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in vitro synthesis of full-length influenza virus complementary rna.influenza virus-specific rna has been synthesized in vitro, using cytoplasmic or microsomal fractions of influenza virus-infected mdck cells. the rna polymerase activity was stimulated 5-30 times by priming with apg. about 20-30% of the product was polyadenylated. most of the in vitro product was of positive polarity, as shown by hybridization to strand specific probes and by t1 fingerprinting of the poly(a)+ and poly(a)- rna segments encoding haemagglutinin and nucleoprotein. the size of poly(a ...19852410253
myxovirus replication in chicken embryos can be suppressed by aprotinin due to the blockage of viral glycoprotein cleavage.injection of the protease inhibitor, aprotinin, into the allantoic cavity of embryonated eggs infected at low m.o.i. with different influenza viruses and paramyxoviruses markedly reduced multiplication by at least 100-fold. under these conditions, most viral particles produced were non-infectious and contained uncleaved glycoproteins, presumably resulting from aprotinin suppression of protease activity.19852410557
a mutant of fowl plague virus (influenza a) with an enhanced electrophoretic mobility of rna segment 8.a temperature-sensitive mutant (ts 1/9) obtained by undiluted passage of fowl plague virus (fpv) at 33 degrees c carried a strong ts defect in rna segment 6 [neuraminidase (na) gene] and a weak ts defect in rna segment 8 [non-structural (ns) protein although the viral proteins have normal migration rates, the ns gene migrated during polyacrylamide gel electrophoresis (page) significantly faster than the ns gene of wild-type fpv, even after denaturation by glyoxal. despite this observation, the n ...19852410558
antigenic determinants of influenza virus haemagglutinin. x. a comparison of the physical and antigenic properties of monomeric and trimeric forms.haemagglutinin prepared from influenza virus a/memphis/1/71 by bromelain digestion was centrifuged through continuous sucrose gradients buffered at ph 7.4 or ph 4.9. from these gradients were isolated two forms of the protein which displayed different equilibrium sedimentation properties. one species behaved as a molecule with a mol. wt. of 190 000, the other with a mol. wt. of 70 000. these results are consistent with the separation of trimeric and monomeric haemagglutinin. a comparison of thei ...19852410559
monoclonal antibodies detect different forms of influenza virus hemagglutinin during viral penetration and biosynthesis.monoclonal antibodies specific for the influenza virus a/pr/8/34 hemagglutinin (ha) were used to examine the structure of the ha glycoprotein by immunofluorescence techniques during infection of mdck cells. one antibody (y8-10c2), shown previously to detect conformational alterations in the ha coinciding with the acid induction of viral fusion activity, bound to internalized virus but not to virus adsorbed to the cell surface. the binding of y8-10c2 was completely inhibited by ammonium chloride ...19852410628
location of antigenic sites on the three-dimensional structure of the influenza n2 virus neuraminidase.sequence analysis of the neuraminidase (na) genes of influenza virus x-7(f1) and of 12 variants selected with monoclonal antibodies has been used to define in physical terms the antigenic structure of this na, which was operationally established by r. g. webster, l. e. brown, and w. g. laver (1984, virology 135, 30-42). x-7(f1) is a reassortant virus containing the na of the early asian (h2n2) isolate a/ri/5+/57, and the results of antigenic and sequence analysis of x-7(f1) and of variants selec ...19852411049
is bivalent binding of monoclonal antibodies to different antigenic areas on the hemagglutinin of influenza virus required for neutralization of viral infectivity?biological activities of fab fragments of monoclonal igg antibodies to each of four nonoverlapping antigenic areas on the hemagglutinin molecule of a/seal/massachusetts/1/80 (h7n7) influenza virus were examined. fab fragments of the antibodies belonging to groups i and ii neutralized viral infectivity. these fab fragments inhibited hemagglutination of the virus and virus-induced hemolysis at ph 5.9. on the other hand, fab fragments of groups iii and iv antibodies showed neither neutralization no ...19852411245
cytotoxic t cells recognize fragments of the influenza nucleoprotein.recent work has shown that a major population of murine influenza a specific cytotoxic t lymphocytes (ctl) recognize the viral nucleoprotein. in order to investigate the mechanism by which this nonglycoprotein component of the virus is recognized by ctl, a series of deletion mutants of an a virus np gene were studied. the results showed that ctl recognize three distinct epitopes of the np molecule. both n- and c-terminal fragments of the protein are transported, independently of each other, to t ...19852411422
fine specificity analysis of human influenza-specific cloned cell lines.influenza-specific human-t-cell clones, proliferating in the presence of virus-infected cells with restriction by class ii molecules and displaying class ii-restricted ctl activity or specific helper activity in antibody synthesis, have been analyzed for antigenic specificities. all of them were obtained by in vitro stimulation against influenza a/texas virus. in all cases the virus specificity appeared identical in cytolytic and proliferative responses. three of the clones were broadly cross-re ...19852411429
immunoenzymatic staining of viral and chlamydial antigens in cell culture.methods are described for the conjugation of antibodies with biotin and for the use of these reagents in an immunoperoxidase staining procedure for infected cell cultures. this technique provides a simple, rapid, and specific approach to the identification and characterization of a number of viral and chlamydial isolates in the diagnostic laboratory.19852411466
homozygous typing cell-defined hla-dw specificities correlate better than serologically defined hla-dr specificities with restriction elements for influenza virus-specific proliferative human t lymphocyte clones.human t lymphocyte clones with specific proliferative response to influenza a virus were derived by limiting dilution from peripheral blood lymphocytes (pbl) after in vitro stimulation with autologous irradiated, virus-infected pbl. four okt3+4+8- t lymphocyte clones (tlc) that showed hla-restricted antigen-specific proliferative responses were used for a detailed analysis of the restriction elements for antigen presentation. none of the clones showed alloreactivity and all required the presence ...19852411701
