Enhanced capacity of antigen presentation of HBc-VLP-pulsed RAW264.7 cells revealed by proteomics analysis

J Proteome Res. 2008 Nov;7(11):4898-903. doi: 10.1021/pr800547v. Epub 2008 Oct 9.

Abstract

Many recent studies have indicated that virus-like particles (VLPs) have many potential applications in the fields of vaccine development and gene therapy. However, we still know little about the subtle mechanisms involved in the presentation of VLPs by antigen presenting cells (APCs). To illustrate the mechanisms, we utilized two-dimensional electrophoresis and tandem MS to compare and identify differentially expressed proteins between hepatitis B virus core antigen VLP (HBc-VLP)-pulsed and control RAW264.7 cells. Of the 25 spots identified as differentially expressed ( p < 0.05) between the two cell lines, 11 (corresponding to 11 unique proteins) were positively identified. Further analysis of two proteins, prohibitin and heat shock protein 70, confirmed that these proteins are expressed at higher levels in HBc-VLP-pulsed RAW264.7 cells compared with control cells. The proteins identified in this study will be useful in revealing the mechanisms that underlie VLP-APC interactions. Overall, this study also provides some useful suggestions for vaccine development and gene therapy.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antigen Presentation / genetics
  • Antigen Presentation / immunology*
  • Blotting, Western
  • Cell Line
  • HSP70 Heat-Shock Proteins / chemistry
  • HSP70 Heat-Shock Proteins / immunology
  • HSP70 Heat-Shock Proteins / metabolism
  • Hepatitis B Core Antigens / genetics
  • Hepatitis B Core Antigens / immunology*
  • Hepatitis B Core Antigens / ultrastructure
  • Isoelectric Point
  • Macrophages / immunology*
  • Macrophages / metabolism
  • Mice
  • Molecular Weight
  • Prohibitins
  • Proteome / analysis*
  • Proteomics / methods*
  • Recombinant Proteins / immunology
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / ultrastructure
  • Repressor Proteins / chemistry
  • Repressor Proteins / immunology
  • Repressor Proteins / metabolism
  • Reproducibility of Results
  • Time Factors

Substances

  • HSP70 Heat-Shock Proteins
  • Hepatitis B Core Antigens
  • Prohibitins
  • Proteome
  • Recombinant Proteins
  • Repressor Proteins