Minor changes in the hemagglutinin of influenza A(H1N1)2009 virus alter its antigenic properties

Background

The influenza A(H1N1)2009 virus has been the dominant type of influenza A virus in Finland during the 2009–2010 and 2010–2011 epidemic seasons. We analyzed the antigenic characteristics of several influenza A(H1N1)2009 viruses isolated during the two influenza seasons by analyzing the amino acid sequences of the hemagglutinin (HA), modeling the amino acid changes in the HA structure and measuring antibody responses induced by natural infection or influenza vaccination.

Methods/Results

Based on the HA sequences of influenza A(H1N1)2009 viruses we selected 13 different strains for antigenic characterization. The analysis included the vaccine virus, A/California/07/2009 and multiple California-like isolates from 2009–2010 and 2010–2011 epidemic seasons. These viruses had two to five amino acid changes in their HA1 molecule. The mutation(s) were located in antigenic sites Sa, Ca1, Ca2 and Cb region. Analysis of the antibody levels by hemagglutination inhibition test (HI) indicated that vaccinated individuals and people who had experienced a natural influenza A(H1N1)2009 virus infection showed good immune responses against the vaccine virus and most of the wild-type viruses. However, one to two amino acid changes in the antigenic site Sa dramatically affected the ability of antibodies to recognize these viruses. In contrast, the tested viruses were indistinguishable in regard to antibody recognition by the sera from elderly individuals who had been exposed to the Spanish influenza or its descendant viruses during the early 20^th century.

Conclusions

According to our results, one to two amino acid changes (N125D and/or N156K) in the major antigenic sites of the hemagglutinin of influenza A(H1N1)2009 virus may lead to significant reduction in the ability of patient and vaccine sera to recognize A(H1N1)2009 viruses.     

Influenza epidemics in the USSR and Czechoslovakia in 1979-1984

In this work materials characterizing the appearance and development of influenza epidemic at the territories of the USSR and the Czech Socialist Republic are presented, the common features and differences of the epidemic process in both countries are recorded. The work shows that in both countries the appearance of this epidemic is caused by the same virus. In most cases the epidemic started earlier and lasted longer in the USSR, but morbidity rate during the epidemic was, on the whole, higher in the Czech Socialist Republic. Similarity in the course of the primary period of the epidemic processes from their appearance to their maximum rise was observed. In both countries the maximum rise of morbidity rate was registered on weeks 3-4 from the beginning of the epidemic.     

Epidemiological analysis of acute respiratory disease (ARD) and characteristics of the influenza epidemic in Bohemia during the season 1986/1987

The authors analyze the findings of epidemiological and virological surveillance of ARD in Bohemia during the season 1986/1987. In all, 57.5% of the Czech population was affected by acute respiratory disease (ARD). There were 5,950,832 cases reported, 124,444 complications (2.1% of the overall morbidity rate) and 5,374 deaths due to influenza, bronchitis, pneumonia and chronic pulmonary affection. The influenza epidemic commenced during the 48-th calendary week (CW) and lasted 5 weeks till the 52-nd CW. The epidemic was due to an influenza virus strain of the subtype A(H1N1) antigenically related to the drift variant A (Singapore) 6/86. Within an extremely short period of the epidemic, 1,094,865 influenza cases were reported and 22,313 cases of complications. 10.7% of the CSR population were affected during the epidemic in whose etiology noninfluenza respiratory viruses were significantly implicated, especially adenoviruses (41.7%) and the RS virus (26.9%). There was no excessive mortality in the course of the epidemic. The authors discuss the atypical nature of this particular influenza epidemic and the etiological role of respiratory viruses.     

Small-scale trial of live-attenuated influenza vaccine (A/Hong Kong/68).

Seventy-one men who were given live-attenuated A/Hong Kong/68 (H3N2) influenza vaccine during November 1973, and 34 men given placebo were examined for changes in antibody level. Overall, 12 of the 71 men (17%) given the vaccine showed a fourfold rise in haemagglutination-inhibition (HI) antibody titre after 14 days. No such rises were seen in the 34 men given placebo. However, 10 of the men showing a fourfold rise were from 19 who had no detectable HI antibody to this virus before vaccination, representing a conversion rate of 53%. The other two had a HI titre of 1/10 before vaccination. The absence of antibody response, at 14 days, in those with an HI titre of 1/20 or greater may indicated that this represents a protective level against infection. However, the vaccine virus was probably overattenuated and may have constituted a weaker challenge than might occur with a wild strain. No influenza virus was isolated from either group in the week after vaccination and no evidence of transmission to the placebo group was seen. Mild symptoms, chills, muscle pain, and stiffness were more frequently seen in the 12 persons showing a fourfold rise in antibody than in the rest of the volunteers.     

