Idiotypic vaccination against human B-cell lymphoma. Rescue of variable region gene sequences from biopsy material for assembly as single-chain Fv personal vaccines

• Published in: Blood 1994-06, Vol.83 (11), p.3279-3288
• Main Authors: HAWKINS, R. E, DELIN ZHU, OVECKA, M, WINTER, G, HAMBLIN, T. J, LONG, A, STEVENSON, F. K
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; Antibodies, Monoclonal - immunology; Base Sequence; Biological and medical sciences; Biopsy; Genes, Immunoglobulin; Hematologic and hematopoietic diseases; Humans; Immunoglobulin Fragments - genetics; Immunoglobulin Fragments - immunology; Immunoglobulin Idiotypes - immunology; Immunoglobulin Variable Region - genetics; Immunoglobulin Variable Region - immunology; Leukemias. Malignant lymphomas. Malignant reticulosis. Myelofibrosis; Lymphoma, B-Cell - immunology; Medical sciences; Molecular Sequence Data; Polymerase Chain Reaction; Vaccination; Vaccines, Synthetic - immunology
• ISSN: 0006-4971; 1528-0020
• DOI: 10.1182/blood.v83.11.3279.3279

Idiotypic determinants on neoplastic B cells could provide tumor antigens for vaccination of patients with B-cell tumors. Because this approach requires an individual vaccine for each patient, simple methods for obtaining idiotypic antigen are desirable. Using polymerase chain reaction (PCR) with family-based V-gene and J-region primers, the variable region genes of heavy and light chains (VH and VL) of Ig have been obtained from biopsy material from 13 patients with B-cell tumors. In each case, analysis of random clones derived from the PCR product showed repeated, clonally-related sequences, whereas normal lymphoid tissue generated no repeated sequences. In 3/3 cases, the repeated sequences were found to be the same as those in a tumor-derived hybridoma. Mutational patterns in the V-genes differed among the tumors, with follicular lymphoma tending to be more highly mutated. The individual VH and VL sequences have been assembled with a flexible linker sequence to encode single-chain Fv (scFv). The scFv sequences can be cloned into bacterial expression vectors to produce protein, or into vectors suitable for direct vaccination using naked DNA. In a model system, expressed scFv protein retained all idiotypic determinants defined by a panel of five anti-idiotypic monoclonal antibodies (MoAbs). Similarly, expressed scFv proteins from two patients were shown to react with anti-idiotypic antibodies. This approach allows production of potential vaccines from surgical biopsies within 2 to 3 weeks.