Showing papers on "Influenza A virus published in 1983"

Cytotoxic T-cell immunity to influenza.

Abstract: In a study designed to determine whether cytotoxic T lymphocytes contribute to immunity against influenza virus infection, we inoculated 63 volunteers intranasally with live unattenuated influenza A/Munich/1/79 virus. Over the next seven days clinical observations were made, and the amount of virus shed was measured. The protective effects of preinfection serum antibody and of cytotoxic T-cell immunity against influenza A virus were assessed for each participant. All subjects with demonstrable T-cell responses cleared virus effectively. This response was observed in volunteers in all age groups, including those born after 1956, who did not have specific antibody and hence had probably not been exposed to this subtype of influenza A virus before. Cytotoxic T cells show cross-reactivity in their recognition of the different subtypes of influenza A virus, in contrast to the antibody response that is specific for each virus subtype. We conclude that cytotoxic T cells play a part in recovery from influenza virus infection.

Construction and characterization of an infectious vaccinia virus recombinant that expresses the influenza hemagglutinin gene and induces resistance to influenza virus infection in hamsters.

Abstract: A DNA copy of the influenza virus hemagglutinin gene, derived from influenza virus A/Jap/305/57 (H2N2) was inserted into the genome of vaccinia virus under the control of an early vaccinia virus promoter. Tissue culture cells infected with the purified recombinant virus synthesized influenza hemagglutinin, which was glycosylated and transported to the cell surface where it could be cleaved with trypsin into HA1 and HA2 subunits. Rabbits and hamsters inoculated intradermally with recombinant virus produced circulating antibodies that inhibited hemagglutination by influenza virus. Furthermore, vaccinated hamsters achieved levels of antibody similar to those obtained upon primary infection with influenza virus and were protected against respiratory infection with the A/Jap/305/57 influenza virus.

The Influenza Virus RNA Segments and Their Encoded Proteins

Abstract: In this chapter the structure of the genes of influenza virus and their encoded proteins will be described. Influenza viruses contain a segmented single-stranded RNA genome which has been called “negative stranded” because the viral messenger RNAs are transcribed from the viral RNA segments. A great deal of new knowledge has been obtained about influenza A, B, and C viruses since the last major reviews of influenza virus in 1975 (Kilbourne, 1975). The complete sequence of the 8 RNA segments of influenza A virus has been obtained and this has led to a greatly increased understanding of the encoded polypeptides and how variation among influenza viruses occurs. A large body of evidence concerning changes in the surface glycoproteins, the hemagglutinin and neuraminidase, has been obtained from studies on the primary sequences, interactions with antibodies and the correlation of these data with the three-dimensional X-ray structure of these proteins.

Dose response of A/Alaska/6/77 (H3N2) cold-adapted reassortant vaccine virus in adult volunteers: role of local antibody in resistance to infection with vaccine virus.

Abstract: An attenuated influenza A candidate vaccine virus, derived from the A/Ann Arbor/6/60 (H2N2) cold-adapted (ca) donor virus and the A/Alaska/6/77 (H3N2) wild-type virus, was evaluated in adult seronegative volunteers (serum hemagglutination-inhibiting antibody titer, less than or equal to 1:8) for level of attenuation, infectivity, antigenicity, and genetic stability. Four groups with similar preinoculation mean titers of serum and nasal wash antibodies were inoculated intranasally with 10(4.5), 10(5.5), 10(6.5), or 10(7.5) 50% tissue culture infectious doses (TCID50) of the ca reassortant virus, and eight other seronegative adult volunteers received the wild-type virus. Only 2 of 66 vaccinees developed fever or mild and brief systemic or upper respiratory tract illness or both. Both volunteers with vaccine-related reactions received the highest dose (10(7.5) TCID50) of ca virus, which indicates that the vaccine retains some mild reactogenicity at a high dosage. In contrast, four of eight volunteers infected with the wild-type virus became ill. Each of the 54 isolates tested retained the temperature-sensitive phenotype of the vaccine virus. Thus, the ca reassortant was genetically stable and attenuated at 10(4.5) to 10(7.5) TCID50 for seronegative adults. The 50% human infective dose of ca virus was approximately 10(5.3) TCID50. Ten and one hundred 50% human infectious doses infected 73 and 83% of vaccinees, respectively, and approximately 75% developed an immunological response at these doses. The failure of the vaccine virus to infect some volunteers was correlated with the presence of pre-inoculation nasal wash immunoglobulin A hemagglutinin antibody.

