2009甲型H1N1流感病毒研究进展

2009年3月在美国和墨西哥爆发的新型甲型H1N1流感在很短的时间内便扩散到世界多个国家,形成了流感的大流行,引起世界卫生组织和各国的高度重视。综述新型甲型H1N1流感病毒的基因组来源、目前主要的检测手段,并对预防和治疗的方法进行简单介绍。     

Keystone Symposium on Antibodies as Drugs: March 27–April 1, 2009, Whistler,BC CA

The symposium on Antibodies as Drugs, organized by Keystone Symposia and chaired by J. Marks, (University of California Los Angeles, USA), E.S. Ward (University of Texas Southwestern Medical Center, USA) and L. Weiner (Georgetown University Medical Center, USA), was held in Whistler, British Columbia. This Canadian Rockies village, which will host the 2010 Olympic Games, served as an enchanting backdrop to the meeting. The more than 350 speakers and attendees included scientists from major pharmaceutical firms, e.g., Abbott, MedImmune/Astra Zeneca, Bristol-Myers Squibb, Merck & Co., Pfizer, Sanofi-Aventis, Schering, GlaxoSmithKline, Eli Lilly, Hoffmann LaRoche, Novartis, Wyeth, and biotechnology companies, e.g., Ablynx, Medarex, Morphosys, GenMab, Amgen, Genentech, ImmunoGen, Agensys, Domantis, Biogen Idec, Centocor, LFB, Micromet, PDL Biopharma, Borean Pharma, Dyax Corp., Symphogen and Syntonix. Academic research groups at Imperial College London, University of Oxford, ETH Zürich, Scripps, Institute Cochin, Karolinska Institute, Utrecht University, Harvard Medical School, Massachusetts Institute of Technology, Baylor College, Paul Ehrlich Institute, University of California San Francisco, University of California San Diego, University of Nantes, University of Tours and Ludwig Institute were also represented, as were regulatory authorities, including the US Food and Drug Administration, National Institutes of Health and the Public Health Agency of Canada). The meeting was very interactive and included thoughtful exchanges during the different sessions and networking events.     

Aflatoxin B1 and zearalenone in dairy feeds in Portugal, 2009–2011

This paper presents 3 years of data (2009–2011) on the occurrence of two mycotoxins, aflatoxin B₁ (AFB₁) and zearalenone (ZEA), in samples of feedstuff for dairy cows (n?=?963), ewes (n?=?42), and goats (n?=?131) produced in Portugal. AFB₁ was found in 15 samples of cow feed (1.6 %), 3 samples of ewe feed (2.3 %) and in 2 samples of goat feed (4.8 %). All but two samples contained AFB₁ at levels below the European Union maximum level (5 μg/kg). Nearly half (45 %) of the samples were contaminated with ZEA, but its levels were relatively low, at 5–136.9 μg/kg, well below the European Union guidance value (500 μg/kg).     

Risk Factors for Death from Influenza A(H1N1)pdm09, State of S?o Paulo,Brazil, 2009

This case-control study aimed to assess the risk factors for death from influenza A(H1N1)pdm09 in patients with laboratory confirmation, who had severe acute respiratory illness-SARI and were hospitalized between June 28^th and August 29^th 2009, in the metropolitan regions of São Paulo and Campinas, Brazil. Medical charts of all the 193 patients who died (cases) and the 386 randomly selected patients who recovered (controls) were investigated in 177 hospitals. Household interviews were conducted with those who had survived and the closest relative of those who had died. 73.6% of cases and 38.1% of controls were at risk of developing influenza-related complications. The 18-to-59-year age group (OR = 2.31, 95%CI: 1.31–4.10 (reference up to 18 years of age)), presence of risk conditions for severity of influenza (OR = 1.99, 95%CI: 1.11–3.57, if one or OR = 6.05, 95%CI: 2.76–13.28, if more than one), obesity (OR = 2.73, 95%CI: 1.28–5.83), immunosuppression (OR = 3.43, 95%CI: 1.28–9.19), and search for previous care associated with the hospitalization (OR = 3.35, 95%CI: 1.75–6.40) were risk factors for death. Antiviral treatment performed within 72 hours of the onset of symptoms (OR = 0.17, 95%CI: 0.08–0.37, if within 48hours, and OR = 0.30, 95%CI: 0.11–0.81, if between 48 and 72 hours) was protective against death. The identification of high-risk patients and early treatment are important factors for reducing morbi-mortality from influenza.     