Epidemiological effectiveness of influenza immunoprophylaxis with Unifluvax vaccine in organized groups of servicemen

The results of vaccination carried out in an organized group with the subunit influenza vaccine "Influvax" are presented. In the immunized group the registered morbidity level exceeded the annual morbidity level by 7-12% only against the expected epidemic rise. Respiratory diseases in this group took a mild course. The morbidity level in the control group corresponded to the predicted value and exceeded the morbidity level in the vaccinated group 4.2-fold. The conclusion was made on the effectiveness of immunoprophylaxis.     

The epidemic spread of the influenza A and B viruses in the USSR in 1985-1986

The data on the spread of influenza A and B in the autumn and winter of 1985-1986 are given. Three epidemics caused by all presently circulating viruses, B, A (H3N2) and A (H1N1), were registered in the USSR. Of these, the greatest one was the epidemic of influenza B; morbidity rate among the adult population during this epidemic was at the level with the morbidity rate characteristic of the epidemics registered at the period of 1962-1972, and morbidity rate among children, especially school children, was even higher.     

Intranasal Vaccination Promotes Detrimental Th17-Mediated Immunity against Influenza Infection

Influenza disease is a global health issue that causes significant morbidity and mortality through seasonal epidemics. Currently, inactivated influenza virus vaccines given intramuscularly or live attenuated influenza virus vaccines administered intranasally are the only approved options for vaccination against influenza virus in humans. We evaluated the efficacy of a synthetic toll-like receptor 4 agonist CRX-601 as an adjuvant for enhancing vaccine-induced protection against influenza infection. Intranasal administration of CRX-601 adjuvant combined with detergent split-influenza antigen (A/Uruguay/716/2007 (H3N2)) generated strong local and systemic immunity against co-administered influenza antigens while exhibiting high efficacy against two heterotypic influenza challenges. Intranasal vaccination with CRX-601 adjuvanted vaccines promoted antigen-specific IgG and IgA antibody responses and the generation of polyfunctional antigen-specific Th17 cells (CD4⁺IL-17A⁺TNFα⁺). Following challenge with influenza virus, vaccinated mice transiently exhibited increased weight loss and morbidity during early stages of disease but eventually controlled infection. This disease exacerbation following influenza infection in vaccinated mice was dependent on both the route of vaccination and the addition of the adjuvant. Neutralization of IL-17A confirmed a detrimental role for this cytokine during influenza infection. The expansion of vaccine-primed Th17 cells during influenza infection was also accompanied by an augmented lung neutrophilic response, which was partially responsible for mediating the increased morbidity. This discovery is of significance in the field of vaccinology, as it highlights the importance of both route of vaccination and adjuvant selection in vaccine development     

Multiple reassortment events in the evolutionary history of H1N1 influenza A virus since 1918

The H1N1 subtype of influenza A virus has caused substantial morbidity and mortality in humans, first documented in the global pandemic of 1918 and continuing to the present day. Despite this disease burden, the evolutionary history of the A/H1N1 virus is not well understood, particularly whether there is a virological basis for several notable epidemics of unusual severity in the 1940s and 1950s. Using a data set of 71 representative complete genome sequences sampled between 1918 and 2006, we show that segmental reassortment has played an important role in the genomic evolution of A/H1N1 since 1918. Specifically, we demonstrate that an A/H1N1 isolate from the 1947 epidemic acquired novel PB2 and HA genes through intra-subtype reassortment, which may explain the abrupt antigenic evolution of this virus. Similarly, the 1951 influenza epidemic may also have been associated with reassortant A/H1N1 viruses. Intra-subtype reassortment therefore appears to be a more important process in the evolution and epidemiology of H1N1 influenza A virus than previously realized.     