Generation of antibody diversity in the immune response of BALB/c mice to influenza virus hemagglutinin. I. Significant variation in repertoire expression between individual mice

Abstract: The paratypic and idiotypic diversity of the BALB/c antibody response to the hemagglutinin (HA) of the influenza A/PR/8/34 virus (PR8) was investigated using a panel of 125 anti-HA hybridoma antibodies derived from 14 BALB/c mice. The paratypic diversity, as assessed by a fine specificity analysis using 51 related influenza viruses, was extensive: 104 distinct paratopes were observed. In three instances, antibodies with indistinguishable paratopes were isolated from two individual mice. A minimum estimate of the size of the adult BALB/c anti-HA paratypic repertoire, calculated from these data, is 1,500. The generation of this diverse repertoire was studied by screening the anti-HA hybridoma panel for the presence of idiotypes (Id) that are markers for variable (V) region sequences derived from related germ line V genes. Three cross-reactive Id (IdX) that are markers for the V(k)21C, V(k)21B, and V(k)21A, D, E, or F L chain subgroups were found, respectively on 16, 1, and 10 anti-HA hybridoma antibodies derived from seven individual BALB/c mice. Thus, the V(k)21 IdX(+) hybridomas constitute 22 percent of the anti-HA hybridoma panel. The V(k)21 IdX are also present on 8.6 percent of K-bearing immunoglobulin in normal BALB/c serum. This suggests that the V(k)21 group is used preferentially in the BALB/c anti-HA immune response. The generation of the anti-HA repertoire was further studied using large panels of anti-HA hybridomas derived from two individual adult BALB/c mice. Anti-idiotypic antisera were raised in rabbits against individual hybridomas from each mouse. One anti-Id serum defined a family of four idiotypically and paratypically related, but not identical, antibodies from mouse 36, which represented 31 percent of the hybridoma antibodies isolated from this mouse. None of the 112 anti-HA hybridoma antibodies derived from 13 other individual mice showed idiotypic cross-reactivity. Furthermore, this Id could not be detected in anti-PR8 antisera from 75 individual BALB/c mice. Another anti-Id serum defined a family of 27 idiotypically related antibodies from mouse 37, which represented 50 percent of the hybridoma antibodies isolated from this mouse. Only 1 of the 71 hybridoma antibodies isolated from 13 other individuals was idiotypically cross-reactive. These results demonstrate that individual adult BALB/c mice express paratypically and idiotypically distinct antibody repertoires to the HA of influenza virus PR8. Based on these observations, we suggest that somatic mutation plays an important role in the generation of the adult anti-HA repertoire. Mechanisms that could account for differences in repertoire expression among individual mice are discussed.

A previously unrecognized influenza B virus glycoprotein from a bicistronic mRNA that also encodes the viral neuraminidase

Abstract: RNA segment 6 of the influenza B virus genome codes for a previously unidentified polypeptide designated NB. The reading frame for this polypeptide begins with the first AUG codon on the mRNA and overlaps the reading frame for the viral neuraminidase by 292 nucleotides. The amino acid sequence of polypeptide NB deduced from the nucleotide sequence of the B/Lee/40 strain consists of 100 amino acids with a molecular weight of 11,242. The sequence contains four potential glycosylation sites, and the protein has been found to be glycosylated in infected cells. NB has not been found in virions. Sucrose gradient sedimentation and analysis of the structure of the mRNA by nuclease S1 mapping and sequence analysis by the primer extension method indicated that polypeptide NB and the neuraminidase are translated from a single bicistronic mRNA. A protein analogous to NB has not been found with influenza A virus, and this represents a major difference between the two virus types.