recognition of viral antigens by human influenza a virus-specific t lymphocyte clones.the nature of the viral antigens recognized by influenza a virus-immune cytotoxic t lymphocytes (ctl) is still a matter of debate. we have used four human influenza a virus-specific t lymphocyte clones with antigen-specific cytotoxic and proliferative activity to investigate the requirements for recognition of viral antigens on infected cells. one clone recognized a cross-reactive determinant on the viral hemagglutinin, and two clones were specific for different epitopes on the viral nucleoprote ...19852411815
impaired measles virus-specific cytotoxic t cell responses in multiple sclerosis.to assess whether an virus-specific immune defect may be associated with multiple sclerosis (ms), we have examined the ability to generate measles virus-and influenza virus-specific cytotoxic t cells (ctl) in patients with ms, normal individuals, and other disease controls (odc). the mean (+/- sem) measles virus-specific ctl response for normal individuals and odc was 26.9 +/- 2.9% (n = 17) and 26.7 +/- 2.8% (n = 13) specific lysis, respectively. in contrast, the capacity of ms patients to gener ...19852411841
inhibition of influenza viral mrna synthesis in cells expressing the interferon-induced mx gene product.interferons alpha and beta induce an efficient antiviral state against influenza virus in mouse cells that possess the mx gene, but not in mouse cells that lack this gene. in mx-containing cells treated with interferon the amount of viral mrna synthesized as a result of primary transcription is drastically reduced. only two viral mrnas could be detected by northern analysis and by translating the poly(a)+ rna from infected cells in wheat germ extracts: a reduced amount of the mrna for nonstructu ...19852411949
influenza virus hemagglutinin-binding t cells in delayed hypersensitivity induced in mice.the ability of spleen t cells from mice inoculated with influenza virus to transfer delayed hypersensitivity increases after 5 days culture in the presence of tcgf and influenza virus or its hemagglutinin. enriched t cells have also a greater ability to bind viral hemagglutinin than cells which were not cultured and restimulated with either antigen. the results suggest the selection in vitro of specific antigen-binding t cells, able to transfer delayed hypersensitivity.19852412403
[synthesis of virus-specific rna in influenza virus-infected cells in the presence of bonafton]. 19852412888
stable expression of cytolytic activity by influenza virus-specific cytotoxic t lymphocytes.influenza-specific immune cytotoxic t lymphocyte (ctl) populations maintain a constant level of in vitro cytolytic activity. this is demonstrable with both heterogeneous populations of anti-viral ctl from immune donors and long-term ctl clones derived from primed ctl precursors. cytolytic machinery is stably expressed by these ctl populations under a variety of in vitro cultivation conditions. this finding is in contrast to results with alloreactive ctl generated by stimulation of primed ctl pre ...19852413123
[neuraminidase antigenic erythrocytic diagnostic agent in determining antibodies to the neuraminidase of influenza a viruses]. 19852413631
virion proteins and the perspectives of gene manipulations in vaccine preparation.the achievements and perspectives of genetic manipulations are described aiming at preparation of first generation subunit vaccines based on the synthesis in bacterial and eukaryotic cells of full-sized virion proteins expressing the main antigenic determinants. the preparation of such vaccines in bacterial cells seems out of perspective in the case of influenza, human hepatitis b, foot- and - mouth disease and some other viruses due to the peculiarities of structure and synthesis as well as low ...19852413750
epizootics of respiratory tract disease in swine in belgium due to h3n2 influenza virus and experimental reproduction of disease.in belgium, influenza virus was isolated from swine in 22 epizootics of respiratory tract disease in swine during 1984. in 8 of the epizootics, h3n2 influenza virus, related to the a/port chalmers/1/73 strain, was isolated. intratracheal inoculation of the isolates induced clinical signs. it was concluded that the a/port chalmers/1/73 strain was established in the belgian swine population and was responsible for the epizootics of respiratory tract disease in swine.19852413789
antigenic variation in the hemagglutinin-neuraminidase protein of human parainfluenza type 3 virus.sixteen monoclonal antibodies directed to the hemagglutinin-neuraminidase (hn) protein of a 1957 isolate of parainfluenza type 3 virus (piv3) were produced and used to examine antigenic variation in clinical strains. analysis of hemagglutination-inhibition reactivity patterns of antigenic variants selected in vitro in the presence of monoclonal antibodies indicated that there were a minimum of six distinct epitopes detectable on the hn molecule. competitive-binding assays indicated that these ep ...19852414910
the antigenic structure of the influenza b virus hemagglutinin: operational and topological mapping with monoclonal antibodies.we have probed the antigenic structure of the influenza b virus hemagglutinin (ha) with monoclonal antibodies specific for the ha of influenza b virus, b/oregon/5/80. seventeen laboratory-selected antigenic variants of this virus were analyzed by hemagglutination-inhibition (hi) assays or elisa and an operational antigenic map was constructed. in addition, the monoclonal antibodies were tested in a competitive binding assay to construct a topological map of the antigenic sites. in contrast to th ...19852414911
characterization of host cell binding variants of influenza virus by monoclonal antibodies.we have previously shown that a plaque-type mutant of influenza virus a/wsn has a growth advantage in mdbk cells because its hemagglutinin (ha) has a greater affinity for host cell receptors than does the ha of the parent virus. we show here that the mutant is also less sensitive than the parent to neutralization by antibodies to epitopes in at least two regions on the ha. wsn-specific monoclonal antibodies which had higher radioimmunoassay (ria) titers against the parent than the mutant virus a ...19852414912
the neuraminidases of the virulent and avirulent a/chicken/pennsylvania/83 (h5n2) influenza a viruses: sequence and antigenic analyses.to define the sequence changes that occurred in an avian influenza virus neuraminidase (na) during the evolution of virulence, we have studied the na of the virulent and avirulent a/chick/penn/83 (h5n2) influenza viruses. a comparison of the deduced amino acid sequence from these viruses shows that the virulent strain, which evolved from the avirulent by the accumulation of point mutations (bean et al., 1985), acquired four amino acid changes in the na: one in the transmembrane segment, one in t ...19852414922
[outlook on the use of interferon and its inducers in preventing influenza]. 19852414924