Whole Genome Characterization,Phylogenetic and Genome Signature Analysis of Human Pandemic H1N1 Virus in Thailand, 2009–2012

Background

Three waves of human pandemic influenza occurred in Thailand in 2009–2012. The genome signature features and evolution of pH1N1 need to be characterized to elucidate the aspects responsible for the multiple waves of pandemic.

Methodology/Findings

Forty whole genome sequences and 584 partial sequences of pH1N1 circulating in Thailand, divided into 1^st, 2^nd and 3^rd wave and post-pandemic were characterized and 77 genome signatures were analyzed. Phylogenetic trees of concatenated whole genome and HA gene sequences were constructed calculating substitution rate and d_N/d_S of each gene. Phylogenetic analysis showed a distinct pattern of pH1N1 circulation in Thailand, with the first two isolates from May, 2009 belonging to clade 5 while clades 5, 6 and 7 co-circulated during the first wave of pH1N1 pandemic in Thailand. Clade 8 predominated during the second wave and different proportions of the pH1N1 viruses circulating during the third wave and post pandemic period belonged to clades 8, 11.1 and 11.2. The mutation analysis of pH1N1 revealed many adaptive mutations which have become the signature of each clade and may be responsible for the multiple pandemic waves in Thailand, especially with regard to clades 11.1 and 11.2 as evidenced with V731I, G154D of PB1 gene, PA I330V, HA A214T S160G and S202T. The substitution rate of pH1N1 in Thailand ranged from 2.53×10⁻³±0.02 (M2 genes) to 5.27×10⁻³±0.03 per site per year (NA gene).

Conclusions

All results suggested that this virus is still adaptive, maybe to evade the host''s immune response and tends to remain in the human host although the d_N/d_S were under purifying selection in all 8 genes. Due to the gradual evolution of pH1N1 in Thailand, continuous monitoring is essential for evaluation and surveillance to be prepared for and able to control future influenza activities.     

Disparities among 2009 Pandemic Influenza A (H1N1) Hospital Admissions: A Mixed Methods Analysis – Illinois,April–December 2009

During late April 2009, the first cases of 2009 pandemic influenza A (H1N1) (pH1N1) in Illinois were reported. On-going, sustained local transmission resulted in an estimated 500,000 infected persons. We conducted a mixed method analysis using both quantitative (surveillance) and qualitative (interview) data; surveillance data was used to analyze demographic distribution of hospitalized cases and follow-up interview data was used to assess health seeking behavior. Invitations to participate in a telephone interview were sent to 120 randomly selected Illinois residents that were hospitalized during April–December 2009. During April–December 2009, 2,824 pH1N1 hospitalizations occurred in Illinois hospitals; median age (interquartile range) at admission was 24 (range: 6–49) years. Hospitalization rates/100,000 persons for blacks and Hispanics, regardless of age or sex were 2–3 times greater than for whites (blacks, 36/100,000 (95% Confidence Interval ([95% CI], 33–39)); Hispanics, 35/100,000 [95%CI,32–37] (; whites, 13/100,000[95%CI, 12–14); p<0.001). Mortality rates were higher for blacks (0.9/100,000; p<0.09) and Hispanics (1/100,000; p<0.04) when compared with the mortality rates for whites (0.6/100,000). Of 33 interview respondents, 31 (94%) stated that they had heard of pH1N1 before being hospitalized, and 24 (73%) did not believed they were at risk for pH1N1. On average, respondents reported experiencing symptoms for 2 days (range: 1–7) before seeking medical care. When asked how to prevent pH1N1 infection in the future, the most common responses were getting vaccinated and practicing hand hygiene. Blacks and Hispanics in Illinois experienced disproportionate pH1N1 hospitalization and mortality rates. Public health education and outreach efforts in preparation for future influenza pandemics should include prevention messaging focused on perception of risk, and ensure community wide access to prevention messages and practices.     