Sequential mutations in hemagglutinins of influenza B virus isolates: definition of antigenic domains

Abstract: Comparative analysis of the amino acid sequences of hemagglutinins (HAs) of influenza B/Lee/40, B/Md/59, and B/HK/73 viruses has allowed examination of the molecular basis of antigenic variation in type B viruses. As seen with influenza type A viruses, antigenic drift in influenza B viruses proceeds mostly through the accumulation of amino acid substitutions within the HA1 portion of the HA molecule. However, the rate of variation observed among the influenza B virus HAs appears to be significantly lower than the observed rate of variation among influenza A virus HAs. The overall rate of amino acid change in the HA1s of the influenza B viruses studied is 2% per 10 years, whereas the HA1s of H3 influenza A viruses vary by 9.2% per 10 years. The sequences of the influenza B HAs were also examined in relation to the three-dimensional model for the A/Aichi/2/68 HA. When the primary amino acid sequences are compared, it appears that most of the important structural features of the type A HAs--such as the sialic acid binding site, the disulfide linkages, and the stem structure of the trimer--are conserved in the influenza B virus HAs. Regions are also identified where extensive amino acid substitutions have occurred among the three antigenically distinct influenza B virus HAs. The locations of these areas in the B HA structure correspond to antigenic regions proposed for the A virus HAs. In addition, modulation of antigenic regions in B virus HAs may also occur through amino acid deletions and variation in glycosylation sites.

Efficient expression of influenza virus NS1 nonstructural proteins in Escherichia coli.

Abstract: RNA segment 8 of the influenza A virus genome codes for two nonstructural proteins, NS1 and NS2, for which the functions are unknown. Cloned cDNA copies of this gene from three different influenza A virus strains were inserted into an Escherichia coli plasmid expression vector, pAS1, carrying the strong regulatable lambda phage promoter, PL. After induction, the NS1 proteins were overproduced to levels of 20-25% of total cellular protein. This was surprising in that the codon composition for these eukaryotic genes is similar to that for weakly expressed proteins in E. coli. Thus, under the appropriate conditions, it appears that high level expression of genes containing a relatively large proportion of minor codons can be obtained. The NS1 protein produced in bacteria from a cloned cDNA copy of the A/PR/8/34 virus NS gene was purified to apparent homogeneity and used to generate a high-titer monospecific rabbit antiserum. Immunoprecipitation studies showed this antibody to be crossreactive against the NS1 proteins produced by several different influenza A virus strains. Immunofluorescence experiments in Madin-Darby canine kidney cells showed the NS1 proteins to be located in the nucleoplasm early in infection for all strains examined. With some of the strains, NS1-specific immunofluorescence was observed predominantly in the nucleoli later in infection. This technology can be used to obtain other viral proteins in pure form for structural, functional, and immunological studies.

Reduction in fever and symptoms in young adults with influenza A/Brazil/78 H1N1 infection after treatment with aspirin or amantadine.

Abstract: During an outbreak of influenza A/Brazil/78 H1N1 infection, 47 volunteers with clinical and virological influenza of less than 2 days duration were treated in a randomized double-blind fashion for 5 days with 100 or 200 mg of amantadine daily or with 3.25 g of aspirin daily. The aspirin treatment group defervesced more rapidly (10.3 h versus 21.5 h and 23.6 h; P less than 0.01), but by the second daily follow-up visit, both groups of amantadine recipients exhibited greater symptomatic improvement. Bothersome side effects resulted in discontinuation of therapy by 35% of the aspirin treatment group but only 3% of the amantadine treatment group (P less than 0.05). Individuals who present to a physician during an influenza A epidemic with characteristic symptoms will experience symptomatic benefit from amantadine treatment, with negligible toxicity.

Influenza virus site recognized by a murine helper T cell specific for H1 strains. Localization to a nine amino acid sequence in the hemagglutinin molecule.

Abstract: The functional helper T cell line Vir-2, derived from a PR8 (H1N1) influenza virus-immunized BALB/c mouse, proliferates in response to syngeneic antigen-presenting cells and naturally occurring strains of subtype H1 human influenza virus from 1934-1957 and 1977-1980 isolates. A conserved region of the hemagglutinin molecule around amino acid position 115 in the heavy chain (HA1) was implicated as being important in this recognition by the lack of stimulatory activity associated with a glutamic acid to lysine substitution at position 115 in the laboratory mutant RV6, derived from wild-type PR8. Characterization of the stimulatory determinant on the wild-type hemagglutinin molecule was then undertaken using cleavage products and synthetic peptides. Vir-2 cells recognized the reduced and alkylated purified HA1 of PR8 virus, and this reactivity was retained after cleavage at methionine and tryptophan residues. High-pressure liquid chromatography separation of cleavage fragments indicated that a short sequence of the HA1 containing residue 115 was being recognized. This recognition was localized to a nine amino acid segment (positions 111-119) by assaying stimulation with synthetic peptide homologues of different lengths from that region. As with native hemagglutinin, Vir-2 cells responded to active peptides when presented by H-2d but not H-2k antigen-presenting cells.