Characteristics of the manifest and latent components of the hepatitis A epidemic process in cities of Russia

Seroepidemiological study of hepatitis A (HA) morbidity was carried out in three Russian cities, with different levels of HA morbidity. The study included the analysis of HA morbidity for 22 years, the determination of antibodies to HA virus (anti-HAV) in 2,958 healthy persons aged 0-12 months to 40 years and older. In one of the cities 7 isolates of HA virus were obtained from unrelated sources and the genotypes of the virus were determined. The study revealed that the frequency of seropositive cases among persons of different ages correlated with the level and prolonged dynamics of HA morbidity. According to the occurrence of anti-HAV, such cities as St. Petersburg, Rostov-on-Don and Yakutsk may be at present classified as territories, moderately endemic in HA. At the same time in the 90 s the epidemic situation in HA was more favorable in Rostov-on-Don than in two other cities. The suggestion was made that a high proportion of seropositive persons among the population of St. Perersburg was linked with an almost twofold rise in HA morbidity in 1993-1995 caused by genotype 1 of the virus. Seroepidemiological studies in HA during the period of a drop in morbidity acquire special importance in the surveillance and control system of this infection.     

Characterization In Vitro and In Vivo of a Pandemic H1N1 Influenza Virus from a Fatal Case

Pandemic 2009 H1N1 (pH1N1) influenza viruses caused mild symptoms in most infected patients. However, a greater rate of severe disease was observed in healthy young adults and children without co-morbid conditions. Here we tested whether influenza strains displaying differential virulence could be present among circulating pH1N1 viruses. The biological properties and the genotype of viruses isolated from a patient showing mild disease (M) or from a fatal case (F), both without known co-morbid conditions were compared in vitro and in vivo. The F virus presented faster growth kinetics and stronger induction of cytokines than M virus in human alveolar lung epithelial cells. In the murine model in vivo, the F virus showed a stronger morbidity and mortality than M virus. Remarkably, a higher proportion of mice presenting infectious virus in the hearts, was found in F virus-infected animals. Altogether, the data indicate that strains of pH1N1 virus with enhanced pathogenicity circulated during the 2009 pandemic. In addition, examination of chemokine receptor 5 (CCR5) genotype, recently reported as involved in severe influenza virus disease, revealed that the F virus-infected patient was homozygous for the deleted form of CCR5 receptor (CCR5Δ32).     

Participation of secretory antibodies in the protection from influenza

Two collective bodies (319 persons in all) were under observation from October, 1974 to March, 1975 for the purpose of studying the problem on the participation of antibodies of the respiratory tract secretion in the protection from influenza virus, type A, infection. During the influenza epidemic outbreak there was revealed a reverse relationship between the antibody concentration in the nasal cavity secretion and the incidence of the disease, and also an interaction between the antibodies of the secretions and the serum in the influenza protection.     

Early Assessment of Anxiety and Behavioral Response to Novel Swine-Origin Influenza A(H1N1)

Background

Since late April, 2009, a novel influenza virus A (H1N1), generally referred to as the “swine flu,” has spread around the globe and infected hundreds of thousands of people. During the first few days after the initial outbreak in Mexico, extensive media coverage together with a high degree of uncertainty about the transmissibility and mortality rate associated with the virus caused widespread concern in the population. The spread of an infectious disease can be strongly influenced by behavioral changes (e.g., social distancing) during the early phase of an epidemic, but data on risk perception and behavioral response to a novel virus is usually collected with a substantial delay or after an epidemic has run its course.

Methodology/Principal Findings

Here, we report the results from an online survey that gathered data (n = 6,249) about risk perception of the Influenza A(H1N1) outbreak during the first few days of widespread media coverage (April 28 - May 5, 2009). We find that after an initially high level of concern, levels of anxiety waned along with the perception of the virus as an immediate threat. Overall, our data provide evidence that emotional status mediates behavioral response. Intriguingly, principal component analysis revealed strong clustering of anxiety about swine flu, bird flu and terrorism. All three of these threats receive a great deal of media attention and their fundamental uncertainty is likely to generate an inordinate amount of fear vis-a-vis their actual threat.

Conclusions/Significance

Our results suggest that respondents'' behavior varies in predictable ways. Of particular interest, we find that affective variables, such as self-reported anxiety over the epidemic, mediate the likelihood that respondents will engage in protective behavior. Understanding how protective behavior such as social distancing varies and the specific factors that mediate it may help with the design of epidemic control strategies.     

