Individualized human scFv vaccines produced in plants: humoral anti-idiotype responses in vaccinated mice confirm relevance to the tumor Ig

J Immunol Methods. 2003 Jul;278(1-2):95-104. doi: 10.1016/s0022-1759(03)00208-4.

Abstract

We have developed a method for rapidly producing in plants the idiotype regions of the tumor-specific Ig as single-chain Fv (scFv) proteins for use in the treatment of non-Hodgkin's lymphoma. Variable region gene sequences were generated from either a tumor hybridoma or human tumor biopsy cells, and idiotype domains were joined by a novel linker and cloned into a modified tobacco mosaic virus (TMV) vector designed to secrete the scFv protein in infected Nicotiana benthamiana plants. Thirty-eight out of 44 human scFv proteins showed Coomassie visible material in crude secretory (interstitial fluid, IF) extracts, 21 of those between 100 and 800 microg/ml. Eight of these proteins were tested for appropriate idiotype responses in vaccinated mice. In all eight cases, anti-idiotype immune responses were induced with minimal cross reactivity to irrelevant Ig or scFv proteins. Four out of four anti-scFv sera were also shown to recognize the Ig on human tumor cells by flow cytometry analysis.

Publication types

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

MeSH terms

  • Animals
  • Antibodies, Anti-Idiotypic / immunology
  • Antigens, Neoplasm / immunology
  • Blotting, Western
  • Cloning, Molecular
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Flow Cytometry
  • Gene Library
  • Genetic Vectors
  • Humans
  • Hybridomas
  • Immunoglobulin Variable Region / chemistry*
  • Immunoglobulin Variable Region / immunology*
  • Lymphoma, Non-Hodgkin / immunology
  • Lymphoma, Non-Hodgkin / therapy
  • Mice
  • Nicotiana / genetics*
  • Polymerase Chain Reaction
  • Receptors, Antigen, B-Cell / immunology
  • Tobacco Mosaic Virus
  • Tumor Cells, Cultured
  • Vaccines / chemical synthesis
  • Vaccines, Synthetic / immunology*

Substances

  • Antibodies, Anti-Idiotypic
  • Antigens, Neoplasm
  • Immunoglobulin Variable Region
  • Receptors, Antigen, B-Cell
  • Vaccines
  • Vaccines, Synthetic