Abstract
The present study reports on the use of gene transfer by retrovirus-derived shuttle vectors in the generation of hybrid hybridomas secreting bispecific monoclonal antibodies. neo- and dhfr- genes were infected into distinct murine hybridomas, thus conferring a dominant resistance trait to geneticin (G418) and to methotrexate. The vectors employed were replication-deficient and dependent on complementation by a helper virus provided by the irradiated packaging lines. After cocultivation with the relevant packaging cell lines, stable hybridoma lines expressing the selectable markers were easily obtained and were then suitable for conventional somatic fusion. This high-efficiency method was used to generate two bispecific monoclonal antibodies simultaneously targeting molecules expressed on cytotoxic cells (i.e., T lymphocytes and natural killer cells) against a human melanoma-associated antigen.
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- Anichini A., Mazzocchi A., Fossati G., Parmiani G. Cytotoxic T lymphocyte clones from peripheral blood and from tumor site detect intratumor heterogeneity of melanoma cells. Analysis of specificity and mechanisms of interaction. J Immunol. 1989 May 15;142(10):3692–3701. [PubMed] [Google Scholar]
- Clark M., Bindon C., Dyer M., Friend P., Hale G., Cobbold S., Calne R., Waldmann H. The improved lytic function and in vivo efficacy of monovalent monoclonal CD3 antibodies. Eur J Immunol. 1989 Feb;19(2):381–388. doi: 10.1002/eji.1830190224. [DOI] [PubMed] [Google Scholar]
- De Berardinis P., Londei M., Carrel S., Feldmann M. Regulation of clonal growth by anti-T-cell receptor antibody-directed lysis. Immunology. 1988 Jul;64(3):439–443. [PMC free article] [PubMed] [Google Scholar]
- Eglitis M. A., Kantoff P., Gilboa E., Anderson W. F. Gene expression in mice after high efficiency retroviral-mediated gene transfer. Science. 1985 Dec 20;230(4732):1395–1398. doi: 10.1126/science.2999985. [DOI] [PubMed] [Google Scholar]
- Giacomini P., Segatto O., Natali P. G. Multiple epitope recognition: an approach to improved radioimmunodetection of tumor-associated antigens. Int J Cancer. 1987 Jun 15;39(6):729–736. doi: 10.1002/ijc.2910390613. [DOI] [PubMed] [Google Scholar]
- Juarez-Salinas H., Ott G. S., Chen J. C., Brooks T. L., Stanker L. H. Separation of IgG idiotypes by high-performance hydroxylapatite chromatography. Methods Enzymol. 1986;121:615–622. doi: 10.1016/0076-6879(86)21060-5. [DOI] [PubMed] [Google Scholar]
- Jung G., Eberhard H. J. An in-vitro model for tumor immunotherapy with antibody heteroconjugates. Immunol Today. 1988 Sep;9(9):257–260. doi: 10.1016/0167-5699(88)91304-7. [DOI] [PubMed] [Google Scholar]
- Kirschmeier P. T., Housey G. M., Johnson M. D., Perkins A. S., Weinstein I. B. Construction and characterization of a retroviral vector demonstrating efficient expression of cloned cDNA sequences. DNA. 1988 Apr;7(3):219–225. doi: 10.1089/dna.1988.7.219. [DOI] [PubMed] [Google Scholar]
- Koolwijk P., Rozemuller E., Stad R. K., De Lau W. B., Bast B. J. Enrichment and selection of hybrid hybridomas by Percoll density gradient centrifugation and fluorescent-activated cell sorting. Hybridoma. 1988 Apr;7(2):217–225. doi: 10.1089/hyb.1988.7.217. [DOI] [PubMed] [Google Scholar]
- Malavasi F., Bellone G., Matera L., Milanese C., Ferrero E., Funaro A., Demaria S., Caligaris-Cappio F., Camussi G., Dellabona P. Murine monoclonal antibodies as probes for the phenotypical, functional, and molecular analysis of a discrete peripheral blood lymphocyte population exerting natural killer activity in vitro. Hum Immunol. 1985 Oct;14(2):87–102. doi: 10.1016/0198-8859(85)90067-9. [DOI] [PubMed] [Google Scholar]
