Skip to main content
Cancer Immunology, Immunotherapy : CII logoLink to Cancer Immunology, Immunotherapy : CII
. 1995 May;40(3):191–200. doi: 10.1007/BF01517351

Construction and characterization of the chimeric monoclonal antibody E48 for therapy of head and neck cancer

Ruud H Brakenhoff 1,, Frank B van Gog 1, James E Looney 2, Marijke van Walsum 1, Gordon B Snow 1, Guus A M S van Dongen 1
PMCID: PMC11037753  PMID: 7728778

Abstract

Data from an ongoing clinical radioimmunoscintigraphy trial indicate that99mTc-labeled monoclonal antibody (mAb) E48 is highly capable of selectively targeting squamous cell carcinoma of the head and neck (HNSCC). The percentage of the injected dose per gram of tumor tissue was found to be high, rendering mAbE48 a promising candidate mAb for therapeutic purposes. We now describe the construction of a chimeric (moouse/human) mAb E48 by recombinant DNA technology. The genes encoding the variable domains of the heavy and light chain were cloned and ligated into experession vectors containing the human ψ1 heavy-chain gene and the human k lightchain gene respectively. Biological properties of the resulting chimeric mAb E48 were compared to the murine form in vitro and in vivo. The reactivities of chimeric (c)mAb and murine (m)mAb E48 with HNSCC, as assessed by immunohistochemical staining as well as immuno-blotting were shown to be similar. The affinity constant appeared to be 0.9×1010 M−1 and 1.6×1010 M−1 for the mmAb and cmAb respectively. The biodistribution of both antibodies was tested by simultaneous injection into nude mice bearing human HNSCC xenografts. cmAb E48 was found to be cleared more rapidly from the blood than mmAb E48, resulting in a 30% lower tumor uptake but similar tumor to non-tumor ratios, 3 days after injection. Moreover, it was shown that cmAb E48 is highly capable of lysing HNSCC targets in ADCC assays in vitro, whereas the mmAb appeared to be almost incative. These data indicate that cmAb E48 has potential as a targeting agent for the eradication of HNSCC in man.

Key Words: Chimeric antibody, Head and neck cancer, Squamous cell carcinoma, Radioimmunotheraphy, Antibody-dependent cellular cytotoxicity

