Since March 2009, pandemic A/H1N1/2009 influenza virus has been spreading throughout many countries including China. The emerged virus caused great harm to human health and social economy. Hemagglutinin (HA) is the most important viral surface glycoprotein, mainly possessing three kinds of functions: (1) binding to host cell receptor, (2) triggering the fusion between viral envelop and target cell membrane, (3) stimulating the body to generate the neutralizing antibody. Advances in the structure, primary function, evolution and antigenicity of pandemic A/H1N1/2009 influenza virus HA protein are reviewed in this paper.
Animals ; Evolution, Molecular ; Hemagglutinin Glycoproteins, Influenza Virus ; chemistry ; genetics ; immunology ; metabolism ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; immunology ; pathogenicity ; physiology ; Influenza, Human ; epidemiology ; virology ; Pandemics

This study aims to investigate the virological impact of the stalk region and cysteine (C) in neuraminidase (NA) of influenza A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1) viruses. The NA of A/ Anhui/1/05 (H5N1), defined as AH N1, lacked 20 amino acids (including C, defined as s20) as compared with NA of A/Ohio/07/2009 (H1N1) (defined as 09N1). We deleted s20 of 09N1 to construct 09N1-s20, and inserted s20 into AH N1 to construct AH N1+s20. To investigate the impact of C on the biological function of NA, we deleted C in 09N1 to construct 09N1-C and inserted C into AH N1 to construct AH N1-C. The pseudo-type viral particle (pp) system was used to evaluate the impact of these mutants on virology. The combination of 09N1-C and 09H1 (defined as 09H1::09N1-C) showed an infectivity 8 times that of the wild type 09H1::09N1, while the infectivity of the combination of AH N1+C and AH H5 (defined as AH H5::AH N1+C) was much lower than that of the wild type AH H5::AH N1. The infectivity of the combination of 09N1-s20 and 09H1 (defined as 09H1::09N1-s20) was 4 times that of the wild type 09H1::09N1; the infectivity of the combination of AH N1+s20 and AH H5 (defined as AH H5:: AH N1+s20) was 1/7 that of the wild type AH H5::AH N1. The co-existence of 09N1-C and AH H5 displayed 6 times the infectivity of AH H5::09N1, while the infectivity of 09H1::AH N1+C was very low. Multimer analysis showed that in the wild type 09N1, the forms of NA were dimer > tetramer > monomer; the major component of NA in 09N1-C was monomer; in 09N1-s20, the forms of NA were monomer > dimer. AH N1 was mainly composed of monomer; in AH N1+s20, the forms of NA were dimer > monomer > tetramer; in AH N1+C, the forms of NA were dimer > tetramer. Deletion of C or s20 from 09N1 did not change the expression of NA. The study suggested that deletion of C from the stalk region of NA in A/Ohio/07/2009 (H1N1) increases infectivity. Insertion of C into NA's stalk region of A/ Anhui/1/05 (H5N1) significantly decreases infectivity. Cysteine deletion in the stalk region is important for the infectivity of A/Anhui/1/05 (H5N1) and A/Ohio/07/2009 (H1N1). It may interfere with the infectivity via changes in NA polymerization.
Amino Acid Motifs ; Humans ; Influenza A Virus, H1N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza A Virus, H5N1 Subtype ; chemistry ; enzymology ; genetics ; pathogenicity ; Influenza, Human ; virology ; Neuraminidase ; chemistry ; genetics ; metabolism ; Viral Proteins ; chemistry ; genetics ; metabolism ; Virulence

BACKGROUNDThe duration of viral shedding and the transmission of 2009 H1N1 influenza among individuals, especially among the younger population with mild illness, are not well understood now. The aim of this study was to determine the viral shedding of the young adult patients with mild 2009 H1N1 influenza in China.