SMi biomarkers summit, 2–3 February 2009, London,UK

As part of a programme to develop biomarker assays for polycyclic aromatic hydrocarbons (PAHs) in marine invertebrates, two species of crabs, Carcinus maenas and Carcinus aestuarii were exposed to benzo(a)pyrene (B(a)P) or crude oil. Microsomes were prepared from the midgut gland (hepatopancreas), examined by gel electrophoresis and Western blotting and assayed for B(a)P monooxygenase activity. In early experiments there was evidence of protein degradation and results were inconsistent and inconclusive. However, when steps were taken to minimize this in subsequent experiments, including the inclusion of four protease inhibitors in the homogenization buffer, there was consistent evidence for an increase of proteins of estimated molecular weight 45-60 kDa, and particularly of a distinct band at c. 48 kDa, following exposure to PAH at levels down to 0.1 ppm in ambient water. In C. aestuarii the increase in this band was found to coincide with an 8-12-fold increaseof B(a)P monooxygenase activity in midgut gland microsomes. These results suggest that one or more forms of cytochrome P450 may be induced by PAHs in these species. However, Western blotting using antibodies raised to vertebrate P450s, and representing four different gene families, failed to recognize any proteins in either the PAH-treated samples or in the controls. The isolation and characterization of induced protein, and the production of antibodies may provide the basis for a biomarker assay to measure a response to environmental PAHs in crabs.     

Twin Peaks: A/H1N1 Pandemic Influenza Virus Infection and Vaccination in Norway, 2009–2010

Background

Vaccination campaigns against A/H1N1 2009 pandemic influenza virus (A/H1N1p) began in autumn 2009 in Europe, after the declaration of the pandemic at a global level. This study aimed to estimate the proportion of individuals vaccinated against A/H1N1p in Norway who were already infected (asymptomatically or symptomatically) by A/H1N1p before vaccination, using a mathematical model.

Methods

A dynamic, mechanistic, mathematical model of A/H1N1p transmission was developed for the Norwegian population. The model parameters were estimated by calibrating the model-projected number of symptomatic A/H1N1p cases to the number of laboratory-confirmed A/H1N1p cases reported to the surveillance system, accounting for potential under-reporting. It was assumed in the base case that the likelihood of vaccination was independent of infection/disease state. A sensitivity analysis explored the effects of four scenarios in which current or previous symptomatic A/H1N1p infection would influence the likelihood of being vaccinated.

Results

The number of model-projected symptomatic A/H1N1p cases by week during the epidemic, accounting for under-reporting and timing, closely matched that of the laboratory-confirmed A/H1N1p cases reported to the surveillance system. The model-projected incidence of symptomatic A/H1N1p infection was 27% overall, 55% in people <10 years old and 41% in people 10–20 years old. The model-projected percentage of individuals vaccinated against A/H1N1p who were already infected with A/H1N1p before being vaccinated was 56% overall, 62% in people <10 years old and 66% in people 10–20 years old. The results were sensitive to assumptions about the independence of vaccination and infection; however, even when current or previous symptomatic A/H1N1p infection was assumed to reduce the likelihood of vaccination, the estimated percentage of individuals who were infected before vaccination remained at least 32% in all age groups.

Conclusion

This analysis suggests that over half the people vaccinated against A/H1N1p in Norway during the 2009 pandemic may already have been infected by A/H1N1p before being vaccinated.     

High Rate of A(H1N1)pdm09 Infections among Rural Thai Villagers, 2009–2010

Background

Pandemic influenza A(H1N1)pdm09 emerged in Thailand in 2009. A prospective longitudinal adult cohort and household transmission study of influenza-like illness (ILI) was ongoing in rural Thailand at the time of emergence. Symptomatic and subclinical A(H1N1)pdm09 infection rates in the cohort and among household members were evaluated.

