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Basic Sciences

Two New Monoclonal Antibodies, Lym-1 and Lym-2, Reactive with Human B-Lymphocytes and Derived Tumors, with Immunodiagnostic and Immunotherapeutic Potential

Alan L. Epstein, Robert J. Marder, Jane N. Winter, Efstathois Stathopoulos, Feng-Ming Chen, John W. Parker and Clive R. Taylor
Alan L. Epstein
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Robert J. Marder
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Jane N. Winter
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Efstathois Stathopoulos
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Feng-Ming Chen
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John W. Parker
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Clive R. Taylor
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DOI:  Published February 1987
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Abstract

Two new monoclonal antibodies (Lym-1 and Lym-2), reactive with the cell surface of B-lymphocytes and derived tumors, have been produced using tumor cell nuclei preparations as immunogens. Specificity screens using live cell radioimmunoassay techniques with 52 well-characterized human lymphoma and leukemia cell lines showed that both Lym-1 and Lym-2 bound to cell lines of B-cell lineage but were unreactive with those of T-cell, myeloid, or erythroid derivation. The B-cell specificity of these reagents was confirmed on 36 lymphoma and 15 leukemia biopsy specimens by using immunoperoxidase or immunofluorescence techniques. Additionally, flow cytometric analysis of 22 lymphoma biopsies showed that the majority of B-cell tumors were Lym-1 and/or Lym-2 positive and that within a given biopsy, a high percentage of the malignant cell population was stained. In both the immunoperoxidase and flow cytometric studies, reactive T-cells or T-cell lymphomas were consistently negative with the exception of Hodgkin's disease tissues which, in some instances, showed a higher than expected positivity with Lym-1 and Lym-2. Approximately 40% of B-cell chronic lymphocytic leukemias were found to be positive with Lym-1 while 80% were positive with Lym-2. Immunoperoxidase staining of frozen sections of human lymphoid tissues showed that both Lym-1 and Lym-2 stained germinal center and mantle zone B-lymphocytes as well as interfollicular histiocytes. Flow cytometric analysis of normal peripheral blood demonstrated specific staining of B-cells which comprised approximately 8% of circulating lymphocytes. Immunoperoxidase staining of nonlymphoid human organs and tissues revealed weak reactivity of Lym-1 with surface colonic epithelium only. Consistent with these findings, 35 solid tumor cell lines of diverse nature were found unreactive with both Lym-1 and Lym-2. Although standard techniques have thus far failed to identify the antigen recognized by Lym-2, the membrane antigen which binds Lym-1 has been shown by immunoprecipitation and competitive radioimmunoassay studies to be a polymorphic variant of the HLA-Dr antigen. Solid-phase radioimmunoassay techniques have shown that the antigens recognized by Lym-1 and Lym-2 are not significantly modulated after antibody exposure nor shed into the circulation of lymphoma patients. Finally, using iodine-125 labeled preparations of purified Lym-1 and Lym-2, we have determined that both reagents have a relatively large number of antibody binding sites per tumor cell and increased avidity for lymphoma cells when compared to normal and reactive lymph node B-cells. Because of the B-cell specificity of these reagents, their increased avidity for lymphoma cells, and their chemical stability after radiolabeling procedures, Lym-1 and Lym-2 appear to be promising reagents for the immunodiagnosis and therapy of the human malignant lymphomas.

Footnotes

  • ↵1 This work was supported in part by USPHS Grants R01-CA30621 and R01-CA40608 awarded by the National Cancer Institute, Department of Health and Human Services.

  • ↵2 To whom requests for reprints should be addressed, at Department of Pathology, University of Southern California School of Medicine, 2025 Zonal Avenue, Los Angeles, CA 90033.

  • Received April 22, 1986.
  • Revision received October 28, 1986.
  • Accepted October 30, 1986.
  • ©1987 American Association for Cancer Research.
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February 1987
Volume 47, Issue 3
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Two New Monoclonal Antibodies, Lym-1 and Lym-2, Reactive with Human B-Lymphocytes and Derived Tumors, with Immunodiagnostic and Immunotherapeutic Potential
Alan L. Epstein, Robert J. Marder, Jane N. Winter, Efstathois Stathopoulos, Feng-Ming Chen, John W. Parker and Clive R. Taylor
Cancer Res February 1 1987 (47) (3) 830-840;

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Two New Monoclonal Antibodies, Lym-1 and Lym-2, Reactive with Human B-Lymphocytes and Derived Tumors, with Immunodiagnostic and Immunotherapeutic Potential
Alan L. Epstein, Robert J. Marder, Jane N. Winter, Efstathois Stathopoulos, Feng-Ming Chen, John W. Parker and Clive R. Taylor
Cancer Res February 1 1987 (47) (3) 830-840;
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