METHODSFrom September 2009 to January 2010, the clinical data and serial nasopharyngeal swabs of 67 patients with 2009 H1N1 influenza and 37 patients with seasonal influenza aged from 18 years to 35 years were collected. The nasopharyngeal swab samples were detected by real time RT-PCR to determine the viral shedding. All the patients did not receive the antiviral therapy but Chinese medicine for detoxicating.

RESULTSAmong the patients with H1N1 virus infection, 82.1% (55/67) patients presented with fever symptom, while more patients with high fever (≥ 39°C) were found in seasonal influenza patients (P < 0.05). For the H1N1 patients, the median interval between the symptom onset and the undetectable RNA was six days (4 - 10 days). But viral shedding was still found in 31.3% patients after 7 days following illness onset. The median interval between disappearance of fever and an undetectable viral RNA level was three days (2 - 8 days), and 17.9% patients were found to be viral shedding 6 days later after normalization of body temperature. For the seasonal influenza patients, 94.6% patients were detected out viral RNA within 7 days. The median interval of seasonal influenza between the symptom onset and the undetectable RNA was four days (3 - 8 days). The median interval between disappearance of fever and an undetectable viral RNA level was three days (2 - 6 days).

CONCLUSIONIt suggests that 7 days isolation period from the illness onset or 24 hours after the resolution of fever and respiratory symptoms are not long enough to cut off the transmission among Chinese young adults with mild illness.

Adult ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; pathogenicity ; Influenza, Human ; epidemiology ; virology ; Male ; Real-Time Polymerase Chain Reaction ; Virus Shedding ; genetics ; physiology ; Young Adult

Animals ; Global Health ; History, 20th Century ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; isolation & purification ; metabolism ; pathogenicity ; Influenza, Human ; epidemiology ; history ; virology ; Spain ; epidemiology ; Viral Proteins ; genetics ; metabolism

The use of antiviral drugs such as influenza neuraminidase (NA) inhibitors is a critical strategy to prevent and control flu pandemic, but this strategy faces the challenge of emerging drug-resistant strains. For a highly pathogenic avian influenza (HPAI) H5N1 virus, biosafety restrictions have significantly limited the efforts to monitor its drug responses and mechanisms involved. In this study, a rapid and biosafe assay based on NA pseudovirus was developed to study the resistance of HPAI H5N1 virus to NA inhibitor drugs. The H5N1 NA pseudovirus was comprehensively tested using oseltamivir-sensitive strains and their resistant mutants. Results were consistent with those in previous studies, in which live H5N1 viruses were used. Several oseltamivir-resistant mutations reported in human H1N1 were also identified to cause decreased oseltamivir sensitivity in H5N1 NA by using the H5N1 NA pseudovirus. Thus, H5N1 NA pseudoviruses could be used to monitor HPAI H5N1 drug resistance rapidly and safely.
Animals ; Birds ; Drug Resistance, Viral ; genetics ; Enzyme Inhibitors ; therapeutic use ; HEK293 Cells ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; genetics ; pathogenicity ; Influenza A Virus, H5N1 Subtype ; drug effects ; genetics ; pathogenicity ; Influenza in Birds ; drug therapy ; genetics ; virology ; Influenza, Human ; drug therapy ; genetics ; virology ; Mutagenesis, Site-Directed ; Neuraminidase ; antagonists & inhibitors ; genetics ; Oseltamivir ; administration & dosage

As the scientific community scrambles to define the ancestry and lineages of the eight segments of new pandemic H1N1 strain, we looked for unique genetic events in this virus's genome to explain the newly found enhanced virulence and transmissibility among humans. Genome annotations of this virus identified a stop mutation replacing serine at codon 12 (S12Stop) of the PB1-F2 protein, a virulence factor in influenza A viruses. Here, we discuss the significance of this finding and how it may contribute to host specialization, explaining the virtual absence of the H1N1 influenza A virus strain in pig populations. This finding is expected to lead to a better understanding of the transmission and pathogenesis of the 2009 pandemic strain.
Amino Acid Sequence ; Gene Expression Regulation, Viral/physiology ; Host-Pathogen Interactions ; Humans ; Influenza A Virus, H1N1 Subtype/*genetics/*pathogenicity ; Influenza, Human/*virology ; Molecular Sequence Data ; Mutation ; Viral Proteins/chemistry/*genetics/metabolism ; Virulence

