Muhammed Babakir-Mina

Identification of Merkel cell polyomavirus in the lower respiratory tract of Italian patients.

Merkel cell polyomavirus (MCPyV) has been found to be integrated monoclonally in a rare skin cancer named Merkel cell carcinoma. More recently, MCPyV has been detected in the upper respiratory tract of pediatric and adult patients. However, the mode of transmission and pathogenic role of MCPyV in the respiratory system has not been determined. In this study, MCPyV was sought in the lower respiratory tract of adult patients admitted to the hospital. MCPyV DNA was detected in 15 (17.24%) out of 87 lower respiratory tract samples. Most of the patients with MCPyV were over 50 years old. Nucleotide sequence of the t‐antigen of MCPyV identified in respiratory secretions showed a homology to those found in Merkel cell carcinoma. In addition, phylogenetic analysis undertaken on the t‐antigen sequences of Italian isolates and other MCPyVs identified in healthy and cancer tissues showed that all these isolates belonged to the same clade. Selective pressure analysis for the t‐antigen revealed the presence of five sites under positive selection (ω = 4.3), with a posterior probabilities above 0.99. The α parameter of the gamma distribution was 0.01, showing that this distribution has a characteristic L‐shape and suggesting a strong nucleotide substitution rate heterogeneity across sites. This study shows that MCPyV can infect the lower respiratory tract, but further investigations are needed to define its pathogenic role in respiratory diseases. J. Med. Virol. 82:505–509, 2010.

Excretion of the novel polyomaviruses KI and WU in the stool of patients with hematological disorders

Infection with human polyomaviruses BKV and JCV is asymptomatic, and lifelong and widespread, among the general population. However, in the setting of immunosuppression, secondary to medications or viral infection, for example, with HIV, reactivation can occur and result in severe disease. In this study, stool specimens from 31 patients with hematological disorders (25 transplanted and 6 non‐transplanted) were examined prospectively to determine whether the novel polyomaviruses KIV and WUV reactivated and were excreted in the gastrointestinal tract. Reactivation was correlated with the appearance of gastrointestinal and respiratory symptoms. Of the 31 patients examined, KIV and WUV were detected in 13 transplanted patients as single infection or in combination with BKV, cytomegalovirus (CMV), and adenovirus (Adv). Because of frequent co‐infections, a clear correlation between novel polyomaviruses and clinical symptoms could not be established. There was no correlation between demographic variables and detection of KIV and WUV. Phylogenetic analysis of the small t‐antigen gene of KIV and WUV isolates showed that the novel polyomaviruses identified in feces clustered with those identified in the respiratory tract suggesting an oral–fecal transmission of these viruses. The novel polyomaviruses KI and WU may have a pathogenic role in immunocompromised patients. J. Med. Virol. 81:1668–1673, 2009.

Identification of the novel KI polyomavirus in paranasal and lung tissues

KI is a novel polyomavirus identified in the respiratory secretions of children with acute respiratory symptoms. Whether this reflects a causal role of the virus in the human respiratory disease remains to be established. To investigate the presence of KIV in the respiratory tissue, we examined 20 fresh lung cancer specimens and surrounding normal tissue along with one paranasal and one lung biopsy from two transplanted children. KIV‐VP1 gene was detected in 9/20 lung cancer patients and 2/2 transplanted patients. However, amplification of the sequence coding for the C‐terminal part of the early region of KIV performed on the 11 positive cases was successful only in two malignant lung tissues, one surrounding normal tissue, and 1/2 biopsies tested. Phylogenetic analysis performed on the early region of KIV (including the four Italian isolates), BKV and JCV revealed the presence of three distinct clades. Within the KIV clade two sub‐clades were observed. A sub‐clade A containing the four Italian strains, and a sub‐clade B comprising the Swedish and Australian isolates. Interestingly, the two Italian strains identified in normal tissue clustered together, whereas those detected in malignant tissue fell outside this cluster. In vitro studies are needed to investigate the transforming potential of KIV strains. J. Med. Virol. 81:558–561, 2009.

Identification of the novel KI and WU polyomaviruses in human tonsils

BACKGROUND Three novel polyomaviruses have been recently discovered: KI, WU and MC polyomaviruses. Their role in human pathology is debated while tissue tropism and site of latency remain unknown. OBJECTIVE To test the hypothesis that KI, WU and MC polyomaviruses can infect human tonsils. STUDY DESIGN Archival paraffin-embedded tonsils from 91 patients affected by different tonsil diseases were screened by polymerase chain reaction to detect viral DNA of KIV, WUV, MCV, BKV and JCV. Phylogenetic and evolutionary analysis of the identified polyomaviruses was carried out. RESULTS Of the 91 tested specimens, 11 contained KIV DNA (12%), 4 WUV DNA (4.4%), 5 BKV DNA (5.5%). MCV and JCV were not detected. Phylogenetic analysis showed that KIVs identified in tonsils fall into a clade distinct from that containing KIVs isolated from respiratory secretions, respiratory tissue and feces. Moreover, four positively selected sites (4.5% of t-Ag sites) were found under strong positive selection (omega=11.4), with posterior probabilities above 0.99. All the sites were located in the N-terminal region of the small t antigen. CONCLUSIONS The results suggest that the novel KI and WU polyomaviruses can infect human tonsils. Future studies are needed to define their role in tonsil diseases.

