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Cancer Immunology, Immunotherapy : CII logoLink to Cancer Immunology, Immunotherapy : CII
. 1991 Jan;34(1):37–42. doi: 10.1007/BF01741322

Characterization of a monoclonal antibody directed against the epidermal growth factor receptor binding site

Rita Pellegrini 1, Filippo Centis 1, Stefania Martignone 1, Antonio Mastroianni 1, Elda Tagliabue 1, Emanuela Tosi 1, Sylvie Ménard 1, Maria I Colnaghi 1,✉
PMCID: PMC11038489  PMID: 1760809

Abstract

In this work a new monoclonal antibody (mAb), designated MGR1, which recognizes the epidermal growth factor receptor (EGF-R) binding site, is described. The main characteristic of this mAb is its ability to discriminate between cells that express normal levels of EGF-R from cells with overexpression, the detectability threshold by immunocytochemical tests being 5 × 104 receptors/cell of 10 µm diameter. MGR1 was found to inhibit EGF binding on the relevant target cells, and vice versa its binding was inhibited by EGF, which indicated that MGR1 recognizes the EGF receptor binding site. MGR1 exerted an inhibitory effect on both the in vitro and in vivo growth of cells with EGF-R overexpression, but had no effect on cells with a normal expression of the receptor. Tumour growth inhibition in athymic mice was also obtained on already implanted tumours. MGR1 therefore seems to be an adequate reagent for the development of immunotherapeutical approaches suitable for the treatment of tumours with EGF-R overexpression.

