been positively associated with the development of VPS. The identification of a constitutive altered local inflammatory profile in VPS women may represent an important point in the characterization of patients{\textquoteright} phenotype as a useful marker influencing the vulvar micro-environment. The aim of this study was to investigative the possible role of the local cytokines production in women with VPS in comparison to healthy women.

STUDY DESIGN: In this study were collected vaginal swabs from 57 healthy women (HC) who never suffered from VPS and from 30 patients diagnosed with vulvodynia (VPS) by at least 3 years and currently symptomatic. All patients included in this study showed the absence of Sexually Transmitted (STD) diseases and Reproductive Tract Infection. Real-time PCR was performed to assess the genomic sequences of ST pathogens. The Luminex Bio-Plex platform was used for the analysis of a panel of 48 immune factors.

RESULTS: Eleven molecules, specifically involved in the pro-inflammatory pathway were significantly modulated in VPS patients in comparison to healthy women, suggesting a persistent inflammatory process.

CONCLUSIONS: Therefore, these inflammatory factors could be possible biological markers involved in this disease. Nevertheless, other studies are needed to consider this specific immune profile as a valid marker of the vulvodynia.

}, keywords = {Adult, Aged, Cytokines, Female, Humans, Inflammation, Middle Aged, Vaginal Smears, Vulva, Vulvodynia, Women{\textquoteright}s Health}, issn = {1872-7654}, doi = {10.1016/j.ejogrb.2018.05.035}, author = {Zanotta, Nunzia and Campisciano, Giuseppina and Scrimin, Federica and Blendi, Ura and Marcuzzi, Annalisa and Vincenti, Ezio and Crovella, Sergio and Comar, Manola} } @article {10782, title = {Fractional CO laser for genitourinary syndrome of menopause in breast cancer survivors: clinical, immunological, and microbiological aspects.}, journal = {Lasers Med Sci}, volume = {33}, year = {2018}, month = {2018 Jul}, pages = {1047-1054}, abstract = {

The composition of vaginal microbiome in menopause and cancer survivor women changes dramatically leading to genitourinary syndrome of menopause (GSM) in up to 70\% of patients. Recent reports suggest that laser therapy may be valuable as a not hormonal therapeutic modality. The aim of the present study was to evaluate the effects of fractional CO laser treatment on the vaginal secretory pathway of a large panel of immune mediators, usually implicated in tissue remodeling and inflammation, and on microbiome composition in postmenopausal breast cancer survivors. The Ion Torrent PGM platform and the Luminex Bio-Plex platform were used for microbiome and immune factor analysis. The significant reduction of clinical symptoms and the non-significant changes in vaginal microbiome support the efficacy and safety of laser treatment. Moreover, the high remodeling status in vaginal epithelium is demonstrated by the significant changes in inflammatory and modulatory cytokine patterns. Laser therapy can be used for the treatment of GSM symptoms and does not show any adverse effects. However, further studies will be needed to clarify its long-term efficacy and other effects.

}, keywords = {Breast Neoplasms, Cancer Survivors, Cytokines, Dyspareunia, Female, Humans, Lasers, Gas, Menopause, Microbiota, Middle Aged, Prospective Studies, Syndrome, Vagina, Vaginal Diseases}, issn = {1435-604X}, doi = {10.1007/s10103-018-2471-3}, author = {Becorpi, Angelamaria and Campisciano, Giuseppina and Zanotta, Nunzia and Tredici, Zelinda and Guaschino, Secondo and Petraglia, Felice and Pieralli, Annalisa and Sisti, Giovanni and De Seta, Francesco and Comar, Manola} } @article {10799, title = {In vivo microbiome and associated immune markers: New insights into the pathogenesis of vaginal dysbiosis.}, journal = {Sci Rep}, volume = {8}, year = {2018}, month = {2018 02 02}, pages = {2307}, abstract = {

The microbiota fulfils a key role in the training and function of the immune system, which contributes to the symbiosis between the host and complex microbial communities. In this study, we characterized the interplay between vaginal bacteria and local immune mediators during dysbiosis in selected women of reproductive age who were grouped according to Nugent{\textquoteright}s criteria. The abundance of Gardnerella vaginalis and Bifidobacterium breve was increased in the intermediate dysbiotic status, while the presence of a plethora of non-resident bacteria characterized the group with overt vaginosis. In response to these increases, the anti-inflammatory IL1ra and pro-inflammatory IL2 increased, while the embryo trophic factors FGFβ and GMCSF decreased compared to the healthy milieu. A specific pattern, including IL1α, IL1β, IL8, MIG, MIP1α and RANTES, distinguished the intermediate group from the vaginosis group, while IL5 and IL13, which are secreted by Th2 cells, were significantly associated with the perturbation of the commensals Lactobacilli, Gardnerella and Ureaplasma. Summarizing, we postulate that although the dysbiotic condition triggers a pro-inflammatory process, the presence of a steady state level of Th2 may influence clinical manifestations. These results raise clinically relevant questions regarding the use of vaginal immunological markers as efficacious tools to monitor microbial alterations.