- Malavasi F., Tetta C., Funaro A., Bellone G., Ferrero E., Franzone A. C., Dellabona P., Rusci R., Matera L., Camussi G. Fc receptor triggering induces expression of surface activation antigens and release of platelet-activating factor in large granular lymphocytes. Proc Natl Acad Sci U S A. 1986 Apr;83(8):2443–2447. doi: 10.1073/pnas.83.8.2443. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mann R., Mulligan R. C., Baltimore D. Construction of a retrovirus packaging mutant and its use to produce helper-free defective retrovirus. Cell. 1983 May;33(1):153–159. doi: 10.1016/0092-8674(83)90344-6. [DOI] [PubMed] [Google Scholar]
- Mezzanzanica D., Canevari S., Ménard S., Pupa S. M., Tagliabue E., Lanzavecchia A., Colnaghi M. I. Human ovarian carcinoma lysis by cytotoxic T cells targeted by bispecific monoclonal antibodies: analysis of the antibody components. Int J Cancer. 1988 Apr 15;41(4):609–615. doi: 10.1002/ijc.2910410422. [DOI] [PubMed] [Google Scholar]
- Miller A. D., Law M. F., Verma I. M. Generation of helper-free amphotropic retroviruses that transduce a dominant-acting, methotrexate-resistant dihydrofolate reductase gene. Mol Cell Biol. 1985 Mar;5(3):431–437. doi: 10.1128/mcb.5.3.431. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller A. D., Trauber D. R., Buttimore C. Factors involved in production of helper virus-free retrovirus vectors. Somat Cell Mol Genet. 1986 Mar;12(2):175–183. doi: 10.1007/BF01560664. [DOI] [PubMed] [Google Scholar]
- Milstein C., Cuello A. C. Hybrid hybridomas and their use in immunohistochemistry. Nature. 1983 Oct 6;305(5934):537–540. doi: 10.1038/305537a0. [DOI] [PubMed] [Google Scholar]
- Mottolese M., Venturo I., Donnorso R. P., Curcio C. G., Rinaldi M., Natali P. G. Use of selected combinations of monoclonal antibodies to tumor associated antigens in the diagnosis of neoplastic effusions of unknown origin. Eur J Cancer Clin Oncol. 1988 Aug;24(8):1277–1284. doi: 10.1016/0277-5379(88)90215-5. [DOI] [PubMed] [Google Scholar]
- Orlandi R., Güssow D. H., Jones P. T., Winter G. Cloning immunoglobulin variable domains for expression by the polymerase chain reaction. Proc Natl Acad Sci U S A. 1989 May;86(10):3833–3837. doi: 10.1073/pnas.86.10.3833. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Perez P., Hoffman R. W., Titus J. A., Segal D. M. Specific targeting of human peripheral blood T cells by heteroaggregates containing anti-T3 crosslinked to anti-target cell antibodies. J Exp Med. 1986 Jan 1;163(1):166–178. doi: 10.1084/jem.163.1.166. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Peruzzi L., Melioli G., De Monte L. B., Alessio M., Zaccolo M., Roggero S., Bellone G., Funaro A., Malavasi F. Microplate selection technique (MPST). A new method for selecting mouse transfectants expressing human gene products. J Immunol Methods. 1989 Sep 29;123(1):113–121. doi: 10.1016/0022-1759(89)90035-5. [DOI] [PubMed] [Google Scholar]
- Segal D. M., Wunderlich J. R. Targeting of cytotoxic cells with heterocrosslinked antibodies. Cancer Invest. 1988;6(1):83–92. doi: 10.3109/07357908809077031. [DOI] [PubMed] [Google Scholar]
- Southern P. J., Berg P. Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982;1(4):327–341. [PubMed] [Google Scholar]
- Staerz U. D., Bevan M. J. Hybrid hybridoma producing a bispecific monoclonal antibody that can focus effector T-cell activity. Proc Natl Acad Sci U S A. 1986 Mar;83(5):1453–1457. doi: 10.1073/pnas.83.5.1453. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Suresh M. R., Cuello A. C., Milstein C. Bispecific monoclonal antibodies from hybrid hybridomas. Methods Enzymol. 1986;121:210–228. doi: 10.1016/0076-6879(86)21019-8. [DOI] [PubMed] [Google Scholar]
- Titus J. A., Perez P., Kaubisch A., Garrido M. A., Segal D. M. Human K/natural killer cells targeted with hetero-cross-linked antibodies specifically lyse tumor cells in vitro and prevent tumor growth in vivo. J Immunol. 1987 Nov 1;139(9):3153–3158. [PubMed] [Google Scholar]