Methods

A cohort of 800 Thai adults underwent active community-based surveillance for ILI from 2008–2010. Acute respiratory samples from ILI episodes were tested for A(H1N1)pdm09 by qRT-PCR; acute and 60-day convalescent blood samples were tested by A(H1N1)pdm09 hemagglutination inhibition assay (HI). Enrollment, 12-month and 24-month follow-up blood samples were tested for A(H1N1)pdm09 seroconversion by HI. Household members of influenza A-infected cohort subjects with ILI were enrolled in household transmission investigations in which day 0 and 60 blood samples and acute respiratory samples were tested by either qRT-PCR or HI for A(H1N1)pdm09. Seroconversion between annual blood samples without A(H1N1)pdm09-positive ILI was considered as subclinical infection.

Results

The 2-yr cumulative incidence of A(H1N1)pdm09 infection in the cohort in 2009/2010 was 10.8% (84/781) with an annual incidence of 1.2% in 2009 and 9.7% in 2010; 83.3% of infections were subclinical (50% in 2009 and 85.9% in 2010). The 2-yr cumulative incidence was lowest (5%) in adults born ≤1957. The A(H1N1)pdm09 secondary attack rate among household contacts was 47.2% (17/36); 47.1% of these infections were subclinical. The highest A(H1N1)pdm09 secondary attack rate among household contacts (70.6%, 12/17) occurred among children born between 1990 and 2003.

Conclusion

Subclinical A(H1N1)pdm09 infections in Thai adults occurred frequently and accounted for a greater proportion of all A(H1N1)pdm09 infections than previously estimated. The role of subclinical infections in A(H1N1)pdm09 transmission has important implications in formulating strategies to predict and prevent the spread of A(H1N1)pdm09 and other influenza virus strains.     

甲型H1N1／2009流感病毒的种属跨越机制

在世界范围流行的甲型H1N1／2009流感病毒具有下述3个重要特征：可寄生于人体,易感人群很多,患者年龄偏低．本研究确定了病毒蛋白中的一块关键区域．该区域对病毒所寄生的物种的种属范围起决定性作用,并且是全球性流感病毒的一个标志性区域．正是该区域氨基酸的特性导致了上述3个特点．具体来说,对宿主的免疫系统而言,病毒蛋白质结构的变化会形成新的标靶结构,并且可以进一步导致宿主范围的变化．基于多肽链发生致病性结构转换的概率,本研究确定了甲型流感病毒中对控制宿主范围起决定性作用的氨基酸的位置．研究发现甲型H1N1／2009流感病毒中处于这些位点的多肽链在本质上可以在寄生于人的毒株中表达,而之前仅在宿主为禽、猪的毒株中被发现．其与另一氨基酸短串的协同构象改变对于甲型H1N1／2009流感病毒的种属跨越具有重要作用．人体对这些关键位点的免疫缺陷导致了甲型H1N1／2009流感病毒宿主人群多和青年人易致病的特点．     

上海2009甲型H1N1流行性感冒病毒全基因组分析

2009年全球暴发2009甲型H1N1流行性感冒(简称流感)疫情,上海于2009年5月出现第1例输入型病例。为了解上海地区输入型2009甲型H1N1流感病毒的生物学特征,以上海较早发现的2例输入型甲型H1N1流感患者作为研究对象,分离出A/Shanghai/37T/2009和A/Shanghai/71T/2009病毒,利用实时定量荧光反转录-聚合酶链反应(RT-PCR)鉴定病毒,通过扫描透射电子显微镜观察、免疫荧光检测、全基因组测序和生物信息软件分析,对这2株流感病毒形态、结构、耐药性、基因特点和病毒型别等进行研究。结果显示,病毒呈现正黏病毒颗粒形态特征;犬肾(MDCK)细胞内的病毒能与患者恢复期血清反应。此2株病毒的全基因核酸序列和氨基酸序列与美国参考株A/California/04/2009(H1N1)有较高同源性,其中第31位氨基酸残基发生改变。对金刚烷胺耐药,而对奥司他韦敏感。基于全基因组的系统发育分析,确认此2株病毒属2009甲型H1N1流感病毒。     

Molecular Evolution of Human H1N1 and H3N2 Influenza A Virus in Thailand, 2006–2009

Background

Annual seasonal influenza outbreaks are associated with high morbidity and mortality.