}, keywords = {Adult, Biomarkers, Cytokines, Dysbiosis, Female, Humans, Microbiota, Th2 Cells, Vagina}, issn = {2045-2322}, doi = {10.1038/s41598-018-20649-x}, author = {Campisciano, Giuseppina and Zanotta, Nunzia and Licastro, Danilo and De Seta, Francesco and Comar, Manola} } @article {10847, title = {Serum Stem Cell Growth Factor Beta for the Prediction of Therapy Response in Hepatocellular Carcinoma.}, journal = {Biomed Res Int}, volume = {2018}, year = {2018}, month = {2018}, pages = {6435482}, abstract = {

Introduction: Chronic inflammatory response is one of major contributors in the development of hepatocellular carcinoma (HCC). Inflammatory molecules, such as cytokines and growth factors in the circulation, can be useful in the diagnosis and prognosis of the patients. The stem cell growth factor beta (SCGF), a newly found protein, is a secreted sulfated glycoprotein and it functions as a growth factor for primitive hematopoietic progenitor cells. The level of SCGF had been reported to be elevated in several cancer types. However, there is very few or even no information on this protein in the study of HCC, even more in clinical studies.

Methods: A multiplex immunoassay panel of 48 cytokines and growth factors were utilized to screen 68 sera from 29 HCC patients at pretreatment (T0), 1 month (T1), and 6 months (T6) after treatment by either radiofrequency ablation (RF) or transarterial chemoembolization (TACE). Treatment response was evaluated according to mRECIST criteria.

Results: Immunoassay screening showed that the levels of IL-17, CTACK, TNF, IL-2R, IL-8, and SCGF were different in Complete Responders (CR) and Nonresponders (NR) groups. At T0 and T1, the SCGF level was significantly the highest in NR (23.8 and 40.7 ng/mL, respectively), followed by early recurrence (25.4 and 25.0 ng/mL), and CR (6.7 and 5.3 ng/mL), independently from HCV, stages, and treatment type. Low basal SCGF level was associated with longer disease-free survival compared to high SCGF.

Conclusion: In this study, for the first time, we demonstrate that the high level of serum SCGF at pre- and posttreatment is associated with HCC nonresponsiveness.

}, keywords = {Aged, Carcinoma, Hepatocellular, Chemoembolization, Therapeutic, Female, Hematopoietic Cell Growth Factors, Humans, Liver Neoplasms, Male, Middle Aged, Neoplasm Recurrence, Local, Retrospective Studies, Stem Cells, Treatment Outcome}, issn = {2314-6141}, doi = {10.1155/2018/6435482}, author = {Sukowati, Caecilia H C and Patti, Riccardo and Pascut, Devis and Ladju, Rusdina B and Tarchi, Paola and Zanotta, Nunzia and Comar, Manola and Tiribelli, Claudio and Croc{\`e}, Lory S} } @article {10439, title = {Vaginal microbiota dysmicrobism and role of biofilm-forming bacteria.}, journal = {Front Biosci (Elite Ed)}, volume = {10}, year = {2018}, month = {2018 06 01}, pages = {528-536}, abstract = {

Bacterial vaginosis involves the presence of a polymicrobial biofilm on the vaginal epithelium, guaranteeing immune escape and spread of antibiotic resistance. To spot known biofilm-forming bacteria, we profiled the vaginal microbiome of sixty-four symptomatic women suffering from a different grade of vaginal disorders and sixty asymptomatic healthy women. Specific microbial profiles distinguished symptomatic from asymptomatic women and characterized the grade of dysmicrobism within the symptomatic group. Lactobacillus crispatus and iners predominated on the healthy vaginal mucosa, while Lactobacillus gasseri predominated in the intermediate dysmicrobism. Furthermore, the intermediate grade of dysmicrobism was characterized by other lactic acid-producers species than Lactobacilli, able to rescue the microbial imbalance, and Ureaplasma parvum-serovar 3. The vaginosis group exhibited the overgrowth of Prevotella bivia, which is known to enhance the biofilm formation by Gardnerella vaginalis, and the presence of Streptococcus anginosus, which is emerging as a new cooperating player of the vaginal biofilm. Identifying specific microorganisms promoting or preventing the biofilm formation could increase the accuracy for a better definition of the vaginal dysmicrobism concept and therapeutic intervention.

