Persistence of immunity to infectious canine hepatitis using a killed vaccine.
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Biomedical subjects
Publications and source records attributed to I G Furminger.
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Influenza virus was centrifuged in a KII rotor through a sucrose gradient containing Triton N101, a non-ionic surfactant. The micelles of surfactant formed a band in the gradient. As virus particles passed through the surfactant, the haemagglutinin and neuraminidase proteins were stripped from the surface and remained near the surfactant micelles. The residual virus particles sedimented into a denser region of the gradient and were thus separated from the haemagglutinin and neuraminidase antigens. Fractions containing the surface antigens were pooled and Triton was removed by phase-separation at the cloud point.
Conventional influenza vaccine containing whole virus particles purified on a zonal centrifuge is pyrogenic and can cause systemic and local adverse side effects. An improved vaccine was therefore prepared which contained only the surface antigens of the virus adsorbed to aluminium hydroxide. The antigenicity of this vaccine was compared with conventional vaccine in chickens. Both vaccines induced similar titres of serum haemagglutination-inhibition and neuraminidase inhibition antibody. The dose response curves, however, were different. The surface antigens at vaccine strength without aluminium hydroxide were of negligible pyrogenicity in rabbits.
Arcton 113 has proved effective in the removal of host membrane fragments which cosediment with influenza virus during differential centrifugation. The virus appeared morphologically undamaged by the procedure and retained its infectivity, neuraminidase, and hemagglutinating activity. Pyrogenicity, however, was considerably reduced.
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Chicks vaccinated with live Hitchner B1 Newcastle disease vaccine at 17 days old and subsequently re-vaccinated with an oil emulsion killed Newcastle disease vaccine at either 38 or 52 days old showed high and persistent HAI antibody levels for at least eight months. Re-vaccination of these birds at 17 weeks old caused a further rise in antibody level to log212 which, even at 38 weeks, had dropped only to log210. Chicken primarily vaccinated with oil emulsion killed vaccine at six weeks old developed HAI antibody levels after four to five weeks of log29 which re-vaccination four weeks later increased to log211. Chicken given killed aluminium hydroxide adjuvant Newcastle disease vaccine were serologically HAI negative 13 weeks after vaccination while those given the oil emulsion vaccine still showed an antibody level of log28. Groups of birds inoculated with oil emulsion vaccine and then, at 20 weeks old, challenged with virulent Newcastle disease showed a 100 per cent survival rate. The particular merits of oil emulsion killed Newcastle disease vaccine for laying and breeding birds are discussed.
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Ferrets were vaccinated with either A/England/42/72 zonal purified influenza queous vaccine containing whole virus particles, or a subunit vaccine containing the purified hemagglutinin and neura-minidase proteins of A/England/42/72 adsorbed onto aluminium hydroxide. The immune status of the ferrets was assessed by reaction to challenge with virulent live virus of the homologous strain. In the preliminary experiment reported, the subunit vaccine conferred greater protection than the zonal purified vaccine.
At present influenza vaccines are standardized on their haemagglutinin content only. Recently it has been shown that both neuraminidase and haemagglutinin antibodies are important in providing protection against the influenza virus. We have, therefore, developed an automated neuraminidase assay, based on the enzymic method of Kendal, but modified to minimise the interference by sucrose. A neuraminidase standard has been established and samples assayed against the standard have a coefficient of variation of plus or minus 6.8 percent. The assay has been adapted to measure the neuraminidase antibody level in serum. The neuraminidase to haemagglutinin ratio has been determined for various influenza strains. Both neuraminidase and haemagglutinin titres are being compared with results obtained by the single radial diffusion method.
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Influenza virus may be precipitated and aggregated by polyethylene glycol into clusters comprising ten to many hundred virions per aggregate. These aggregates are sparingly soluble and may be freed of contaminating polymer by washing in the appropriate buffer at room temperature or by precipitation in 30% (v/v) ethanol at subzero temperatures.Immunogenic studies in guinea-pigs of the virus in different states of dispersion revealed that the aggregated virus is the superior antigen to the virus in the monomeric form or in the dissociated state following treatment with ether.
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