The study of antibodies and antigens dissociated from the immune complexes extracted from ovarian carcinoma ascitic fluid

Abstract

In previous studies in our laboratory, heteroantisera were produced in rabbits by immunization with extracts of ascitic fluids from ovarian epithelial carcinomas. Reverse passive hemagglutination (RPHA) assay of these sera was used for the immunodiagnosis of ovarian cancer. Antibodies attached to lysed sheep red blood cells were able to detect tumor antigens in 63% of ovarian carcinoma sera. In the present study we have shown that the antigens detected in these assays are in fact contained in circulating immune complexes and are recoverable by disassociation with 8 M urea and separation of antibody and antigen by ion exchange chromatography. The antibodies thus obtained from immune complexes (IC) were assayed by indirect immunofluorescence against a variety of paraffin-embedded tissue sections. Poorly differentiated ovarian epithelial carcinomas and papillary serous adenocarcinomas, as well as the tumors grown by implantation of the latter in nude mice, were all positively stained, while the Krukenberg tumor metastatic to the ovary, the benign ovarian cyst, the normal ovary, and the normal fallopian tube were all negative. Complete blocking of immunofluorescence staining by prior absorption of antibody with the dissociated antigen was achieved. The present study demonstrates that the antibodies and antigens dissociated from the immune complexes isolated from the ascitic fluids provide useful reagents for immunodiagnosis.