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Purified and highly aggregated chimeric protein DIIIC-2 induces a functional immune response in mice against dengue 2 virus

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Abstract

It was previously reported that DIIIC-2 (a fusion protein composed of domain III of the envelope protein and the capsid protein from dengue 2 virus), as an aggregate antigen from a partially purified preparation, induced a functional protective immune response against dengue 2 virus in the mouse encephalitis model. In the present work, a purification procedure was developed for DIIIC-2, and soluble and aggregated fractions of the purified protein were characterized and evaluated in mice. The purification process rendered a protein preparation of 91 % purity, and the remaining 9 % consisted of fragments and aggregates of the same recombinant protein. After the in vitro aggregation process, upon addition of oligodeoxynucleotides, 80 % of the protein formed aggregates, whereas 20 % remained as soluble protein. An immunological evaluation revealed the proper immunogenicity of the aggregated purified protein in terms of induction of antiviral and neutralizing antibodies, cell-mediated immunity and protection upon dengue 2 virus challenge in the mouse encephalitis model. Based on these results, we can assert that the purified protein DIIIC-2 is functional and could be used for further scalable steps and preclinical studies in non-human primates.

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Acknowledgments

The authors are very grateful to Dr. Ricardo Silva (CIGB) and Dr. Jorge Martín (CIGB) for their critical reading and useful comments in the revision of the manuscript.

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Authors and Affiliations

  1. Vaccine Division, Center for Genetic Engineering and Biotechnology (CIGB), Avenue 31, P.O. Box 6162, Havana 6, 10 600, Cuba

    Ernesto Marcos, Lázaro Gil, Laura Lazo, Enma Brown, Edith Suzarte, Iris Valdés, Lissandra Méndez, Gerardo Guillén & Lisset Hermida

  2. Department of Virology, PAHO/WHO Collaborating Center for the Study of Dengue and Its Vector, Pedro Kourí Tropical Medicine Institute (IPK), P.O. Box 601, Havana, Cuba

    Alienys Izquierdo, Angélica García & María G. Guzmán

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  1. Ernesto Marcos
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  2. Lázaro Gil
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  3. Laura Lazo
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Correspondence to Lisset Hermida.

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Marcos, E., Gil, L., Lazo, L. et al. Purified and highly aggregated chimeric protein DIIIC-2 induces a functional immune response in mice against dengue 2 virus. Arch Virol 158, 225–230 (2013). https://doi.org/10.1007/s00705-012-1471-z

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  • DOI: https://doi.org/10.1007/s00705-012-1471-z

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