纸坊病病毒病因的研究

1985年4～10月与1986年6～8月,在贵州省沿河县的纸坊村和崔家坨村先后发生了病因不明的传染病。纸坊村约有1/5的村民发病,病死率为12％,崔家坨村有1/10的村民发病,病死率高达30％。发病波及各年龄组,以青壮年为多,有家庭集聚现象。 本病起病急,轻症者只有头晕、乏力、肌痛、多汗、心悸伴以低热,有的初期有短暂的腹泻。重症者有高热(40℃以上)、大汗、心悸、游走性肌肉痉挛伴有明显疼痛和触痛,以腰骶部及四肢肌肉为好发部位。病人烦燥不安,2～5天内死亡。经实验室检查,排除了食物中毒、农药中毒、钩端螺旋体病和弓形体感染。从病人和接触者的粪便中分离到9株病毒,性状一致,为RNA型25nm的球形颗粒,耐酸,耐乙醚,能凝集人“O”型血球。经血清学鉴定为ECHO3型病毒。16份病人双份血清的检测结果表明,恢复期血清对该病毒中和抗体有4倍以上升高者共8例(纸坊村和崔家坨各4例)。病人单份血清也都有较高的抗体。有理由认为两年中先后在两个村庄发生的传染病与ECHO3型病毒有密切关系。查阅文献,尚未见有关ECHO3型病毒引起以肌痛、游走性肌痉挛为特征的疾病的报道。     

Epidemiology of the Hong Kong/68 Variant of Influenza A2 in Britain

Two influenza epidemics in Britain in 1968-9 and 1969-70, were due to the Hong Kong/68 variant of influenza A2 virus. The first epidemic was prolonged with low morbidity and mortality rates; the second was sharp with high rates. The difference between total morbidity and mortality in the two epidemics, however, was less than it appeared to be—the estimated excess morbidity and mortality due to all causes in 1969-70 was only about 50% greater than in 1968-9.Antibody studies showed that about one-quarter of two groups of adults investigated were infected in the first epidemic and about one-third in the second. After the two epidemics about one-third still had no antibody to the A2/Hong Kong/68 virus.     

Influenza activity in Czechoslovakia and the USSR, 1980-1985

Between 1980 and 1985, Czechoslovakia had experienced 4 and the USSR 3 major influenza outbreaks. Of the 3 epidemic outbreaks in the USSR, 2 were associated with influenza B virus (in the 1980/81 and 1983/84 seasons) and 1 with influenza A virus of the H3N2 subtype. In the USSR, influenza A (H1N1) virus never predominated as a cause of epidemic during the 5 years period. In Czechoslovakia, 2 epidemics (in the 1980/81 and 1983/84 seasons) were due to influenza A (H1N1) virus. The epidemic in the 1981/82 season had two waves of unequal heights and a mixed type B and subtype A (H3N2) etiology; a two-wave epidemic associated with isolates of influenza A (H1N1) and influenza B viruses was also recorded in the 1983/84 season. The influenza A (H3N2) epidemic in 1983 was of explosive character. All influenza viruses circulating in the two countries between 1980 and 1985 were of the same antigenic profile, but were isolated from the epidemics that occurred in different influenza seasons. The virological surveillance revealed strains of virus closely related to drift variants detected from outbreaks in 1977-1979 and the new variants A/Chile 1/83, A/Philippines 2/82, A/Caen 1/84 and B/USSR 100/83.     

Results of virological and serological studies of three influenza A Hong-Kong epidemics in Leningrad.

An analysis of morbidity of the population in the course of 3 influenza A/Hong-Kong epidemics showed a pronounced decrease in influenza affection of adult population in the last epidemic in 1971--1972. Comparative studies of the diagnostic value of CFR and HIT demonstrated identical sensitivity of CFR as a method of influenza diagnostics in both the epidemic and interepidemic periods. HIT was suitable for the detection of influenza only in the epidemic period. In the interepidemic period, the percentage of influenza infection diagnosed by means of HIT ammounted to only 23--24 of all serologically confirmed cases of influenza. The highest percentage of virus isolation was observed when material from patients with serologically confirmed influenza was used. All strains of influenza A virus isolated in 1969 and 1970 were similar in their sensitivity to inhibitors of animal sera. During the last influenza epidemic, 2 of the 136 isolated strains were found to be resistant to gamma inhibitors and highly sensitive to the inhibitors showed their close relationship to gamma inhibitors. Antigenic analysis of the influenza A strains isolated during the 3 influenza epidemics revealed changes in the antigenic structure of the agents of the influenza epidemic in Leningrad in comparison with the standard strain A/Hong-Kong/I/68 (H3N2).     