[detection of a heterogeneous population of influenza virus type a/h1n1 with monoreceptor sera to hemagglutinin antigenic determinants].studies of the antigenic composition of hemagglutinins of influenza h1n1 virus variants isolated in 1981 using monoreceptor antibody to individual antigenic determinants obtained by the selective adsorption method showed some variants to represent heterogeneous populations manifesting antigenic properties of different h1n1 viruses. passages of the heterogeneous virus in the presence of antiinfluenza serum resulted in cloning a subpopulation with the antigenic properties of the hemagglutinin simi ...19852414925
biological and immunological properties of haemagglutinin and neuraminidase expressed from cloned cdnas in prokaryotic and eukaryotic cells.to study the biological and immunological properties of influenza virus surface glycoproteins, cdna copies of the haemagglutinin (ha) and the neuraminidase (na) genes of a/wsn/33 influenza virus were cloned and expressed in prokaryotic and eukaryotic cells. in escherichia coli, maximum expression of ha is obtained only as a fusion protein in which the nh2-terminal portion is provided by a bacterial protein (i.e. beta gal or trple'). the ha expressed in bacteria (bacterial ha) is recognized by po ...19852414936
expression of haemagglutinin gene.antigenic drift of the haemagglutinin (ha) molecule of type a influenza viruses has been thought to occur by the accumulation of a series of point mutations in the antigenically important regions of the molecule. in order to study the antigenic sites on the ha molecule, an attempt was made to use the site-specific mutagenesis method.19852414937
conformational changes in influenza virus haemagglutinin and its monomer detected by monoclonal antibodies.exposure of influenza virus haemagglutinin to ph 5 results in conformational changes occurring in the molecule which are accompanied by antigenic modifications. furthermore, isolated haemagglutinin (ha) at a concentration of 0.1 nm undergoes dissociation from the trimeric to a monomeric form when exposed to ph 5. whether present on intact virus or as the isolated monomer, each form of haemagglutinin from ph 5 exhibits similar alterations in antigenic characteristics. these forms of ha show modif ...19852414938
the influence of norakin on the reproduction of influenza a and b viruses.the action of the anticholinergic drug norakin (triperiden) on the reproduction of influenza virus a and b strains was studied. in cell culture, primary transcription of influenza a/fpv/weybridge and of influenza b/japan/73 is strongly inhibited by norakin (20 micrograms/ml). when present simultaneously, norakin and rimantadine exert an additive effect. rimantadine-resistant mutants of influenza a exhibit a definite, though limited, cross-resistance to norakin, and vice versa. the results indica ...19852415085
expression of h-2i region-restricted cytolytic activity by an lyt-2+ influenza virus-specific t lymphocyte clone.the expression of lyt-2 on t lymphocytes has been postulated to correlate closely with restriction by, or alloreactivity to, class i mhc gene products, whereas i region-restricted or alloreactive populations appear to be associated with lyt-1 and l3t4 expression. however, exceptions to this axiom among alloreactive t cells have been shown to exist. in this report we describe a clonal population of influenza virus-specific t lymphocytes that bears the lyt-2+, l3t4- phenotype. notably, this clone ...19852415581
the nucleoprotein as a possible major factor in determining host specificity of influenza h3n2 viruses.in an attempt to assess the importance of the nucleoprotein (np) in the determination of host specificity, a series of experiments was performed on influenza a viruses of the h3n2 subtype. we have examined rescue of mutants of a/fpv/rostock/34 with temperature-sensitive (ts) lesions in the nucleoprotein (np) gene by double infection of chick embryo cells with h3n2 strains isolated from different species. the ts mutants could be rescued by all avian h3n2 strains but not by any of the human h3n2 i ...19852416114
isolation and characterization of influenza c virus inhibitor in rat serum.two hemagglutination inhibitors for influenza c virus were isolated from pooled sera of normal rats by sequential chromatography on blue sepharose cl 6b, ultrogel aca 22, and deae-cellulose. the two inhibitors were identified as alpha 1-macroglobulin and murinoglobulin by comparison with the authentic samples. these inhibitors abolished the hemagglutination by influenza c virus strains but did not affect the hemagglutination by influenza a and b virus strains. hemagglutination inhibition activit ...19852416143
isolation and characterization of human t cell lines and clones reactive to rabies virus: antigen specificity and production of interferon-gamma.by using a preparation of inactivated rabies virus, the blood mononuclear cells from five rabies vaccine recipients were stimulated in vitro in the presence of interleukin 2. t cell lines that displayed significant proliferative responses to whole rabies virus and to preparations of rabies glycoprotein and nucleocapsid were obtained from all the individuals. other antigens, such as diphtheria and tetanus toxoids, influenza a virus, hepatitis b surface antigen, and serum albumin, failed to induce ...19862416820
practical considerations of the ability of monoclonal antibodies to detect antigenic differences between closely related variants.this report describes an investigation of the abilities of different immunoassays to detect differences in affinities between related antigens for particular monoclonal antibodies. nine different monoclonal antibodies were used and 6 strains of influenza virus represented closely related antigens. parameters defining assay sensitivity were estimated experimentally for each antibody in 4 different immunoassays. predicted failures of particular assays to detect differences in antigen affinities, b ...19852416849
the antiviral activity of tetrazole phosphonic acids and their analogues.5-(phosphonomethyl)-1h-tetrazole and a number of related tetrazoles have been prepared and their effects on the replication of herpes simplex viruses-1 and -2 have been investigated as well as their abilities to inhibit the dna polymerases induced by these viruses and the rna transcriptase activity of influenza virus a. contrary to an earlier report, 5-(phosphonomethyl)-1h-tetrazole was not an efficient inhibitor of the replication of hsv-1 and hsv-2 in tissue culture. analogues of 5-(phosphonom ...19852417198
loss of enzyme activity in a site-directed mutant of influenza neuraminidase compared to expressed wild-type protein.full-length double-stranded dna copies of the neuraminidase (na) gene of influenza virus a/tokyo/3/67 (n2) and a mutant generated in vitro by site-specific, oligonucleotide-directed mutagenesis with a substitution of leucine for tryptophan at position 178 were cloned into an sv40 late replacement expression vector. indirect immunofluorescence of cells infected with these recombinant vectors showed the presence of na protein in the cytoplasm and on the surface of infected cells. cells expressing ...19862417413
