Human Syncytial Virus Research Articles

HPV related p16INK4A and HSV in benign and potentially malignant oral mucosa pathologies

BackgroundThe association of Human Papilloma Virus (HPV) and Human Syncytial Virus (HSV) infection with inflammatory and potentially malignant disorders of the oral cavity (OPMD) is unknown. The aim of this cross-sectional study was to stablish the expression of the p16INK4A and HSV proteins, to test potential correlation between those parameters in biopsies from clinically diagnosed oral lesions.MethodsImmunochemical analysis of 211 formalin-fixed, paraffin-embedded (FFPE) blocks from 211 individuals was provided. The clinical diagnosis included in the research were Oral lichen planus (N = 30), Oral Leukoplakia (N = 13) Mucocele (N = 25), Erosion/ulceration/ inflammation of mucosa (N = 8), Overgrowth of mucosa (N = 135).ResultsTwo hundred eleven analyzed FFPE samples resulted with the median age of 58.5 years (the average age 54.0 years and SD ± 17 years). The female/male ratio was 2.3 (69.7% vs 30.3% respectively). All the samples positive for HSV also expressed p16INK4A (p = 0.000), that’s showed various levels of association with the diverse clinical diagnosis reaching the higher level in OM 49.1% (29 positive samples) and OLP 30.5% (18). p16INK4A was associated with OLP at 30.5% (18), and fibroma 30.5%. HSV expression was mostly present in fibroma at 47.6% (10 positive samples).ConclusionHSV and p16INK4A positivity in relation to diagnosis of the biopsies showed statistically most often p16INK4A in OLP and fibroma. The results of co-expression of p16INK4A and HSV in mucocele and fibroma in oral mucosa suggest a cooperation between the molecular alterations induced by these two viruses. Squamous papilloma samples positive for p16INK4A were also positive for HSV, suggesting that the putative pro-oncogenic action of HSV could be an early event.

Duplex real-time RT-PCR assay for detection and subgroup-specific identification of human respiratory syncytial virus

Human respiratory syncytial virus (HRSV) is a leading cause of acute respiratory illness in young children worldwide. Reliable detection and identification of HRSV subgroup A and B infections are essential for accurate disease burden estimates in anticipation of licensure of novel HRSV vaccines and immunotherapies. To ensure continued reliability, molecular assays must remain current with evolving virus strains. We have developed a HRSV subgroup-specific real-time RT-PCR (rRT-PCR) assay for detection and subgroup identification using primers and subgroup-specific probes targeting a conserved region of the nucleoprotein gene combined in a single duplex reaction using all genome sequence data currently available in GenBank. The assay was validated for analytical sensitivity, specificity, reproducibility, and clinical performance with a geographically diverse collection of viral isolates and respiratory specimens in direct comparison with an established pan-HRSV rRT-PCR reference test. The assay was sensitive, reproducibly detecting as few as 5–10 copies/reaction of target RNA. The assay was specific, showing no amplification with a panel of 16 other common respiratory pathogens or predicted by in silico primer/probe analysis. The duplex rRT-PCR assay based on the most current available genome sequence data permits rapid, sensitive and specific detection and subgroup identification of HRSV.

Molecular Epidemiology of Human Respiratory Syncytial Virus in Iranian ≥60 Years Old Hospitalized Patients with Acute Respiratory Symptoms.

Human respiratory syncytial virus (HRSV) is an important cause of acute respiratory infection (ARI) and mortality in ≥60 years old population around the world. The aim of this study was to determine the frequency and circulating genotypes of HRSV in Iranian patients ≥ 60 years old. In this cross-sectional molecular epidemiology study, we examined the C-terminal located hypervariable domain of G glycoprotein of HRSV in throat swabs from Tehran, Hormozgan, Boushehr, West Azarbayjan, Ghom and Alborz provinces of Iran which were addressed to national influenza center between October 2013 and March 2015. During these two consecutive years, a total of 225 samples collected from patients older than 60 years were tested using RT hemi-nested PCR and sequencing and the acquired sequences were phylogenetically analyzed. Sixteen out of 225 samples (7.1%) yielded a positive result. Among the positive samples, 13 cases (81%) pertained to antigenic group A and the remaining 3 cases (19%) belonged to group B. Three genotypes including GA1, GA2 and BA9 were identified in the first year of survey whereas during the second year, only GA1 and GA2 genotypes were detected. Our study indicates that HRSV genotypes from both A and B antigenic groups which were discovered in pediatric population previously, are circulating among Iranian ≥60 years old population.

Respiratory syncytial virus receptor expression in the mouse and viral tropism.

Human respiratory syncytial virus (RSV) infects airway epithelium and can cause serious illnesses such as bronchiolitis and pneumonia. With the discovery of cell-surface nucleolin as a fusion receptor for RSV, the question arose as to whether nucleolin could explain RSV tropism in vivo. Here, we report the distribution of cell-surface nucleolin expression in tissues of normal mice and how this distribution of expression relates to what is known about RSV tropism and its clinical manifestations. Our results show evidence of cell-surface nucleolin expression in the respiratory tract. In addition, cell-surface nucleolin is expressed in tissues outside of the respiratory tract, many of which correspond to previous reports of tissue-specific RSV infection, and others that may allude to additional potential sites for RSV infection in vivo. Furthermore, our work provides a foundation for the investigation of nucleolin's physiological function in various healthy mammalian tissues.

