Summary
We describe a slide assay that allows the demonstration of antigens localized in the nucleus from isolated white blood cells as well as from single tumor cells derived from malignant effusions. With the antibodies Ki-67 and anti-p-145 an increased rate of nuclear and nucleolar staining resulted in cells from highly malignant lymphomas. An almost identical reaction was obtained when tumor cells from malignant effusions were tested. Cells isolated from the blood of patients with leukemic spread of lymphomas of low malignancy yielded a weak staining comparable to that of normal mesothelial cells from non-tumorous cavity fluids. The detection of estrogen and progesterone receptors (ER and PR) localized in the cell nucleus can be achieved by the same assay. The reaction is enhanced by incubation of the tumor cells for 30 min at 37‡ C prior to fixation. Pleural effusions from 20 patients with breast cancer were tested. ER was positive in 13 and PR was positive in 12 of the 20 samples. In 5 cases there was a divergent reaction with ER and PR antibody. The hormone receptors of the primary tumor were known in 15 (ER) and 14 (PR) patients, respectively. In each cohort there was only one case with a negative reaction of the primary tumor and a positive reaction with the isolated tumor cells from the pleural effusions. These results indicate that the demonstration of hormone receptor proteins in cells from malignant effusions is possible and that there is a correlation with the status of the primary site of cancer.
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Abbreviations
- ER:
-
estrogen receptor
- PR:
-
progesterone receptor
References
BØyum A (1968) Isolation of mononuclear cells and granulocytes from human blood. Scand J Clin Lab Invest 21 [Suppl 97]:77–89
Bross KJ, Pangalis GA, Staatz CG, Blume KG (1978) Demonstration of cell surface antigens and their antibodies by the peroxidase-anti-peroxidase method. Transplantation 25:331–334
Deibert P, Bross KJ, Löhr GW, Hirsch FW (1988) Demonstration of intracellular antigens in single cells or cell suspensions. Blut 57:227
Di Lorenzo D, Zaniboni A, Simoncini E, Marpicati P, Montini E, Alghisi A, Gorni F, Marini B (1986) Estrogen and progesterone receptors in neoplastic cells of metastatic pleural effusion of breast carcinoma before and after tamoxifen therapy. Correlation with clinical response. Chemiotherapia 5:232–236
Foon KA, Todd RF (1986) Immunologic classification of leukemia and lymphoma. Blood 68:1–31
Freeman JW, McRorie DK, Busch RK, Gyorkey F, Gyorkey P, Ross BE, Spohn WH, Busch H (1986) Identification and partial characterization of a nucleolar antigen with a molecular weight of 145 000 found in a broad range of human cancers. Cancer Res 46:3593–3598
Freeman JW, Dowell BL, Ochs RL, Ross BE, Busch H (1987) Effect of differentiation on the expression of a nucleolar antigen with a molecular weight of 145000 in HL-60 cells. Cancer Res 47:586–591
Gerdes J, Schwab W, Lemke H, Stein H (1983) Production of a mouse monoclonal antibody reactive with a human nuclear antigen associated with cell proliferation. Int J Cancer 31:13–20
Gerdes J, Lemke H, Baisch H, Wacker HH, Schwab U, Stein H (1984a) Cell cycle analysis of a cell proliferation-associated human nuclear antigen defined by the monoclonal antibody Ki-67. J Immunol 133:1710–1715
Gerdes J, Dallenbach F, Lennert K, Lemke H, Stein H (1984) Growth fractions in malignant non-Hodgkin's lymphomas (NHL) as determined in situ with the monoclonal antibody Ki-67. Hematol Oncol 2:365–371
Greene GL, Nolan C, Engler JP, Jensen EV (1980) Monoclonal antibodies to human estrogen receptor. Proc Natl Acad Sci USA 77:5115–5119
Guzman J, Bross KJ, Würtemberger G, Costabel U (1989) Immunocytology in malignant pleural mesothelioma. Chest 95:590–595
Hirsch FW, Deibert P, Löhr GW, Bross KJ (1989) Demonstration of estrogen (ER) and progesterone (PR) receptor proteins in single cells from malignant effusions. Blut 59:323
King WJ, Greene GL (1984) Monoclonal antibodies localize estrogen receptor in the nuclei of target cells. Nature 307:745–747
Kranz BR, Thierfelder S (1983) Increased sensitivity in the immunocytochemical detection of TdT by the use of detergents on glutaraldehyde fixed cells. Immunobiology 165:305
Levine MN, Young JEM, Ryan ED, Newhouse MT (1986) Pleural effusion in breast cancer. Thoracoscopy for hormone receptor determination. Cancer 57:324–327
Nagasawa T, Nagasawa S (1984) Enrichment of malignant cells from pleural effusions of patients with malignant and non-malignant diseases. Cancer Lett 22:1–9
Ochs RL, Freeman JW, Reilly MT, Busch H (1985) Identification of a growth related 145 kD nucleolar protein. J Cell Biol 101:211
Perrot-Applanat M, Logeat F, Groyer-Picard MT, Milgrom E (1985) Immunocytochemical study of mammalian progesterone receptor using monoclonal antibodies. Endocrinology 116:1473–1484
Redard M, Vassilakos P, Weintraub J (1989) A simple method for estrogen receptor antigen preservation in cytologic specimens containing breast carcinoma cells. Diagn Cytopathol 5:188–193
Sasaki K, Murakami T, Kawasaki M, Takahashi M (1987) The cell cycle associated change of the Ki-67 reactive nuclear antigen expression. J Cell Physiol 133:579–584
Schrape S, Jones DB, Wright DH (1987) A comparison of three methods for the determination of the growth fraction in non-Hodgkin's lymphoma. Br J Cancer 55:283–286
Sternberger LA (1974) The unlabeled antibody enzyme method. Immunocytochemistry, foundation of immunology series. Prentice Hall, Englewood NY, pp 129–172
Wittliff JL (1984) Steroid receptors in breast cancer. Cancer 53:630–643
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Hirsch, F.W., Deibert, P., Busch, H. et al. Demonstration of estrogen and progesterone receptors as well as Ki-67 and p-145 antigens in single tumor cells from blood and pleural effusions using a slide assay. J Cancer Res Clin Oncol 117, 217–222 (1991). https://doi.org/10.1007/BF01625427
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DOI: https://doi.org/10.1007/BF01625427