ABSTRACT
Inactivated (whole-virion, split, subunit, and adjuvanted) vaccines and live attenuated vaccine were tested in parallel to compare their immunogenicity and protective efficacy. Homologous and heterosubtypic protection against the challenge with influenza H5N1 and H1N1 viruses in a mouse model were studied. Single immunization with live or inactivated whole-virion H5N1 vaccine elicited a high level of serum antibodies and provided complete protection against the challenge with the lethal A/Chicken/Kurgan/3/05 (H5N1) virus, whereas application of a single dose of the split vaccine was much less effective. Adjuvants increased the antibody levels. Addition of the Iso-SANP adjuvant to the split vaccine led to a paradoxical outcome: it increased the antibody levels but reduced the protective effect of the vaccine. All tested adjuvants shifted the ratio between IgG1 and IgG2a antibodies. Immunization with any of the tested heterosubtypic live viruses provided partial protection against the H5N1 challenge and significantly reduced mouse mortality, while inactivated H1N1 vaccine offered no protection at all. More severe course of illness and earlier death were observed in mice after immunization with adjuvanted subunit vaccines followed by the challenge with the heterosubtypic virus compared to challenged unvaccinated animals.
Subject(s)
Immunity , Influenza A Virus, H1N1 Subtype/immunology , Influenza A Virus, H5N1 Subtype/immunology , Influenza Vaccines/pharmacology , Influenza, Human/immunology , Influenza, Human/prevention & control , Vaccines, Inactivated/immunology , Adjuvants, Immunologic/pharmacology , Animals , Humans , Influenza A Virus, H1N1 Subtype/isolation & purification , Influenza A Virus, H5N1 Subtype/isolation & purification , Influenza, Human/virology , Mice , Mice, Inbred BALB C , VaccinationABSTRACT
Macroporous glasses, chemically modified by tris-hydroxymethyl-aminomethane, were used for gel-permeation chromatography of tick-borne encephalitis (TBE) virus suspension. The highest yield of purified virus was observed in the carriers containing not less than 0.01 micrograms-equivalent/m2 surface-attached tris-hydroxymethyl-aminomethane groups at pH of the eluent higher than 7.6. The obtained modified porous carriers appeared to be suitable for the purification of both infectious and inactivated preparations of TBE virus.
Subject(s)
Encephalitis Viruses, Tick-Borne/isolation & purification , Animals , Brain/microbiology , Cells, Cultured , Encephalitis Viruses, Tick-Borne/growth & development , Encephalitis, Tick-Borne/microbiology , Glass , Humans , Hydrogen-Ion Concentration , Kidney , Kinetics , Mice , Swine , TromethamineABSTRACT
The structure and properties of infectious and formaldehyde-treated particles of tick-borne encephalitis virus, concentrated and purified by chromatography on macroporous glass, were studied. In addition to complete virions, such preparations contain some incomplete forms that differ in density, morphology and protein composition (incomplete forms do not contain nucleocapsid protein). The physico-chemical analysis of complete virions showed that formaldehyde treatment causes a) the formation of glycoprotein dimers and b) a portion of nucleocapsid protein to become tightly cross-linked with viral RNA. Formaldehyde treatment of incomplete forms resulted only in the formation of a small amount of glycoprotein dimers. Incomplete forms and glycoprotein extracted from inactivated preparations had protective and antigenic activity.
Subject(s)
Encephalitis Viruses, Tick-Borne/drug effects , Formaldehyde/pharmacology , Virion/drug effects , Antibodies, Viral/biosynthesis , Antigens, Viral/immunology , Capsid/analysis , Centrifugation, Isopycnic , Electrophoresis, Polyacrylamide Gel , Encephalitis Viruses, Tick-Borne/analysis , Encephalitis Viruses, Tick-Borne/immunology , Isoelectric Point , RNA, Viral/analysis , Viral Proteins/analysis , Viral Proteins/immunology , Viral Structural ProteinsABSTRACT
Tissue culture rabies virus was purified by gel chromatography on chemically modified macroporous glasses (MPG). Modification of the MPG surface by egg or human albumin prevented rabies virions from interaction with the glass surface so that all purified virus was eluted from the column. Chemically modified MPG could be used on a preparative scale for the purification of both infectious and inactivated rabies virus, the immunogenicity of the latter having been preserved. Protein content of the purified virus fractions was up to 4 microgram/ml, of which virus protein represented up to 5%.
Subject(s)
Rabies virus/isolation & purification , Antigens, Viral/isolation & purification , Chromatography, Gel , Ovalbumin , Rabies virus/immunology , Serum Albumin , Silicic AcidABSTRACT
The capacity of some chemically treated porous glass to adsorb tissue culture virus of tick-borne encephalitis was studied. Chemical binding of albumin and succinanhydride with the porous glass matrix was found to eliminate virion adsorption on the carrier surface. This permitted to carry out gel filtration chromatography of several virus strains and to obtain highly purified preparations of tick-borne encephalitis virus. Gel filtration on modified porous glass with different pore sizes was used for the determinations of cromatographic radius of virus particle.