Secretory immunological response after intranasal inactivated influenza A virus vaccinations: evidence for immunoglobulin A memory.

Abstract: An intranasal, inactivated trivalent influenza A vaccine containing 7 micrograms of A/Bangkok/1/79 (H3N2) hemagglutinin was administered to 20 children aged 1 to 6 years to assess the local and systemic immune responses to antigen delivered to the respiratory tract. Six children without prior influenza virus infection exhibited no local immune response and manifested only a minimal systemic response to the intranasal vaccine. In contrast, five individuals who were previously infected with a live attenuated influenza A H3N2 virus vaccine, although having no residual secretory antibody at the time of challenge, promptly developed a local antibody response to intranasal, inactivated antigen. Therefore, the live influenza A virus vaccine had induced memory in the local immunoglobulin A (IgA) immune system. The third group of nine children had previously been infected with wild-type H3N2 influenza virus. A majority of these children had residual local and systemic antibody at the time of challenge but they demonstrated some boosting of local IgA antibody with administration of intranasal inactivated vaccine. The competence of the secretory IgA immune system in young children in mounting primary and secondary responses to influenza antigens has important implications for approaches to prevention of influenzal illness.

Sequential mutations in the NS genes of influenza virus field strains.

Abstract: The complete nucleotide sequences of the NS genes from three human influenza viruses, A/FM/1/47 (H1N1), A/FW/1/50 (H1N1), and A/USSR/90/77 (H1N1), were determined. Only five single-base differences were found within the sequences of the A/FW/1/50 and A/USSR/90/77 NS genes, thus confirming earlier data suggesting that the 1977 H1N1 viruses are closely related to virus strains that were circulating around 1950. Comparison of all three sequences with those from A/PR/8/34 and A/Udorn/72 viruses illustrates that these genes (with the exception of that of the A/USSR/90/77 strain) evolve through cumulative base changes along a single common lineage. A nucleotide sequence variation of approximately 2.2 to 3.4% per 10 years was determined for the NS gene segments. Extensive size variation was also observed among the NS1 proteins of the various human viruses. The A/FM/1/47 NS1 protein, which consists of 202 amino acids, is 15% shorter than the A/Udorn/72 NS1 protein, which consists of 237 amino acids.

Hemagglutinin-specific antibody responses in immunoglobulin G, A, and M isotypes as measured by enzyme-linked immunosorbent assay after primary or secondary infection of humans with influenza A virus.

Abstract: The isotype-specific antibody responses to purified hemagglutinin of adults undergoing either primary or secondary infection with an influenza A virus were characterized by using an enzyme-linked immunosorbent assay. Twenty-eight military recruits undergoing primary infection with A/USSR/92/77 (H1N1)-like virus had serum antibody rises in the immunoglobulin M (IgM) (86%), IgG (100%), and IgA (96%) isotypes. In contrast, 19 adult volunteers undergoing secondary infection with A/Peking/2/79 (H3N2) wild-type virus had serum antibody titer rises largely restricted to the IgG (68%) and IgA (74%) classes, with only 1 volunteer having a serum IgM antibody titer rise. Nasal wash hemagglutinin-specific antibody responses in the adults undergoing secondary infection were predominantly in the IgA class (74%). There was a correlation between the presence of and the magnitude of nasal wash and serum hemagglutinin-specific IgA antibody responses in these adults. This suggested that there was a common source for the hemagglutinin-specific local IgA antibody and serum IgA antibody produced after infection. The recruits undergoing primary H1N1 influenza virus infection had H1 hemagglutinin-specific enzyme-linked immunosorbent assay antibody in each of the IgA, IgG, and IgM isotypes in their acute-phase serum. However, no role for this cross-reactive antibody in modifying the severity of illness experienced by the recruits could be demonstrated.

