Regulation of HIV-Gag expression and targeting to the endolysosomal/ secretory pathway by the luminal domain of Lysosomal-Associated Membrane Protein (LAMP-1) enhance Gag-specific immune response

Rodrigo Maciel Da Costa Godinho, Flavio Lemos Matassoli, Carolina Gonçalves De Oliveira Lucas, Paula Ordonhez Rigato, Jorge Luiz Santos Gonçalves, Maria Notomi Sato, Milton Maciel, Ligia Maria Torres Peçanha, J. Thomas August, Ernesto Torres De Azevedo Marques, Luciana Barros De Arruda

Research output: Contribution to journalArticle

Abstract

We have previously demonstrated that a DNA vaccine encoding HIV-p55gag in association with the lysosomal associated membrane protein-1 (LAMP-1) elicited a greater Gag-specific immune response, in comparison to a DNA encoding the native gag. In vitro studies have also demonstrated that LAMP/Gag was highly expressed and was present in MHCII containing compartments in transfected cells. In this study, the mechanisms involved in these processes and the relative contributions of the increased expression and altered traffic for the enhanced immune response were addressed. Cells transfected with plasmid DNA constructs containing p55gag attached to truncated sequences of LAMP-1 showed that the increased expression of gag mRNA required p55gag in frame with at least 741 bp of the LAMP-1 luminal domain. LAMP luminal domain also showed to be essential for Gag traffic through lysosomes and, in this case, the whole sequence was required. Further analysis of the trafficking pathway of the intact LAMP/Gag chimera demonstrated that it was secreted, at least in part, associated with exosome-like vesicles. Immunization of mice with LAMP/gag chimeric plasmids demonstrated that high expression level alone can induce a substantial transient antibody response, but targeting of the antigen to the endolysosomal/secretory pathways was required for establishment of cellular and memory response. The intact LAMP/gag construct induced polyfunctional CD4+ T cell response, which presence at the time of immunization was required for CD8 + T cell priming. LAMP-mediated targeting to endolysosomal/secretory pathway is an important new mechanistic element in LAMP-mediated enhanced immunity with applications to the development of novel anti-HIV vaccines and to general vaccinology field.

Original languageEnglish (US)
Article numbere99887
JournalPloS one
Volume9
Issue number6
DOIs
StatePublished - Jun 16 2014

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • General

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    Godinho, R. M. D. C., Matassoli, F. L., Lucas, C. G. D. O., Rigato, P. O., Gonçalves, J. L. S., Sato, M. N., Maciel, M., Peçanha, L. M. T., August, J. T., Marques, E. T. D. A., & De Arruda, L. B. (2014). Regulation of HIV-Gag expression and targeting to the endolysosomal/ secretory pathway by the luminal domain of Lysosomal-Associated Membrane Protein (LAMP-1) enhance Gag-specific immune response. PloS one, 9(6), [e99887]. https://doi.org/10.1371/journal.pone.0099887