Individualized human scFv vaccines produced in plants: humoral anti-idiotype responses in vaccinated mice confirm relevance to the tumor Ig.
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.
McCormick
Alison A
AA
Large Scale Biology Corporation, Vacaville, CA 95688, USA. alison.mccormick@lsbc.com
Reinl
Stephen J
SJ
Cameron
Terri I
TI
Vojdani
Fakhrieh
F
Fronefield
Michele
M
Levy
Ronald
R
Tusé
Daniel
D
D000818:Animals; D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000951:Antigens, Neoplasm / Q000276:immunology; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005434:Flow Cytometry; D015723:Gene Library; D005822:Genetic Vectors; D006801:Humans; D006825:Hybridomas; D007135:Immunoglobulin Variable Region / Q000737:chemistry* / Q000276:immunology*; D008228:Lymphoma, Non-Hodgkin / Q000276:immunology / Q000628:therapy; D051379:Mice; D016133:Polymerase Chain Reaction; D011947:Receptors, Antigen, B-Cell / Q000276:immunology; D014026:Nicotiana / Q000235:genetics*; D014027:Tobacco Mosaic Virus; D014407:Tumor Cells, Cultured; D014612:Vaccines / Q000138:chemical synthesis; D014614:Vaccines, Synthetic / Q000276:immunology*
D000888:Antibodies, Anti-Idiotypic; D000951:Antigens, Neoplasm; D007135:Immunoglobulin Variable Region; D011947:Receptors, Antigen, B-Cell; D014612:Vaccines; D014614:Vaccines, Synthetic