Int. J. Cancer: 21, 675-682 (1978)
CHARACTERIZATION OF IMMUNE COMPLEXES FROM THE PLEURAL EFFUSION OF A BREAST CANCER PATIENT L. D. PAPSIDERO, S. R. HARVEY, M. C. SNYDERMAN, T. NEMOTO,L. VALENZUELA and T. M. CHU Departments of Diagnostic Immunology Research and Biochemistry and Breast Surgery, Roswell Park Memorial Institute, Bufalo, N . Y. 14263, USA
Pleural effusion from a patient with adenoin the circulation of patients with breast cancer carcinoma of the breast was precipitated by 503/, suggesting a humoral response to the tumor (Chiu ammonium sulfate and chromatographed on Sephaet al., 1977; Nordquist et al., 1977a, b). Anti-tumor dex G-2OO. The resulting elution profile consisted antibodies may form complexes with tumor antigens of three protein peaks. Fractions were monitored by immunodiffusion and assayed for ['251]Clq to produce the high levels of immune complexes seen in breast cancer patients (Hoffken et al., 1977). binding activity (Clq B.A.). The f i r s t protein peak (eluted at the void volume) showed appreciable Our recent observations have suggested that Clq B.A. and contained immunoglobulin G (IgG) extracellular fluids surrounding tumors may provide and complement 3 (C3). These data suggested that a convenient source for the isolation of tumorantigen-antibody complexes were present i n this associated antigens and antibody (Cortes et al., macromolecular peak. IgG and C3 were also present 1974; Chu et al., 1977; Harvey and Chu, 1977). i n the second peak of Sephadex G-200 gel filtration In an attempt to isolate breast tumor-associated while the third peak contained the albumin fraction. antigen from the pleural fluid of a patient, we The known tumor-associated antigens, carcinofound the presence of high molecular weight IgG embryonic antigen, alpha-fetoprotein and ferritin, and complement component 3 (C3) in gel filtration were not detected in any of the protein fractions as determined by immunodiffusion analysis. The f i r s t fractions which also showed complement, [1251]Clq, Sephadex protein peak which contained high molebinding activity. Since several workers have reported cular weight IgG was chromatographed on Sephathe presence of IgG in the form of immune complexes rose 68. The second protein peak obtained by sedimenting from 10s to 19s (Heimer and Klein, Sepharose filtration contained IgG and C3 and was 1976; Samoyoa et al., 1977; Schrohenloher and further applied t o a column of protein A-Sepharose. Volankis, 1977), it was felt that similar complexes Proteins which bound t o the immobilized promay have been present in the pleural effusion we tein A were dissociated- with 2.5 M KSCN and chromatographed on Sepharose CL-68 under disso- examined. In this communication we report on the ciating conditions. IgM rheumatoid factor, IgG and nature of the high molecular weight IgG isolated by four putative antigens were obtained by these dis- gel filtration and Protein A-Sepharose affinity sociation and chromatographic procedures. The chromatography. Since it has been shown to specifiisolated IgG exhibited a molecular weight of mono- cally bind Fc domains of IgG molecules (Forsgren meric IgC and bound t o saline extracts of malignant and Sjoquist, 1966; Kronvall and Frommel, 1970), tissue at a greater amount than t o normal or protein A was utilized to isolate IgG-containing benign breast extracts (p
CHARACTERIZATION OF IMMUNE COMPLEXES FROM THE PLEURAL EFFUSION OF A BREAST CANCER PATIENT L. D. PAPSIDERO, S. R. HARVEY, M. C. SNYDERMAN, T. NEMOTO,L. VALENZUELA and T. M. CHU Departments of Diagnostic Immunology Research and Biochemistry and Breast Surgery, Roswell Park Memorial Institute, Bufalo, N . Y. 14263, USA
Pleural effusion from a patient with adenoin the circulation of patients with breast cancer carcinoma of the breast was precipitated by 503/, suggesting a humoral response to the tumor (Chiu ammonium sulfate and chromatographed on Sephaet al., 1977; Nordquist et al., 1977a, b). Anti-tumor dex G-2OO. The resulting elution profile consisted antibodies may form complexes with tumor antigens of three protein peaks. Fractions were monitored by immunodiffusion and assayed for ['251]Clq to produce the high levels of immune complexes seen in breast cancer patients (Hoffken et al., 1977). binding activity (Clq B.A.). The f i r s t protein peak (eluted at the void volume) showed appreciable Our recent observations have suggested that Clq B.A. and contained immunoglobulin G (IgG) extracellular fluids surrounding tumors may provide and complement 3 (C3). These data suggested that a convenient source for the isolation of tumorantigen-antibody complexes were present i n this associated antigens and antibody (Cortes et al., macromolecular peak. IgG and C3 were also present 1974; Chu et al., 1977; Harvey and Chu, 1977). i n the second peak of Sephadex G-200 gel filtration In an attempt to isolate breast tumor-associated while the third peak contained the albumin fraction. antigen from the pleural fluid of a patient, we The known tumor-associated antigens, carcinofound the presence of high molecular weight IgG embryonic antigen, alpha-fetoprotein and ferritin, and complement component 3 (C3) in gel filtration were not detected in any of the protein fractions as determined by immunodiffusion analysis. The f i r s t fractions which also showed complement, [1251]Clq, Sephadex protein peak which contained high molebinding activity. Since several workers have reported cular weight IgG was chromatographed on Sephathe presence of IgG in the form of immune complexes rose 68. The second protein peak obtained by sedimenting from 10s to 19s (Heimer and Klein, Sepharose filtration contained IgG and C3 and was 1976; Samoyoa et al., 1977; Schrohenloher and further applied t o a column of protein A-Sepharose. Volankis, 1977), it was felt that similar complexes Proteins which bound t o the immobilized promay have been present in the pleural effusion we tein A were dissociated- with 2.5 M KSCN and chromatographed on Sepharose CL-68 under disso- examined. In this communication we report on the ciating conditions. IgM rheumatoid factor, IgG and nature of the high molecular weight IgG isolated by four putative antigens were obtained by these dis- gel filtration and Protein A-Sepharose affinity sociation and chromatographic procedures. The chromatography. Since it has been shown to specifiisolated IgG exhibited a molecular weight of mono- cally bind Fc domains of IgG molecules (Forsgren meric IgC and bound t o saline extracts of malignant and Sjoquist, 1966; Kronvall and Frommel, 1970), tissue at a greater amount than t o normal or protein A was utilized to isolate IgG-containing benign breast extracts (p
