Skip to main content
NCBI home page
British Journal of Cancer logoLink to British Journal of Cancer
. 1991 Dec;64(6):1001–1010. doi: 10.1038/bjc.1991.455

Detection by a human monoclonal antibody of a glycoprotein associated with malignant proliferation of mammary epithelial cells.

S A Imam 1, L A Mills 1, C R Taylor 1
PMCID: PMC1977871  PMID: 1662531

Abstract

A tumour-associated antigen (TAA.62) with an apparent mol. wt. of 62 kd, identified by a human monoclonal antibody (IgG2, kappa-light chain), was found to be expressed at elevated levels in the cytoplasmic compartment of malignant as compared with normal mammary epithelial cells in both tissues and cultured cells. Increased levels of cytoplasmic expression of the antigen were also observed in malignant cells of cervix, colon, kidney, lung, and stomach. The patterns of expression of TAA.62 in cultured cells mirrored those of tissues and the antigen was expressed at elevated levels in the established breast cancer lines or oncogenically transformed mammary carcinoma cell line (tumourigenic) compared with the immortalised mammary epithelial cell line (non-tumourigenic). Aliquots of TAA.62 were purified to homogeneity from the conditioned-medium of malignant and immortalised breast cells by immunoaffinity chromatography using immobilised anti-TAA.62 antibody, and gel filtration. Both preparations of TAA.62 yielded a single band with an apparent molecular weight of 62 kd under reducing condition on sodium dodecyl sulphate-polyacrylamide gel electrophoresis, and both were identical in terms of size and immunoreactivity to anti-TAA.62 antibody. However, TAA.62(T) isolated from tumourigenic cell lines itself interacted with a cell surface molecule having an apparent molecular weight of 160 kd on both the malignant and immortalised cells: TAA.62(I) isolated from immortalized cell lines, showed no comparable interaction. Scatchard analysis of the concentration-dependent binding of TAA.62(T) to 160 kd-receptor molecule revealed a 2.6 x 10(4) binding sites per cell. The association constant of such binding was determined to be approximately 16.6 nM. Finally, addition of anti-TAA.62 antibody to culture medium resulted in the inhibition of proliferation of the malignant cells, but showed no effect on the normal cells. The results suggest that TAA.62 may interact as a ligand with its 160 kd cell surface receptor with a possible growth related function.

Full text

PDF
1001

Images in this article

1004Figure 1
on p.1004
1005Figure 2
on p.1005
1005Figure 3
on p.1005
1006Figure 4
on p.1006
1008Figure 10
on p.1008