clinical applications of antiviral agents for chemophrophylaxis and therapy of respiratory viral infections.table iii summarizes clinical applications of antiviral agents in respiratory viral infections. (table: see text) for influenza a virus infections, both oral amantadine and rimantadine are effective when used for seasonal prophylaxis and for prophylaxis in institutional populations. both of these drugs, as well as aerosolized ribavirin, have antiviral and therapeutic effects in uncomplicated influenza. it remains to be determined whether any of these modalities or possibly their combined use [44 ...19852417551
the anticholinergic anti-parkinson drug norakin selectively inhibits influenza virus replication. 19852417553
selection of influenza a virus adsorptive mutants by growth in the presence of a mixture of monoclonal antihemagglutinin antibodies.the influenza virus hemagglutinin contains four major regions that are recognized by antibodies able to neutralize viral infectivity. to investigate the effect of an antibody response directed against each of these sites on viral evolution, influenza virus a/pr/8/34 (h1n1) was grown in allantois-on-shell cultures in the presence of a mixture of monoclonal antihemagglutinin antibodies. this selection mixture contained antibodies (two or three antibodies per antigenic site) whose concentrations we ...19862418215
abnormalities of the in vitro cellular and humoral responses to tetanus and influenza antigens with concomitant numerical alterations in lymphocyte subsets in down syndrome (trisomy 21).peripheral blood leukocytes (pbl) from noninstitutionalized individuals with trisomy 21, paired with closely age-matched and/or family members as controls, were analyzed for different aspects of their cellular and humoral immune responses, and were phenotypically characterized by means of various monoclonal antibodies. both the in vitro pbl proliferative and antibody responses to a bacterial antigen (tetanus toxoid) and to viral antigens (influenza a/bangkok and b/singapore) were significantly d ...19862419411
arrest of daudi cell growth by inactive influenza virus in-vitro.we investigated the destructive capability of three influenza a strains, victoria, pr8, and their recombinant x47, against the human lymphoma cell line daudi. both victoria and x47 strains share the same envelope glycoproteins (h3n2), while pr8, the second parental strain of x47, differs in its envelope glycoproteins (hon1). the h3n2 strains and particularly the x47 recombinant were cytotoxic to daudi cells while the hon1 strain was not. to reduce the virulence of the oncolytic viruses, we inact ...19852419962
[isolation of the influenza virus from the tree sparrow and a study of the infectivity of this virus in wild birds of the central dnieper river area].an influenza virus belonging to the serovariant a/h3n2 and registered as a/sparrow/ukraine/83 was isolated from a member of synanthropic birds, a tree sparrow, near kanev. this virus showed low pathogenicity and immunologic activity in experimental infection of sparrows and other birds. sera from a number of avian and mammal species had antibodies to this virus which indicates that synanthropic and semi-synanthropic birds may be a connecting link in spread of influenza virus.19852420068
biological characteristics of a cold-adapted influenza a virus mutation residing on a polymerase gene.the biological function of a cold-adapted (ca) mutation residing on the pb2 gene of an influenza a/ann arbor/6/60 (a/aa/6/60) ca variant virus in the viral replication cycle at 25 degrees c was studied. the viral polypeptide synthesis of a/aa/6/60 ca variant at 25 degrees c was evident approximately 6 hours earlier than the wild type (wt) virus and yielded twice as many products. the quantitative analysis of viral complementary rna (crna), synthesized in the presence of cycloheximide, revealed t ...19862420313
in vitro biologic responses to native and surface-modified asbestos.a comparative study was made of in vitro biologic responses to native chrysotile, amosite, and crocidolite and corresponding asbestos fibers whose surfaces were modified by metal oxides. interferon induction by influenza virus was depressed by approximately 50% by all native asbestos whereas corresponding surface modified asbestos minimally affected this nonspecific cellular defense mechanism. the release of the cytoplasmic enzyme, lactate dehydrogenase (ldh), and lysosomal enzymes, beta-n-acety ...19862420583
conformational-dependent recognition of influenza virus hemagglutinin by murine t helper clones.a panel of h-2k class ii-restricted t helper (th) clones were established from individual cba mice intranasally infected with a/x31 influenza virus. in a study of their fine specificity, it was revealed that recognition of hemagglutinin (ha) by certain th clones was dependent on conformational features of the molecule. five ha-specific th clones failed to recognize x31 mutant viruses r19 and r20 each with a his to arg substitution at position 17 of ha1. this amino acid substitution affects the c ...19862420611
influence of antigen structure on the activation and induction of unresponsiveness in cloned human t lymphocytes.using cloned human helper t lymphocytes reactive with a 24 amino acid peptide (p20) of the carboxyl terminal of the ha-1 molecule of influenza haemagglutinin we have investigated the influence of antigen structure on the activation and in the induction of antigen specific unresponsiveness of t cells. for this analysis stereoisomers and structural isomers of p20 have been constructed. p20 in the form of a single loop created by a disulphide based between residue 306 and an additional cysteine at ...19862420700
peptide maps of t mutant of influenza virus.peptide maps were prepared by high-voltage electrophoresis and chromatography for t 30 degrees c mutant and the original strain a port chalmers 1/73 h3n2 of influenza virus. clear differences in the distribution and number of spots between the mutant and the original strain were found. the differences are due presumably to changes in the amino-acid sequences.19852421538
the influence of isoprinosine (iso) on some cell immunity parameters in mice infected with influenza viruses and in mixed viral-bacterial infections. i. the release of the migration inhibiting factor from spleen leukocytes.the influence of iso, the antiviral drug of immunomodulating activity, on the course of experimental influenza infections and mixed, viral-bacterial infections was studied. spleen leukocytes migration inhibition test, performed in vitro in the presence of specific antigens stimulating influence of the drug administered to the infected animals was observed.19852421684