}, keywords = {Adult, Biofilms, Female, Humans, Microbiota, Vagina, Vaginosis, Bacterial}, issn = {1945-0508}, author = {Campisciano, Giuseppina and Zanotta, Nunzia and Petix, Vincenzo and Corich, Lucia and De Seta, Francesco and Comar, Manola} } @article {10494, title = {Influence of vaginal lactoferrin administration on amniotic fluid cytokines and its role against inflammatory complications of pregnancy.}, journal = {J Inflamm (Lond)}, volume = {14}, year = {2017}, month = {2017}, pages = {5}, abstract = {

BACKGROUND: An altered amniotic cytokine profile has been reported in inflammatory pregnancy complications with a leading role for IL-6, a marker of the foetal systemic inflammatory response. Up to this date there is no exhaustive information neither on the foetal cytokine balance nor on the best method for its modulation. We aimed to evaluate the influence of vaginal lactoferrin administration on amniotic fluid concentration of 47 cytokines, chemokines and growth factors.

METHODS: Sixty women undergoing genetic amniocentesis were enrolled in an open-label clinical trial. 300~mg of vaginal lactoferrin (Florence, Italy) were randomly administered to obtain 3 groups: A, 20 untreated patients; B and C (20 patients in each) respectively treated 4 and 12~h before amniocentesis. Cytokines, chemokines and growth factors concentrations were quantified by a magnetic bead Luminex multiplex immunoassays panel technology. Data analysis was performed with the software Stata (v. 13.1) and GraphPad Prism (v. 5). Group comparisons were performed using Kruskal-Wallis followed by Mann-Whitney U tests, with Bonferroni correction for multiple comparisons. A < 0.05 was considered significant.

RESULTS: Among the 47 tested mediators, 24 (51.06\%) were influenced by lactoferrin. 11 (23.4\%), showed a highly significant difference (p <0.001); among these IL-9, IL-15, IFN-γ, IP-10, TNF-α, IL-1α and MCP-3 underwent a down-regulation, while IL-17 and FGF-basic, G-CSF, GM-CSF an up-regulation. Difference between group C and both B and A was small for IL-15, IP-10, IL-1α, MCP-3, while it was negligible for IL-9, IFN-γ and TNF-α. IL-17 and the 3 growth factors were strongly enhanced in B and C groups. IL-17, FGF-basic and GM-CSF showed increasing concentrations in both B and C groups, while G-CSF resulted up-regulated only in group C. Significance was intermediate ( < 0.01) for the down regulated IL-2RA, IL-12p40 and IFNα2 (6.38\%) while it was small for 10 mediators (21.27\%) 7 of which (IL-2, IL-4, eotaxin, PDGF-BB, RANTES, IL-18 and MIF) down-regulated and 3 (MCP-1, IL-3, and SDF-1α) up-regulated.

CONCLUSION: Lactoferrin down-regulates 17 pro-inflammatory amniotic mediators while up-regulating 7 anti-inflammatory amniotic mediators, 5 of which definitively belonging to an anti-inflammatory profile. These findings open to clinical investigation on its use against inflammatory complications of pregnancy.

}, issn = {1476-9255}, doi = {10.1186/s12950-017-0152-9}, author = {Maritati, Martina and Comar, Manola and Zanotta, Nunzia and Seraceni, Silva and Trentini, Alessandro and Corazza, Fabrizio and Vesce, Fortunato and Contini, Carlo} } @article {10571, title = {SV40 Infection of Mesenchymal Stromal Cells From Wharton{\textquoteright}s Jelly Drives the Production of Inflammatory and Tumoral Mediators.}, journal = {J Cell Physiol}, volume = {232}, year = {2017}, month = {2017 Nov}, pages = {3060-3066}, abstract = {

The Mesenchymal Stromal Cells from umbilical cord Wharton{\textquoteright}s jelly (WJSCs) are a source of cells with high potentiality for the treatment of human immunological disorders. Footprints of the oncogenic viruses Simian Virus 40 (SV40) and JC Virus (JCPyV) have been recently detected in human WJSCs specimens. The aim of this study is to evaluate if WJSCs can be efficiently infected by these Polyomaviruses and if they can potentially exert tumoral activity. Cell culture experiments indicated that WJSCs could sustain both SV40 and JCPyV infections. A transient and lytic replication was observed for JCPyV, while SV40 persistently infected WJSCs over a long period of time, releasing a viral progeny at low titer without evident cytopathic effect (CPE). Considering the association between SV40 and human tumors and the reported ability of the oncogenic viruses to drive the host innate immune response to cell transformation, the expression profile of a large panel of immune mediators was evaluated in supernatants by the Bioplex platform. RANTES, IL-3, MIG, and IL-12p40, involved in chronic inflammation, cells differentiation, and transformation, were constantly measured at high concentration comparing to control. These findings represent a new aspect of SV40 biological activity in the humans, highlighting its interaction with specific host cellular pathways. In view of these results, it seems to be increasingly urgent to consider Polyomaviruses in the management of WJSCs for their safely use as promising therapeutic source. J. Cell. Physiol. 232: 3060-3066, 2017. {\textcopyright} 2016 Wiley Periodicals, Inc.

