Search tips
Search criteria 


Logo of jcinvestThe Journal of Clinical InvestigationCurrent IssueArchiveSubscriptionAbout the Journal
J Clin Invest. 1993 January; 91(1): 69–76.
PMCID: PMC329996

Characterization of the internalization of bacillus Calmette-Guerin by human bladder tumor cells.


Adjuvant intravesical Mycobacterium bovis BCG is the treatment of choice for recurrent superficial bladder cancer. Fibronectin (FN) was previously demonstrated to be necessary for the retention of BCG within the bladder and for the expression of antitumor activity. Recent studies have demonstrated that BCG attach and are ingested by bladder epithelial cells, suggesting the existence of a second bacterial attachment mechanism. We report the characterization of the molecules involved in BCG attachment and internalization by the human bladder transitional cell carcinoma cell line T-24. Pretreatment of T-24 cells with monoclonal antibodies to either alpha 5 or beta 1 integrin subunits significantly inhibited both BCG attachment and ingestion. Exogenous FN was observed to enhance both attachment and ingestion of BCG, and anti-FN was observed to inhibit both phenomena. Latex beads precoated with either FN or laminin (LN) but not BSA were ingested by T-24 cells, but only FN-coated beads inhibited BCG attachment and ingestion. Pretreatment of BCG with FN augmented both attachment and ingestion. The role of bacterial FN binding proteins was evaluated. A monoclonal antibody to a 55-kD FN-binding protein was observed to abrogate attachment and ingestion. These results demonstrate that attachment and ingestion of BCG are mediated in part by the alpha 5 beta 1 integrin receptor and are dependent on FN. These studies demonstrate a mechanism of entrance of mycobacteria into epithelial cells and suggest a second role for FN in the adjuvant antitumor effect of BCG.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (2.0M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Morales A, Eidinger D, Bruce AW. Intracavitary Bacillus Calmette-Guerin in the treatment of superficial bladder tumors. J Urol. 1976 Aug;116(2):180–183. [PubMed]
  • Herr HW, Pinsky CM, Whitmore WF, Jr, Oettgen HF, Melamed MR. Effect of intravesical Bacillus Calmette-Guerin (BCG) on carcinoma in situ of the bladder. Cancer. 1983 Apr 1;51(7):1323–1326. [PubMed]
  • Brosman SA. Experience with bacillus Calmette-Guerin in patients with superficial bladder carcinoma. J Urol. 1982 Jul;128(1):27–30. [PubMed]
  • Mori K, Lamm DL, Crawford ED. A trial of bacillus Calmette-Guérin versus adriamycin in superficial bladder cancer: a South-West Oncology Group Study. Urol Int. 1986;41(4):254–259. [PubMed]
  • Lamm DL, Blumenstein BA, Crawford ED, Montie JE, Scardino P, Grossman HB, Stanisic TH, Smith JA, Jr, Sullivan J, Sarosdy MF, et al. A randomized trial of intravesical doxorubicin and immunotherapy with bacille Calmette-Guérin for transitional-cell carcinoma of the bladder. N Engl J Med. 1991 Oct 24;325(17):1205–1209. [PubMed]
  • Kavoussi LR, Brown EJ, Ritchey JK, Ratliff TL. Fibronectin-mediated Calmette-Guerin bacillus attachment to murine bladder mucosa. Requirement for the expression of an antitumor response. J Clin Invest. 1990 Jan;85(1):62–67. [PMC free article] [PubMed]
  • Ratliff TL, Gillen D, Catalona WJ. Requirement of a thymus dependent immune response for BCG-mediated antitumor activity. J Urol. 1987 Jan;137(1):155–158. [PubMed]