Biologics and molecular recognition: Mike Brigham‐Burke memorial issue

Biologics and molecular recognition: Mike Brigham‐Burke memorial issue

Construction and identification of replication deficient recombinant adenovirus encoding F gene of subgroup A human respiratory syncytial virus

A strain of replication deficient recombinant adenvirus encoding fusion glycoprotein (F) of subgroup A human respiratory syncytial virus (RSV) was constructed and the expression of F was identified. The F gene was obtained from pGEM3zf-F with Xho I and Hind III, cloned into adenoviruse shuttle vector pShuttle-CMV,and then the resulting pShuttle-CMV/F was transformed into E. coli BJ5183/p with pAdeasy-1 to produce pre-adenoviral plasmid encoding F by homologous recombination. This resultant plasmid was linearized by digestion with Pac I and transfected into 293 packaging cells to generate FGAd-F. Finally, the expression of F protein was identified by Western Blot analysis. FGAd/F was successfully constructed, and the expression of RSV F protein was identified by Western Blot. We have obtained a strain of replication-defective adenovirus FGAd/F encoding RSV F protein, which can be used further to investigate its protective efficacy against RSV infection in vivo.

Naturally occurring antibodies to the human syncytial virus in West Africa.

AbstractThe seroepidemiology of the human syncytial virus has been investigated by means of an indirect immunofluorescence test on 113 sera from normal antenatal women in Nigeria and 233 sera from normal individuals of both sexes and all ages in the Ivory Coast. It was found that 3.5% of samples from Nigeria and 19.7% of those from the Ivory Coast were positive. The findings are discussed.

The prevalence of naturally occurring antibodies to human syncytial virus in East African populations.

A seroepidemiological study of naturally occurring antibodies to the human syncytial virus has been carried out by means of an indirect immunofluorescence test on 639 East Africans, consisting of 493 normal Ugandans, 66 Kenyan patients with nasopharyngeal carcinoma (NPC), and 80 Kenyan and Tanzanian patients with various other tumours or non-cancerous conditions. It was found that 3.4% of the normal individuals had antibodies to the virus and serial serum samples were available from 14 of these, permitting the study of antibody class in seroconversion and antibody persistence. As in an earlier survey, a significantly higher incidence of antibodies was found amongst NPC patients. Blocking and indirect immunofluorescence test with simian foamy viruses (SFV) showed some cross-reactivity between SFV 6 and the human syncytial virus, but not identity. The results are discussed in relation to the very real occurrence of natural infection by human syncytial virus in certain geographical regions.

Pathogenesis of respiratory syncytial virus infection in ferret and fetal human tracheas in organ culture.

The pathogenesis of human respiratory syncytial virus infection was studied in ferret and fetal human tracheas in organ culture. Although the patterns of virus growth were similar in these species, the sites and morphologic consequences of virus replication differed markedly. In human trachea, synthesis of respiratory syncytial virus occurred in a population of ciliated epithelial cells, whereas other cells in the epithelial layer were spared. Virus replication was associated with cell injury characterized by ballooning degeneration and syncytium formation. In ferret trachea, virus growth occurred in fibroblasts of the lamina propria and serosa. Ciliated epithelial cells did not contain viral antigen and remained histologically normal. These observations are relevant to understanding the pathogenesis of human disease and the evaluation of animal models of respiratory syncytial virus bronchiolitis.

Preliminary Seroepidemiological Studies on the Human Syncytial Virus

The seroepidemiology of the human syncytial virus was investigated by means of an indirect immunofluorescence test on 241 sera from Kenya, Tunisia, Singapore and Britain. These included sera from patients with nasopharyngeal carcinoma, other tumours of the oro-nasopharynx, tumours of other parts of the body, and from normal donors. In this study, the virus was found to infect only Kenyan Africans and all but one of these seropositive subjects had tumours, particularly of the oro-nasopharyngeal spaces. The significance of these findings is discussed.

Further Biological Properties of the Human Syncytial Virus

Some biological properties of the human syncytial virus have been examined. A 13-day plaque assay in whole human embryo fibroblasts (HEF) has been developed using a liquid (growth medium) overlay. The plaques were 0.7-2 mm in diameter and often showed a clear central zone with irregular edges. Pretreatment of HEF monolayers with the polycation DEAE-dextran for either 30 min or 1 h was found to enhance plaque formation by a factor of from 2- to 7-fold. The plaque assay procedure required cell cultures undergoing active cell division. Adsorption kinetics and growth cycle studies in HEF indicated a relatively long adsorption period (3 h) and a relatively prolonged latent period of 24 h. Even under optimal conditions, virus yields were low and did not exceed 1 PFU per infected cell. Like other animal syncytium-forming 'foamy' viruses, the human virus induced both intranuclear and cytoplasmic antigens detectable by immmunofluorescence and was also markedly labile to freezing and thawing.