References

  • 1.Bader CC, Krohn KA, Bernstein ID. In vitro measurement of avidity of radioiodinated antibodies. Int J Radiat Appl Instrum. 1987;14:605. doi: 10.1016/0883-2897(87)90033-x. [DOI] [PubMed] [Google Scholar]
  • 2.Brakenhoff RH, Geurts van Kessel AHM, Oldenburg M, Wijnen JTh, Bloemendal H, Meera Khan P, Schoenmakers JGG. Human αB-crystallin (CRYA2) gene mapped to chromosome 11q12-q23. Hum Genet. 1990;85:237. doi: 10.1007/BF00193203. [DOI] [PubMed] [Google Scholar]
  • 3.Caton AJ, Brownlee GG, Staudt LM, Gerhard W. Structural and functional impications of a restricted antibody response to a defined antigenic redgion on the influenza virus hemagglutinin. EMBO J. 1986;56:1577. doi: 10.1002/j.1460-2075.1986.tb04399.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4.Cheung NKV, Lazarus H, Miraldi FD, Abramowsky CR, Kallick S, Saarinen UM, Spitser T, Strandjord SE, Coccia PF, Berger NA. Ganglioside GD2 specific monoclonal antibody 3F8: a phase I study in patients with neuroblastoma and malignant melanoma. J. Clin Oncol. 1987;5:1430. doi: 10.1200/JCO.1987.5.9.1430. [DOI] [PubMed] [Google Scholar]
  • 5.Courtenay-Luck NS, Epenetos AA, Moore R, Larche M, Pectaside D, Dhokia B, Ritter MA. Development of primary and secondary immune responses to mouse monoclonal antibodies used in diagnosis and therapy of malignant neoplasms. Cancer Res. 1986;46:6489. [PubMed] [Google Scholar]
  • 6.De Bree R, Roos JC, Quak JJ, Den Hollander W, Van den Brekel MWM, Van de Wal JE, Tobi H, Snow GB, Van Dongen GAMS. Clinical imaging of head and neck cancer with99mTc-labeled monoclonal antibody E48 IgG or F(ab')2 . J Nucl Med. 1994;35:775. [PubMed] [Google Scholar]
  • 7.Feinberg AP, Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1988;132:6. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  • 8.Gerretsen M, Quak JJ, Suh JS, Van Walsum, Meijer CJLM, Snow GB, Van Dongen GAMS. Superior localisation of monoclonal antibody E48 F(ab')2 fragment in xenografts of human squamous cell carcinoma of the head and neck and of the vulva as compared to monoclonal antibody E48 IgG. Br J Cancer. 1991;63:37. doi: 10.1038/bjc.1991.9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9.Gerretsen M, Schrijvers AHGJ, Van Walsum M, Braakhuis BJM, Snow GB, Van Dongen GAMS. Radioimmunothraphy of head and neck squamous cell carcinoma with131I-labeled monoclonal antibody E48. Br J Cancer. 1992;66:496. doi: 10.1038/bjc.1992.302. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Gerretsen M, Visser GWM, van Walsum M, Meijer CJLM, Snow GB, Van Dongen GAMS. 186Re-labeled monoclonad antibody E48 IgG mediated therapy of human head and neck squamous cell carcinoma xenografts. Cancer Res. 1993;53:3524. [PubMed] [Google Scholar]
  • 11.Gerretsen M, Visser GWM, Brakenhoff RH, Snow GB, Van Dongen GAMS. Complete ablation of small human head and neck squamous cell carcinoma xenografts with186Relabeled monoclonal antibody E48 IgG. Cell Biophys. 1994;24:1. doi: 10.1007/BF02789224. [DOI] [PubMed] [Google Scholar]
  • 12.Gough NM. Rapid and quantitative preparation of cytoplasmic RNA from small numbers of cells. Anal Biochem. 1988;173:93. doi: 10.1016/0003-2697(88)90164-9. [DOI] [PubMed] [Google Scholar]
  • 13.Herlyn D, Koprowski H. IgG2a monoclonal antibodies inhibit human tumor growth through interaction with effector cells. Proc Natl Acad Sci USA. 1982;79:4761. doi: 10.1073/pnas.79.15.4761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Huse WD, Sastry L, Iverson SA, Kang AS, Alting-Mees M, Burton DR, Benkovic SJ, Lerner RA. Generation of a large combinatorial library of the immunoglobulin repretoire in phage lambda. Science. 1989;246:1275. doi: 10.1126/science.2531466. [DOI] [PubMed] [Google Scholar]