Objective

To index and document evolutionary changes among influenza A H1N1 and H3N2 viruses isolated from Thailand during 2006–2009, using complete genome sequences.

Methods

Nasopharyngeal aspirates were collected from patients diagnosed with respiratory illness in Thailand during 2006–2009. All samples were screened for Influenza A virus. A total of 13 H1N1 and 21 H3N2 were confirmed and whole genome sequenced for the evolutionary analysis using standard phylogenetic approaches.

Results

Phylogenetic analysis of HA revealed a clear diversification of seasonal from vaccine strain lineages. H3N2 seasonal clusters were closely related to the WHO recommended vaccine strains in each season. Most H1N1 isolates could be differentiated into 3 lineages. The A/Brisbane/59/2007 lineage, a vaccine strain for H1N1 since 2008, is closely related with the H1N1 subtypes circulating in 2009. HA sequences were conserved at the receptor-binding site. Amino acid variations in the antigenic site resulted in a possible N-linked glycosylation motif. Recent H3N2 isolates had higher genetic variations compared to H1N1 isolates. Most substitutions in the NP protein were clustered in the T-cell recognition domains.

Conclusion

In this study we performed evolutionary genetic analysis of influenza A viruses in Thailand between 2006–2009. Although the current vaccine strain is efficient for controlling the circulating outbreak subtypes, surveillance is necessary to provide unambiguous information on emergent viruses. In summary, the findings of this study contribute the understanding of evolution in influenza A viruses in humans and is useful for routine surveillance and vaccine strain selection.     

Estimating Influenza Deaths in Canada, 1992–2009

Background

Poisson regression modelling has been widely used to estimate the disease burden attributable to influenza, though not without concerns that some of the excess burden could be due to other causes. This study aims to provide annual estimates of the mortality and hospitalization burden attributable to both seasonal influenza and the 2009 A/H1N1 pandemic influenza for Canada, and to discuss issues related to the reliability of these estimates.

Methods

Weekly time-series for all-cause mortality and regression models were used to estimate the number of deaths in Canada attributable to influenza from September 1992 to December 2009. To assess their robustness, the annual estimates derived from different parameterizations of the regression model for all-cause mortality were compared. In addition, the association between the annual estimates for mortality and hospitalization by age group, underlying cause of death or primary reason for admission and discharge status is discussed.

Results

The crude influenza-attributed mortality rate based on all-cause mortality and averaged over 17 influenza seasons prior to the 2009 A/H1N1 pandemic was 11.3 (95%CI, 10.5 - 12.1) deaths per 100 000 population per year, or an average of 3,500 (95%CI, 3,200 - 3,700) deaths per year attributable to seasonal influenza. The estimated annual rates ranged from undetectable at the ecological level to more than 6000 deaths per year over the three A/Sydney seasons. In comparison, we attributed an estimated 740 deaths (95%CI, 350–1500) to A(H1N1)pdm09. Annual estimates from different model parameterizations were strongly correlated, as were estimates for mortality and morbidity; the higher A(H1N1)pdm09 burden in younger age groups was the most notable exception.

Interpretation

With the exception of some of the Serfling models, differences in the ecological estimates of the disease burden attributable to influenza were small in comparison to the variation in disease burden from one season to another.     

Estimated HIV Incidence in California, 2006–2009

Introduction

Accurate estimates of HIV incidence are crucial for prioritizing, targeting, and evaluating HIV prevention efforts. Using the methodology the CDC used to estimate national HIV incidence, we estimated HIV incidence in Los Angeles County (LAC), San Francisco (SF), and California’s remaining counties.

