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Characterization of Cytolytic Effector Cells in Peripheral Blood of Healthy Individuals and Cancer Patients

I. Surface Markers and K Cell Activity After Separation of B Cells and Lymphocytes with Fc-Receptors by Column Fractionation¹

Department of Immunology, University of Stockholm, Fack, S-10691 Stockholm 1, Sweden

Abstract

To characterize the effector cells involved in ADCC in a model system (lysis of ⁵¹Cr-labeled chicken erythrocytes induced by rabbit IgG antibodies), in "spontaneous", and in tumor-associated cytotoxicity, we prepared subpopulations of peripheral blood lymphocytes from healthy individuals and from cancer patients by fractionation on columns according to three different principles. Columns with anti-immunoglobulin activity but lacking Fc-structures (F(ab')₂/anti-F(ab')₂ columns) removed 99% of the SIg⁺ cells (B cells), but only 20% of the Fc-receptor-bearing cells, as characterized by rosette formation with IgG-coated bovine erythrocytes (EA⁺ cells), and 25% of the complement receptor-bearing cells (EAC⁺ cells). Cells with receptor for sheep erythrocytes (E⁺ cells) were enriched. The K cell activity of this fraction was unchanged. Columns charged with immune complexes (OA/anti-OA columns) which display no anti-immunoglobulin activity retained 70% of the EA⁺ cells but only 20% of the SIg⁺ cells. About 50% of the EAC⁺ cells and 10% of the E⁺ cells were also removed. The K cell activity of this fraction was strongly reduced. IgG/anti-IgG columns very efficiently remove SIg⁺ cells, EA⁺ cells, and EAC⁺ cells whereas E⁺ cells are enriched. Again K cell activity is strongly reduced. The results indicate that B cells with high density SIg are inactive in the K cell assay and that the majority of mature B cells either have Fc-receptors of low avidity or have receptors for immunoglobulin which are qualitatively different from the Fc-receptors of SIg^- cells. Furthermore, a significant fraction of the lymphocytes with complement receptors is different from these B cells. It can also be inferred that a significant subpopulation of the cells with receptors for sheep erythrocytes also have Fc-receptors for IgG. The cytotoxicity of these different lymphocyte fractions against a variety of human tumor cells will be reported in the following paper.

Footnotes

¹ This work was supported by Grants 365-B76-08XB from the Swedish Cancer Society and B 2032-047 from the Swedish Natural Science Research Council.

² G. R. Pape is a holder of a fellowship from Deutsche Forschungs-gemeinschaft, F. R. G. Permanent address: Medizinische Klinik (II), Universität Tübingen, F.R.G.