}, keywords = {Cell Line, Transformed, Cell Separation, Cell Transformation, Viral, Chemokine CCL5, Chemokine CXCL9, Cytopathogenic Effect, Viral, DNA, Viral, Host-Pathogen Interactions, Humans, Inflammation Mediators, Interleukin-12 Subunit p40, Interleukin-3, JC Virus, Mesenchymal Stem Cells, Real-Time Polymerase Chain Reaction, Simian virus 40, Time Factors, Up-Regulation, Viral Load, Virus Replication, Wharton Jelly}, issn = {1097-4652}, doi = {10.1002/jcp.25723}, author = {Cason, Carolina and Campisciano, Giuseppina and Zanotta, Nunzia and Valencic, Erica and Delbue, Serena and Bella, Ramona and Comar, Manola} } @article {8321, title = {Candidate Soluble Immune Mediators in Young Women with High-Risk Human Papillomavirus Infection: High Expression of Chemokines Promoting Angiogenesis and Cell Proliferation.}, journal = {PLoS One}, volume = {11}, year = {2016}, month = {2016}, pages = {e0151851}, abstract = {

BACKGROUND: The causal interpretation of cervical immune response to Human Papillomavirus (HPV) infection is complex and poorly characterized mainly due to the delicate balance that exists between viral infection, increase of inflammatory cytokines and host risk factors. This study aims to explore the significance of cervical immune mediators associated to cell survival, angiogenesis and interaction with immune response, in predicting the risk to develop HPV-related intraepithelial lesions.

METHODS: A panel of 48 cytokines and growth factors were explored in a selected cohort of 168 immunocompetent women including 88 diagnosed with low (LSIL) or high (HSIL) squamous intraepithelial lesions of the cervix and 80 with normal cervical cytology (NIL). HPV genotyping was performed by Linear Array HPV test and the soluble concentration of 48 immune molecules was analyzed using the Bio-Plex platform.

RESULTS: The prevalence of single HR-HPV infection was 30\% in NIL and 100\% in LSIL and HSIL women. The expression of 13 cytokines, including interleukins IL-6, IL-3, IL-12p40, IL-12p70, IL-16, IL-18, LIF, of chemokines CCL7 (MCP-3), CXCL9 (MIG), CXCL12 (SDF-1α) and of the tropic factors VEGF, G-CSF, M-CSF were significantly associated with the presence of infection, with levels being higher in women with precancerous lesions compared to NIL HPV negative women. Only the growth factor GM-CSF was positively associated with the cytological abnormalities.

CONCLUSIONS: The ability of HR-HPV to escape from innate immune recognition and to orchestrate the production of specific inflammatory and growth factors, involved in early inflammatory response and in the cell-proliferating phase of intraepithelial damage, was documented in women before the development of cervical lesions.

}, keywords = {Adult, Cell Survival, Cervix Uteri, Chemokines, Cohort Studies, Female, Humans, Neovascularization, Pathologic, Papillomaviridae, Papillomavirus Infections}, issn = {1932-6203}, doi = {10.1371/journal.pone.0151851}, author = {Zanotta, Nunzia and Tornesello, Maria Lina and Annunziata, Clorinda and Stellato, Giovanni and Buonaguro, Franco Maria and Comar, Manola} } @article {3503, title = {In vivo detection of polyomaviruses JCV and SV40 in mesenchymal stem cells from human umbilical cords.}, journal = {Pediatr Blood Cancer}, volume = {61}, year = {2014}, month = {2014 Aug}, pages = {1347-9}, abstract = {

BACKGROUND: Multipotent stromal cells are present in the Wharton{\textquoteright}s jelly matrix (WJSC) of the umbilical cord and can be used as an allogeneic source of cells to treat immunological disorders. Recently it was demonstrated that adult bone marrow (BM)-derived mesenchimal stromal cells (MSC) are susceptible to infection with viruses showing potential oncogenic properties, such as the polyomavirus JC (JCV). The aim of this study was to investigate the presence of human polyomaviruses (JCV, BK Virus-BKV, SV40, and Merkel cell polyomavirus-MCPyV) in WJSC, and explore the risk of infection.

