Skip to main content
PMC home page
Cancer Immunology, Immunotherapy : CII logoLink to Cancer Immunology, Immunotherapy : CII
. 1986 Dec;23(3):185–191. doi: 10.1007/BF00205648

A monoclonal antibody with binding and inhibiting activity towards human pancreatic carcinoma cells

I. Immunohistological and immunochemical characterization of a murine monoclonal antibody selecting for well differentiated adenocarcinomas of the pancreas

K Bosslet 1,✉, H F Kern 2, E J Kanzy 1, A Steinstraesser 3, A Schwarz 3, G Lüben 1, H U Schorlemmer 1, H H Sedlacek 1
PMCID: PMC11038228  PMID: 3539336

Abstract

The murine monoclonal antibody (MAb) BW 494 was characterized in relation to its tissue specificity, the epitope recognized, in vitro and in vivo radiolocalization and its potential to mediate antibody dependent cellular cytotoxicity (ADCC) and complement mediated cytolysis (CMC). The MAb defined carbohydrate epitope located on a >200 k daltons glycoprotein was mainly expressed on the majority of well differentiated adenocarcinomas of the pancreas. Furthermore, the epitope is accessible to MAb BW 494 in vivo, allowing an enrichment of radioactive antibody at the tumor site in nude mice. Additionally, MAb BW 494 is able to use human peripheral blood lymphocytes as effector cells for ADCC reactions against appropriate tumor target cells in vitro. In contrast, the antibody does not mediate human or rabbit CMC.

Keywords: Adenocarcinoma, Nude Mouse, Effector Cell, Tumor Site, Tissue Specificity

