Apoptosis-mediated enhancement of DNA-raised immune responses by mutant caspases

Nat Biotechnol. 2001 Jun;19(6):543-7. doi: 10.1038/89289.

Abstract

Apoptotic bodies can be used to target delivery of DNA-expressed immunogens into professional antigen-presenting cells (APCs). Here we show that antigen-laden apoptotic bodies created by vectors co-expressing influenza virus hemagglutinin (HA) or nucleoprotein (NP) genes and mutant caspase genes markedly increased T-cell responses. Both CD8 and CD4 T-cell responses were affected. The adjuvant activity was restricted to partially inactivated caspases that allowed immunogen expression before the generation of apoptotic bodies. Active-site mutants of murine caspase 2 and an autocatalytic chimera of murine caspase 2 prodomain and human caspase 3 induced apoptosis that did not interfere with immunogen expression. The adjuvant activity also enhanced B-cell responses, but to a lesser extent than T-cell responses. The large increases in T-cell responses represent one of the strongest effects to date of a DNA adjuvant on cellular immunity.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • 3T3 Cells
  • Animals
  • Annexin A5 / pharmacology
  • Antigen-Presenting Cells
  • Apoptosis*
  • Binding Sites
  • CD4-Positive T-Lymphocytes / metabolism
  • CD8-Positive T-Lymphocytes / metabolism
  • Caspases / genetics*
  • Caspases / immunology*
  • DNA / metabolism*
  • Dose-Response Relationship, Drug
  • Green Fluorescent Proteins
  • Hemagglutinins / biosynthesis
  • Humans
  • Luminescent Proteins / metabolism
  • Mice
  • Mice, Inbred BALB C
  • Mutation
  • Nucleoproteins / biosynthesis
  • Orthomyxoviridae / metabolism
  • Plasmids / metabolism
  • Time Factors
  • Transfection

Substances

  • Annexin A5
  • Hemagglutinins
  • Luminescent Proteins
  • Nucleoproteins
  • Green Fluorescent Proteins
  • DNA
  • Caspases