Key words: Monoclonal antibody, Epidermal growth factor receptor

References

  • 1.Abound-Pirak E, Hurwitz E, Pirak ME, Bellot F, Schlessinger J, Sela M. Efficacy of antibodies to epidermal growth factor receptor against KB carcinoma in vitro and in nude mice. J Natl Cancer Inst. 1988;80:1605. doi: 10.1093/jnci/80.20.1605. [DOI] [PubMed] [Google Scholar]
  • 2.Abound-Pirak E, Hurwitz E, Bellot F, Schlessinger J, Sela M. Inhibition of human tumour growth in nude mice by a conjugate of doxorubicin with monoclonal antibodies to epidermal growth factor receptor. Proc Natl Acad Sci USA. 1989;86:3778. doi: 10.1073/pnas.86.10.3778. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3.Canevari S, Orlandi R, Ripamonti M, Tagliabue E, Aguanno S, Miotti S, Ménard S, Colnaghi MI. Ricin A chain conjugated with monoclonal antibodies selectively killing human carcinoma cells in vitro. J Natl Cancer Inst. 1985;75:831. doi: 10.1093/jnci/75.5.831. [DOI] [PubMed] [Google Scholar]
  • 4.Carpenter G. Receptors for epidermal growth factor and other polypeptide mitogens. Annu Rev Biochem. 1987;56:881. doi: 10.1146/annurev.bi.56.070187.004313. [DOI] [PubMed] [Google Scholar]
  • 5.DePotter CR, VanDaele S, VanDeVijver MJ, Pauwels C, Maertens G, DeBoever J, Vandekerckhove D, Roels H. The expression of the neu oncogene product in breast lesions and in normal fetal and adult human tissues. Histopathology. 1989;15:362. doi: 10.1111/j.1365-2559.1989.tb01587.x. [DOI] [PubMed] [Google Scholar]
  • 6.Ey PL, Prowse SJ, Jenken CR. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-Sepharose. Immunochemistry. 1978;15:429. doi: 10.1016/0161-5890(78)90070-6. [DOI] [PubMed] [Google Scholar]
  • 7.Fendly BM, Winget M, Hudziak RM, Lipari MT, Napier MA, Ullrich A. Characterization of murine monoclonal antibodies reactive to either the human epidermal growth factor receptor or HER2/neu gene product. Cancer Res. 1990;50:1550. [PubMed] [Google Scholar]
  • 8.Foon KA. Biological response modifiers: the new immunotherapy. Cancer Res. 1989;49:1621. [PubMed] [Google Scholar]
  • 9.Gill GN, Kawamoto T, Cochet C, Le A, Sato JD, Masui H, McLeod C, Mendelsohn J. Monoclonal anti-epidermal growth factor receptor antibodies which are inhibitors of epidermal growth factor binding and antagonists of epidermal growth factor-stimulated tyrosine protein kinase activity. J Biol Chem. 1984;259:7755. [PubMed] [Google Scholar]
  • 10.Gullick WJ, Marsden JJ, Whittle N, Ward B, Bobrow L, Waterfield MD. Expression of epidermal growth factor receptors on human cervical ovarian and vulvar carcinomas. Cancer Res. 1986;46:285. [PubMed] [Google Scholar]
  • 11.Harris AL. The epidermal growth factor receptor as a target for therapy. Cancer Cells. 1990;2:321. [PubMed] [Google Scholar]
  • 12.Hendler FJ, Ozane BW. Human squamous cell lung cancers express increased epidermal growth factor receptors. J Clin Invest. 1984;74:647. doi: 10.1172/JCI111463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Kalofonos HP, Pawlikowska TR, Hemingway A, Courtenay-Luck N, Dhokia B, Snook D, Sivolapenko GB, Hooker GR, McKenzie CG, Lavender PJ, Thomas DGT, Epenetos AA. Antibody guided diagnosis and therapy of brain gliomas using radiolabeled monoclonal antibodies against epidermal growth factor receptor and placental alkaline phosphatase. J Nucl Med. 1989;30:1636. [PubMed] [Google Scholar]
  • 14.Kamata N, Chida K, Rikimaru K, Horikoshi M, Enomoto S, Kuroki T. Growth-inhibitory effects of epidermal growth factor and overexpression of its receptors on human squamous cell carcinomas in culture. Cancer Res. 1986;46:1648. [PubMed] [Google Scholar]
  • 15.Kawamoto T, Sato JD, Le A, Polikoff J, Sato GH, Mendelsohn J. Growth stimulation of A431 cells by epidermal growth factor: Identification of high-affinity receptors for epidermal growth factor by an anti-receptor monoclonal antibody. Proc Natl Acad Sci USA. 1983;80:1337. doi: 10.1073/pnas.80.5.1337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Knowles AF. Inhibition of growth and induction of enzyme activities in a clonal human hepatoma cell line (Li-7A): comparison of the effects of epidermal growth factor receptor antibody. J Cell Physiol. 1988;134:109. doi: 10.1002/jcp.1041340113. [DOI] [PubMed] [Google Scholar]
  • 17.Laemmli VK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970;227:680. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  • 18.Libermann TA, Nusbaum HR, Razon N, Kris R, Lax I, Soreq H, Whittle N, Waterfield MD, Ullrich A, Schlessinger J. Amplification, enhanced expression and possible rearrangement of EGF receptor gene in primary human brain tumours of glial origin. Nature. 1985;313:144. doi: 10.1038/313144a0. [DOI] [PubMed] [Google Scholar]
  • 19.Masui H, Kawamoto T, Sato JD, Wolf B, Sato G, Mendelsohn J. Growth inhibition of human tumour cells in athymic mice by anti-epidermal growth factor receptor monoclonal antibodies. Cancer Res. 1984;44:1002. [PubMed] [Google Scholar]