References

  • 1.Atkinson BF, Ernst CS, Herlyn M, Steplewski Z, Sears HF, Koprowski H. Gastrointestinal cancer-associated antigen in immunoperoxidase assay. Cancer Res. 1982;42:4820. [PubMed] [Google Scholar]
  • 2.Borowitz MJ, Tuck FL, Sindelar WF, Fernsten PD, Metzgar RS. Monoclonal antibodies against human pancreatic adenocarcinoma: distribution of DU-PAN-2 antigen on glandular epithelia and adenocarcinomas. J Natl Cancer Inst. 1984;72:999. [PubMed] [Google Scholar]
  • 3.Bosslet K, Lüben G, Stark M, Sedlacek HH. Molecular characteristics of two lung carcinoma cell line associated membrane antigens. Behring Inst Mitt. 1984;74:27. [PubMed] [Google Scholar]
  • 4.Bosslet K, Lüben G, Schwarz A, Hundt E, Harthus HP, Seiler FR, Muhrer C, Klöppel G, Kayser K, Sedlacek HH. Immunohistochemical localization and molecular characteristics of three monoclonal antibody-defined epitopes detectable on carcinoembryonic antigen (CEA) Int J Cancer. 1985;36:75. doi: 10.1002/ijc.2910360113. [DOI] [PubMed] [Google Scholar]
  • 5.Brown JP, Klitzman JM, Hellström KE. A microassay for antibody binding to tumour cell surface antigens using 125I-labelled protein-A from Staphylococcus aureus. J Immunol Methods. 1977;15:57. doi: 10.1016/0022-1759(77)90017-5. [DOI] [PubMed] [Google Scholar]
  • 6.Dippold WG, Dienes HP, Knuth A, Meyer zum Büschenfelde K-H. Immunohistochemical localization of ganglioside GD3 in human malignant melanoma, epithelial tumors and normal tissues. Cancer Res. 1985;45:3699. [PubMed] [Google Scholar]
  • 7.Edwards PAW. Heterogenous expression of cell-surface antigens in normal epithelia and their tumours, revealed by monoclonal antibodies. Br J Cancer. 1985;51:149. doi: 10.1038/bjc.1985.24. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 8.Ey PL, Prowse SJ, Jenkin CR. Isolation of pure IgG1, IgG2a, IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemisty. 1978;15:429. doi: 10.1016/0161-5890(78)90070-6. [DOI] [PubMed] [Google Scholar]
  • 9.Folch J, Lees M, Sloane Stanley GH. A simple method for the isolation and purification of total lipids from animal tissue. J Biol Chem. 1957;226:497. [PubMed] [Google Scholar]
  • 10.Fraker PJ, Speck JC. Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1, 3, 4, 6-tetrachloro 3, 6-diphenylglycoluri. Biochem Biophys Res Commun. 1978;80:849. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
  • 11.Harpin ML, Coulon-Morelec MJ, Yeni P, Danon F, Baumann N. Direct sensitive immunocharacterization of gangliosides on plastic thin-layer plates using the peroxidase staining. J Immunol Methods. 1985;78:135. doi: 10.1016/0022-1759(85)90336-9. [DOI] [PubMed] [Google Scholar]
  • 12.Hellström I, Brankovan V, Hellström KE. Strong antitumor activities of IgG3 antibodies to a human melanoma-associated ganglioside. Proc Natl Acad Sci USA. 1985;82:1499. doi: 10.1073/pnas.82.5.1499. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Heyderman E. Immunoperoxidase technique in histology: Applications, methods and controls. J Clin Pathol. 1979;32:971. doi: 10.1136/jcp.32.10.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Hirohashi S, Ino Y, Kodama T, Shimosato Y. Distribution of blood group antigens A, B, H and I (Ma) in mucusproducing adenocarcinoma of human lung. J Natl Cancer Inst. 1984;72:1299. [PubMed] [Google Scholar]
  • 15.Hoogenraad N, Helman T, Hoogenraad J. The effect of preinjection of mice with pristane on ascites tumour formation and monoclonal antibody production. J Immunol Methods. 1983;61:317. doi: 10.1016/0022-1759(83)90225-9. [DOI] [PubMed] [Google Scholar]
  • 16.Houghton AN, Mintzer D, Cordon-Cardo C, Welt S, Fliegel B, Vadhan S, Carswell E, Melamed MR, Oettgen HF, Old LJ (1986) Mouse monoclonal antibody detecting GD3 ganglioside: A phase I trial in patients with malignant melanoma. In press. Proc Natl Acad Sci USA [DOI] [PMC free article] [PubMed]
  • 17.Imai K, Moriya Y, Fujita H, Tsujisaki M, Kawaharada M, Yachi A. Immunological characterization and molecular profile of carcinoembryonic antigen detected by monoclonal antibodies. J Immunol. 1984;132:2992. [PubMed] [Google Scholar]
  • 18.Klöppel G, Lingenthal G, von Bülow M, Kern HF. Histological and fine structural features of pancreatic ductal adenocarcinoma in relation to growth and prognosis: studies in xenografted tumors and clinico-histopathological correlation in a series of 75 cases. Histopathology. 1985;9:841. doi: 10.1111/j.1365-2559.1985.tb02870.x. [DOI] [PubMed] [Google Scholar]
  • 19.Köhler G, Howe SC, Milstein C. Fusion between immunoglobulin secreting and nonsecreting myeloma cell lines. Eur J Immunol. 1976;6:292. doi: 10.1002/eji.1830060411. [DOI] [PubMed] [Google Scholar]
  • 20.Ledeen RW, Yu RK. Gangliosides: Structure, Isolation and Analysis. Methods Enzymol. 1982;83:139. doi: 10.1016/0076-6879(82)83012-7. [DOI] [PubMed] [Google Scholar]
  • 21.Magnani JL, Brockhaus M, Smith DF, Ginsburg V. A monosialoganglioside is a monoclonal antibody-defined antigen of colon carcinoma. Science. 1982;212:55. doi: 10.1126/science.7209516. [DOI] [PubMed] [Google Scholar]
  • 22.Magnani JL, Nilsson B, Brockhaus M, Zopf D, Steplewski Z, Koprowski H, Ginsburg V. A monoclonal antibody defined antigen associated with gastrointestinal cancer is a ganglioside containing sialylated lacto-N-fucopentaose II. J Biol Chem. 1982;257:14365. [PubMed] [Google Scholar]
  • 23.Magnani JL, Steplewski Z, Koprowski H, Ginsburg V. Identification of the gastrointestinal and pancreatic cancer associated antigen detected by monoclonal antibody 199 in the sera of patients as a mucin. Cancer Res. 1983;43:5489. [PubMed] [Google Scholar]
  • 24.Ouchterlony Ö. Antigen antibody reactions in gels. Arkiv Kemi Mineral Geol. 1948;26:1. [Google Scholar]
  • 25.Pimm MV, Baldwin RW. Quantitative evaluation of the localization of a monoclonal antibody (791T36) in human osteogenic sarcoma xenografts. Eur J Cancer Clin Oncol. 1984;20:515. doi: 10.1016/0277-5379(84)90238-4. [DOI] [PubMed] [Google Scholar]
  • 26.Pukel CS, Lloyd KO, Travassos LR, Dippold WG, Oettgen HF, Old LJ. GD3 — a prominent ganglioside of human melanoma: detection and characterization by mouse monoclonal antibody. J Exp Med. 1982;155:1133. doi: 10.1084/jem.155.4.1133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27.Reinherz EL, Kung PC, Goldstein G, Schlossman SF. Separation of functional subsets of human T-cells by a monoclonal antibody. Proc Natl Acad Sci USA. 1979;76:4061. [Google Scholar]
  • 28.Rusckowski M, Doherty PW, Virzi F, Hnatowich DJ. A simple in vitro method of screening panels of monoclonal antibodies for tumor binding. J Immunol Methods. 1985;76:375. doi: 10.1016/0022-1759(85)90314-x. [DOI] [PubMed] [Google Scholar]
  • 29.Schulz G, Bumol TF, Reisfeld RA. Monoclonal antibody directed effector cells selectively lyse human melanoma cells in vitro and in vivo. Proc Natl Acad Sci USA. 1983;80:5407. doi: 10.1073/pnas.80.17.5407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Shulman M, Wilde CD, Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978;276:269. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  • 31.Suzuki K. The pattern of mammalian brain gangliosides-II. Evaluation of the extraction procedures, post-mortem changes and the effect of formalin preservation. J Neurochem. 1965;12:629. doi: 10.1111/j.1471-4159.1965.tb04256.x. [DOI] [PubMed] [Google Scholar]
  • 32.Svennerholm L. Quantitative estimation of sialic acids. II. A colorimetric resorcinol-hydrochloric acid method. Biochim Biophys Acta. 1957;24:604. doi: 10.1016/0006-3002(57)90254-8. [DOI] [PubMed] [Google Scholar]
  • 33.Schmiegel WH, Kalthoff H, Arndt R, Gieseking J, Greten H, Klöppel G, Kreiker Ch, Ladak A, Lampe V, Ulrich S. Monoclonal antibody-defined human pancreatic cancer-associated antigens. Cancer Res. 1985;45:1402. [PubMed] [Google Scholar]
  • 34.Yachi A, Imai K, Fujita H, Moriya Y, Tanda M, Endo T, Tsujisaki M, Kawaharada M. Immunohistochemical distribution of the antigenic determinants detected by monoclonal antibodies to carcinoembryonic antigen. J Immunol. 1984;132:2998. [PubMed] [Google Scholar]
  • 35.Yu RK, Ledeen RW. Gangliosides of human, bovine, and rabbit plasma. J Lipid Res. 1972;13:680. [PubMed] [Google Scholar]

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

RESOURCES