Isolation of lung carcinoma-associated antibodies from immune complexes and production of heterologous antisera

Abstract

Immune complexes isolated from pleural effusions of lung carcinomas were dissociated by ion exchange chromatography in the presence of 8 M urea. The antibodies thus obtained from 2 lung adenocarcinomas and 2 squamous cell carcinomas were assayed by indirect immunofluorescence against a variety of target cells in fresh suspensions, tissue cultures, and paraffin-embedded sections. Strong cytoplasmic fluorescence was obtained with greater than 90% of the cell populations of all major histological types of lung carcinomas, and negative fluorescence was obtained with the cells of normal lung tissues and most of the nonpulmonary carcinomas. Antigen fractions prepared from the same lung carcinoma-associated immune complexes were used to immunize rabbits for the production of antisera directed against human lung carcinoma. The rabbit antibody preparations after proper absorptions were assayed in indirect immunofluorescence showing patterns of reactivity with normal and tumor cells of various derivations similar to those obtained with the human antibodies. Complete blocking of immunofluorescence staining with both allogeneic and xenogeneic antibody preparations was achieved by prior absorption with lung tumor tissue extracts or with various lung tumor antigen preparations. The present study demonstrates the isolation from lung tumor effusion immune complexes of antibodies with high affinity for lung carcinoma cells and of antigens that can be used to produce tumor-directed heterologous antibodies. The immunofluorescence staining of lung carcinoma-associated antigens in situ on paraffin-embedded sections provides new, topographical information.