Immunogenicity and the protective effectiveness of the subunit trivalent influenza vaccine Grippovac

In the process of the immunological approbation of several experimental batches of the triyalent subunit influenza vaccine Grippovac, the pronounced immunogenicity of the antigens of all strains contained in the vaccine was established; most of the vaccinees were found to retain sufficiently high antibody titers for a year, and the essential total increase of antibody titers was found to occur after a single booster immunization. The serological checking of the diagnosed cases of influenza among immunized and nonimmunized children, carried out in two boarding schools during the influenza epidemic in the winter of 1984-1985 provided the proofs of the high effectiveness of Grippovac in preventing viral influenza, types A and B: the decrease of influenza morbidity among the immunized children reached 79,5-67,2-66,5% and the total morbidity rate in influenza in these two boarding scholls dropped, on the average, 3,0-3,5 times.     

The role of neutrophils in the upper and lower respiratory tract during influenza virus infection of mice

Background

Neutrophils have been shown to play a role in host defence against highly virulent and mouse-adapted strains of influenza virus, however it is not clear if an effective neutrophil response is an important factor moderating disease severity during infection with other virus strains. In this study, we have examined the role of neutrophils during infection of mice with influenza virus strain HKx31, a virus strain of the H3N2 subtype and of moderate virulence for mice, to determine the role of neutrophils in the early phase of infection and in clearance of influenza virus from the respiratory tract during the later phase of infection.

Methods

The anti-Gr-1 monoclonal antibody (mAb) RB6-8C5 was used to (i) identify neutrophils in the upper (nasal tissues) and lower (lung) respiratory tract of uninfected and influenza virus-infected mice, and (ii) deplete neutrophils prior to and during influenza virus infection of mice.

Results

Neutrophils were rapidly recruited to the upper and lower airways following influenza virus infection. We demonstrated that use of mAb RB6-8C5 to deplete C57BL/6 (B6) mice of neutrophils is complicated by the ability of this mAb to bind directly to virus-specific CD8⁺ T cells. Thus, we investigated the role of neutrophils in both the early and later phases of infection using CD8⁺ T cell-deficient B6.TAP^-/- mice. Infection of B6.TAP^-/- mice with a low dose of influenza virus did not induce clinical disease in control animals, however RB6-8C5 treatment led to profound weight loss, severe clinical disease and enhanced virus replication throughout the respiratory tract.

Conclusion

Neutrophils play a critical role in limiting influenza virus replication during the early and later phases of infection. Furthermore, a virus strain of moderate virulence can induce severe clinical disease in the absence of an effective neutrophil response.     

Assessment of the effectiveness of inactivated influenza vaccines for adults

In 9 controlled epidemiological observations (1977-1984) the effectiveness of modern Soviet whole-virion vaccines was studied in organized groups of adults and at industrial enterprises. During the epidemic outbreaks of influenza of different etiology and intensity morbidity rate in influenza and acute respiratory diseases was shown to decrease 1.1-2.2 times among the vaccinees, depending on the correspondence of epidemic and vaccine influenza strains. The absence of influenza virus B in inactivated influenza vaccines was the reason for their low effectiveness during influenza outbreaks of mixed etiology B + A (H1N1).     

Early outbreak of 2009 influenza A (H1N1) in Mexico prior to identification of pH1N1 virus

Background

In the aftermath of the global spread of 2009 influenza A (pH1N1) virus, still very little is known of the early stages of the outbreak in Mexico during the early months of the year, before the virus was identified.

Methodology/Main Findings

We fit a simple mathematical model, the Richards model, to the number of excess laboratory-confirmed influenza cases in Mexico and Mexico City during the first 15 weeks in 2009 over the average influenza case number of the previous five baseline years of 2004-2008 during the same period to ascertain the turning point (or the peak incidence) of a wave of early influenza infections, and to estimate the transmissibility of the virus during these early months in terms of its basic reproduction number. The results indicate that there may have been an early epidemic in Mexico City as well as in all of Mexico during February/March. Based on excess influenza cases, the estimated basic reproduction number R₀ for the early outbreak was 1.59 (0.55 to 2.62) for Mexico City during weeks 5–9, and 1.25 (0.76, 1.74) for all of Mexico during weeks 5–14.

Conclusions

We established the existence of an early epidemic in Mexico City and in all of Mexico during February/March utilizing the routine influenza surveillance data, although the location of seeding is unknown. Moreover, estimates of R₀ as well as the time of peak incidence (the turning point) for Mexico City and all of Mexico indicate that the early epidemic in Mexico City in February/March had been more transmissible (larger R₀) and peaked earlier than the rest of the country. Our conclusion lends support to the possibility that the virus could have already spread to other continents prior to the identification of the virus and the reporting of lab-confirmed pH1N1 cases in North America in April.