Urological Research

, Volume 20, Issue 3, pp 219–228 | Cite as

Morphological aspects of the interaction of Bacillus Calmette-Guérin with urothelial bladder cells in vivo and in vitro: relevance for antitumor activity?

  • J. S. Teppema
  • E. C. de Boer
  • P. A. Steerenberg
  • A. P. M. van der Meijden
Original Articles


Intravesical administration of Bacillus Calmette-Guérin (BCG) has been shown to be effective in the treatment of patients with superficial bladder cancer. For a better understanding of the mechanism of this antitumor activity, scanning and transmission electron microscope (SEM, TEM) studies were carried out to investigate morphological aspects of the interaction of BCG with the bladder wall in vivo and in vitro. Adherence of BCG to the bladder wall in vivo was studied 1 and 24 h after single or multiple (6×) BCG instillations in intact and in electrocauterized guinea pig bladders. Despite extensive search with SEM for its presence, virtually no BCG was found on the intact urothelium, and BCG was only occasionally observed in the coagulation lesions. SEM and TEM studies revealed adherence and phagocytosis of BCG by the T24 human bladder carcinoma cell line in vitro. Time sequence studies on the phagocytosis and fate of BCG showed that T24 cells are capable of progressively degrading the mycobacteria in phagolysosomes. However, BCG did not alter MHC class II antigen expression on T24 cells in vitro. In contrast, 54 urine sediments and bladder washings of 11 bladder cancer patients, taken prior to or after several intravesical BCG instillations, failed to demonstrate urothelial (tumor) cells showing evidence of BCG phagocytosis (682 cells screened by TEM), while BCG was phagocytized avidly by leukocytes. These data suggest that a direct interaction of BCG with urothelial bladder cells in vivo can be called in question.

