RESUMO
INTRODUCTION: The COVID-19 pandemic caused by SARS-CoV-2 has created serious health problems worldwide. The most effective way to prevent the occurrence of new epidemic outbreaks is vaccination. One of the modern and effective approaches to vaccine development is the use of virus-like particles (VLPs). The aim of the study is to develop a technology for production of VLP based on recombinant SARS-CoV-2 proteins (E, M, N and S) in insect cells. MATERIALS AND METHODS: Synthetic genes encoding coronavirus proteins E, M, N and S were used. VLP with various surface proteins of strains similar to the Wuhan virus, Delta, Alpha and Omicron were developed and cloned into the pFastBac plasmid. The proteins were synthesized in the baculovirus expression system and assembled into VLP in the portable Trichoplusia ni cell. The presence of insertion in the baculovirus genome was determined by PCR. ELISA and immunoblotting were used to study the antigenic activity of VLP. VLP purification was performed by ultracentrifugation using 20% sucrose. Morphology was assessed using electron microscopy and dynamic light scattering. RESULTS: VLPs consisting of recombinant SARS-CoV-2 proteins (S, M, E and N) were obtained and characterized. The specific binding of antigenic determinants in synthesized VLPs with antibodies to SARS-CoV-2 proteins has been demonstrated. The immunogenic properties of VLPs have been studied. CONCLUSION: The production and purification of recombinant VLPs consisting of full-length SARS-CoV-2 proteins with a universal set of surface antigens have been developed and optimized. Self-assembling particles that mimic the coronavirus virion induce a specific immune response against SARS-CoV-2.
Assuntos
Baculoviridae , COVID-19 , SARS-CoV-2 , Glicoproteína da Espícula de Coronavírus , Vacinas de Partículas Semelhantes a Vírus , Animais , SARS-CoV-2/genética , SARS-CoV-2/imunologia , SARS-CoV-2/metabolismo , Vacinas de Partículas Semelhantes a Vírus/imunologia , Vacinas de Partículas Semelhantes a Vírus/genética , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/imunologia , Glicoproteína da Espícula de Coronavírus/química , Glicoproteína da Espícula de Coronavírus/metabolismo , Humanos , COVID-19/virologia , COVID-19/imunologia , Baculoviridae/genética , Baculoviridae/metabolismo , Vacinas contra COVID-19/imunologia , Anticorpos Antivirais/imunologia , Proteínas M de Coronavírus/genética , Proteínas M de Coronavírus/imunologia , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Proteínas do Nucleocapsídeo de Coronavírus/genética , Proteínas do Nucleocapsídeo de Coronavírus/química , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Proteínas da Matriz Viral/genética , Proteínas da Matriz Viral/imunologia , FosfoproteínasAssuntos
Agonistas Adrenérgicos/farmacologia , Glândulas Endócrinas/efeitos dos fármacos , Epinefrina/farmacologia , Hipertensão/fisiopatologia , Aldosterona/sangue , Animais , Corticosterona/sangue , Glândulas Endócrinas/fisiopatologia , Hipertensão/sangue , Insulina/sangue , Ratos , Ratos Endogâmicos , Ratos Wistar , Especificidade da Espécie , Estresse Fisiológico/sangue , Estresse Fisiológico/fisiopatologia , Tiroxina/sangue , Tri-Iodotironina/sangueAssuntos
1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/farmacologia , Comportamento Animal/efeitos dos fármacos , Condicionamento Operante/efeitos dos fármacos , Dopaminérgicos/farmacologia , Doenças dos Macacos/induzido quimicamente , Doença de Parkinson Secundária/induzido quimicamente , Animais , Comportamento Animal/fisiologia , Condicionamento Operante/fisiologia , Macaca mulatta , Masculino , Doenças dos Macacos/fisiopatologia , Doença de Parkinson Secundária/fisiopatologia , Fatores de TempoRESUMO
Behavioral changes and eye movements were studied through chronic low-dose MPTP exposure (ten intramuscular injections of 0.2 mg/kg per day) in four monkeys (Macaca rhesus). The monkeys developed the deficit of spontaneous saccades and decreased oculomotor range. The vertical component of saccades were hardly reduced. The examination of vestibuloocular reflex revealed no changes (r = approximately 1.0), but the fast phase of nystagmus was blocked.
Assuntos
Movimentos Oculares/fisiologia , Músculos Oculomotores/fisiologia , Reflexo Vestíbulo-Ocular/fisiologia , 1-Metil-4-Fenil-1,2,3,6-Tetra-Hidropiridina/administração & dosagem , Adolescente , Animais , Criança , Feminino , Humanos , Macaca mulatta/fisiologia , MasculinoRESUMO
Development of behavioral disturbances was investigated during N-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) administration (0,2 mg/kg, i/m, at 48 h. intervals, total dose 11.2-13.2 mg/kg) in experiments on two monkeys Macaca rhesus. MPTP induced the complex of symptoms, which are typical for idiopathic Parkinson's disease. Administration of amiridine (0,25-0,4 mg/kg) to MPTP-treated monkeys caused gradual decline of Parkinson syndrome. Mechanisms of action of amiridine are discussed.
