Breast Cancer Research and Treatment

, Volume 24, Issue 2, pp 103–113 | Cite as

Epitope expression on the breast epithelial mucin

  • Roberto L. Ceriani
  • Jerry A. Peterson
  • Edward W. Blank
  • Derek T. A. Lamport


Multiple epitope expression on the breast epithelial mucin was explored using a panel of monoclonal antibodies (MoAbs) created against milk and breast tissue preparations, against blood group determinants, and against other non-breast epithelial mucins. Since the breast epithelial mucin is now used in both diagnostic and therapeutic modalities for breast cancer, and also because altered or incomplete glycosylation in varying degrees is expected in breast carcinoma tissue, the antigenic target used here was the native mucin and sequential stages of deglycosylation introduced to it by HF treatment. Partial deglycosylation increased exposure of core peptide amino acid sequences increasing MoAb binding generally, while it either decreased or occasionally increased binding of blood group oligosaccharides. Cross reactivity of MoAbs to other mucins was low with the breast epithelial mucin (BEM). The study of the affinity binding constants of some of the anti-BEM peptide MoAbs predicted carbohydrate participation in their epitope structure. The identification of different epitopes on the BEM, investigations on their possible epitopic structure, and the study of MoAb binding during different stages of glycosylation of the molecule leads to knowledge on the contribution of carbohydrates to their epitopes and strengthens the ability to understand their performance in their diverse possible applications in breast cancer diagnosis, prognosis, and therapy.

Key words

breast epithelial mucin epitopes glycoproteins monoclonal antibodies protein glycosylation tandem repeat peptides 