To establish the mouse-lethal model for pandemic H1N1 influenza virus, provide an animal model for studying the pathogenicity and host adaptation of 2009 pandemic H1N1 influenza virus, and find out the key amino acid mutations which may affect viral virulence and replication. A pandemic H1N1 influenza virus strain, A/Sichuan/SWL1/2009 (H1N1, SC/1) was passaged in mouse lung by 15 cycles with intranasal infection. The passaged viruses were all propagated in MDCK cells and sequenced. Based on the sequencing results, four mice in each group were inoculated with 6 selected viruses and their weight and survival rate were monitored during the following 14 days after infection. Additionally, SC/1-MA P14 and P15 viruses were sequenced after purification by Plague Assay. Viral virulence was increased after serial passages and the mortality of 100% was detected after 7 passages. Several amino acid residue mutations of passaged viruses which may contribute to the enhanced virulence were observed. The increased virulence of passaged viruses and mammalian host adaptation maybe associated with amino acid mutations in viral functional proteins. Finally, we established a mouse-lethal model.
Amino Acid Substitution ; Animals ; Base Sequence ; Cell Line ; China ; epidemiology ; Disease Models, Animal ; Dogs ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; growth & development ; pathogenicity ; physiology ; Influenza, Human ; epidemiology ; virology ; Mice ; Mice, Inbred BALB C ; Pandemics ; Survival Analysis ; Viral Plaque Assay ; Virulence ; Virus Replication

OBJECTIVETo perform gene expression profiles comparison so that to identify and understand the potential differences in pathogenesis between the pandemic and seasonal A (H1N1) influenza viruses.

METHODSA549 cells were infected with A/California/07/09 (H1N1) and A/GuangdongBaoan/51/08 (H1N1) respectively at the same MOI of 2 and collected at 2, 4, 8, and 24 h post infection (p.i.). Gene expression profiles of A549 cells were obtained using the 22 K Human Genome Oligo Array, and differentially expressed genes were analyzed at selected time points.

RESULTSMicroarrays results indicated that both of the viruses suppressed host immune response related pathways including cytokine production while pandemic H1N1 virus displayed weaker suppression of host immune response than seasonal H1N1 virus. Observation on similar anti-apoptotic events such as activation of apoptosis inhibitor and down-regulation of key genes of apoptosis pathways in both infections showed that activities of promoting apoptosis were different in later stage of infection.

CONCLUSIONSThe immuno-suppression and anti-apoptosis events of pandemic H1N1 virus were similar to those seen by seasonal H1N1 virus. The pandemic H1N1 virus had an ability to inhibit biological pathways associated with cytokine responses, NK activation and macrophage recognition.

Apoptosis ; genetics ; Cell Line, Tumor ; Cytopathogenic Effect, Viral ; Disease Outbreaks ; Down-Regulation ; Epithelial Cells ; metabolism ; virology ; Gene Expression ; Gene Expression Profiling ; Humans ; Immunity, Innate ; genetics ; Influenza A Virus, H1N1 Subtype ; classification ; pathogenicity ; Influenza, Human ; epidemiology ; genetics ; immunology ; virology ; Oligonucleotide Array Sequence Analysis ; Pandemics ; Seasons ; Up-Regulation ; Virulence