Micro neuraminidase-inhibition assay for classification of influenza A virus neuraminidases.

Abstract: SUMMARY A neuraminidase-inhibition (NI) assay performed in microtiter plates is described. This micro-NI assay is a modification of the NI assay recommended by the World Health Organization. It reduces the quantity of reagents required and permits antigenic classification of many isolates simultaneously. To determine the accuracy and sensitivity of this micro-NI assay, 110 influenza A viruses, representing all subtypes, based upon the nine known neuraminidases (NAs), were classified by both the micro-NI and macro-NI assays in two separate laboratories. The NAs were identified accurately by the micro-NI assay. Virus mixtures were detected by both assays, although the macro-NI was clearly more sensitive. The micro-NI assay was also suitable for testing sera for the presence of antibodies to the NAs. Although the micro-NI assay did not provide the quantitation of the macro-NI assay, it did prove to be a rapid method for virus classification and antibody studies on influenza A viruses.

Effect of influenza A virus on leukocyte histamine release.

Abstract: Viral respiratory infections provoke asthma in many patients. In the following study we examined the effect of an in vitro incubation of influenza A on leukocyte histamine release. After incubation with a live influenza A (H3N2) virus, calcium ionophore A23187 (0.5, 1.0, and 1.5 microgram/ml)-induced leukocyte histamine release (HR) was enhanced (p less than 0.05). This effect was also found with heat- or ether-inactivated virus. Similarly, influenza A-exposed leukocytes had augmented leukocyte HR during subsequent incubation with ragweed AgE. Incubation of the leukocyte suspension with interferon (800 IU/ml) for 24 hr was also associated with enhanced HR to ragweed AgE. In contrast, interferon did not alter the calcium ionophore A23187 HR. Therefore, although interferon may mediate the enhanced leukocyte HR when ragweed AgE is the inciting stimulus, it does not change HR to the calcium ionophore.

Studies with inactivated equine influenza vaccine. 2. Protection against experimental infection with influenza virus A/equine/Newmarket/79 (H3N8).

Abstract: Forty ponies immunized with inactivated virus vaccine containing A/equine/Miami/63 (H3N8) virus and six unvaccinated, seronegative ponies were experimentally challenged with a representative of recent equine H3N8 virus isolates, A/equine/Newmarket/79. All unvaccinated ponies became infected as judged by virus excretion, febrile responses and antibody responses, but only two of the vaccinated ponies were fully protected. Pre-challenge antibody levels to A/Newmarket/79 virus detected by single radial haemolysis (SRH) correlated well with the degree of clinical protection but the levels required for complete protection (SRH zones greater than 65 mm2) were high. The importance of these results in relation to conventional vaccination procedures against equine influenza is discussed.

Localization, synthesis, and activity of an antigenic site on influenza virus hemagglutinin

Abstract: This paper reports the antigenicity of the fusion region of the influenza virus hemagglutinin (HA). Two peptides, comprising the fusion region (residues 1-11 of the HA2 part of HA) of strain A and strain B influenza virus, were synthesized and their abilities to bind rabbit, goat, and human anti-influenza antibodies were determined. In addition, 30 anti-HA monoclonal antibodies were examined for their ability to bind the synthetic peptides. In quantitative immunoadsorbent titrations, the two peptides bound considerable amounts of antibodies in rabbit and goat antisera against virus or HA of the A or B strain as well as in several human sera from patients recovering from influenza A. Of the 30 anti-HA monoclonal antibodies, 5 bound completely and 4 bound partially to the peptides. Antibodies were raised in rabbits against the peptides by immunizing with peptide-bovine serum albumin conjugates or with the free peptides. Anti-peptide antibodies were bound by HA and by the intact virus of the respective strain. However, these antisera failed to exhibit significant virus neutralizing activity. In contrast, the monoclonal antibodies that reacted with these peptides inhibited viral infectivity. The results clearly show that residues 1-11 of HA2 represent an important antigenic site on influenza virus.

Swine influenza-like viruses in turkeys: potential source of virus for humans?