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BLOOM H. J., RICHARDSON W. W. Histological grading and prognosis in breast cancer; a study of 1409 cases of which 359 have been followed for 15 years. Br J Cancer. 1957 Sep;11(3):359–377. doi: 10.1038/bjc.1957.43. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Borup-Christensen P., Erb K., Jensenius J. C., Nielsen B., Svehag S. E. Human-human hybridomas for the study of anti-tumor immune response in patients with colorectal cancer. Int J Cancer. 1986 May 15;37(5):683–688. doi: 10.1002/ijc.2910370507. [DOI] [PubMed] [Google Scholar]
  3. Ceriani R. L., Thompson K., Peterson J. A., Abraham S. Surface differentiation antigens of human mammary epithelial cells carried on the human milk fat globule. Proc Natl Acad Sci U S A. 1977 Feb;74(2):582–586. doi: 10.1073/pnas.74.2.582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Clark R., Stampfer M. R., Milley R., O'Rourke E., Walen K. H., Kriegler M., Kopplin J., McCormick F. Transformation of human mammary epithelial cells by oncogenic retroviruses. Cancer Res. 1988 Aug 15;48(16):4689–4694. [PubMed] [Google Scholar]
  5. Colcher D., Hand P. H., Nuti M., Schlom J. A spectrum of monoclonal antibodies reactive with human mammary tumor cells. Proc Natl Acad Sci U S A. 1981 May;78(5):3199–3203. doi: 10.1073/pnas.78.5.3199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cote R. J., Morrissey D. M., Houghton A. N., Beattie E. J., Jr, Oettgen H. F., Old L. J. Generation of human monoclonal antibodies reactive with cellular antigens. Proc Natl Acad Sci U S A. 1983 Apr;80(7):2026–2030. doi: 10.1073/pnas.80.7.2026. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hammond S. L., Ham R. G., Stampfer M. R. Serum-free growth of human mammary epithelial cells: rapid clonal growth in defined medium and extended serial passage with pituitary extract. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5435–5439. doi: 10.1073/pnas.81.17.5435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Haspel M. V., McCabe R. P., Pomato N., Janesch N. J., Knowlton J. V., Peters L. C., Hoover H. C., Jr, Hanna M. G., Jr Generation of tumor cell-reactive human monoclonal antibodies using peripheral blood lymphocytes from actively immunized colorectal carcinoma patients. Cancer Res. 1985 Aug;45(8):3951–3961. [PubMed] [Google Scholar]
  9. Herlyn M., Steplewski Z., Herlyn D., Koprowski H. Colorectal carcinoma-specific antigen: detection by means of monoclonal antibodies. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1438–1442. doi: 10.1073/pnas.76.3.1438. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Imam A., Drushella M. M., Taylor C. R. A novel immunoperoxidase procedure of using human monoclonal antibodies for the detection of cellular antigens in tissue sections. J Immunol Methods. 1986 Jan 22;86(1):17–20. doi: 10.1016/0022-1759(86)90259-0. [DOI] [PubMed] [Google Scholar]
  11. Imam A., Drushella M. M., Taylor C. R., Tökés Z. A. Generation and immunohistological characterization of human monoclonal antibodies to mammary carcinoma cells. Cancer Res. 1985 Jan;45(1):263–271. [PubMed] [Google Scholar]
  12. Imam A., Stephanian E. Use of xenograft tissue for the initial screening of human monoclonal antibodies by immunohistological technique. J Immunol Methods. 1988 Nov 10;114(1-2):69–72. doi: 10.1016/0022-1759(88)90155-x. [DOI] [PubMed] [Google Scholar]
  13. Imam A., Taylor C. R. Biochemical and immunological characterizations of antigens recognised by human monoclonal antibodies. Br J Cancer. 1989 Jun;59(6):922–928. doi: 10.1038/bjc.1989.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Lowe D. H., Handley H. H., Schmidt J., Royston I., Glassy M. C. A human monoclonal antibody reactive with human prostate. J Urol. 1984 Oct;132(4):780–785. doi: 10.1016/s0022-5347(17)49872-7. [DOI] [PubMed] [Google Scholar]
  17. Minna J. D., Cuttitta F., Rosen S., Bunn P. A., Jr, Carney D. N., Gazdar A. F., Krasnow S. Methods for production of monoclonal antibodies with specificity for human lung cancer cells. In Vitro. 1981 Dec;17(12):1058–1070. doi: 10.1007/BF02618603. [DOI] [PubMed] [Google Scholar]
  18. Parham P. On the fragmentation of monoclonal IgG1, IgG2a, and IgG2b from BALB/c mice. J Immunol. 1983 Dec;131(6):2895–2902. [PubMed] [Google Scholar]
  19. Schlom J., Wunderlich D., Teramoto Y. A. Generation of human monoclonal antibodies reactive with human mammary carcinoma cells. Proc Natl Acad Sci U S A. 1980 Nov;77(11):6841–6845. doi: 10.1073/pnas.77.11.6841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sikora K., Wright R. Human monoclonal antibodies to lung-cancer antigens. Br J Cancer. 1981 May;43(5):696–700. doi: 10.1038/bjc.1981.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Soule H. R., Linder E., Edgington T. S. Membrane 126-kilodalton phosphoglycoprotein associated with human carcinomas identified by a hybridoma antibody to mammary carcinoma cells. Proc Natl Acad Sci U S A. 1983 Mar;80(5):1332–1336. doi: 10.1073/pnas.80.5.1332. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Starling J. J., Cote R. J., Marder P., Borowitz M. J., Johnson D. A. Tissue distribution and cellular location of the antigens recognized by human monoclonal antibodies 16.88 and 28A32. Cancer Res. 1988 Dec 15;48(24 Pt 1):7273–7278. [PubMed] [Google Scholar]
  23. Staros J. V. N-hydroxysulfosuccinimide active esters: bis(N-hydroxysulfosuccinimide) esters of two dicarboxylic acids are hydrophilic, membrane-impermeant, protein cross-linkers. Biochemistry. 1982 Aug 17;21(17):3950–3955. doi: 10.1021/bi00260a008. [DOI] [PubMed] [Google Scholar]

Articles from British Journal of Cancer are provided here courtesy of Cancer Research UK

RESOURCES