Rapid immune responses to a botulinum neurotoxin Hc subunit vaccine through in vivo targeting to antigen-presenting cells

Infect Immun. 2011 Aug;79(8):3388-96. doi: 10.1128/IAI.00166-11. Epub 2011 May 16.

Abstract

The clostridial botulinum neurotoxins (BoNTs) are the most potent protein toxins known. The carboxyl-terminal fragment of the toxin heavy chain (Hc) has been intensively investigated as a BoNT vaccine immunogen. We sought to determine whether targeting Hc to antigen-presenting cells (APCs) could accelerate the immune responses to vaccination with BoNT serotype A (BoNT/A) Hc. To test this hypothesis, we targeted Hc to the Fc receptors for IgG (FcγRs) expressed by dendritic cells (DCs) and other APCs. Hc was expressed as a fusion protein with a recombinant ligand for human FcγRs (R4) to produce HcR4 or a similar ligand for murine FcγRs to produce HcmR4. HcR4, HcmR4, and Hc were produced as secreted proteins using baculovirus-mediated expression in SF9 insect cells. In vitro receptor binding assays showed that HcR4 effectively targets Hc to all classes of FcγRs. APCs loaded with HcR4 or HcmR4 are substantially more effective at stimulating Hc-reactive T cells than APCs loaded with nontargeted Hc. Mice immunized with a single dose of HcmR4 or HcR4 had earlier and markedly higher Hc-reactive antibody titers than mice immunized with nontargeted Hc. These results extend to BoNT neutralizing antibody titers, which are substantially higher in mice immunized with HcmR4 than in mice immunized with Hc. Our results demonstrate that targeting Hc to FcγRs augments the pace and magnitude of immune responses to Hc.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood
  • Antibodies, Neutralizing
  • Antigen-Presenting Cells / immunology*
  • Antigen-Presenting Cells / metabolism
  • Antitoxins / blood
  • Bacterial Vaccines / administration & dosage
  • Bacterial Vaccines / immunology*
  • Bacterial Vaccines / metabolism
  • Baculoviridae / genetics
  • Botulinum Toxins, Type A / immunology*
  • Botulism / prevention & control*
  • Cell Line
  • Female
  • Genetic Vectors
  • Mice
  • Mice, Inbred BALB C
  • Receptors, Fc / metabolism*
  • Recombinant Fusion Proteins / administration & dosage
  • Recombinant Fusion Proteins / immunology
  • Recombinant Fusion Proteins / metabolism
  • Spodoptera
  • Vaccines, Subunit / administration & dosage
  • Vaccines, Subunit / immunology
  • Vaccines, Subunit / metabolism
  • Vaccines, Synthetic / administration & dosage
  • Vaccines, Synthetic / immunology
  • Vaccines, Synthetic / metabolism

Substances

  • Antibodies, Bacterial
  • Antibodies, Neutralizing
  • Antitoxins
  • Bacterial Vaccines
  • Receptors, Fc
  • Recombinant Fusion Proteins
  • Vaccines, Subunit
  • Vaccines, Synthetic
  • Botulinum Toxins, Type A