PURPOSE: Pneumonia was an important cause of death in 2009 H1N1 influenza pandemic (pH1N1). Clinical characteristics of pH1N1 have been described well, but discriminative characteristics suggesting pH1N1 infection in pneumonia patients are not evident today. We evaluated differences between clinical and radiologic characteristics for those associated and not associated with pH1N1 influenza during the pandemic period. MATERIALS AND METHODS: We reviewed all patients with pneumonia who visited the Armed Forces Capital Hospital between July 2009 and February 2010. During this period, all pneumonia patients were tested for pH1N1 by reverse transcription-polymerase chain reaction (RT-PCR) using nasopharyngeal specimens. RESULTS: In total, 98 patients with pneumonia were enrolled. Their median age was 20 years and all patients were males. Forty-nine (50%) of patients had pH1N1 infection and the others (50%) had negative results in pH1N1 RT-PCR. Patients with pH1N1 infection complained of dyspnea more commonly (83.3% vs. 29.0%; p<0.001), had higher Acute Physiology and Chronic Health Evaluation (APACHE) II scores [5 (range, 0-12) vs. 3 (range, 0-11); p<0.01], fewer days of prehospital illness [2 (range, 0-10) vs. 4 (range, 0-14); p=0.001], and a higher chance of bilateral infiltrates on chest X-ray (CXR) (67.3% vs. 14.3%; p<0.001) and ground-glass opacity (GGO) lesions on computed tomography (CT; 48.9% vs. 22.0%; p<0.001) than patients without pH1N1 infection. CONCLUSION: Dyspnea, bilateral infiltrates on CXR, and GGO on CT were dominant features in pH1N1-associated pneumonia. Understanding these characteristics can help selection of patients who require prompt antiviral therapy.
Adolescent ; Adult ; Antiviral Agents/therapeutic use ; Dyspnea/virology ; Humans ; Influenza A Virus, H1N1 Subtype/genetics/*pathogenicity ; Influenza, Human/*complications/radiography/virology ; Male ; Middle Aged ; Pneumonia/etiology/radiography ; Pneumonia, Viral/drug therapy/etiology/*radiography/*virology ; Radiography, Thoracic ; Tomography, X-Ray Computed ; Young Adult

BACKGROUNDThe first case of pandemic influenza A (H1N1) virus infection in Macau Special Administrative Region (SAR) of the People's Republic of China was documented on June 18, 2009. Subsequently, persons with suspected infection or of contact with suspected cases received screening. All the confirmed cases were hospitalized and treated with oseltamivir. Their clinical features were observed. This may help for better management for later patients and be of benefit to the government of Macau SAR to adjust its strategy to combat the pandemic influenza A (H1N1) virus infection more efficiently.

METHODSFrom June to July 2009, the initial 72 cases of influenza A (H1N1) in Macau were hospitalized in Common Hospital Centre S. Januario (CHCSJ). The infection was confirmed by real-time reverse-transcriptase polymerase chain reaction (RT-PCR). The clinical features of the disease were closely observed and documented. Oseltamivir was given to all patients within 48 hours after the onset of disease and maintained for 5 days.

RESULTSThe mean age of the 72 patients was 21 years old. Forty of them were men and 32 were women. The median incubation of the virus was 2 days (1 to 7 days). The most common symptoms were fever (97.2%) and cough (77.8%). The rate of gastrointestinal symptoms including nausea, vomiting, and diarrhea was 2.8%. Fever typically lasted for 3 days (1 to 9 days). The median time from the onset to positive results of real-time RT-PCR was 6 days (3 to 13 days). After treatment with oseltamivir, most patients became afebrile within 48 hours. Only one aged patient with a history of glaucoma and hypothyroidism was found to have lung infiltration on chest X-ray.

CONCLUSIONSThe initial cases of pandemic influenza A (H1N1) virus infection in Macau SAR showed that most of the infected persons had a mild course. The virus could be detected by real-time RT-PCR within a median of 6 days from the onset. Oseltamivir was effective.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Child ; Child, Preschool ; China ; Female ; Humans ; Influenza A Virus, H1N1 Subtype ; drug effects ; genetics ; pathogenicity ; Influenza, Human ; diagnosis ; drug therapy ; pathology ; Macau ; Male ; Middle Aged ; Oseltamivir ; therapeutic use ; Reverse Transcriptase Polymerase Chain Reaction ; Young Adult