The human polyomaviruses KI and WU: virological background and clinical implications

In 2007, two novel polyomaviruses KI and WU were uncovered in the respiratory secretions of children with acute respiratory symptoms. Seroepidemiological studies showed that infection by these viruses is widespread in the human population. Following these findings, different biological specimens and body compartments have been screened by real‐time PCR in the attempt to establish a pathogenetic role for KI polyomavirus (KIPyV) and WU polyomavirus (WUPyV) in human diseases. Although both viruses have been found mainly in respiratory tract samples of immunocompromised patients, a clear causative link with the respiratory disease has not been established. Indeed, the lack of specific clinical or radiological findings, the frequent co‐detection with other respiratory pathogens, the detection in subjects without signs or symptoms of respiratory disease, and the variability of the viral loads measured did not allow drawing a definitive conclusion. Prospective studies carried out on a large sample size including both immunocompromised and immunocompetent patients with and without respiratory symptoms are needed. Standardized quantitative real‐time PCR methods, definition of a clear clinical cutoff value, timing in the collection of respiratory samples, are also crucial to understand the pathogenic role, if any, of KIPyV and WUPyV in human pathology.

Viral causes of influenza-like illness: insight from a study during the winters 2004-2007.

Limited information is available on the viral etiology of influenza‐like illness in southern European countries, and it is still a matter of debate whether certain symptoms can be used to distinguish among the specific viruses that cause influenza‐like illness. The main objective of the present study was to identify the demographic and clinical predictors of influenza‐like illness due to specific viral agents. The study, which was observational in design, was conducted in Rome and Naples, Italy. Cases of influenza‐like illness were defined as individuals with fever >37.5°C and at least one systemic and one respiratory symptom, recruited during the winters of 2004–2005, 2005–2006, and 2006–2007. Influenza and other respiratory viruses were identified using the polymerase chain reaction (PCR), performed on throat swabs. Basic individual information was collected using a standard form. A total of 580 persons were included in the analysis. Viral pathogens were identified in fewer than 50% of the cases. Overall, 240 viral agents were detected: 22.8% were positive for influenza viruses, 10.9% for adenoviruses, 6.0% for parainfluenza viruses, and 1.7% for respiratory syncytial virus. The month of diagnosis, and muscle and joint pain were associated with influenza virus, though the positive predictive value (PPV) was low. Abdominal pain was associated with adenovirus infection. Although the PPV of symptoms for influenza virus infection was low, especially in low activity periods, these findings may help clinicians to improve their ability to perform diagnoses. J. Med. Virol. 81:2066–2071, 2009.

Origin of the 2009 Mexico influenza virus: a comparative phylogenetic analysis of the principal external antigens and matrix protein

Triple-reassortant swine influenza A (H1) viruses, containing genes from avian, human, and swine influenza viruses, emerged and became an outbreak among humans worldwide. Over a 1,000 cases were identified within the first month, chiefly in Mexico and the United States. Here, the phylogenetic analysis of haemagglutin (HA), neuraminidase (NA), and matrix protein (MP) was carried out. The analysis showed that the H1 of this reassortant originated from American pigs, while NA and MP were more likely from European pigs. All of the 2009 isolates appear homogeneous and cluster together, although they are distinct from classical human A (H1N1) viruses.

Identification of the novel KI polyomavirus in the respiratory tract of an Italian patient.

Recently, a new human polyomavirus, KIV, was detected in respiratory specimens of patients with acute respiratory tract infection. Whether this reflects a causal role of the virus in the respiratory tract is still debated. To investigate the presence of KIV in respiratory samples of Italian patients and to determine the degree of similarity with other known polyomaviruses, 222 respiratory specimens collected by general practitioners between 2006 and 2007 were screened. The entire VP1 gene region was amplified and sequenced. Maximum Likelihood tree was generated by PAUP* software. One out of 222 samples tested was positive for KIV. Phylogenetic analysis indicated that this isolate clustered with other KIV isolates, while the WUV isolates seem to belong to a different lineage. The phylogenetic tree also showed that all other known polyomaviruses are quite distant from this isolate. This is the first report describing the presence of KIV in the respiratory tract of a 5‐year‐old Italian child with acute respiratory symptoms. Further investigations are needed to establish an etiological link of KIV with acute respiratory illness. J. Med. Virol. 80:2012–2014, 2008.

Real Time PCR TaqMan assays for detection of polyomaviruses KIV and WUV in clinical samples.

Abstract Recently, polyomaviruses KI and WU were identified in the airways of patients with acute respiratory symptoms. The epidemiology and pathogenesis of these two viruses are not fully understood, and the development of molecular assays, such as Real Time PCR, was useful for examining their biology and role in different clinical syndromes. The evaluation of different target regions for the amplification of polyomaviruses KI and WU, comparing published primer/probe sets and sets designed in the laboratory is described and was used for testing 175 clinical specimens (84 stools and 91 tonsils). The results showed that the laboratory designs were more sensitive for the detection of polyomaviruses KI and WU DNA in clinical samples. The choice of the primer/probe set, and primarily of the region for amplification, may be relevant for understanding the pathogenic role of viruses such as polyomaviruses KI and WU.

KI and WU polyomaviruses and CD4+ cell counts in HIV-1- infected patients , Italy

To investigate an association between KI and WU polyomavirus (KIPyV and WUPyV) infections and CD4+ cell counts, we tested HIV-1–positive patients and blood donors. No association was found between cell counts and virus infections in HIV-1–positive patients. Frequency of KIPyV infection was similar for both groups. WUPyV was more frequent in HIV-1–positive patients.