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Monoclonal antibodies specific for non-small cell human lung cancers
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Kyoizumi 1 ~ S., Kounol'2~ N., Akiyama I, M., Noumi , K. Yamakido , M.1. Radiation Effects Research Foundation, Hiroshima, Japan. 2. Hiroshima University, Hiroshima, Japan. We produced murine monoclonal antibodies to human lung squamous carcinoma and adenocarcinoma cell lines, and examined their specificity to lung carcinoma tissues as well as the biochemical characteristics of target antigens. (a) LuCa2 (y 1 ) and LuCa4 (y 1 ) antibodies were reactive with lung squamous cell carcinoma, but not with other types of lung cancer. LuCa3 and LuCa4 immunoprecipitated MW230K and 240K proteins and a MWI50K protein respectively from a squamous carcinoma cell line. (b) LuCa6 (~) antibody reacted with all types of l u ~ _ c a r c i n o m a tissue, and was shown by a bI-CEA binding assay to be reactive with CEA. (c) LuCa9 (¥ ]) antibody was only reactive with lung adenocarcinoma and precipitated a 140K protein from an adenocarcinoma cell line. (d) LuCa9 (~) and LuCal0 (~) antibodies were reactive with some of lung carcinoma tissues. Antigens reactive with these two antibodies were resistant to protease treatment but sensitive to NaIO, treatment. Monoclonal Antibodies Specific for NonSmall Cell Human Lung Cancers. Chen, F.A., Repasky, E.A., Zylstra, S., Saji, S., Takita, H., Bankert, R.B. Roswell Part Memorial Institute, Buffalo, N.Y., U.S.A. Monoclonal antibodies have been produced in the mouse against primary human lung tumors of different histological types (squamous cell carcinoma, adenocarcinoma, and large cell carcinoma) which were obtained from surgery. The hybridoma culture supernatants were screened with an immunocytoadherence assay using autologous tumor cells and autologous B-lymphoblasts. Six monoclonal antibodies 2C6, 5D7, 5E8 anti-squamous; 5C7, 2B7 anti-adenocarcinoma; and 1 F10 anti-large cell carcinoma were identified. The isotype of the representative antibodies (5E8, 5C7, and 1 FI0) was determined to be G , K. Cultured human adeno1 carcinoma (PC-9) was surface labelled with radioactive I125, immunoprecipitated with 5E8 and run on SDS polyacrylamide gel. By this procedure, a membrane-associated protein with a molecular weight of approximately 160,000 daltons was identified. The surface labelling of human large cell lung carcinoma, immunoprecipitation with 1 FI0, and gel electrophoresis ten-
tatively identified a protein with a molecular weight of 68,000 daltons. Specificity of the antibodies are being tested by immunofluorescent and immunohistochemical staining of various human tumors. H~nan ~bnoclonal Antibody from Lymphocyte of Lymph Node Adjoining Lung Cancer Recognizing in Antigen. Hirohashi, S., Shimosato, Y., Ishikawa, Y., Hakomori, S. National Cancer Center Research Institute, Tokyo, Japan. Fred Hutchinson Cancer Research Center, Seattle, U.S.A. A monoclonal antibody has been produced'by the hybridoma technique using a suspension of lymphocytes from a lymph node adjoining moderately differentiated adenocarcinoma of the lung and mouse myeloma cells (P3-X63-Ag8-UI). The hybridomas were selected by the enzyme immunohistochemical method using paraffin sections of the original tumor and various normal tiseues. T} ~n, those that produced the antibody reactive wit" the tumor were cloned by limiting dilution, and finally one clone was obtained (1004-5). This monoclonal antibody (IgM,)~) reacted immunohistochemically with 6 cases of 21 adenocarcinomas, 12 of 17 squamous cell carcinomas, 5 of 19 large cell carcinomas and none of 20 small cell carcinomas of the lung. It also reacted with bronchial basal cells, some alveolar lining cells and red blood cells in some instances. The antibody is an autoantibody against i antigen. From these results together with results using other monoclonal antibodies obtained in our laboratory with lung cancer as immunogen, which recognize A-like antigen and Lewis X, respectively, it was demonstrated that many monoclonal antibodies obtained by these methods were found to recognize sugar chains, particularly those related to the blood group. They are considered to be useful for the study of incomplete or abnormal synthesis of the sugar chain in cancer cells. Characterization of Monoclonal Antibody to Human Lung Adenocarcinoma Protein Antigen. Nakamura, H., Niitsuma, M., Yamada, T., Kawamura, I., Suzuta, T., Hayata, Y. Tokyo Medical College, Tokyo, Japan. Monoclonal antibody ID5 (IgMk, pi7.65) was developed in our laboratory from a fusion of the murine myeloma cell line SI94/5XXOBUI with splenocytes of BALB/c mice previously primed with a human lung adenocarcinoma cell line PC7. Specificity was tested by indirect immunofluorescence method against cells from freshly isolated tumors and by Avidin Biotin peroxidase Complex(ABC) method against paraffin embedded tissue sections. Lung Cancer Tested Positive(%) Squamous ii/ii (i00.0) Adeno 6/13 (46.1) Malig. Mesothelioma i/i (i00.0) Large 0/3 (0.0) Small 0/9 (0.0) This monoclonal antibody was unreactive with different lymphoblastoid cell lines, erythrocytes of several blood groups, liver, kidney, spleen,