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  1. 1.
    Ceriani RL, Peterson JA, Lee JY, Moncada R, Blank EW: Characterization of cell surface antigens of human mammary epithelial cells with monoclonal antibodies prepared against human milk fat globule. Somatic Cell Genet 9:415–427, 1983CrossRefPubMedGoogle Scholar
  2. 2.
    Ceriani RL, Sasaki M, Sussman H, Wara WM, Blank EW: Circulating human mammary epithelial antigens in breast cancer. Proc Natl Acad Sci USA 79:5420–5424, 1982PubMedGoogle Scholar
  3. 3.
    Peterson JA, Zava DT, Duwe AK, Blank EW, Battifora H, Ceriani RL: Biochemical and histological characterization of antigens preferentially expressed on the surface and cytoplasm of breast carcinoma cells identified by monoclonal antibodies against the human milk fat globule. Hybridoma 9:221–235, 1990PubMedGoogle Scholar
  4. 4.
    Linsley PS, Ochs V, Laska S, Horn D, Ring DB, Frankel AE, Brown JP: Elevated levels of high molecular weight antigen detected by antibody W1 in sera from breast cancer patients. Cancer Res 46:5444–5450, 1986PubMedGoogle Scholar
  5. 5.
    Taylor-Papadimitriou J, Peterson JA, Arklie J, Burchell J, Ceriani RL, Bodmer WF: Monoclonal antibodies to epithelium-specific components of the human milk fat globule membrane: production and reaction with cells in culture. Int J Cancer 28:17–21, 1981PubMedGoogle Scholar
  6. 6.
    Ceriani RL, Blank EW, Peterson JA, Battifora H, Singh H: Immunotherapeutic preclinical evaluation of antihuman milk fat globule MoAbs Mc5 and BrE-1. Antibody Immunoconj Radiopharm 3:181–198, 1990Google Scholar
  7. 7.
    Foster CS, Edwards PA, Dinsdale EA, Neville AM: Monoclonal antibodies to the human mammary and extramammary tissues. Virchows Arch (Pathol Anat) 394:279–293, 1982CrossRefGoogle Scholar
  8. 8.
    Hilkens J, Kroezen V, Bonfrer JMG, De Jong-Bakker M, Bruning PF: MAM-6 antigen, a new serum marker for breast cancer monitoring. Cancer Res 46:2582–2587, 1986PubMedGoogle Scholar
  9. 9.
    Colcher D, Hand PH, Nuti M, Schlom J: A spectrum of monoclonal antibodies reactive with human mammary tumor cells. Proc Natl Acad Sci USA 78:3199–3203, 1981PubMedGoogle Scholar
  10. 10.
    Papsidero LD, Croghan GA, O'Connell MF, Valenzuela LA, Nemoto T, Chu TM: Monoclonal antibodies (F36/22 and M7/105) to human breast carcinoma. Cancer Res 43:1741–1747, 1983PubMedGoogle Scholar
  11. 11.
    Stacker SA, Thompson CH, Riglar C, McKenzie IFC: A new breast carcinoma antigen defined by a monoclonal antibody. J Natl Cancer Inst 75:801–811, 1985PubMedGoogle Scholar
  12. 12.
    Ligtenber MJL, Vos HL, Gennissen AMC, Hilkens J: Episialin, a carcinoma-associated mucin, is generated by a polymorphic gene encoding splice variants with alternative amino termini. J Biol Chem 265:5573–5578, 1990PubMedGoogle Scholar
  13. 13.
    Gendler SJ, Lancaster CA, Taylor-Papadimitriou J, Duhig T, Peat N, Burchell J, Pemberton L, Lalani EN, Wilson D: Molecular cloning and expression of human tumor-associated polymorphic epithelial mucin. J Biol Chem 265:15286–15293, 1990PubMedGoogle Scholar
  14. 14.
    Burchell J, Taylor-Papadimitriou J, Boshell M, Gendler S, Duhig T: A short sequence, within the amino acid tandem repeat of a cancer-associated mucin, contains immunodominant epitopes. Int J Cancer 44:691–696, 1989PubMedGoogle Scholar
  15. 15.
    Peterson JA, Larocca D, Walkup G, Amiya R, Zava DT, Ceriani RL: Monoclonal antibodies to overlapping polypeptide epitopes of the breast mucin have different specificities to the native antigen. (Submitted for publication)Google Scholar
  16. 16.
    Xing PX, Tjandra JJ, Reynolds K, McLaughlin PJ, Purcell DFJ, McKenzie IFC: Reactivity of anti-human milk fat globule antibodies with synthetic peptides. J Immunol 142:3503–3509, 1990Google Scholar
  17. 17.
    Geysen HM, Rodda S, Mason T, Tribbick G, Schoofs P: Strategies for epitope analysis using peptide synthesis. J Immunol Methods 102:259–274, 1987CrossRefPubMedGoogle Scholar
  18. 18.
    Laver WG, Air GMG, Webster RG, Smith-Gill SJ: Epitopes on protein antigens: misconceptions and realities. Cell 61:553–556, 1990CrossRefPubMedGoogle Scholar
  19. 19.
    Hanisch FG, Uhlenbruck G, Peter-Katalinic J, Egge H, Dabrowski J, Dabrowski U: Structures of neutral O-linked polylactosaminoglycans on human skim milk mucins. A novel type of linearly extended poly-N-acetyllactosamine backbones with Gal β(1–4) GlcNAc β(1–6) repeating units. J Biol Chem 264:872–883, 1989PubMedGoogle Scholar
  20. 20.
    Parodi AJ, Blank EW, Peterson JA, Ceriani RL: Dolichol-bound oligosaccharides and the transfer of distal monosaccharides in the synthesis of glycoproteins by normal and tumor mammary epithelial cells. Breast Cancer Res Treat 2:227–237, 1982CrossRefPubMedGoogle Scholar
  21. 21.
    Hull SR, Bright A, Carraway KL, Abe M, Hayes DF, Kufe DW: Oligosaccharide differences in the DF3 sialomucin antigen from normal human milk and the BT-20 human breast carcinoma cell line. Cancer Commun 1:261–267, 1989PubMedGoogle Scholar
  22. 22.