PROCEDURE: MSC samples from 35 umbilical cords were investigated by quantitative Real Time PCRs for the presence of DNA sequences of JCV, BKV, SV40, and MCPyV.

RESULTS: JCV DNA was detected in 1/35 (2.8\%) of MSC samples, while SV40 DNA was found in 3/35 (8.6\%) of the examined samples. None of the samples showed sequences of BKV and MCPyV.

CONCLUSIONS: The present study demonstrates the in vivo ability of polyomaviruses to infect WJSC. Since the therapeutic approach with the WJSC has high potentiality and a more intensive use can be easily hypothesized, the need to develop consensus guidelines to detect rare viral infections in MSC is pressing.

}, keywords = {DNA, Viral, Female, Fetal Blood, Humans, JC Virus, Male, Mesenchymal Stromal Cells, Polyomavirus Infections, Simian virus 40, Tumor Virus Infections}, issn = {1545-5017}, doi = {10.1002/pbc.24943}, author = {Comar, Manola and Delbue, Serena and Zanotta, Nunzia and Valencic, Erica and Piscianz, Elisa and Del Savio, Rossella and Tesser, Alessandra and Tommasini, Alberto and Ferrante, Pasquale} } @article {3589, title = {Increased levels of C-C chemokine RANTES in asbestos exposed workers and in malignant mesothelioma patients from an hyperendemic area.}, journal = {PLoS One}, volume = {9}, year = {2014}, month = {2014}, pages = {e104848}, abstract = {

BACKGROUND: Asbestos-induced mesothelial inflammatory processes are thought to be the basic mechanisms underlying Malignant Mesothelioma (MM) development. Detection of MM often occurs at late stage due to the long and unpredictable latent period and the low incidence in asbestos exposed individuals. The aim of this study was to investigate early immunological biomarkers to characterize the prognostic profile of a possible asbestos-induced disease, in subjects from a MM hyperendemic area.

METHODS: The Luminex Multiplex Panel Technology was used for the simultaneous measurement of serum levels of a large panel of 47 analytes, including cytokines and growth factors, from workers previously exposed to asbestos (Asb-workers), asbestos-induced MM patients and healthy subjects. In addition, to explore the influence on serum cytokines profile exerted by SV40 infection, a cofactor in MM development, a quantitative real time PCR was performed for sequences detection in the N-terminal and intronic regions of the SV40 Tag gene. Statistical analysis was done by means of the Mann-Whitney test and the Kruskall-Wallis test for variance analysis.

RESULTS: A variety of 25 cytokines linked to pulmonary inflammation and tumor development were found significantly associated with Asb-workers and MM patients compared with healthy controls. A specific pattern of cytokines were found highly expressed in Asb-workers: IFN-alpha (p<0.05), EOTAXIN (p<0.01), RANTES (p<0.001), and in MM patients: IL-12(p40), IL-3, IL-1 alpha, MCP-3, beta-NGF, TNF-beta, RANTES (p<0.001). Notably, the chemokine RANTES measured the highest serum level showing an increased gradient of concentration from healthy subjects to Asb-workers and MM patients (p<0.001), independently of SV40 infection.

CONCLUSION: This study shows that, in subjects from an hyperendemic area for MM, the C-C chemokine RANTES is associated with the exposure to asbestos fibres. If validated in larger samples, this factor could have the potential to be a critical biomarker for MM prognosis as recently reported for breast tumor.

}, keywords = {Aged, Aged, 80 and over, Asbestos, Biomarkers, Tumor, Case-Control Studies, Chemokine CCL5, Cytokines, Endemic Diseases, Gene Expression, Gene Expression Profiling, Humans, Lung Neoplasms, Mesothelioma, Middle Aged, Occupational Exposure, Simian virus 40, Viral Proteins}, issn = {1932-6203}, doi = {10.1371/journal.pone.0104848}, author = {Comar, Manola and Zanotta, Nunzia and Bonotti, Alessandra and Tognon, Mauro and Negro, Corrado and Cristaudo, Alfonso and Bovenzi, Massimo} } @article {3483, title = {No evidence of Polyomavirus and EBV infections in Italian patients with mixed cryoglobulinemia infected chronically with HCV.}, journal = {J Med Virol}, volume = {86}, year = {2014}, month = {2014 Apr}, pages = {666-71}, abstract = {