Methods

We estimated new HIV infections in 2006–2009 among adults and adolescents in LAC, SF and the remaining California counties using the Serologic Testing Algorithm for Recent Seroconversion (STARHS). STARHS methodology uses the BED HIV-1 capture enzyme immunoassay to determine recent HIV infections by testing remnant serum from persons newly diagnosed with HIV. A population-based incidence estimate is calculated using HIV testing data from newly diagnosed cases and imputing for persons unaware of their HIV infection.

Results

For years 2007–2009, respectively, we estimated new infections in LAC to be 2426 (95% CI 1871–2982), 1669 (CI 1309–2029) and 1898 (CI 1452–2344) (p<0.01); in SF for 2006–2009, 492 (CI 327–657), 490 (CI 335–646), 458 (CI 342–574) and 367 (CI 261–473) (p = 0.14); and in the remaining California counties in 2008–2009, 2526 (CI 1688–3364) and 2993 (CI 2141–3846) respectively. HIV infection rates among men who have sex with men (MSM) in LAC were 100 times higher than other risk populations; the SF MSM rate was 3 to 18 times higher than other demographic groups. In LAC, incidence rates among African-Americans were twice those of whites and Latinos; persons 40 years or older had lower rates of infection than younger persons.

Discussion

We report the first HIV incidence estimates for California, highlighting geographic disparities in HIV incidence and confirming national findings that MSM and African-Americans are disproportionately impacted by HIV. HIV incidence estimates can and should be used to target prevention efforts towards populations at highest risk of acquiring new HIV infections, focusing on geographic, racial and risk group disparities.     

Prevalence,Awareness, Treatment,and Control of Hypertension in United States Counties, 2001–2009

Hypertension is an important and modifiable risk factor for cardiovascular disease and mortality. Over the last decade, national-levels of controlled hypertension have increased, but little information on hypertension prevalence and trends in hypertension treatment and control exists at the county-level. We estimate trends in prevalence, awareness, treatment, and control of hypertension in US counties using data from the National Health and Nutrition Examination Survey (NHANES) in five two-year waves from 1999–2008 including 26,349 adults aged 30 years and older and from the Behavioral Risk Factor Surveillance System (BRFSS) from 1997–2009 including 1,283,722 adults aged 30 years and older. Hypertension was defined as systolic blood pressure (BP) of at least 140 mm Hg, self-reported use of antihypertensive treatment, or both. Hypertension control was defined as systolic BP less than 140 mm Hg. The median prevalence of total hypertension in 2009 was estimated at 37.6% (range: 26.5 to 54.4%) in men and 40.1% (range: 28.5 to 57.9%) in women. Within-state differences in the county prevalence of uncontrolled hypertension were as high as 7.8 percentage points in 2009. Awareness, treatment, and control was highest in the southeastern US, and increased between 2001 and 2009 on average. The median county-level control in men was 57.7% (range: 43.4 to 65.9%) and in women was 57.1% (range: 43.0 to 65.46%) in 2009, with highest rates in white men and black women. While control of hypertension is on the rise, prevalence of total hypertension continues to increase in the US. Concurrent increases in treatment and control of hypertension are promising, but efforts to decrease the prevalence of hypertension are needed.     

Environmental Lead Exposure Among Children in Chengdu, China, 2007–2009

Lead poisoning is one of the most significant environmental health threats which children face. The aim of the study was to evaluate the relationship between the blood lead level (BLL) of children and environmental pollution, as well as the effect of BLL on children's heath, in the biggest city in south-west China. A questionnaire survey was administered to 3,624 children's (6 months-7 years old) guardian in urban Chengdu from 2007 to 2009, collecting data on inhabited environment, living habits, and health status. All blood samples were collected for BLL test when the children came to the hospital. The analysis was conducted in West China Second University Hospital, Chengdu, China in December, 2009. A total of 3,624 children were included. The mean BLL was 62.31 μg/L. Three hundred thirty-three (9.2%) of 3,624 children's BLL were above 100 μg/L. Taking Chinese medicinal herbs usually, puffed foods, and substitutes of breast milk were risk factors (P < 0.05).The study shows that Chinese children's lead poisoning prevention and treatment lags far behind developed countries. The findings have implications for environmental health policy.