  • 20.Masui H, Moroyama T, Mendelsohn J. Mechanism of antitumour activity in mice for anti-epidermal growth factor receptor monoclonal antibodies with different isotypes. Cancer Res. 1986;46:5592. [PubMed] [Google Scholar]
  • 21.Masui H, Kamrath H, Apell G, Houston LL, Mendelsohn J. Cytotoxicity against human tumor cells mediated by the conjugate of anti-epidermal growth factor receptor monoclonal antibody to recombinant ricin A chain. Cancer Res. 1989;49:3482. [PubMed] [Google Scholar]
  • 22.Ménard S, Tagliabue E, Canevari S, Fossati G, Colnaghi MI. Generation of monoclonal antibodies reacting with normal and cancer cells of human breast. Cancer Res. 1983;43:1295. [PubMed] [Google Scholar]
  • 23.Miotti S, Canevari S, Ménard S, Mezzanzanica D, Porro G, Pupa SM, Regazzoni M, Tagliabue E, Colnaghi MI. Characterization of human ovarian carcinoma-associated antigens defined by novel monoclonal antibodies with tumour-restricted specificity. Int J Cancer. 1987;39:297. doi: 10.1002/ijc.2910390306. [DOI] [PubMed] [Google Scholar]
  • 24.Natali PG, Nicotra MR, Bigotti A, Venturo I, Slamon DJ, Fendly BM, Ullrich A. Expression of the p185 encoded by HER2 oncogene in normal and transformed human tissues. Int J Cancer. 1990;45:457. doi: 10.1002/ijc.2910450314. [DOI] [PubMed] [Google Scholar]
  • 25.Ozawa S, Ueda M, Ando N, Abe O, Shimizu N. High incidence of EGF receptor hyperproduction in esophageal squamous-cell carcinomas. Int J Cancer. 1987;39:333. doi: 10.1002/ijc.2910390311. [DOI] [PubMed] [Google Scholar]
  • 26.Ozawa S, Ueda M, Ando N, Abe O, Minoshima S, Shimizu N. Selective killing of squamous carcinoma cells by an immunotoxin that recognizes the EGF receptor. Int J Cancer. 1989;43:152. doi: 10.1002/ijc.2910430128. [DOI] [PubMed] [Google Scholar]
  • 27.Rodeck U, Herlyn M, Herlyn D, Molthoff C, Atkinson B, Varello M, Steplewski Z, Koprowski H. Tumour growth modulation by a monoclonal antibody to the epidermal growth factor receptor: immunologically mediated and effector cell-independent effects. Cancer Res. 1987;47:3692. [PubMed] [Google Scholar]
  • 28.Sakai K, Mori S, Kawamoto T, Taniguchi S, Kobori O, Morioka Y, Kuroki T, Kano K. Expression of epidermal growth factor receptors on normal human gastric epithelia and gastric carcinomas. J Natl Cancer Inst. 1986;77:1047. [PubMed] [Google Scholar]
  • 29.Sato GH, Sato JD. Growth factor receptor monoclonal antibodies and cancer immunotherapy. J Natl Cancer Inst. 1989;81:1600. doi: 10.1093/jnci/81.21.1600. [DOI] [PubMed] [Google Scholar]
  • 30.Schreiber AB, Lax I, Yarden Y, Eshhar Z, Schlessinger J. Monoclonal antibodies against receptor for epidermal growth factor induce early and delayed effect of epidermal growth factor. Proc Natl Acad Sci USA. 1981;78:7535. doi: 10.1073/pnas.78.12.7535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Slamon DJ, Press MF, Godolphin W, Ramos L, Haran P, Shek L, Stuart SG, Ullrich A. Studies of the HER2/neu proto-oncogene in human breast cancer. Cancer Cells. 1989;7:371. [Google Scholar]
  • 32.Sobol RE, Astarita RW, Hofeditz C, Masui H, Fairshter R, Royston I, Mendelsohn J. Epidermal growth factor receptor expression in human lung carcinomas defined by a monoclonal antibody. J Natl Cancer Inst. 1987;79:403. [PubMed] [Google Scholar]
  • 33.Taetle R, Honeysett JM, Houston LL. Effects of anti-epidermal growth factor (EGF) receptor antibodies and an anti-EGF receptor recombinant-ricin A chain immunoconjugate on growth of human cells. J Natl Cancer Inst. 1988;80:1053. doi: 10.1093/jnci/80.13.1053. [DOI] [PubMed] [Google Scholar]
  • 34.Tagliabue E, Ménard S, Della Torre G, Barbanti P, Mariani-Costantini R, Porro G, Colnaghi MI. Generation of monoclonal antibodies reacting with human epithelial ovarian cancer. Cancer Res. 1985;45:379. [PubMed] [Google Scholar]
  • 35.Vollmar AM, Banker DE, Mendelsohn J, Herschman HR. Toxicity of ligand and antibody-directed ricin A chain conjugates recognizing the epidermal growth factor receptor. J Cell Physiol. 1987;131:418. doi: 10.1002/jcp.1041310314. [DOI] [PubMed] [Google Scholar]
  • 36.Waterfield MD, Mayes EL, Stroobant P, Bennet LPB, Young S, Goodfellow PN, Banting GS, Ozanne B. A monoclonal antibody to the human epidermal growth factor receptor. J Cell Biochem. 1982;20:149. doi: 10.1002/jcb.240200207. [DOI] [PubMed] [Google Scholar]
  • 37.Weber W, Gill GN. Production of an epidermal growth factor receptor related-protein. Science. 1984;224:294. doi: 10.1126/science.6324343. [DOI] [PubMed] [Google Scholar]
  • 38.Weiner DB, Nordberg J, Robinson R, Nowell PC, Gazdar A, Greene MI, Williams Wv, Cohen JA, Kern JA. Expression of theneu gene-encoded protein (p185neu) in human non-small cell carcinomas of the lung. Cancer Res. 1990;50:421. [PubMed] [Google Scholar]
  • 39.Yamamoto T, Kamata N, Kawano H, Shimizu S, Kuroki T, Tayoshima K, Rikimaru K, Nomura N, Ishizaki R, Pastan I, Gamou S, Shimizu N. High incidence of amplification of the epidermal growth factor receptor gene in human squamous carcinoma cell lines. Cancer Res. 1986;46:414. [PubMed] [Google Scholar]

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