Abstract: Influenza A viruses (subtype H1N1), recently isolated from turkeys in different areas of the United States, were determined to be closely related to strains typically associated with pigs. This conclusion was based on comparisons of H1N1 isolates from pigs, humans, ducks, and turkeys with polyclonal and monoclonal antibodies, RNA-RNA competitive hybridization, and replication studies. One of the H1N1 isolates from turkeys caused influenza in a laboratory technician, who displayed fever, respiratory illness, virus shedding, and seroconversion. These results suggest that turkeys as well as pigs are involved in the maintenance of these viruses and their transmission to humans.

Production of human monoclonal antibody to X31 influenza virus nucleoprotein.

Abstract: In vitro stimulation of human peripheral blood mononuclear cells with X31 influenza virus antigen has been used to enrich for specific anti-X31 antibody-producing cells. Following Epstein-Barr virus transformation of these stimulated cells, a cell line which produces human antibody to X31 virus was derived and subsequently cloned. The cloned cells secrete and IgGl kappa antibody which is directed against the nucleoprotein of A type influenza virus. Culture supernatants contain 10 to 20 micrograms/ml of specific antibody which is now used as a standard for the ELISA assay used in our laboratory to detect antibodies to influenza virus.

Analysis of the antigen specificity of influenza haemagglutinin-immune human T lymphocyte clones: identification of an immunodominant region for T cells.

Abstract: Human T lymphocyte clones specific for the haemagglutinin (HA) molecule of A/Texas/1/77 were maintained in long-term culture with T cell-growth factor. The clones were analysed for their viral antigen specificity using serologically defined type A influenza subtypes and chemically synthesized peptides of the HA-1 molecule. With the exception of one T cell clone that recognized only the HA molecule used for immunization, the clones responded to determinants that showed partial or complete cross-reactivity amongst the strain A subtypes. The cross-reactive population of T cell clones were specific for peptides distinct from the antibody binding sites of HA. Furthermore, one peptide located at the carboxyl terminus of the HA-1 molecule appeared to be immunodominant, although the critical residues for T cell antigen recognition within that could not be identified.

Analysis of antigenic variation in equine 2 influenza A viruses.

Abstract: Influenza outbreaks involving viruses of the H3N8 subtype (equine 2) often occur in vaccinated horses. For this reason, a series of influenza viruses of the H3N8 subtype were examined to determine if antigenic variation could be detected in isolates during the period 1963-81. Antigenic analyses with post-infection ferret sera and monoclonal antibodies showed that the haemagglutinins of recent isolates were antigenically distinguishable from the prototype A/eq/Miami/1/63 and that antigenically distinguishable groups of equine 2 viruses co-circulate in the horse population. Based on these studies, it is recommended that a recent equine strain, A/equine/Fontainebleu/1/79 or A/equine/Kentucky/1/81, serve as an additional prototype strain for this subtype.Antigenic variation in equine 2 viruses may be of epidemiological significance, yet the overall conservation of these strains makes it unlikely that vaccine failures can be attributed solely to antigenic changes in these viruses. A sufficiently potent vaccine, containing a current representative of the most prevalent equine 2 strain, may improve the protection afforded by equine vaccines.

Different virulence of influenza A virus strains and susceptibility to pneumococcal otitis media in chinchillas.

Abstract: We have previously shown that chinchillas infected with a multiply passaged laboratory strain of influenza A/NWS/33 (H1N1) develop negative middle-ear pressure; polymorphonuclear leukocyte oxidative, bactericidal, and chemotactic dysfunction; and increased susceptibility to pneumococcal otitis media. Because influenza A virus strains show different virulence in humans, three such strains were compared in the chinchilla model. Negative middle-ear pressure and tympanic membrane inflammation developed significantly more often in chinchillas infected with wild-type H3N2 virus than with either wild-type H1N1 virus or an attenuated, cold-adapted H3N2 vaccine strain, CR29. Marked depression in polymorphonuclear leukocyte chemiluminescent activity also developed significantly more often in H3N2 infected animals than in H1N1- or CR29-infected animals. Intranasal challenge of influenza virus-infected animals with type 7 Streptococcus pneumoniae resulted in a significantly greater occurrence of pneumococcal otitis media in H3N2-infected animals than in H1N1-, CR29-, or non-influenza-infected control animals. Clearance of pneumococci from nasal washings of animals infected with wild-type H3N2 was significantly delayed in comparison with the other groups. Thus, the previously demonstrated increased susceptibility to otitis media among children infected with H3N2 influenza virus may relate to the capacity of this strain to induce negative middle-ear pressure, polymorphonuclear leukocyte dysfunction, and alteration in the mucosal clearance of pneumococci.