  • Becich MJ, Carroll S, Ratliff TL. Internalization of bacille Calmette-Guerin by bladder tumor cells. J Urol. 1991 Jun;145(6):1316–1324. [PubMed]
  • Saito H, Tomioka H, Watanabe T, Sato K. Mechanisms of phagocytosis of Mycobacterium leprae and other mycobacteria by human oligodendroglial cells. Infect Immun. 1986 Jan;51(1):163–167. [PMC free article] [PubMed]
  • Band AH, Chitamber SD, Bhattacharya A, Talwar GP. Mechanism of phagocytosis of mycobacteria by Schwann cells and their comparison with macrophages. Int J Lepr Other Mycobact Dis. 1986 Jun;54(2):294–299. [PubMed]
  • Hamill RJ, Vann JM, Proctor RA. Phagocytosis of Staphylococcus aureus by cultured bovine aortic endothelial cells: model for postadherence events in endovascular infections. Infect Immun. 1986 Dec;54(3):833–836. [PMC free article] [PubMed]
  • Mukherjee R, Antia NH. Host-parasite interrelationship between M. leprae and Schwann cells in vitro. Int J Lepr Other Mycobact Dis. 1986 Dec;54(4):632–638. [PubMed]
  • Isberg RR, Leong JM. Multiple beta 1 chain integrins are receptors for invasin, a protein that promotes bacterial penetration into mammalian cells. Cell. 1990 Mar 9;60(5):861–871. [PubMed]
  • Leininger E, Roberts M, Kenimer JG, Charles IG, Fairweather N, Novotny P, Brennan MJ. Pertactin, an Arg-Gly-Asp-containing Bordetella pertussis surface protein that promotes adherence of mammalian cells. Proc Natl Acad Sci U S A. 1991 Jan 15;88(2):345–349. [PubMed]
  • Rook GA, Champion BR, Steele J, Varey AM, Stanford JL. I-A restricted activation by T cell lines of anti-tuberculosis activity in murine macrophages. Clin Exp Immunol. 1985 Feb;59(2):414–420. [PubMed]
  • Hall DE, Reichardt LF, Crowley E, Holley B, Moezzi H, Sonnenberg A, Damsky CH. The alpha 1/beta 1 and alpha 6/beta 1 integrin heterodimers mediate cell attachment to distinct sites on laminin. J Cell Biol. 1990 Jun;110(6):2175–2184. [PMC free article] [PubMed]
  • Fradet Y, Cordon-Cardo C, Whitmore WF, Jr, Melamed MR, Old LJ. 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]
  • Kennel SJ, Foote LJ, Falcioni R, Sonnenberg A, Stringer CD, Crouse C, Hemler ME. Analysis of the tumor-associated antigen TSP-180. Identity with alpha 6-beta 4 in the integrin superfamily. J Biol Chem. 1989 Sep 15;264(26):15515–15521. [PubMed]
  • Basara ML, McCarthy JB, Barnes DW, Furcht LT. Stimulation of haptotaxis and migration of tumor cells by serum spreading factor. Cancer Res. 1985 Jun;45(6):2487–2494. [PubMed]
  • Pommier CG, Inada S, Fries LF, Takahashi T, Frank MM, Brown EJ. Plasma fibronectin enhances phagocytosis of opsonized particles by human peripheral blood monocytes. J Exp Med. 1983 Jun 1;157(6):1844–1854. [PMC free article] [PubMed]
  • Cantarero LA, Butler JE, Osborne JW. The adsorptive characteristics of proteins for polystyrene and their significance in solid-phase imunoassays. Anal Biochem. 1980 Jul 1;105(2):375–382. [PubMed]
  • Coplen DE, Brown EJ, McGarr J, Ratliff TL. Characterization of fibronectin attachment by a human transitional cell carcinoma line, T24. J Urol. 1991 Jun;145(6):1312–1315. [PubMed]
  • Portnoy DA, Jacks PS, Hinrichs DJ. Role of hemolysin for the intracellular growth of Listeria monocytogenes. J Exp Med. 1988 Apr 1;167(4):1459–1471. [PMC free article] [PubMed]
