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. 1987 Oct;56(4):439–444. doi: 10.1038/bjc.1987.220

Cellular heterogeneity in normal and neoplastic human urothelium: a study using murine monoclonal antibodies.

G Dotsikas 1, T Konowalchuk 1, P P Major 1, P E Kovac 1, G K Ward 1, S S Stewart 1, G B Price 1, M M Elhilali 1, W J Mackillop 1
PMCID: PMC2001815  PMID: 3689660

Abstract

To assist in the description of the cellular heterogeneity present in normal and neoplastic urothelium, a panel of monoclonal antibodies (MoAbs) was raised against human transitional cell carcinoma (TCC) of the urinary bladder. All immunizations were carried out using whole cells and membrane preparations from well differentiated human TCCs. Two fusions produced 145 hybridomas. Following primary screening by ELISA and secondary screening with immunohistochemistry, three useful antibodies were identified. MoAb 35.48 binds to all cell layers of the normal urothelium and well differentiated tumours, but not to the majority of poorly differentiated tumours. MoAb 21.48 binds preferentially to the basal cell layer of normal urothelium and to some well differentiated papillary TCCs, but poorly differentiated tumours exhibit diffusely positive staining. MoAb 21.48 also shows cross-reactivity with basal cell layers of other epithelia. MoAb 5.48 binds preferentially to the superficial cell layers of normal urothelium and well differentiated TCCs, but exhibits less binding in poorly differentiated tumours with loss of the preferential superficial staining. Quantitative flow cytometric studies indicate that MoAb 5.48 binds to a cell-surface antigen which is present on significantly fewer cells of poorly differentiated tumours than on either normal urothelium (P less than 0.05), or well differentiated tumours (P = 0.05).