Mixed cryoglobulinemia is a lymphoproliferative disorder associated with hepatitis C virus (HCV). In patients chronically affected by HCV the prevalence of mixed cryoglobulinemia is variable ranging from 0\% to 56\%. To verify whether polyomaviruses (PyV) play a role in this disorder a total of 222 blood samples from 63 HCV chronic patients, 43 with mixed cryoglobulinemia, 59 chronic lymphocytic leukemia, 50 polytransfused patients, and 50 blood donors were evaluated for Merkel (MCPyV), BKV, JCV, and SV40. EBV was additionally included in the analysis since association with this disorder has been reported. Mixed cryoglobulinemia patients infected chronically with HCV resulted negative for both PyV and EBV. MCPyV was found in 1 subject with Merkel Cell Carcinoma, in 10\% of polytransfused and in 10\% of blood donors while EBV was detected in 22\% of polytransfused, 10\% of B-cell lymphatic leukemia patients and 4\% of blood donors (P < 0.01). Taken together, the absence of PyV and EBV in HCV-mixed cryoglobulinemia patients seems to exclude a direct involvement of these viruses in the pathogenesis of this disease while the presence of MCPyV in healthy individuals, at the same rate as in polytransfused patients, may reinforce data on a minimal role of this virus in other human pathologies.

}, keywords = {Adult, Base Sequence, Cryoglobulinemia, DNA, Viral, Epstein-Barr Virus Infections, Female, Hepacivirus, Hepatitis C, Chronic, Herpesvirus 4, Human, Humans, Italy, Leukemia, B-Cell, Male, Middle Aged, Polyomavirus, Polyomavirus Infections, Sequence Analysis, DNA, Vaginal Smears, Young Adult}, issn = {1096-9071}, doi = {10.1002/jmv.23867}, author = {Comar, Manola and Zanotta, Nunzia and Del Savio, Rossella and Vascotto, Fulvia and Calabrese, Nadia and Zorat, Francesca and Pozzato, Gabriele} } @article {1916, title = {Asbestos and SV40 in malignant pleural mesothelioma from a hyperendemic area of north-eastern Italy.}, journal = {Tumori}, volume = {98}, year = {2012}, month = {2012 Mar-Apr}, pages = {210-4}, abstract = {

AIMS AND BACKGROUND: Malignant mesothelioma is a fatal cancer of increasing incidence in north-eastern Italy. Together with asbestos, the polyomavirus SV40 was hypothesized to contribute to the onset of malignant mesothelioma. To investigate the putative role of SV40 in the individual susceptibility to asbestos-induced malignant mesothelioma, we conducted a molecular epidemiological study on a series of malignant mesothelioma patients from an area in north-eastern Italy hyperendemic for malignant pleural mesothelioma.

METHODS AND STUDY DESIGN: We collected 63 mesothelioma samples from incidence cases of patients diagnosed with malignant pleural mesothelioma in the period 2009-2010. DNA was extracted from patients{\textquoteright} tissue biopsies using the BioRobot EZ1 Qiagen workstation. SV40 sequence detection and quantification was performed by specific real time PCR. The 74.6\% of the 63 enrolled patients had a history of asbestos exposure. The epithelioid histotype was more prevalent in males (64.0\%) and the mixed in females (61.5\%) who showed significantly higher cancer co-morbidity (46.1\% vs 12\%, P = 0.005). SV40 was detected in 22\% of MM tumors, with a low viral load. In SV40-positive patients, a threefold increased risk of asbestos exposure was observed, more evident in females (OR 4.32) than in males (OR 1.20).

CONCLUSIONS: Our findings indicate that a high prevalence of SV40 was present in malignant mesothelioma incident cases from an area hyperendemic for malignant mesothelioma in north-eastern Italy. Although asbestos is considered the main risk factor in malignant mesothelioma onset, a role for SV40 could be hypothesized.

}, keywords = {Adult, Aged, Asbestos, Carcinogens, Disease Susceptibility, DNA, Viral, Endemic Diseases, Female, Humans, Italy, Male, Mesothelioma, Middle Aged, Pleural Neoplasms, Polyomavirus Infections, Real-Time Polymerase Chain Reaction, Risk Factors, Simian virus 40, Tumor Virus Infections, Viral Load}, issn = {2038-2529}, doi = {10.1700/1088.11931}, author = {Comar, Manola and Zanotta, Nunzia and Pesel, Giuliano and Visconti, Patrizia and Maestri, Iva and Rinaldi, Rosa and Crovella, Sergio and Cortale, Maurizio and De Zotti, Renata and Bovenzi, Massimo} } @article {1965, title = {Association between the JC polyomavirus infection and male infertility.}, journal = {PLoS One}, volume = {7}, year = {2012}, month = {2012}, pages = {e42880}, abstract = {