an influenza a virus-specific and hla-drw8-restricted t cell clone cross-reacting with a transcomplementation product of the hla-dr2 and dr4 haplotypes.the clone ta10 is a t3+ t4+ t8- proliferative and cytolytic human t cell clone. this clone has been shown to be specific for the hemagglutinin of influenza a texas virus and restricted by an hla class ii molecule associated with the drw8-dw8.1 phenotype. here we show that ta10 and all of its subclones can also react with eight hla-drw8 negative, epstein-barr virus (ebv)-transformed cell lines or phytohemagglutinin blasts in the absence of influenza antigens. all of these cell lines are hla-dr2/d ...19862422266
protection of mice from lethal influenza: evidence that defective interfering virus modulates the immune response and not virus multiplication.intranasal administration of defective interfering (di) influenza virus (a/wsn) ensured the survival of 80% of c3h mice otherwise lethally infected with wsn by the intranasal route, whereas a control group which received beta-propiolactone-inactivated di virus in place of di virus died at 7.4 days post-infection. di virus-treated mice developed significantly less lung consolidation than controls although qualitatively the cellular pathology in the two groups was indistinguishable. surprisingly, ...19862422317
[the nature of amino acid substitution in antigenic drift of hemagglutinin n3 and neuraminidase n2 from the influenza virus].the nature of amino acid replacements in 16 drift variants of hemagglutinin h3 subtype and 5 drift variants of neuraminidase n2 subtype of the influenza a virus were studied. the dependences of relative replacement frequencies and relative quantities of frequent replacements upon differences of properties of substituted residues are plotted. in contrast to most of the known proteins, amino acid replacements in hemagglutinin and neuraminidase depend weakly on the physico-chemical parameters of am ...19862422540
analysis of antigenic determinants on internal and external proteins of influenza virus and identification of antigenic subpopulations of virions in recent field isolates using monoclonal antibodies and immunogold labelling.an electron microscopic immunogold labelling technique employing monoclonal antibodies has been applied to the antigenic analysis of influenza a and b viruses. reassortant influenza a h3n2 viruses containing haemagglutinin molecules from viruses isolated between 1968 and 1982 were analysed with a panel of monoclonal antibodies raised against viruses which appeared over the same period. the immunogold labelling technique clearly demonstrated the antigenic drift in the haemagglutinin molecule that ...19862423056
rapid rna sequencing using double-stranded template dna, sp6 polymerase, and 3'-deoxynucleotide triphosphates.a simple and efficient nucleic acid sequencing method is described in which rna transcription by the sp6 polymerase is specifically terminated using 3'-deoxynucleotide triphosphates. initial difficulties in resolving the rna ladder were overcome by replacing guanosine triphosphate by inosine triphosphate in the reaction mixture and electrophoresing gels at high temperature (50 degrees c). this method presents advantages over current sequencing techniques: unprocessed plasmid dna is the template ...19862423310
extensive diversity in the recognition of influenza virus hemagglutinin by murine t helper clones.a panel of h-2k class ii-restricted th clones were established from individual cba mice primed by infection with x31 influenza virus. 27 clones, which showed specific recognition of the ha surface glycoprotein, were all h3n2 subtype specific, in contrast to a t cell line which was crossreactive and which may have other specificities. 20 distinct ha-specific clones recognized a tryptic cleavage fragment of x31 consisting of residue 28-328 of ha1 (tops) which includes all the ab-combining regions ...19862423631
antigenic mapping of an avian h1 influenza virus haemagglutinin and interrelationships of h1 viruses from humans, pigs and birds.monoclonal antibodies to the haemagglutinin (ha) of the avian h1 influenza virus a/duck/alberta/35/76 were used to construct an operational antigenic map of the ha molecule and to study the interrelationships of h1 viruses from different hosts. haemagglutination inhibition tests between the monoclonal antibodies and variants selected by them provided evidence of four antigenic regions which overlap to varying degrees. avian h1 influenza viruses displayed a spectrum of reactivities to the monoclo ...19862423640
[structural rearrangements in influenza virus hemagglutinin during crossing the inter-species barrier]. 19862424700
functional definition of hla-dr and -dq epitopes specific for dr2-short, dwfjo haplotype.t-lymphocyte clones specific for the influenza a/texas virus were obtained by limiting dilution of activated t cells from an hla a2/3,b7/39,cw -/-,dr2-short/2 short,dqw1,dwfjo/fjo donor. among the proliferating clones studied, and irrespective of their antigenic specificities, most of them were restricted by epitope(s) on hla-dr molecules present only on dr2-short/dwfjo cells but not on dr2-negative or dr2-long positive (dw2,dw12,dw-) cells. two clones were restricted by epitopes borne by dq pro ...19862424833
fine specificity and antigen receptor expression among influenza virus-specific cytolytic t lymphocyte clones.influenza virus stimulates a vigorous cytolytic t lymphocyte (ctl) response in the mouse that is directed to several virion polypeptides. this report examines the fine specificity of a panel of murine influenza-specific ctl clones restricted by mhc class i products of the h-2d haplotype. ten of 22 a/japan/305/57-specific ctl clones analyzed were directed to the a/japan/305/57 hemagglutinin protein as detected by using target cells infected with a recombinant vaccinia virus containing hemagglutin ...19862424996
[radioimmunological analysis of viral antigens on a flat adsorbent of polyethylene film]. 19862425137
[use of monospecific antibodies for determining the antigenic determinant composition in the hemagglutinins of the h3n2 influenza viruses of 1979-1983].the study of the antigenic composition of hemagglutinins of influenza h3n2 viruses isolated during epidemic outbreaks in 1979-1983, carried out by means of monospecific antibodies to individual antigenic determinants, showed heterogeneity of the antigenic structure of epidemically active variants. some variants were antigenically identical to a/bangkok/1/79 virus. hemagglutinin structure of a/shanghai/31/80, a/philippines/2/82 and the like was found to contain a qualitatively new antigenic deter ...19862425492
[antigenic markers of the hemagglutinins of the influenza a and b viruses in the process of variability and the rapid diagnosis of influenza]. 19862425495
[antigenic drift of influenza a1 virus]. 19852425981