  • 15.Janson CH, Tehrani M, Wigzell Rational use of biological response modifiers in hematological malignancies: a review of treatment with interfron, cytotoxic cells and antibodies. Leuk Res. 1989;13:1039. doi: 10.1016/0145-2126(89)90148-3. [DOI] [PubMed] [Google Scholar]
  • 16.Jones PT, Dear PH, Foote J, Neuberger MS, Winter G. Replacing the complementarity-determining regions in a human antibody with those from a mouse. Nature. 1986;321:522. doi: 10.1038/321522a0. [DOI] [PubMed] [Google Scholar]
  • 17.Kavaler J, Caton AJ, Staudt LM, Schwartz D, Gerhard W. A set of closely related antibodies dominates the primary antibody response to the antigenic site CB of the A/PR/8/34 influenza virus hemagglutinin. J Immunol. 1990;145:2312. [PubMed] [Google Scholar]
  • 18.Khazaeli MB, Conry R, LoBuglio AF. Human immune response to monoconal antibodies. J Immunother. 1994;15:42. doi: 10.1097/00002371-199401000-00006. [DOI] [PubMed] [Google Scholar]
  • 19.Laemmli UK. 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]
  • 20.LoBuglio AF, Wheeler RH, Trang J, Haynes A, Rogers K, Harvey EB, Sun L, Grayeb J, Khazaeli MB. Mouse/human chimeric monoclonal antibody in man: kinetics and immune response. Proc. Natl Acad Sci USA. 1989;86:4220. doi: 10.1073/pnas.86.11.4220. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.LoBuglio AF, Khazaeli MB, Meredith R, Saleh MN. Chimeric monoclonal antibodies in cancer theraphy. In: NDDO EORTC, editor. Proceedings of the 7th NCI-EORTC symposium on new drugs in cancer therapy. Amsterdam, March: Free Univ. Press; 1992. p. 196. [Google Scholar]
  • 22.Looney JE, Knight DM, Arevalo-Moore M, Trinh H, Pak K-Y, Dalesandro MR, Rieber EP, Riethmüller G, Daddona PE, Ghrayeb J. High-level expression and characterization of a mouse-human chimeric CD4 antibody with therapeutic potential. Hum Antibody Hybridomas. 1992;3:191. [PubMed] [Google Scholar]
  • 23.Massagur LFAG, Thomas CMG, Segers MFG, Corstens FHM, Verheijen RHM, Kenemans P, Poels LG. Specific and nonspecific immunoassays to detect HAMA after administration of indium-111-labeled OV-TL-3 F(ab')2 monoclonal antibody to patients with ovarian cancer. J Nucl Med. 1992;33:1958. [PubMed] [Google Scholar]
  • 24.Meredith RF, LoBuglio AF, Plott WE, Orr RA, Brezovich IA, Russell CD, Harvey EB, Yester MV, Wagner AJ, Spencer SA. Pharmacokinetics, immune response, and biodistribution of iodine-131-labeled chimeric mouse/human IgG, κ 17-1A monoclonal antibody. J Nucl Med. 1991;32:1162. [PubMed] [Google Scholar]
  • 25.Morrison SL, Johson MJ, Herzenberg LA, Oi VT. Chimeric human antibody molecules: mouse antigen-binding domains with human constant region domains. Proc Natl Acad Sci USA. 1984;81:6851. doi: 10.1073/pnas.81.21.6851. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 26.Quak JJ, Van Dongen GAMS, Balm AJM, Brakkee JPG, Scheper RJ, Snow GB, Meijer CJLM. A 22-kDa surface antigen detected by monoclonal antibody E48 is exclusively expressed in stratified squamous and transitional epithelia. Am J Pathol. 1990;136:191. [PMC free article] [PubMed] [Google Scholar]
  • 27.Quak JJ, Van Dongen GAMS, Gerretsen M, Hayashida D, Balm AJM, Brakkee JPG, Snow GB, Meijer CJLM. Production of monoclonal antibody (K931) to a squamous cell carcinoma antigen identified as the 17-1A antigen. Hybridoma. 1990;9:377. doi: 10.1089/hyb.1990.9.377. [DOI] [PubMed] [Google Scholar]
  • 28.Quak JJ, Schrijvers AHGJ, Brakkee JGP, Davis HD, Scheper RJ, Balm AJM, Meijer CJLM, Snow GB, Van Dongen GAMS. Expression and characterization of two differentiation antigens in stratified squamous epithelia and carcinomas. Int J Cancer. 1992;50:507. doi: 10.1002/ijc.2910500402. [DOI] [PubMed] [Google Scholar]
  • 29.Ragnhammer P, Fagerberg J, Frödin J-E, Hjelm A-L, Lindemalm C, Magnusson I, Masucci G, Mellstedt H. Effect of monoclonal antibody 17-1A and GM-CSF in patients with advanced colorectal carcinoma: long lasting, complete remissions can be induced. Int J Cancer. 1993;53:751. doi: 10.1002/ijc.2910530508. [DOI] [PubMed] [Google Scholar]
