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

@article{citeulike:3187117, abstract = {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.}, address = {Medical Research Council Laboratory of Molecular Biology, Cambridge, UK.}, author = {Hawkins, R. E. and Zhu, D. and Ovecka, M. and Winter, G. and Hamblin, T. J. and Long, A. and Stevenson, F. K.}, citeulike-article-id = {3187117}, citeulike-linkout-0 = {http://view.ncbi.nlm.nih.gov/pubmed/8193363}, citeulike-linkout-1 = {http://www.hubmed.org/display.cgi?uids=8193363}, day = {1}, issn = {0006-4971}, journal = {Blood}, keywords = {nhl}, month = {June}, number = {11}, pages = {3279--3288}, posted-at = {2008-09-03 16:56:50}, priority = {2}, title = {Idiotypic vaccination against human B-cell lymphoma. Rescue of variable region gene sequences from biopsy material for assembly as single-chain Fv personal vaccines.}, url = {http://view.ncbi.nlm.nih.gov/pubmed/8193363}, volume = {83}, year = {1994} }

TY - JOUR ID - citeulike:3187117 L3 - citeulike-article-id:3187117 N2 - 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. IS - 11 JF - Blood SN - 0006-4971 AD - Medical Research Council Laboratory of Molecular Biology, Cambridge, UK. EP - 3288 TI - Idiotypic vaccination against human B-cell lymphoma. Rescue of variable region gene sequences from biopsy material for assembly as single-chain Fv personal vaccines. VL - 83 SP - 3279 KW - nhl AU - Hawkins, RE AU - Zhu, D AU - Ovecka, M AU - Winter, G AU - Hamblin, TJ AU - Long, A AU - Stevenson, FK PY - 1994/06/01/ UR - http://view.ncbi.nlm.nih.gov/pubmed/8193363 ER -