influenza nucleoprotein-specific cytotoxic t-cell clones are protective in vivo.influenza nucleoprotein (np) serves as a target antigen on abortively infected cells for cytotoxic t cells (tc) cross-reactive for all type a influenza viruses, and it can also prime mice for such tc. it is important to test the protective ability of np-specific tc clones in vivo in a productive influenza infection. in this report, we show that tc clones of this antigenic specificity protect mice against a lethal influenza infection on transfer to syngeneic recipients, and also that they reduce ...19862426185
an antigen-binding assay to determine the specificity of monoclonal antibodies against influenza virus and mapping of epitopes.a simple antigen-binding assay is described for determining the specificity of monoclonal antibodies against a/victoria/3/75 influenza virus. monoclonal antibodies were bound to polyvinylchloride microtitre wells via protein a and anti-immunoglobulin serum. radiolabelled cell extracts were then added and allowed to adsorb to the antigen. the bound material was eluted and analyzed by polyacrylamide gel electrophoresis and autoradiography. the method can also be used to study competitive binding o ...19862426296
characterization of monoclonal antibodies specific for sequential influenza a/pr/8/34 virus variants.a panel of monoclonal antibodies specific for a corresponding panel of sequentially selected variants of influenza a/pr/8/34 virus has been established. although the monoclonal antibodies are paratypically distinct, idiotypic relatedness has been observed. two cross-reactive idiotypes have been defined that are associated with the 7183 and s107 vh gene families, respectively. three of the four monoclonal antibodies utilize the vk21 group of light chains, and three vh genes belong to the vh7183 f ...19862426361
influenza viral glycoproteins induced cell-mediated cytotoxicity by an interferon-independent mechanism.human natural killer (nk) cells exposed to the influenza surface antigen neuraminidase (na) show high cytotoxic activity, as evaluated using chromium-labeled k562 target cells in a standard overnight cytotoxicity assay. the role of interferon (ifn) in the stimulation of nk cells was examined by using three separate approaches. the use of appropriate antibodies to check ifn- and na-specific cell-mediated cytotoxicity (cmc) stimulation showed that antibodies to ifn (-alpha, -gamma) did not alter n ...19862427216
structural and functional implications of a restricted antibody response to a defined antigenic region on the influenza virus hemagglutinin.a group of hybridoma antibodies that recognize structurally overlapping epitopes on the influenza virus hemagglutinin have been analyzed for the sequence of their immunoglobulin heavy and light chain variable regions. all vh regions derive from the same gene family, and only two vk genes, from different families, are involved. the repetitive and restricted use of these variable region genes indicates that considerable structural requirements influence the generation of antibodies specific for th ...19862427335
[dynamics of change in the antibacterial activity of mouse peritoneal cells during the development of experimental influenzal infection].the method of the radiometric determination of the antibacterial activity of peritoneal cells in the process of the development of viral infection has been approved. the genetically determined specific level of the antibacterial activity of these cells in different strains of mice has been established. the development of the lethal form of experimental influenza a in mice has been shown to be accompanied by the mobilization of nonspecific protective factors, including the system of interferons a ...19862428190
the molecular basis of antigenic variation in influenza virus. 19862428216
immunogenicity of loop-structured short synthetic peptides mimicking the antigenic site a of influenza virus hemagglutinin.in order to assess the relevance of conformation for the antigenic site a of the hemagglutinin of influenza virus we synthesized two peptides, comprising two variant sequences of the central part of site a (amino acids 140 - 146 of subunit ha1) inserted into an artificial peptide skeleton, which imposes a loop-like structure on the respective sequence stretch. assuming that the loop structure in the synthetic peptides would roughly approximate to the structure of the cognate protein sequence we ...19862428615
highly polymorphic products of both hla-dr and hla-dq genes contribute to the polymorphism of the hla-drw13 haplotype.we studied the polymorphisms of hla-dr and hla-dq products from hla-drw13 haplotypes by analyzing the restriction of influenza a-specific cloned t cells from an hla-drw13,dqw1,dw19 homozygous individual. the results show that some functional epitopes, which can be borne by either hla-dr or hla-dq molecules, are strictly correlated with the hla-dw19 subtype of hla-drw13. this clearly indicates that both hla-dr and hla-dq products contribute to the hla-dw19 subdivision of hla-drw13. at least two d ...19862428744
[immunological study of antigenic determinants in the isolated hemagglutinin of the influenza a virus (h1n1)].a preparation of isolated subunits of influenza a (h1n1) virus hemagglutinin was examined for antigenic properties by four serological tests: radioimmunoassay, radial immunodiffusion test, complement fixation test, and hemagglutination inhibition test with hyperimmune polyclonal serum to intact virion as well as with monospecific antibodies to individual antigenic determinants obtained by adsorption technique. in isolated hemagglutinin subunits, radioimmunoassay identified three main virus-speci ...19862429447
[genomic and antigenic analysis of strains of influenza a virus (h1n1) isolated in 1982-1983: approaches to the choice of the optimal vaccine strain].features of the genome and antigenic specificity of hemagglutinin of some influenza a (h1n1) virus strains circulating in the epidemic period of 1982-1983 were studied comparatively. analysis of the genome of the isolates under study in comparison with that of the reference a/england/333/80 strain and with each other has established changes not only in the genes coding for hemagglutinin and neuraminidase but also most of the genes coding for unglycolysed proteins. the antigenic specificity of he ...19862429448
[synergic therapeutic action of aprotinin and remantadine in experimental influenzal infection].chemotherapeutic treatment of influenza infection is possible with aprotinine, an inhibitor of proteinases blocking proteolytic shearing of virion hemagglutinin which is necessary for multicycle virus infection. the chemotherapeutic effect of antiinfluenza drug remantadine is due to inhibition of intracellular deproteinization of virions in the course of their penetration into cells. in mice infected with a highly lethal dose (about 400 mld50/animal) of influenza a/aichi/2/68 (h3n2) virus treatm ...19862429449