    Burchell J, Gendler S, Taylor-Papadimitriou J, Girling A, Lewis A, Millis R, Lamport D: Development and characterization of breast cancer reactive monoclonal antibodies directed to the core protein of the human milk mucin. Cancer Res 47:5476–5482, 1987PubMedGoogle Scholar
  23. 23.
    Ceriani RL, Thompson K, Peterson JA, Abraham S: Surface differentiation antigens of human mammary epithelial cells carried on the human milk fat globule. Proc Natl Acad Sci USA 74:582–586, 1977PubMedGoogle Scholar
  24. 24.
    Taylor-Papadimitriou J: Report on the First International Workshop on Carcinoma Associated Mucins. Int J Cancer 49:1–5, 1991PubMedGoogle Scholar
  25. 25.
    Ceriani RL, Larocca D, Peterson JA, Enloe S, Amiya R, Walkup G, Blank EW: A novel serum assay for breast epithelial antigen using a fusion protein. Analyt Biochem 201:178–184, 1992CrossRefPubMedGoogle Scholar
  26. 26.
    Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC: Measurement of protein using bicinchoninic acid. Analyt Biochem 150:76–85, 1985CrossRefPubMedGoogle Scholar
  27. 27.
    Mort A, Lamport D: Anhydrous hydrogen fluoride deglycosylates glycoproteins. Analyt Biochem 82:289–309, 1977CrossRefPubMedGoogle Scholar
  28. 28.
    Ceriani RL: Solid phase identification and molecular weight determination of cell membrane antigens with monoclonal antibodies.In: Bechtol KB, McKern TJ, Kennett R (eds) Monoclonal Antibodies and Functional Cell Lines — Progress and Applications. Plenum Press, New York, 1984, pp 398–402Google Scholar
  29. 29.
    Sheldon K, Marks A, Baumal R: Characterization of binding of four monoclonal antibodies to the human ovarian adenocarcinoma cell line HEY. Biochem Cell Biol 65:423–428, 1991Google Scholar
  30. 30.
    Walter P, Green S, Greene GL, Krust A, Bornert JM, Heltsch JM, Staub A, Jensen EV, Scarce G, Waterfield M, Chambon P: Cloning of the human estrogen receptor cDNA. Proc Natl Acad Sci USA 82:7889–7893, 1985PubMedGoogle Scholar
  31. 31.
    Hull SR, Carraway KL: Mechanism of expression of Thomsen-Friedenreich (T) antigen at the cell surface of a mammary carcinoma. FASEB J 2:2380, 1988PubMedGoogle Scholar
  32. 32.
    Kjeldsen TK, Clausen H, Hirohashi S, Ogawa T, Iijima H, Hakomori S: Preparation and characterization of monoclonal antibodies directed to the tumor-associated O-linked sialosyl-2→6 α-N-acetylgalactosaminyl(sialosyl-Tn) epitope. Cancer Res 48:2214–2220, 1988PubMedGoogle Scholar
  33. 33.
    Ceriani RL: Circulating breast epithelial antigens.In Herberman RB, DW Mercer (eds) Immunodiagnosis of Cancer, II. Marcel Dekker, New York, 1988, pp 223–241Google Scholar
  34. 34.
    Ceriani RL, Sasaki M, Orthendahl D, Kaufman L: Localization of human breast tumors grafted in nude mice with a monoclonal antibody directed against a defined cell surface antigen of human antigen of human mammary epithelial cells. Breast Cancer Res Treat 12:177–189, 1988CrossRefPubMedGoogle Scholar
  35. 35.
    Gonzalez R, Deinhart D, Kasliwal R, Johnson T, Lear J, Bloedow D, Hartmann C, Glenn S, Butchko G, Miller G, Ceriani RL, Bunn P: A Phase I study of radiolabeled monoclonal antibody (Mc5) in breast cancer. (Submitted for publication)Google Scholar
  36. 36.
    Kalofonos HP, Epenetos AA: Antibody guided diagnosis and therapy of patients with breast cancer.In Ceriani RL (ed) Immunological Approaches to the Diagnosis and Therapy of Breast Cancer. Plenum Press, New York, 1987, pp 245–257Google Scholar
  37. 37.
    DeNardo SJ, Warhoe KA, O'Grady LF, Gobuty AH, Macey DJ, Helstrom I, Helstrom KE, Kroger LEL, DeNardo GL: Radioimmunotherapy of breast cancer: MoAb biokinetics to treatment protocols [abstract]. 4th Int Workshop on Monoclonal Antibodies and Breast Cancer, San Francisco, 1990Google Scholar
  38. 38.
    Castagna M, Nuti M, Squartini F: Mammary cancer antigen recognized by monoclonal antibody B72.3 in apocrine metaplasia of the breast. Cancer Res 47:902–906, 1987PubMedGoogle Scholar
  39. 39.
    Russo J: Immunocytochemical markers in breast cancer.In Russo J (ed) Immunocytochemistry in Tumor Diagnosis. M Nijhoff, Boston, 1985, pp 207–232Google Scholar
  40. 40.
    Duwe AK, Ceriani RL: Human milk fat globule membrane derived mucin is a disulfide-linked heteromer. Biochem Biophys Res Commun 165:1305–1311, 1989CrossRefPubMedGoogle Scholar
  41. 41.
    Gum JR, Hicks JW, Swallow DM, Lagace RL, Byrd JC, Lamport DT, Siddiki B, Kim YS: Molecular cloning of cDNAs derived from a novel human intestinal mucin gene. Biochem Biophys Res Commun 171:407–415, 1990CrossRefPubMedGoogle Scholar
  42. 42.
    Lan MS, Batra SK, Qi WN, Metzgar RS, Hollingsworth MA: Cloning and sequencing of a human pancreatic tumor mucin cDNA. J Biol Chem 265:15294–15299, 1990PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • Roberto L. Ceriani
    • 1
  • Jerry A. Peterson
    • 1
  • Edward W. Blank
    • 1
  • Derek T. A. Lamport
    • 2
  1. 1.John Muir Cancer and Aging Research InstituteWalnut Creek
  2. 2.DOE Plant Research LabMichigan State UniversityEast LansingUSA

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