Key words

Intravestical BCG Bladder urothelium Morphological interaction Man Guinea pig 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Becich MJ, Carroll S, Ratliff TL (1991) Internalization of bacille Calmette-Guérin by bladder tumor cells. J Urol 145:1316Google Scholar
  2. 2.
    Böhle A, Gerdes J, Ulmer AJ, Hofstetter AG, Flad F (1990) Effects of local BCG therapy in patients with bladder carcinoma on immunocompetent cells of the bladder wall. J Urol 144:53Google Scholar
  3. 3.
    Brown AE, Holzer TJ, Anderson BR (1987) Capacity of human neutrophils to kill Myobacterium tuberculosis. J Infect Dis 156:985Google Scholar
  4. 4.
    Bubeník J, Baresová M, Viklicky V, Jakoubková J, Sainerová H, Donner J (1973) Established cell line of urinary bladder carcinoma (T24) containing tumour-specific antigen. Int J Cancer 11:765Google Scholar
  5. 5.
    Burchard GD, Bierther M (1985) Study on the micromorphology of Mycobacterium leprae. Arch Dermatol Res 277:220Google Scholar
  6. 6.
    Davies M (1982) Bacillus Calmette-Guérin as an antitumor agent. The interaction with cells of the mammalian immune system. Biochim Biophys Acta 651:143Google Scholar
  7. 7.
    De Boer EC, de Jong WH, van der Meijden APM, Steerenberg PA, Witjes F, Vegt PDJ, Debruyne FMJ, Ruitenberg EJ (1991) Leukocytes in the urine after intravesical BCG treatment for superficial bladder cancer. A flow cytofluorometric analysis. Urol Res 19:45Google Scholar
  8. 8.
    De Jong WH, Steerenberg PA, Ruitenberg EJ (1987) Bacillus Calmette-Guérin (BCG) and its use for cancer immunotherapy. In: den Otter W, Ruitenberg EJ (eds) Tumor immunology-mechanisms, diagnosis, therapy. Elsevier, Amsterdam, p 283Google Scholar
  9. 9.
    El-Demiry MIM, Smith G, Ritchie AWS, James K, Cumming JA, Hargreave TB, Chisholm GD (1987) Local immune responses after intravesical bacille Calmette-Guérin treatment for carcinoma in situ. Br J Urol 60:543Google Scholar
  10. 10.
    Johnson SC, Zwilling BS (1985) Continous expression of I-A antigen by peritoneal macrophages from mice resistant to Mycobacterium bovis (BCG). J Leukocyte Biol 38:635Google Scholar
  11. 11.
    Lage JM, Bauer WC, Kelley DR, Ratliff TL, Catalona WJ (1986) Histological parameters and pitfalls in the interpretation of bladder biopsies in bacillus Calmette-Guérin treatment of superficial bladder cancer. J Urol 135:916Google Scholar
  12. 12.
    Leake ES, Ockers JR, Myrvik QN (1977) In vitro interactions of the BCG and Ravenel strains of Mycobacterium bovis with rabbit macrophages: adherence of the phagosomal membrane to the bacterial cell wall and the problem of the perbacillary space. J Reticuloendothel Soc 22:129Google Scholar
  13. 13.
    Leong AS-Y, Wannakrairot P, Jose J, Milios J (1990) Bacillus Calmette Guérin-treated superficial bladder cancer: correlation of morphology with immunophenotyping. J Pathol (London) 162:35Google Scholar
  14. 14.
    Mitchell MS, Murahata RI (1979) Modulation of immunity by Bacillus Calmette-Guérin (BCG). Pharmacol Ther 4:329Google Scholar
  15. 15.
    Morales A (1989) BCG in the treatment of bladder cancer: state of the art. In: Debruyne FMJ, Denis L, van der Meijden APM (eds) BCG in superficial bladder cancer. (EORTC Genitourinary Group, monography 6) Liss, New York, p 3Google Scholar
  16. 16.
    Morales A, Nickel JC (1986) Immunotherapy of superficial bladder cancer with BCG. World J Urol 3:209Google Scholar
  17. 17.
    Morales A, Eidinger D, Bruce AW (1976) Intracavitary bacillus Calmette-Guérin in the treatment of superficial bladder tumors. J Urol 116:180Google Scholar
  18. 18.
    Moulder JM (1985) Comparative biology of intracellular parasitism. Micribiol Rev 49:298Google Scholar
  19. 19.
    Mshana RN, Hastings RC, Krahenbuhl JL (1988) Infection with live mycobacteria inhibits in vitro detection of Ia antigen on macrophages. Immunobiology 177:40Google Scholar
  20. 20.
    Myrvik QN, Leake ES, Tenner-Racz K (1983) Interaction of mycobacteria with normal and immunlogically activated alveolar macrophages. Adv Exp Med Biol 162:83Google Scholar
  21. 21.
    Newling D (1990) Intravesical therapy in the management of superficial transitional cell carcinoma of the bladder: the experience of the EORTC GU group. Br J Cancer 61:497Google Scholar
  22. 22.
    Pode D, Alon Y, Horowitz AT, Vlodavsky I, Biran S (1986) The mechanism of human bladder tumor implantation in an in vitro model. J Urol 136:482Google Scholar
  23. 23.
    Prescott S, James K, Busuttil A, Hargreave TB, Chisholm GD, Smyth JF (1989) HLA-DR expression by high-grade superficial bladder cancer treated with BCG. Br J Urol 63:264Google Scholar
  24. 24.
    Prescott S, James K, Hargreave TB, Chisholm GD, Smyth JF (1989) Immunopathological effects of intravesical BCG therapy. In: Debruyne FMJ, Denis L, van der Meijden APM (eds) BCG in superficial bladder cancer. (EORTC Genitourinary Group, monograph 6) Liss, New York, p 93Google Scholar
  25. 25.
    Ratliff TL (1989) Mechanisms of action of intravesical BCG for bladder cancer. In: Debruyne FMJ, Denis L, van der Meijden APM (eds) BCG in superficial bladder cancer. (EORTC Genitourinary Group, monograph 6) Liss, New York, p 107Google Scholar
  26. 26.
    Ratliff TL, Palmer JO, McGarr JA, Brown EJ (1987) Intravesical Bacillus Calmette-Guérin therapy for murine bladder tumors: initiation of the response by fibronectin-mediated attachment of Bacillus Calmette-Guérin. Cancer Res 47:1762Google Scholar
  27. 27.
    Ratliff TL, Kavoussi LR, Catalona WJ (1988) Role of fibronectin in intravesical BCG therapy for superficial bladder cancer. J Urol 139:410Google Scholar
  28. 28.
    Rigatti P, Colombo R, Montorsi F, Centemero A, Guazzoni G, Di Gorolamo V, Galli L, Trabucchi E (1990) Local Bacillus Calmette-Guérin therapy for superficial bladder cancer: clinical, histological and ultrastructural patterns. Scand J Urol Nephrol 24:191Google Scholar
  29. 29.
    Ryter A, Frehel C, Rastogi N, David HL (1984) Macrophage interaction with mycobacteria including M. leprae. Acta Leprol 2:211Google Scholar
  30. 30.
    Sibley LD, Franzblau SG, Krahenbuhl JL (1987) Intracellular fate of Mycobacterium leprae in normal and activated mouse macrophages. Infect Immun 55:680Google Scholar
  31. 31.
    Silva MT, Macedo PM (1984) Ultrastructural characterization of normal and damaged membranes of Mycobacterium leprae and of cultivable mycobacteria. J Gen Microbiol 130:369Google Scholar
  32. 32.
    Silva MT, Appelberg R, Silva MNT, Macedo PM (1987) In vivo killing and degradation of Mycobacterium aurum within mouse peritoneal macrophages. Infect Immun 55:2006Google Scholar
  33. 33.
    Stefanini GF, Bercovich E, Mazzeo V, Grigioni WF, Emili E, D'Errico A, Lo Cigno M, Tamagnini N, Mazetti M (1989) Class I and class II HLA antigen expression by transitional cell carcinoma of the bladder: correlation with T-cell infiltration and BCG treatment. J Urol 141:1449Google Scholar
  34. 34.
    Van der Meijden APM, Debruyne FMJ, Steerenberg PA, de Jong WH (1989) Aspects of non-specific immunotherapy with BCG in superficial bladder cancer: an overview. In: Debruyne FMJ, Denis L, van der Meijden APM (eds) BCG in superficial bladder cancer. (EORTC Genitourinary Group, monograph 6) Liss, New York, p 11Google Scholar
  35. 35.
    Van der Meijden APM, de Jong WH, Steerenberg PA, Walvoort HC, de Boer EC, Debruyne FMJ, Ruitenberg EJ (1988) Intravesical BCG administration in the guinea pig. A histomorphological study. Virchows Arch [B] 55:207Google Scholar

Copyright information

© Springer-Verlag 1992

Authors and Affiliations

  • J. S. Teppema
    • 1
  • E. C. de Boer
    • 1
  • P. A. Steerenberg
    • 1
  • A. P. M. van der Meijden
    • 2
    • 3
  1. 1.Laboratory for PathologyNational Institute of Public Health and Environmental ProtectionBilthovenThe Netherlands
  2. 2.Department of UrologyGrootziekengasthuis's-HertogenboschThe Netherlands
  3. 3.Department of UrologyAcademic Hospital, Catholic UniversityNijmegenThe Netherlands

Personalised recommendations