enhancement of histamine release from human basophils pretreated with different sialidases.histamine release from human basophils was investigated in vitro after removal of cell membrane sialic acid by three different sialidases. pretreatment of the cells with sialidases from cl. perfringens, v. cholera or influenza virus a2 enhanced histamine release induced by subsequent stimulation of the cells with anti-ige or the plant lectin concanavalin a and caused a shift to the left of the dose-response curve for anti-ige. the enhanced histamine release was reflected in a increased calcium s ...19862429528
laboratory characterization of influenza a viruses (h3n2)--pathogens of leningrad 1983 epidemic.seventy strains of influenza a virus (h3n2) isolated in leningrad during the influenza epidemic in january--march 1983 were studied. the virus isolates appeared to be markedly heterogeneous with respect to antigenic characteristics of surface glycoproteins, biological properties (haemagglutinating and neuraminidase activities, sensitivity to gamma-inhibitors, etc.) as well as genome structure. the identity of the virus isolates with the reference influenza a virus was as follows: 10% were simila ...19862429534
assembly of influenza hemagglutinin trimers and its role in intracellular transport.the hemagglutinin (ha) of influenza virus is a homotrimeric integral membrane glycoprotein. it is cotranslationally inserted into the endoplasmic reticulum as a precursor called ha0 and transported to the cell surface via the golgi complex. we have, in this study, investigated the kinetics and cellular location of the assembly reaction that results in ha0 trimerization. three independent criteria were used for determining the formation of quaternary structure: the appearance of an epitope recogn ...19862429970
hla b37 determines an influenza a virus nucleoprotein epitope recognized by cytotoxic t lymphocytes.human influenza a virus-specific, cytotoxic t cells have been shown previously to recognize the virus nucleoprotein on infected cells. ctl preparations from four hla b37-positive donors were shown to recognize a synthetic peptide that corresponded to amino acids 335-349 of the nucleoprotein sequence. influenza-specific ctl from 10 donors of other hla types failed to recognize this epitope. cd8+ ctl lines were derived from lymphocytes of two hla b37-positive donors and used to show that the pepti ...19862430041
[influenza a(h1n1) virus drift; alteration of the influenza vaccine for the 1986/'87 season]. 19862430197
synthesis of interferon in human leukocyte cells stimulated with influenza virus.strains of influenza a virus were tested for their ability to induce interferon synthesis in suspension of human leukocytes. both, active as well as uv-inactivated strains were shown to be weak interferon inducers as compared to newcastle virus. it appeared that interferon synthesis in human leukocytes was conditioned not by infectivity of the virus but by the presence of its surface antigens--hemagglutinin and neuraminidase.19862430545
influenza virus infection induces functional alterations in peripheral blood lymphocytes.this report describes alterations in functional responses to lectin-induced stimulation of peripheral blood lymphocytes and in the natural killer cell (nkc) activity, of college students, obtained during an outbreak of influenza a/philippines/2/82(h3n2) virus infection. these results are compared with similar observations in college students with an acute, febrile, noninfluenzal respiratory illness that occurred during the same outbreak. the lymphopenia typical of influenza during acute illness ...19862431043
influenza virus-induced encephalopathy in mice: interferon production and natural killer cell activity during acute infection.mice injected intracerebrally with infectious influenza virus (60 hemagglutinin units) developed lethargy, seizures, comas, and died 2 to 5 days postinfection. as early as 6 h after infection, the cerebrospinal fluid (csf) in these animals was infiltrated with polymorphonuclear cells, mononuclear leukocytes, and large granular lymphocytes. potent natural killer (nk) cell activity was observed for both csf and spleen cell populations over the same period. this nk cell activity correlated with int ...19862431159
[effect of influenza virus type a and interferon on various pathogenetic factors in bronchial asthma]. 19862431398
antigenic determinants of influenza virus hemagglutinin. xii. the epitopes of a synthetic peptide representing the c-terminus of ha1.a synthetic peptide comprising the c-terminal 24 amino acids of the heavy chain (ha1) of influenza virus hemagglutinin was constructed and examined for antigenic and immunogenic activity. monoclonal antibodies as well as polyclonal antisera raised against the synthetic peptide were able to bind to intact virus. this binding was greatly enhanced if the virus was first subjected to ph 5, suggesting that this treatment exposes the c-terminus of ha1. using synthetic analogs of the native sequence it ...19862431541
[the biological effects of coordination compounds of transitional metals. 6. effect of 4-methyl-2-aminopyridine-palladium chloride and cis-dichlorodiammine-platinum(ii) on retroviruses and the virus-associated rna polymerase of the influenza virus].the effect on retroviruses of two transition metal complexes of known antiviral activity, 4-methyl-2-amino-pyridine-palladium-chloride (map) and cis-dichloro-diammine-platinum(ii) (cis-ddp) has been investigated. the experiments included the evaluation of the action of compounds on virus particle-associated reverse transcriptase in exogenous assays, on virus propagation in persistently infected cell cultures and on virus infectivity in mice. in disrupted viruses and in the absence of excess prot ...19862431560
[molecular basis of the variability of epidemic strains of human influenza viruses].the data from literature and authors own studies are reviewed on variability of human influenza viral strains, isolated during the same epidemic season in different periods of pandemic cycle. the data obtained indicate that variability of epidemic strains of human influenza virus deals with the genes coding for outer membrane proteins (hemagglutinin and neuraminidase) as well as nonglycosylated proteins. circulation of a number of viral variants of the same serotype, differing in antigenic speci ...19852432423
characterization and in vivo distribution of influenza-virus-specific t-lymphocytes in the murine respiratory tract.cloned populations of antigen-specific, murine t-lymphocytes, maintained in long-term continuous culture, have been reported to exhibit functional activity upon adoptive transfer. few data are currently available on the in vivo distribution of these homogeneous, functional cell populations. such information is important to understanding the mechanisms by which t-lymphocytes exert their effector activity. we examined 2 cloned populations of influenza virus-specific t-lymphocytes. one clone was a ...19872432811
