Optimized production and purification of Coxsackievirus B1 vaccine and its preclinical evaluation in a mouse model

Autor: Pär G. Larsson, Olli H. Laitinen, Vesa P. Hytönen, Juha A. E. Määttä, Virginia M. Stone, Malin Flodström-Tullberg, Minna M. Hankaniemi, Varpu Marjomäki, Amir-Babak Sioofy-Khojine, Heikki Hyöty
Rok vydání: 2017
Předmět:
0301 basic medicine
formalin inactivation
virukset
viruses
Drug Evaluation
Preclinical
Polysorbates
medicine.disease_cause
Antibodies
Viral
Mice
0302 clinical medicine
Multiplicity of infection
Immunogenicity
Vaccine
vaccine
Chlorocebus aethiops
030212 general & internal medicine
Immunogenicity
Vaccination
Infectious Diseases
coxsackievirus B1
Molecular Medicine
Female
Ultracentrifuge
Virus Cultivation
Coxsackievirus Infections
Biology
Coxsackievirus
ta3111
Virus
Microbiology
03 medical and health sciences
Formaldehyde
medicine
Animals
CVB1
Vero Cells
coxsackievirus
General Veterinary
General Immunology and Microbiology
rokotteet
ta1182
Public Health
Environmental and Occupational Health
Viral Vaccines
biology.organism_classification
Virology
Antibodies
Neutralizing
Enterovirus A
Human
Disease Models
Animal
030104 developmental biology
Vaccines
Inactivated
virus purification
Enterovirus
Zdroj: Vaccine. 35(30)
ISSN: 1873-2518
Popis: Coxsackie B viruses are among the most common enteroviruses, causing a wide range of diseases. Recent studies have also suggested that they may contribute to the development of type 1 diabetes. Vaccination would provide an effective way to prevent CVB infections, and the objective of this study was to develop an efficient vaccine production protocol for the generation of novel CVB vaccines. Various steps in the production of a formalin-inactivated Coxsackievirus B1 (CVB1) vaccine were optimized including the Multiplicity Of Infection (MOI) used for virus amplification, virus cultivation time, type of cell growth medium, virus purification method and formulation of the purified virus. Safety and immunogenicity of the formalin inactivated CVB1 vaccine was characterized in a mouse model. Two of the developed methods were found to be optimal for virus purification: the first employed PEG-precipitation followed by gelatin-chromatography and sucrose cushion pelleting (three-step protocol), yielding 19-fold increase in virus concentration (0.06µg/cm2) as compared to gold standard method. The second method utilized tandem sucrose pelleting without a PEG precipitation step, yielding 83-fold increase in virus concentration (0.24µg/cm2), but it was more labor-intensive and cannot be efficiently scaled up. Both protocols provide radically higher virus yields compared with traditional virus purification protocols involving PEG-precipitation and sucrose gradient ultracentrifugation. Formalin inactivation of CVB1 produced a vaccine that induced a strong, virus-neutralizing antibody response in vaccinated mice, which protected against challenge with CVB1 virus. Altogether, these results provide valuable information for the development of new enterovirus vaccines.
Databáze: OpenAIRE
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