In recent years the incidence of male infertility has increased. Many risk factors have been taken into consideration, including viral infections. Investigations into viral agents and male infertility have mainly been focused on human papillomaviruses, while no reports have been published on polyomaviruses and male infertility. The aim of this study was to verify whether JC virus and BK virus are associated with male infertility. Matched semen and urine samples from 106 infertile males and 100 fertile males, as controls, were analyzed. Specific PCR analyses were carried out to detect and quantify large T (Tag) coding sequences of JCV and BKV. DNA sequencing, carried out in Tag JCV-positive samples, was addressed to viral protein 1 (VP1) coding sequences. The prevalence of JCV Tag sequences in semen and urine samples from infertile males was 34\% (72/212), whereas the BKV prevalence was 0.94\% (2/212). Specifically, JCV Tag sequences were detected in 24.5\% (26/106) of semen and 43.4\% (46/106) of urine samples from infertile men. In semen and urine samples from controls the prevalence was 11\% and 28\%, respectively. A statistically significant difference (p<0.05) in JCV prevalence was disclosed in semen and urine samples of cases vs. controls. A higher JC viral DNA load was detected in samples from infertile males than in controls. In samples from infertile males the JC virus type 2 strain, subtype 2b, was more prevalent than ubiquitous type 1. JCV type 2 strain infection has been found to be associated with male infertility. These data suggest that the JC virus should be taken into consideration as an infectious agent which is responsible for male infertility.

}, keywords = {Adult, Amino Acid Sequence, Amino Acid Substitution, Base Sequence, BK Virus, Capsid Proteins, DNA, Viral, Humans, Infertility, Male, JC Virus, Male, Molecular Sequence Data, Polyomavirus Infections, Semen, Sequence Analysis, DNA, Tumor Virus Infections}, issn = {1932-6203}, doi = {10.1371/journal.pone.0042880}, author = {Comar, Manola and Zanotta, Nunzia and Croci, Eleonora and Murru, Immacolata and Marci, Roberto and Pancaldi, Cecilia and Dolcet, Ornella and Luppi, Stefania and Martinelli, Monica and Giolo, Elena and Ricci, Giuseppe and Tognon, Mauro} } @article {1607, title = {Secondary lymphoid tissue as an important site for WU polyomavirus infection in immunocompetent children.}, journal = {J Med Virol}, volume = {83}, year = {2011}, month = {2011 Aug}, pages = {1446-50}, abstract = {

The polyomaviruses KI and WU (KIPyV and WUPyV) have been identified in respiratory specimens from children with acute respiratory infections, which suggests the respiratory tract as a possible site of infection. However, the persistence of infection in the lymphoid system is unknown. Fresh samples (n = 211) of tonsils, adenoids, and peripheral blood mononuclear cells (PBMCs) from 83 immunocompetent children (mean age 4.8 years) were tested for amplification of the KIPyV VP1 and WUPyV VP2 genes. The known BK and JC polyomaviruses and the lymphotropic human herpesvirus (HHV)-6 were also investigated by quantitative real-time PCR and direct sequencing. In addition, 98 nasopharyngeal swabs collected from children (mean age 6.2 years) affected by seasonal influenza-like illness were tested. Of the lymphoid tissues, 34.9\% were positive for WUPyV, 4.8\% for BK virus, and 33.8\% for HHV-6. KIPyV and JC virus were not detected in these specimens. None of the polyomaviruses were detected in PBMCs. Among the nasopharyngeal samples, the prevalence of WUPyV was 27.5\%, although 70\% of the positive samples were co-infected with at least one of the following respiratory viruses: influenza virus, adenovirus, and respiratory syncytial virus. Phylogenetic analysis revealed high sequence homology (99\%) between lymphoid- and nasopharynx-derived WUPyV strains. These results suggest that the tonsils and adenoids of immunocompetent children are a reservoir for WUPyV infection; probably due to the respiratory route of transmission. In addition, the prevalence of WUPyV was high among the children, and the virus was identified more frequently in older children than during the first years of life.