enhancement of host resistance against virus infections by mtp-pe, a synthetic lipophilic muramyl peptide--i. increased survival in mice and guinea pigs after single drug administration prior to infection, and the effect of mtp-pe on interferon levels in sera and lungs.muramyl tripeptide-phosphatidylethanolamine (mtp-pe, cgp 19835) displays prophylactic antiviral activity in mice infected with influenza viruses a and b, parainfluenza 1 virus or herpes simplex type 1 viruses (hsv/1) and in guinea pigs infected with herpes simplex type 2 viruses (hsv/2). mtp-pe is effective when given in a single intranasal dose as early as 1-4 weeks before infection. in the case of hsv/2 infections, prophylactic effectiveness can be demonstrated after a single topical applicati ...19862433236
monoclonal antibodies for characterizing h3n2 influenza viruses that persist in pigs in china. 19872433358
the lack of effect of inoculation with equine influenza vaccine on theophylline pharmacokinetics in the horse.several studies conducted during the past few years have shown that the pharmacokinetics of a variety of drugs may be altered following viral infection or vaccination. the elimination of drugs which are extensively metabolized, such as theophylline, may be prolonged, especially following exposure to rna viruses such as type a influenza or similar orthomyxoviruses. the purpose of this study was to determine whether vaccination of horses with equine influenza virus affected pharmacokinetic paramet ...19862433461
functional and antigenic domains of the matrix (m1) protein of influenza a virus.the membrane- and ribonucleocapsid (rnp)-binding domains of the matrix (m1) protein of influenza a virus (wsn strain) were partially mapped and characterized by reactivity with monoclonal antibodies (mab) as well as by proteolytic cleavages and amino acid sequencing of the resulting peptides. of two peptides formed by formic acid hydrolysis, a 9-kilodalton fragment at the amino-terminal third of the m1 protein was recognized by mab m2-1c6 (to epitope 1), and a 15-kilodalton fragment at the carbo ...19872433462
a single amino acid substitution in influenza hemagglutinin abrogates recognition by monoclonal antibody and a spectrum of subtype-specific l3t4+ t cell clones.a fine specificity analysis of influenza hemagglutinin-specific iak-restricted t cell clones using natural virus variants of the h3n2 subtype, monoclonal antibody-selected variants and a synthetic peptide corresponding to a variable region of the ha1 polypeptide has provided insight on the structural basis for t cell recognition. a glycine to arginine substitution at ha1 135 abrogates recognition by a panel of t cell clones which, according to their reactivity for natural virus variants, have di ...19872434337
analysis by plaque reduction neutralization assay of intertypic rotaviruses suggests that gene reassortment occurs in vivo.the sb-1a rotavirus recovered from a diarrheic piglet in the united states is a naturally occurring intertypic rotavirus. when studied by reciprocal neutralization tests, the sb-1a virus was similar, if not identical, to the porcine gottfried virus (serotype 4) and the porcine osu virus (serotype 5). analysis of reassortant viruses prepared from the sb-1a virus and the serotype 2 human ds-1 virus revealed that the antigenic specificity of the outer capsid protein vp3 of sb-1a was shared with the ...19872434522
high protection of animals lethally infected with influenza virus by aprotinin-rimantadine combination.the successful therapeutic synergism of aprotinin and rimantadine, which are known to attack different viral targets, was demonstrated in influenza-virus-infected animals. combined treatment with these drugs of mice infected with a highly lethal dose of mouse-adapted influenza virus prevented the development of fatal haemorrhagic pneumonia and protected about 75% of animals; whereas the separate administration of aprotinin and rimantadine induced 35% and 15% protection, respectively.19872434613
hemagglutination-inhibiting antibodies to equine influenza viruses in japanese horses and antigenic variation of the viruses. 19862435948
[comparative study of interferon inducers obtained from natural sources].interferon-inducing and antiviral effects of natural dsrna preparations of phage phi 6 and yeast cells were studied in the culture of murine cells l-929 and on random bred albino mice. both the preparations showed interferon inducing activity in the cell culture. however, for realization of their effect modification of the surface cell membrane by polycation exchange resin (deae-dextran) was required. the interferon-inducing activity of both of the natural dsrna in the mice was high. the maximum ...19872436574
modulation of t cells by cyclic amp in murine influenza virus infection. i. in vitro inhibition of cytotoxic t lymphocytes by exogenous cyclic amp analogues and by agents which increase intracellular cyclic amp concentrations.the effects of 8-bromo-cyclic amp and cyclic amp agonists (cholera toxin plus hydrocortisone and prostaglandin e2 plus 3-isobutyl-1-methylxanthine) on the cytotoxic activity of t cells generated during murine influenza virus infection have been examined. treatment of influenza a/wsn virus primed primary immune spleen cells and their cultured secondary effector t cells from balb/c mice with these agonists resulted in increases in cyclic amp levels. these agents also had a marked inhibitory effect ...19872437075
[study of the host glycosyl component of the influenza virus using monoclonal antibodies].the crossreacting glycosylated host component (cohc) of influenza virus propagated in chick embryo has been studied using the monoclonal antibodies (ma). the specificity of ma to cohc has been demonstrated by different methods. both ma of clones a9 and b7 react with the spatially overlapping antigenic sites of oligosaccharide chain, but only ma of b7 are active in hemagglutination inhibition test. the cohc is shown to be associated with both surface glycoprotein adsorbing on the surface of the v ...19872437446
cytotoxic t lymphocytes recognize a fragment of influenza virus matrix protein in association with hla-a2.both human and murine cytotoxic t cells (ctl) elicited in response to infection with influenza a viruses have been shown to be specific for internal viral proteins, such as the matrix and nucleoprotein. individual ctl epitopes have been identified in the nucleoprotein by successfully substituting short synthetic peptides for the intact virus in the preparation of target cells in cytotoxicity assays. the defined peptide epitopes have each been recognized by ctl in association with individual clas ...19872437457
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