Controlled organization of multimolecular complexes of enveloped virus glycoproteins: study of immunogenicity.

Some technological and immunological problems facing the preparation of subunit viral vaccines are discussed. Solubilization of enveloped virus glycoproteins with various detergents has been studied. It has been demonstrated that a novel non-ionic detergent, MESK, can be used to prepare the glycoproteins of enveloped viruses in defined supramolecular forms: monomers, micelles, liposomes and multimeric complexes. These preparations have been tested for immunogenicity. It has been shown that the immunogenicity of glycoproteins in micellar form or in liposomes is comparable with that of the whole virus. The immunogenicity of the glycoprotein complex with the glycoside Quil A appeared to be significantly higher in comparison with the whole virus and was similar to the immunogenicity of glycoproteins mixed with Freund's complete adjuvant.

[Inhibition of the infectivity of enveloped viruses while retaining the specific biological activity of the viral proteins]. / Podavlenie infektsionnosti obolochechnykh virusov s sokhraneniem spetsificheskoi biologicheskoi aktivnosti virusnykh belkov.

A method of inactivation of enveloped viruses by treatment of virus-containing material with a nonionic detergent MESK was studied. Treatment with the detergent of allantoic or culture fluids containing influenza, parainfluenza, herpes simplex, and Venezuelan equine encephalomyelitis viruses was shown to result in significant (to 10 lg ID50) decrease of the infectivity of the viruses. At that, the specific biological activity of viral proteins: the hemagglutinating and neuraminidase activities in the case of influenza and parainfluenza viruses, and the hemagglutinating activity in the case of Venezuelan equine encephalomyelitis virus, remained unchanged. Treatment of the viruses with the detergent in the presence of 2% BSA, 5% gamma-globulin, or 50% blood serum also inactivated the infectivity of virions. The inactivating effect of the MESK detergent is associated with solubilization of external glycoproteins of the virus envelope. The method of mild nondenaturating inactivation of enveloped viruses with the MESK detergent may prove to be useful in laboratory practice.

[Solubilization of glycoproteins from enveloped viruses by detergents]. / Soliubilizatsiia glikoproteidov obolochechnykh virusov detergentami.

The effects of some known ionic and nonionic detergents as well as that of a novel nonionic detergent MESK on various enveloped viruses were investigated. It was found that nonionic detergens (MESK, Triton X-100, octyl-beta-D-glucopyranoside) selectively solubilize glycoproteins of enveloped viruses. The most mild selective action is exerted by the nonionic detergent MESK. Using this detergent, pure preparations of glycoproteins of influenza, parainfluenza, equine Venezuela encephalomyelitis, rabies, vesicular stomatitis and herpes viruses were obtained. The procedure of isolation of purified glycoproteins includes incubation of viral suspensions with MESK, removal of subviral structures by centrifugation and purification of glycoproteins from detergent admixtures by dialysis. Purified glycoproteins retain their native structure and a high biological activity and immunogenicity. MESK seems to be due a perspective tool in the production of subunit vaccines.

[Isolation of glycoproteins of the Venezuelan equine encephalomyelitis virus and an evaluation of their immunogenic activity]. / Vydelenie glikoproteidov virusa venesuél'skogo éntsefalomielita loshadei i otsenka ikh immunogennoi aktivnosti.

Isolation of purified Venezuelan equine encephalomyelitis virus glycoproteins by means of a new Soviet nonionic detergent, MESK, is described. The MESK detergent was shown to permit isolation of approximately 70% of virus particle glycoproteins. The resulting preparation had a high hemagglutinating activity, contained no admixtures of foreign proteins and was not infectious. The study of the immunogenicity of purified glycoproteins in experimental mice and rabbits showed them to be capable of inducing high levels of serum antibodies. The immunogenicity of the isolated glycoproteins was comparable to their immunogenicity as components of virus particles. Treatment of the virus with MESK detergent also yielded preparations with predominant content of capsid protein. The described procedure for disintegration and purification of viral proteins is technologically simple and may be the basis for manufacture of a subunit vaccine. The resulting material may also be used for preparation of diagnosticums and in laboratory studies.

Isolation and studies of myxovirus glycoproteins.

The isolation of ortho- and paramyxovirus glycoproteins using a new nonionic detergent (MESK) is reported. MESK was shown to solubilize most of the viral envelope glycoproteins without decreasing their biologic activity. Solubilized glycoproteins are not contaminated by any internal viral proteins or by appreciable quantities of viral envelope lipids. The removal of MESK by dialysis resulted in the formation of glycoprotein micelles. The immunogenic activity of isolated glycoproteins was compared to that of virus particles. Immunization with isolated glycoproteins was shown to protect mice against a lethal influenza infection. Virions were treated with MESK in the presence of exogenous egg phosphatidylcholine, detergent was removed by dialysis and the glycoprotein was reconstituted in the vesicles. This reconstitution was accompanied by restoration of the haemolytic activity of Sendai virus proteins up to that of native virus particles. The level of activity, also the morphology and buoyant density of the vesicle were dependent on the protein/lipid ratio. MESK proved to be of value for the selective solubilization of the surface glycoproteins of animal enveloped viruses and their reconstitution in liposomes.

[Isolation of purified glycoproteins of influenza virus and analysis of their antigenic and immunogenic activity]. / Poluchenie ochishchennykh glikoproteidov virusa grippa i issledovanie ikh antigennoi i immunogennoi aktivnosti.

Investigations carried out by the authors have demonstrated that a new specially synthesized nonionic detergent, MECK, can be successfully used for obtaining pure influenza virus surface antigens in preparative amounts. Hemagglutinin and neuraminidase isolated by means of MECK retain their structure and biological activity. Special attention has been devoted to the study of the immunological characteristics of the preparations thus obtained. The high immunogenicity of solubilized hemagglutinin and the complete retention of its antigenic activity have been confirmed. The simplicity of the method for the isolation of influenza virus surface antigens and their purification from internal viral proteins and the detergent make it possible to recommend the above method for the preparative isolation of external antigens.

[Effect of naphthenic acids on the permeability of ascitic tumor cells to potassium ions]. / Vliianie naftenovykh kislot na pronitsaemost' astsitnykh opukholevykh kletok dlia ionov kaliia

Ascite cells were treated with naphthenic acid preparation, containing a mixture of naphthenic acids with average molecular weight equal to 240, and with a fraction of this preparation differing in the boiling temperature. A 5-10(-7)M and higher concentrations of the preparation and its fraction brought about an increased permeability in ascite cells for K-ions. The effectiveness of naphthenic acids is associated with their hydrophobic properties, which was characterized in the present communication by the value of hydrophilic-lipophilic balance calculated according to the Davies formula.