}, keywords = {Adenoids, Child, Child, Preschool, DNA, Viral, Female, Humans, Infant, Leukocytes, Mononuclear, Male, Nasopharynx, Palatine Tonsil, Phylogeny, Polymerase Chain Reaction, Polyomavirus, Polyomavirus Infections, Prevalence, Sequence Analysis, DNA, Sequence Homology}, issn = {1096-9071}, doi = {10.1002/jmv.22124}, author = {Comar, Manola and Zanotta, Nunzia and Rossi, Tatiana and Pelos, Giorgio and D{\textquoteright}Agaro, Pierlanfranco} } @article {1632, title = {Detection of SV40 in colon cancer: a molecular case-control study from northeast Italy.}, journal = {J Med Virol}, volume = {82}, year = {2010}, month = {2010 Jul}, pages = {1197-200}, abstract = {

To explore the involvement of the simian polyomavirus SV40 in human colon cancer, a molecular case-control study was undertaken in patients and in their relatives living in an area where the spread of SV40 has already been documented. From 2006 to 2008, 94 colon cancer patients (age: 37-90 years) and 91 subjects (age: 32-70 years) relatives of each index case were enrolled. A blood sample and a specimen of cancer tissue or biopsy were collected, from each patient or control, respectively. Samples were analyzed twice for Polyomavirus (i.e., SV40, JCV, and BKV) by PCR and by quantitative real-time PCR (RT-qPCR) with reproducible results. No BKV/JCV was detected either in normal or pathological tissues. SV40 was not present in control subjects, either normal tissue or in biopsies from adenomas or polyps. All blood samples were negative. Conversely, six adenocarcinoma specimens were positive for SV40 sequences (overall prevalence 6.4\%, P = 0.03 in comparison with controls). Nevertheless, the SV40-associated colon cancer risk proved statistically not significant (OR = 3.91; P = 0.115) when adjusted for age. Quantitation of SV40 DNA performed by RT-qPCR showed a low viral load ranging from 6.2 x 10(1) to 9 x 10(3) copies per reaction. This molecular case-control survey showed, for the first time in fresh samples and by RT-qPCR, that SV40 can be detected in colon cancer tissue. However, the finding was not statistically significant when compared with a well-structured community control group. Thus, the role of SV40 and other polyomavirus in colon cancer genesis deserves further investigation.

}, keywords = {Adenocarcinoma, Adult, Aged, Aged, 80 and over, BK Virus, Case-Control Studies, Colonic Neoplasms, DNA, Viral, Female, Humans, Italy, JC Virus, Male, Middle Aged, Polymerase Chain Reaction, Polyomavirus Infections, Simian virus 40, Tumor Virus Infections}, issn = {1096-9071}, doi = {10.1002/jmv.21798}, author = {Campello, Cesare and Comar, Manola and Zanotta, Nunzia and Minicozzi, Anna and Rodella, Luca and Poli, Albino} } @article {1640, title = {JCV/BKV and SV40 viral load in lymphoid tissues of young immunocompetent children from an area of north-east Italy.}, journal = {J Med Virol}, volume = {82}, year = {2010}, month = {2010 Jul}, pages = {1236-40}, abstract = {

Polyomavirus infection occurring during childhood is followed by a lifelong latency in immunocompetent subjects. The major site of polyomavirus persistence are the uroepithelial cells which leads to oral transmission. It has recently been hypothesized that tonsils could be a possible reservoir. The role of tonsil, adenoid, and peripheral blood mononuclear cells (PBMCs) as possible sites of JCV, BKV, and SV40 latency in young healthy children was assessed. Two hundred fifteen fresh specimens, including 57 tonsil, 80 adenoid, and 78 PBMC samples from 80 immunocompetent children (mean age 4.8 years) were analyzed to determine the viral load by quantitative real-time PCR. The human herpes virus 6 (HHV-6) was tested as a lymphotropic reference virus. Polyomavirus was detected in 5/80 (6.2\%) children while HHV-6 infection affected 27/80 children (33.7\%) (P < 0.001). SV40 was detected in one adenoid sample, while footprints of BKV were found in one adenoid and three tonsil samples. JCV was never found in all samples. Polyomavirus sequences were not detected in the 78 blood samples. One adenoid and two tonsils from three children (1.4\%) were positive for both polyomavirus and HHV-6. Infections were characterized by low replication rates ranging typically from 1 x 10e(2)/5.5 x 10e(4) to 6.8 x 10e(3)/8.5 x 10e(4) viral copies/number of cells. In conclusion, tonsils and adenoids of children could effectively harbor BKV and SV40, although only very few cells proved to be infected. Nevertheless, the low prevalence of polyomavirus, in comparison with the lymphotropic HHV-6, suggests that these tissues are unlikely to be the preferred site of polyomavirus latency, at least in younger children.

}, keywords = {Adenoids, BK Virus, Carrier State, Child, Child, Preschool, Female, Herpesvirus 6, Human, Humans, Immunocompetence, Italy, JC Virus, Male, Neutrophils, Palatine Tonsil, Polyomavirus Infections, Simian virus 40, Viral Load, Virus Latency}, issn = {1096-9071}, doi = {10.1002/jmv.21786}, author = {Comar, Manola and Zanotta, Nunzia and Bovenzi, Massimo and Campello, Cesare} }