  • Staquet MJ, Levarlet B, Dezutter-Dambuyant C, Schmitt D, Thivolet J. Identification of specific human epithelial cell integrin receptors as VLA proteins. Exp Cell Res. 1990 Apr;187(2):277–283. [PubMed]
  • Takada Y, Huang C, Hemler ME. Fibronectin receptor structures in the VLA family of heterodimers. Nature. 1987 Apr 9;326(6113):607–609. [PubMed]
  • Ruoslahti E. Fibronectin and its receptors. Annu Rev Biochem. 1988;57:375–413. [PubMed]
  • Ratliff TL, Palmer JO, McGarr JA, Brown EJ. Intravesical Bacillus Calmette-Guérin therapy for murine bladder tumors: initiation of the response by fibronectin-mediated attachment of Bacillus Calmette-Guérin. Cancer Res. 1987 Apr 1;47(7):1762–1766. [PubMed]
  • Ratliff TL, Kavoussi LR, Catalona WJ. Role of fibronectin in intravesical BCG therapy for superficial bladder cancer. J Urol. 1988 Feb;139(2):410–414. [PubMed]
  • Aslanzadeh J, Brown EJ, Quillin SP, Ritchey JK, Ratliff TL. Characterization of soluble fibronectin binding to Bacille Calmette-Guérin. J Gen Microbiol. 1989 Oct;135(10):2735–2741. [PubMed]
  • Bohnsack JF, Takahashi T, Brown EJ. Interaction of culture-derived macrophages with the fibroblast-binding domain of fibronectin is a necessary but inefficient signal for fibronectin enhancement of CR1-mediated phagocytosis. J Immunol. 1986 May 15;136(10):3793–3798. [PubMed]
  • Bohnsack JF, O'Shea JJ, Takahashi T, Brown EJ. Fibronectin-enhanced phagocytosis of an alternative pathway activator by human culture-derived macrophages is mediated by the C4b/C3b complement receptor (CR1). J Immunol. 1985 Oct;135(4):2680–2686. [PubMed]
  • Bevilacqua MP, Amrani D, Mosesson MW, Bianco C. Receptors for cold-insoluble globulin (plasma fibronectin) on human monocytes. J Exp Med. 1981 Jan 1;153(1):42–60. [PMC free article] [PubMed]
  • Hamill RJ, Vann JM, Proctor RA. Phagocytosis of Staphylococcus aureus by cultured bovine aortic endothelial cells: model for postadherence events in endovascular infections. Infect Immun. 1986 Dec;54(3):833–836. [PMC free article] [PubMed]
  • Van de Water L, Destree AT, Hynes RO. Fibronectin binds to some bacteria but does not promote their uptake by phagocytic cells. Science. 1983 Apr 8;220(4593):201–204. [PubMed]
  • Proctor RA, Prendergast E, Mosher DF. Fibronectin mediates attachment of Staphylococcus aureus to human neutrophils. Blood. 1982 Apr;59(4):681–687. [PubMed]
  • Isberg RR. Discrimination between intracellular uptake and surface adhesion of bacterial pathogens. Science. 1991 May 17;252(5008):934–938. [PubMed]
  • Band AH, Chitamber SD, Bhattacharya A, Talwar GP. Mechanism of phagocytosis of mycobacteria by Schwann cells and their comparison with macrophages. Int J Lepr Other Mycobact Dis. 1986 Jun;54(2):294–299. [PubMed]
  • Cornette J, Capron A, Ouaissi MA. Trypanosoma cruzi: fibronectin promotes uptake of epimastigote culture forms by human neutrophils and monocytes. Int Arch Allergy Appl Immunol. 1988;86(2):139–146. [PubMed]
  • Griffin FM, Jr, Griffin JA, Leider JE, Silverstein SC. Studies on the mechanism of phagocytosis. I. Requirements for circumferential attachment of particle-bound ligands to specific receptors on the macrophage plasma membrane. J Exp Med. 1975 Nov 1;142(5):1263–1282. [PMC free article] [PubMed]
  • Stoner GL. Importance of the neural predilection of Mycobacterium leprae in leprosy. Lancet. 1979 Nov 10;2(8150):994–996. [PubMed]

Articles from The Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation