Cancer Immunology, Immunotherapy

, Volume 52, Issue 6, pp 367–377 | Cite as

A new small cell lung cancer (SCLC)-specific marker discovered through antigenic subtraction of neuroblastoma cells

  • Pamela Krueger
  • Christina Nitz
  • Randi Foster
  • Colleen MacDonald
  • Oren Gelber
  • Guita Lalehzadeh
  • Robert Goodson
  • Jill Winter
  • Cohava Gelber
Original Article


Small cell lung cancer (SCLC) is an aggressive form of lung cancer associated with cigarette smoking and presently accounts for approximately 20% of all lung cancer cases. SCLC cells derive from a neuroendocrine origin and therefore their antigenic profile coincides, to a great extent, with that of neuroendocrine cells. Multiple attempts to generate SCLC-specific MoAbs during the past decade have failed because all SCLC-specific MoAbs isolated also react against neuroendocrine tissues or normal immune cells. Cross-reactivity with normal antigens raises safety concerns due to the inevitable toxicity of such interactions and the dreaded effects. The concept of DIAAD™ (Differential Immunization for Antigen and Antibody Discovery) provides for an immune response that can be effectively focused on cancer antigens. The object is to overcome obstacles resulting from an antigenic hierarchical pattern biased towards a response to dominant antigens in order to induce a robust immune response to cancer antigens. Cancer antigens are weak or nonimmunogenic molecules. Due to the fact that the immune system responds more strongly to immunodominant antigens than to weak immunogenic antigens, cancer cell proliferation is unencumbered. DIAAD employs protocols of induction of tolerance and immunity, conducted in sequential order to "biologically subtract" the immune response of dominant antigens expressed by normal cells. This biological subtraction is achieved in a laboratory animal by first eliminating the immune response to the normal cells or closely related cancer cells, followed by immunization of the same laboratory animal with diseased cells. This procedure directs the immune response exclusively towards antigens expressed by the diseased and not the normal cells. Our objective was to use DIAAD to generate monoclonal antibodies specific to SCLC antigens that are not shared by neuroendocrine cells by contrasting a pool of human SCLC cell lines with a pool of human neuroendocrine cancer cell lines. Four monoclonal antibodies reacted strongly and exclusively with SCLC cells and identified a membrane molecule comprising a single chain glycoprotein. Two of four antibodies were selected for a detailed analysis that revealed a narrow tissue specificity of antigen expressed by colon, lung, and pancreatic cancers (less than 20% staining was found on breast, ovarian and prostate cancer). These antibodies did not bind to various other cancers such as kidney, carcinoid, lymphoma, sarcoma, adrenal, liver, melanoma, seminoma, leiomyoma, basal cell cancer, or undifferentiated cancer. The epitope recognized by the selected MoAbs was destroyed with the removal of carbohydrates from SCLC cells. This result does not exclude the possibility of protein–carbohydrate cooperation in epitope recognition. However, it strongly suggests the pivotal role of carbohydrates in antibody binding to this molecule. Upon binding to the extracellular molecule on SCLC cells, the antibodies were shown to internalize. A low or insignificant level of internalization was recorded following incubation of the antibodies with neuroendocrine-derived tumors. The capacity of these antibodies to internalize upon binding the extracellular receptors renders them potential candidates for prodrug or immunotoxin-targeted therapeutics. In a qualitative experiment involving immunoaffinity purification, the SCLC antigen was shown to be differentially detected in sera of SCLC patients. Plans are being generated to explore the possible utility of this novel SCLC-specific antigen recognized by the above MoAbs as a new biomarker for early diagnosis of the disease, as well as for therapeutic intervention for SCLC.


Tolerance Subtractive immunization MoAbs Cancer antigens SCLC (small cell lung cancer) 


  1. 1.
    Beaumier PL, Venkatesan P, Vanderheyden JL, Burgua WD, Kunz LL, Fritzberg,AR, AbramsPG, Morgan AC (1991)186Re radiommunotherapy of small cell lung carcinoma xenografts in nude mice. Cancer Res 51:676PubMedGoogle Scholar
  2. 2.
    Brigham BA, Bunn PA, Minna JD, Cohen MH, Ihde DC, Shackney SE (1978) Growth rates of small cell bronchogenic carcinomas. Cancer 42:2880PubMedGoogle Scholar
  3. 3.
    de Leij L, Poppema S, Nulend JK, ter Haar A, Schwander E, Ebbens F, Postmus PE, The TH (1985) Neuroendocrine differentiation antigen of human lung carcinoma and Kulchitski cells. Cancer Res 45:2192PubMedGoogle Scholar
  4. 4.
    Embleton MJ, Garnett MC (1985) Antibody targeting of anti-cancer agents: In: Baldwin RW, Byer VS (eds) Monoclonal antibodies for cancer detection and therapy. Academic Press, New York, p 317Google Scholar
  5. 5.
    Gazdar AF (1994) The molecular and cellular basis of human lung cancer. Anticancer Res 13:261Google Scholar
  6. 6.
    Heike Y, Sone S, Yano S, Seimiya H, Tsuruo T, Ogura T (1993) M-CSF gene transduction in multidrug-resistant human cancer cells to enhance anti-P-glycoprotein antibody-dependent macrophage-mediated cytotoxicity. Int J Cancer 54:851PubMedGoogle Scholar
  7. 7.
    Hekman A, Honselaar A Vuist W, et al. (1991) Initial experience with treatment of human B cell lymphoma with anti-CD19 monoclonal antibody. Cancer Immunol Immunother 32:364PubMedGoogle Scholar
  8. 8.
    Hosono M, Endo K, Hososno MN, Kobayashi H, Sakahra H, Ryuzo U, Kunishi J (1994) Treatment of small cell lung cancer xenografts with iodine-131-anti-neural cell adhesion molecules monoclonal antibody and evaluation of adsorbed dose in tissue. J Nucl Med 35:296PubMedGoogle Scholar
  9. 9.
    Jackson D, Waibel R, Weber E, Bell J, Stahel RA (1992) CD24, a signal-transducing molecule expressed on human B cells, is a major surface antigen on small cell lung carcinomas. Cancer Res 52:5264PubMedGoogle Scholar
  10. 10.
    Jurcic JG, Scheinberg DA, Houghton A (1996) Monoclonal antibody therapy of cancer: In: Pinedo HM, Chabner BA, Longo DL (eds) Cancer chemotherapy and biological response modifiers, Annual 16. Elsevier Science, New York, p 168Google Scholar
  11. 11.
    Köhler G, Milstein C (1975) Continuous cultures of fused cells secreting antibody of predefined specificity. Nature 256:495PubMedGoogle Scholar
  12. 12.
    Kwak LW, Grossbard ML, Urba WJ (1995) Clinical applications of monoclonal antibodies in cancer: In: DeVita Jr VT, Hellman S, Rosenberg SA (eds) Biologic therapy of cancer, 2nd edn. Lippincott, Philadelphia, p 553Google Scholar
  13. 13.
    Livingston RB (1994) Small cell lung cancer: from the laboratory to the clinic. Anticancer Res 14:255Google Scholar
  14. 14.
    Livingston RB (1997) Combined modality therapy of lung cancer. Clin Cancer Res 3:2638PubMedGoogle Scholar
  15. 15.
    Miller RA, Levy R (1981) Response of cutaneous T cell lymphoma to therapy with hybridoma monoclonal antibody. Lancet II:226Google Scholar
  16. 16.
    Olabiran Y, Ledermann JA, Martson NJ, Boxer GM, Souhami RL, Spiro SG, Stahel RA (1994) The selection of antibodies targeted therapy of small cell lung cancer (SCLC) using a human tumor spheroid model to compare the uptake of cluster 1 and cluster w4 antibodies. Br J Cancer 69:247PubMedGoogle Scholar
  17. 17.
    Smith A, Waibel R, Stahel RA (1991) Adoptive immunotherapy of small cell cancer xenografts using131I-labeled SWA11 antibody. Oncology 64:263Google Scholar
  18. 18.
    Souhami RL, Beverley PCL, Bobrow LG, Ledermann JA (1991) Antigens of lung cancer: results of the second international workshop on lung cancer antigens. J Natl Cancer Inst 83:609PubMedGoogle Scholar
  19. 19.
    Stahel RA (1989) Monoclonal antibodies to lung cancer. Chest 96:27SPubMedGoogle Scholar
  20. 20.
    Stahel RA, Ginsburg R, Haveman K, et al. (1989) Staging and prognostic factors in small cell lung cancer: a consensus report. Lung Cancer 5:119Google Scholar
  21. 21.
    Takahashi T, Ueda R, Song X, Nishida K, Shinzato M, Namikawa R, Ariyoshi Y, Ota K, Kato K, Nagatsu T (1986) Two novel cell surface antigens on small cell lung carcinoma defined by mouse monoclonal antibodies NE-25 and PE-35. Cancer Res 46:4770PubMedGoogle Scholar
  22. 22.
    Waldman TA (1994) Lymphokine receptors: a target for immunotherapy of lymphomas. Ann Oncol 5:13Google Scholar
  23. 23.
    Watanabe J, Okaba T, Fujiasawa T, Takaku F, Hirohashi S, Shimosato Y (1987) Monoclonal antibody that distinguishes small cell lung cancer from non-small cell lung cancer. Cancer Res 47:826PubMedGoogle Scholar
  24. 24.
    Wolpaw DR (1996) Early detection in lung cancer. Case findings and screening. Med Clin North Am 80:63PubMedGoogle Scholar
  25. 25.
    Zeuthen J, Vangsted AJ (1993) Monoclonal antibodies for diagnosis and potential therapy of small cell lung cancer—the ganglioside antigen fucosyl-GM1. Acta Oncol 32:845PubMedGoogle Scholar

Copyright information

© Springer-Verlag 2003

Authors and Affiliations

  • Pamela Krueger
    • 1
  • Christina Nitz
    • 1
  • Randi Foster
    • 1
  • Colleen MacDonald
    • 1
  • Oren Gelber
    • 1
  • Guita Lalehzadeh
    • 2
  • Robert Goodson
    • 2
  • Jill Winter
    • 2
  • Cohava Gelber
    • 1
  1. 1.Molecular DiscoveriesLLCNew YorkUSA
  2. 2.Chiron CorporationUSA

Personalised recommendations