Carboxy-terminally truncated Dengue 4 virus envelope glycoprotein expressed in Pichia pastoris induced neutralizing antibodies and resistance to Dengue 4 virus challenge in mice

We have expressed a recombinant Dengue 4 virus envelope glycoprotein (E4rec), truncated at its C-terminus by 53 amino acids, in Pichia pastoris. The presence of E4rec was confirmed by Western-blot using anti-DEN 4 hyper immune mouse ascitic fluid. E4rec migrated during SDS-PAGE as a 64 kDa protein....

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Bibliographic Details
Published in:Archives of virology 2003-11, Vol.148 (11), p.2267-2273
Main Authors: MUNE, M, RODRIGUEZ, R, RAMIREZ, R, SOTO, Y, SIERRA, B, RODRIGUEZ ROCHE, R, MARQUEZ, G, GARCIA, J, GUILLEN, G, GUZMAN, M. G
Format: Article
Language:English
Subjects:
Amino acids
Animals
Antibodies
Antibodies, Viral - blood
Biological and medical sciences
Dengue - prevention & control
Dengue fever
Dengue Virus - immunology
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genomes
Glycoproteins
Human viral diseases
Immunization
Infectious diseases
Medical sciences
Mice
Mice, Inbred BALB C
Microbiology
Miscellaneous
Mycology
Neutralization Tests
Pichia - genetics
Proteins
Recombinant Proteins - immunology
Vaccines, Synthetic - immunology
Viral diseases
Viral Envelope Proteins - immunology
Viral Vaccines - immunology
Virology
Viruses
Yeast
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Description
Summary:We have expressed a recombinant Dengue 4 virus envelope glycoprotein (E4rec), truncated at its C-terminus by 53 amino acids, in Pichia pastoris. The presence of E4rec was confirmed by Western-blot using anti-DEN 4 hyper immune mouse ascitic fluid. E4rec migrated during SDS-PAGE as a 64 kDa protein. Treatment with endoglycosidases showed that the E protein was modified by the addition of short mannose chains and the absence of hyperglycosylation. When administered to BALB-C mice, E4rec elicited a DEN 4 neutralizing antibody response haemagglutination inhibition antibodies and specific memory T cell response. Mice immunized were also significantly protected against lethal DEN 4 virus challenge (86.6%, p < 0.001).
ISSN:0304-8608
1432-8798
DOI:10.1007/s00705-003-0167-9