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Selected References

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  1. Barnes R. W., Dick A. L., Hadley H. L., Johnston O. L. Survival following transurethral resection of bladder carcinoma. Cancer Res. 1977 Aug;37(8 Pt 2):2895–2897. [PubMed] [Google Scholar]
  2. Bates D. A., Le Grimellec C., Bates J. H., Loutfi A., Mackillop W. J. Effects of thermal adaptation at 40 degrees C on membrane viscosity and the sodium-potassium pump in Chinese hamster ovary cells. Cancer Res. 1985 Oct;45(10):4895–4899. [PubMed] [Google Scholar]
  3. Bizzari J. P., Mackillop W. J. The estimation of self-renewal in the clonogenic cells of human solid tumours: a comparison of secondary plating efficiency and colony size. Br J Cancer. 1985 Aug;52(2):189–195. doi: 10.1038/bjc.1985.177. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Fradet Y., Cordon-Cardo C., Thomson T., Daly M. E., Whitmore W. F., Jr, Lloyd K. O., Melamed M. R., Old L. J. Cell surface antigens of human bladder cancer defined by mouse monoclonal antibodies. Proc Natl Acad Sci U S A. 1984 Jan;81(1):224–228. doi: 10.1073/pnas.81.1.224. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fradet Y., Cordon-Cardo C., Whitmore W. F., Jr, Melamed M. R., Old L. J. Cell surface antigens of human bladder tumors: definition of tumor subsets by monoclonal antibodies and correlation with growth characteristics. Cancer Res. 1986 Oct;46(10):5183–5188. [PubMed] [Google Scholar]
  6. Grossman H. B. Hybridoma antibodies reactive with human bladder carcinoma cell surface antigens. J Urol. 1983 Sep;130(3):610–614. doi: 10.1016/s0022-5347(17)51324-5. [DOI] [PubMed] [Google Scholar]
  7. Horibata K., Harris A. W. Mouse myelomas and lymphomas in culture. Exp Cell Res. 1970 Apr;60(1):61–77. doi: 10.1016/0014-4827(70)90489-1. [DOI] [PubMed] [Google Scholar]
  8. Hsu S. M., Raine L., Fanger H. Use of avidin-biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem. 1981 Apr;29(4):577–580. doi: 10.1177/29.4.6166661. [DOI] [PubMed] [Google Scholar]
  9. Koho H., Paulie S., Ben-Aissa H., Jónsdóttir I., Hansson Y., Lundblad M. L., Perlmann P. Monoclonal antibodies to antigens associated with transitional cell carcinoma of the human urinary bladder. I. Determination of the selectivity of six antibodies by cell ELISA and immunofluorescence. Cancer Immunol Immunother. 1984;17(3):165–172. doi: 10.1007/BF00205481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  11. Mackillop W. J., Bizarri J. P., Ward G. K. Cellular heterogeneity in normal and neoplastic human urothelium. Cancer Res. 1985 Sep;45(9):4360–4365. [PubMed] [Google Scholar]
  12. Mackillop W. J., Buick R. N. Cellular heterogeneity in human ovarian carcinoma studied by density gradient fractionation. Stem Cells. 1982;1(6):355–366. [PubMed] [Google Scholar]
  13. Mackillop W. J., Ciampi A., Till J. E., Buick R. N. A stem cell model of human tumor growth: implications for tumor cell clonogenic assays. J Natl Cancer Inst. 1983 Jan;70(1):9–16. [PubMed] [Google Scholar]
  14. Mackillop W. J., Stewart S. S., Buick R. N. Density/volume analysis in the study of cellular heterogeneity in human ovarian carcinoma. Br J Cancer. 1982 Jun;45(6):812–820. doi: 10.1038/bjc.1982.132. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Major P. P., Kovac P. E., Lavallée M. L., Kovalik E. C. Monoclonal antibodies to antigens abnormally expressed in breast cancer. J Histochem Cytochem. 1987 Feb;35(2):139–148. doi: 10.1177/35.2.3540102. [DOI] [PubMed] [Google Scholar]
  16. Masuko T., Yagita H., Hashimoto Y. Monoclonal antibodies against cell surface antigens present on human urinary bladder cancer cells. J Natl Cancer Inst. 1984 Mar;72(3):523–530. [PubMed] [Google Scholar]
  17. Messing E. M., Bubbers J. E., Whitmore K. E., deKernion J. B., Nestor M. S., Fahey J. L. Murine hybridoma antibodies against human transitional carcinoma-associated antigens. J Urol. 1984 Jul;132(1):167–172. doi: 10.1016/s0022-5347(17)49512-7. [DOI] [PubMed] [Google Scholar]
  18. Pierce G. B., Wallace C. Differentiation of malignant to benign cells. Cancer Res. 1971 Feb;31(2):127–134. [PubMed] [Google Scholar]
  19. Sasaki M. Production and characterization of monoclonal antibodies to the established human bladder cancer cell lines. Keio J Med. 1984 Apr;33(1):39–56. doi: 10.2302/kjm.33.39. [DOI] [PubMed] [Google Scholar]
  20. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  21. Stewart S. S., Price G. B. Realtime acquisition, storage, and display of correlated three-parameter flow cytometric data. Cytometry. 1986 Jan;7(1):82–88. doi: 10.1002/cyto.990070112. [DOI] [PubMed] [Google Scholar]
  22. Summerhayes I. C., McIlhinney R. A., Ponder B. A., Shearer R. J., Pocock R. D. Monoclonal antibodies raised against cell membrane components of human bladder tumor tissue recognizing subpopulations in normal urothelium. J Natl Cancer Inst. 1985 Dec;75(6):1025–1038. [PubMed] [Google Scholar]
  23. Trejdosiewicz L. K., Southgate J., Donald J. A., Masters J. R., Hepburn P. J., Hodges G. M. Monoclonal antibodies to human urothelial cell lines and hybrids: production and characterization. J Urol. 1985 Mar;133(3):533–538. doi: 10.1016/s0022-5347(17)49048-3. [DOI] [PubMed] [Google Scholar]
  24. Ward G. K., Stewart S. S., Price G. B., Mackillop W. J. Cellular heterogeneity in normal human urothelium: an analysis of optical properties and lectin binding. J Histochem Cytochem. 1986 Jul;34(7):841–846. doi: 10.1177/34.7.3754881. [DOI] [PubMed] [Google Scholar]
  25. Whitmore W. F., Jr Surgical management of low stage bladder cancer. Semin Oncol. 1979 Jun;6(2):207–216. [PubMed] [Google Scholar]
  26. Young D. A., Prout G. R., Jr, Lin C. W. Production and characterization of mouse monoclonal antibodies to human bladder tumor-associated antigens. Cancer Res. 1985 Sep;45(9):4439–4446. [PubMed] [Google Scholar]

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