  • 30.Reynolds JC, Del Vecchio S, Sakahara H, Lora ME, Carrasquillo JA, Neumann RD, Larson SM. Anti-murine respponse to mouse monoclonal antibodies: clinical findings and implications. Nucl Med Biol. 1989;16:121. doi: 10.1016/0883-2897(89)90182-7. [DOI] [PubMed] [Google Scholar]
  • 31.Riethmüller G, Schneider-Gadicke E, Schlimok G, Schmieldel W, Raab R, Hofken K, Gruber R, Pichlmaier H, Hirche H, Pichlmaier R, Buggisch P, Witte J. Randomised trial of monoclonal antibody for adjuvant theraphy of resected Dukes'C colorectal carcinoma. Lancet. 1994;343:1177. doi: 10.1016/s0140-6736(94)92398-1. [DOI] [PubMed] [Google Scholar]
  • 32.Sambrook J, Fritsch EF, Maniatis T. Molecular cloning: a laboratory manual. 2nd edn. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory; 1989. [Google Scholar]
  • 33.Schrijvers AHGJ, Gerretsen M, Fritz J, Van Walsum M, Quak JJ, Snow GB, Van Dongen GAMS. Evidence for a role of the monoclonal antibody E48 defined antigen in cell-cell adhesion in squamous epithelia and head and neck squamous cell carcinomas. Exp Cell Res. 1991;196:264. doi: 10.1016/0014-4827(91)90260-2. [DOI] [PubMed] [Google Scholar]
  • 34.Seybold K, Trinkler M, Frey LD, Locher JT. Long-term HAMA-follow up after immunoscintigraphy using antigranulocyte and anti-tumour mAbs: advanced experiences in 210 patients. In: Lind P, editor. Proceedings of the 9th Annual Meeting of the International Research Group on Immunoscintigraphy and Immunotheraphy. Austria: Klagen furt; 1994. p. 28. [Google Scholar]
  • 35.Steplewski Z, Sun LK, Shearman CW, Ghrayeb J, Donnadona P, Koprowski H. Biological activity of human-mouse IgG1, IgG2, IgG3, and IgG4 chimeric monoclonal antibodies with antitumor specificity. Proc Natl Acad Sci USA. 1988;85:4852. doi: 10.1073/pnas.85.13.4852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 36.Steplewski Z, Lubeck MD, Scholz H, Loibner J, McDonald Smith, Koprowski H. Tumor cell lysis and tumor growth inhibiton by the isotype variants of mAb Br 55-2 directed against Y oloigosaccharide. In Vivo. 1991;5:79. [PubMed] [Google Scholar]
  • 37.Van Dongen GAMS, Leverstein H, Roos JC, Quak JJ, van den Brekel M, van Lingen A, Martens HJM, Castelijns J, Visser GWM, Meijer CJLM, Teule JJ, Snow GB. Radioimmujnoscintigraphy of head and neck tumors using99mTclabeled monoclonal antibody E48 F(ab')2 . Cancer Res. 1992;52:2569. [PubMed] [Google Scholar]
  • 38.Van Dongen GAMS, Brakenhoff RH, De Bree R, Gerretsen M, Quak JJ, Snow GB. Progress in radioimmuotherapy of head and neck cancer: review. Oncol Rep. 1994;1:259. [PubMed] [Google Scholar]
  • 39.Visser G, Gerretsen M, Herscheid J, Snow GB, Van Dongen GAMS. Labeling of monoclonal antibodies with186Re using the MAG3 chelate for radioimmunotherapy of cancer: a technical protocol. J Nucl Med. 1993;34:1953. [PubMed] [Google Scholar]
  • 40.Wanebo HJ, Blackinton D, Kouttab Mehta S. Contribution of serum inhibitory factors and immune cellular defects to the depressed cell-mediated immunity in patients in patients with head and neck cancer. Am J Surg. 1993;166:389. doi: 10.1016/s0002-9610(05)80339-3. [DOI] [PubMed] [Google Scholar]
  • 41.Wessels BW, Harisiadis L, Carabell SC. Dosimetry and radiobiological efficacy of clincial radioimmunotherapy. J Nucl Med. 1989;30:827. [Google Scholar]
  • 42.Woodcock D, Crowther PJ, Doherty J, DeCruz E, Noyer-Weidner M, Smith SS, Michal MZ, Graham MW. Quantitative evaluation ofEscherichia coli host strains for tolerance to cytosine methylation in plasmid and phage recombinants. Nucl Acids Res. 1989;17:3469. doi: 10.1093/nar/17.9.3469. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Cancer Immunology, Immunotherapy : CII are provided here courtesy of Springer

RESOURCES