Alginic acid-coated chitosan nanoparticles loaded with legumain DNA vaccine: effect against breast cancer in mice

PLoS One. 2013;8(4):e60190. doi: 10.1371/journal.pone.0060190. Epub 2013 Apr 5.

Abstract

Legumain-based DNA vaccines have potential to protect against breast cancer. However, the lack of a safe and efficient oral delivery system restricts its clinical application. Here, we constructed alginic acid-coated chitosan nanoparticles (A.C.NPs) as an oral delivery carrier for a legumain DNA vaccine. First, we tested its characteristic in acidic environments in vitro. DNA agarose electrophoresis data show that A.C.NPs protected DNA better from degradation in acidic solution (pH 1.5) than did chitosan nanoparticles (C.NPs). Furthermore, size distribution analysis showed that A.C.NPs tended to aggregate and form micrometer scale complexes in pH<2.7, while dispersing into nanoparticles with an increase in pH. Mice were intragastrically administrated A.C.NPs carrying EGFP plasmids and EGFP expression was detected in the intestinal Peyer's patches. Full-length legumain plasmids were loaded into different delivery carriers, including C.NPs, attenuated Salmonella typhimurium and A.C.NPs. A.C.NPs loaded with empty plasmids served as a control. Oral vaccination was performed in the murine orthotopic 4T1 breast cancer model. Our data indicate that tumor volume was significantly smaller in groups using A.C.NPs or attenuated Salmonella typhimurium as carriers. Furthermore, splenocytes co-cultured them with 4T1 cells pre-stimulated with CoCl2, which influenced the translocation of legumain from cytoplasm to plasma membrane, showed a 4.7 and 2.3 folds increase in active cytotoxic T lymphocytes (CD3(+)/CD8(+)/CD25(+)) when treated with A.C.NPs carriers compared with PBS C.NPs. Our study suggests that C.NPs coated with alginic acid may be a safe and efficient tool for oral delivery of a DNA vaccine. Moreover, a legumain DNA vaccine delivered orally with A.C.NPs can effectively improve autoimmune response and protect against breast cancer in mice.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Alginates / chemistry*
  • Animals
  • Antigen-Presenting Cells / immunology
  • Biological Transport
  • Cancer Vaccines / chemistry
  • Cancer Vaccines / genetics
  • Cancer Vaccines / immunology*
  • Cancer Vaccines / metabolism
  • Cell Proliferation
  • Chitosan / chemistry*
  • Cysteine Endopeptidases / genetics*
  • Cysteine Endopeptidases / immunology
  • Drug Carriers / chemistry
  • Female
  • Gastric Mucosa / metabolism
  • Glucuronic Acid / chemistry
  • Hexuronic Acids / chemistry
  • Hydrogen-Ion Concentration
  • Macrophages / immunology
  • Mammary Neoplasms, Experimental / immunology
  • Mammary Neoplasms, Experimental / pathology
  • Mammary Neoplasms, Experimental / prevention & control*
  • Mice
  • Mice, Inbred BALB C
  • Nanoparticles / chemistry*
  • Peyer's Patches / immunology
  • T-Lymphocytes / immunology
  • Vaccines, DNA / chemistry
  • Vaccines, DNA / genetics
  • Vaccines, DNA / immunology*
  • Vaccines, DNA / metabolism

Substances

  • Alginates
  • Cancer Vaccines
  • Drug Carriers
  • Hexuronic Acids
  • Vaccines, DNA
  • Glucuronic Acid
  • Chitosan
  • Cysteine Endopeptidases
  • asparaginylendopeptidase

Grants and funding

This work was supported by the National Basic Research Program of China (973 Program) number 2013CB967201, National Natural Science Foundation of China 81273331 (to R. Xiang) and National Natural Science Foundation of China 81270774 (to X. Tan). Funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.