Individuals Infected with Two Subtypes of Influenza A Virus in the Same Season

Abstract: Participants in the Houston Family Study were observed during a period of two mixed outbreaks due to two subtypes of influenza A virus: H3N2 and H1N1 (1977-1981) Virus specimens, serum samples, and clinical records were obtained to identify and characterize infections In 1977-1978, 40% of 238 persons in 59 families were infected by influenza A virus (H3N2), 11% by influenza A virus (H1N1), and 4% by both In 1980-1981, for 319 persons in 79 families, the corresponding rates were 27%, 20%, and 5% Interference between subtypes was not detected Both subtypes were isolated from six children (range of intervals between isolations, six to 55 days), and five of the six were ill with both infections Nineteen persons had two infections with one or both detected serologically; illnesses were associated with 77% of isolates and up to 56% of seroconversions in these persons Infection of the same individual with two subtypes in the same season is a newly observed phenomenon that may affect the future epidemiology of influenza A virus as well as preventive measures

Modulation of glycosylation and transport of viral membrane glycoproteins by a sodium ionophore.

Abstract: Analysis of viral glycoprotein expression on surfaces of monensin-treated cells using a fluorescence-activated cell sorter (FACS) demonstrated that the sodium ionophore completely inhibited the appearance of the vesicular stomatitis virus (VSV) G protein on (Madin-Darby canine kidney) MDCK cell surfaces. In contrast, the expression of the influenza virus hemagglutinin (HA) glycoprotein on the surfaces of MDCK cells was observed to occur at high levels, and the time course of its appearance was not altered by the ionophore. Viral protein synthesis was not inhibited by monensin in either VSV- or influenza virus-infected cells. However, the electrophoretic mobilities of viral glycoproteins were altered, and analysis of pronase-derived glycopeptides by gel filtration indicated that the addition of sialic acid residues to the VSV G protein was impaired in monensin-treated cells. Reduced incorporation of fucose and galactose into influenza virus HA was observed in the presence of the ionophore, but the incompletely processed HA protein was cleaved, transported to the cell surface, and incorporated into budding virus particles. In contrast to the differential effects of monensin on VSV and influenza virus replication previously observed in monolayer cultures of MDCK cells, yields of both viruses were found to be significantly reduced by high concentrations of monensin in suspension cultures, indicating that cellular architecture may play a role in determining the sensitivity of virus replication to the drug. Nigericin, an ionophore that facilitates transport of potassium ions across membranes, blocked the replication of both influenza virus and VSV in MDCK cell monolayers, indicating that the ion specificity of ionophores influences their effect on the replication of enveloped viruses.

Laboratory-based surveillance of influenza A(H1N1) and A(H3N2) viruses in 1980-81: antigenic and genomic analyses

Abstract: During 1981, the A/Brazil/11/78-like strains of influenza virus that had been prevalent from 1978 to 1980 were displaced by a new set of heterogeneous, but closely related, variants (reference strain, A/England/333/80). Genomic analysis revealed that these new variants were almost exclusively nonrecombinant H1N1 viruses, i.e., they contained no genes of H3N2 origin. However, a few recombinant viruses containing the new variant HA and genes of H3N2 origin were identified. Antigenic analysis of H3N2 viruses indicated that they were also heterogeneous. The majority of these virus isolates were antigenically intermediate between A/Texas/1/77 and A/Bangkok/1/79, but additional variants were detected. Genomic analysis revealed that the H3N2 viruses isolated in the winter of 1980-81 were quite similar to H3N2 viruses isolated from 1977-79 in their T1 oligonucleotide maps. No H1N1 genes were detected in H3N2 virus isolates. Comparison of pairs of oligonucleotide maps of total virus RNA indicated that a similar rate of genetic change had occurred for nonrecombinant H1N1 viruses, for recombinant H1N1 viruses, and for H3N2 viruses and that, in general, pairs of viruses exhibited increasing numbers of changes in their oligonucleotide maps as the time interval between isolation of the viruses increased.