Kyoizumi 1 ~ S., Kounol'2~ N., Akiyama I, M., Noumi , K. Yamakido , M.1. Radiation Effects Research Foundation, Hiroshima, Japan. 2. Hiroshima University, Hiroshima, Japan. We produced murine monoclonal antibodies to human lung squamous carcinoma and adenocarcinoma cell lines, and examined their specificity to lung carcinoma tissues as well as the biochemical characteristics of target antigens. (a) LuCa2 (y 1 ) and LuCa4 (y 1 ) antibodies were reactive with lung squamous cell carcinoma, but not with other types of lung cancer. LuCa3 and LuCa4 immunoprecipitated MW230K and 240K proteins and a MWI50K protein respectively from a squamous carcinoma cell line. (b) LuCa6 (~) antibody reacted with all types of l u ~ _ c a r c i n o m a tissue, and was shown by a bI-CEA binding assay to be reactive with CEA. (c) LuCa9 (¥ ]) antibody was only reactive with lung adenocarcinoma and precipitated a 140K protein from an adenocarcinoma cell line. (d) LuCa9 (~) and LuCal0 (~) antibodies were reactive with some of lung carcinoma tissues. Antigens reactive with these two antibodies were resistant to protease treatment but sensitive to NaIO, treatment. Monoclonal Antibodies Specific for NonSmall Cell Human Lung Cancers. Chen, F.A., Repasky, E.A., Zylstra, S., Saji, S., Takita, H., Bankert, R.B. Roswell Part Memorial Institute, Buffalo, N.Y., U.S.A. Monoclonal antibodies have been produced in the mouse against primary human lung tumors of different histological types (squamous cell carcinoma, adenocarcinoma, and large cell carcinoma) which were obtained from surgery. The hybridoma culture supernatants were screened with an immunocytoadherence assay using autologous tumor cells and autologous B-lymphoblasts. Six monoclonal antibodies 2C6, 5D7, 5E8 anti-squamous; 5C7, 2B7 anti-adenocarcinoma; and 1 F10 anti-large cell carcinoma were identified. The isotype of the representative antibodies (5E8, 5C7, and 1 FI0) was determined to be G , K. Cultured human adeno1 carcinoma (PC-9) was surface labelled with radioactive I125, immunoprecipitated with 5E8 and run on SDS polyacrylamide gel. By this procedure, a membrane-associated protein with a molecular weight of approximately 160,000 daltons was identified. The surface labelling of human large cell lung carcinoma, immunoprecipitation with 1 FI0, and gel electrophoresis ten-
tatively identified a protein with a molecular weight of 68,000 daltons. Specificity of the antibodies are being tested by immunofluorescent and immunohistochemical staining of various human tumors. H~nan ~bnoclonal Antibody from Lymphocyte of Lymph Node Adjoining Lung Cancer Recognizing in Antigen. Hirohashi, S., Shimosato, Y., Ishikawa, Y., Hakomori, S. National Cancer Center Research Institute, Tokyo, Japan. Fred Hutchinson Cancer Research Center, Seattle, U.S.A. A monoclonal antibody has been produced'by the hybridoma technique using a suspension of lymphocytes from a lymph node adjoining moderately differentiated adenocarcinoma of the lung and mouse myeloma cells (P3-X63-Ag8-UI). The hybridomas were selected by the enzyme immunohistochemical method using paraffin sections of the original tumor and various normal tiseues. T} ~n, those that produced the antibody reactive wit" the tumor were cloned by limiting dilution, and finally one clone was obtained (1004-5). This monoclonal antibody (IgM,)~) reacted immunohistochemically with 6 cases of 21 adenocarcinomas, 12 of 17 squamous cell carcinomas, 5 of 19 large cell carcinomas and none of 20 small cell carcinomas of the lung. It also reacted with bronchial basal cells, some alveolar lining cells and red blood cells in some instances. The antibody is an autoantibody against i antigen. From these results together with results using other monoclonal antibodies obtained in our laboratory with lung cancer as immunogen, which recognize A-like antigen and Lewis X, respectively, it was demonstrated that many monoclonal antibodies obtained by these methods were found to recognize sugar chains, particularly those related to the blood group. They are considered to be useful for the study of incomplete or abnormal synthesis of the sugar chain in cancer cells. Characterization of Monoclonal Antibody to Human Lung Adenocarcinoma Protein Antigen. Nakamura, H., Niitsuma, M., Yamada, T., Kawamura, I., Suzuta, T., Hayata, Y. Tokyo Medical College, Tokyo, Japan. Monoclonal antibody ID5 (IgMk, pi7.65) was developed in our laboratory from a fusion of the murine myeloma cell line SI94/5XXOBUI with splenocytes of BALB/c mice previously primed with a human lung adenocarcinoma cell line PC7. Specificity was tested by indirect immunofluorescence method against cells from freshly isolated tumors and by Avidin Biotin peroxidase Complex(ABC) method against paraffin embedded tissue sections. Lung Cancer Tested Positive(%) Squamous ii/ii (i00.0) Adeno 6/13 (46.1) Malig. Mesothelioma i/i (i00.0) Large 0/3 (0.0) Small 0/9 (0.0) This monoclonal antibody was unreactive with different lymphoblastoid cell lines, erythrocytes of several blood groups, liver, kidney, spleen,