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Characterization of Functional Fc-Receptor Material from Human Lymphoblastoid Cell Lines

II. Serologic and Cellular Analysis

From the Cancer Research Division, Department of Pathology, Botterell Hall, Queen's University, Kingston, Ontario, Canada, K7L 3N6, and Department F/Konzern/PAI, F. Hoffman-LaRoche and Co., CH-4002 Basel, Switzerland

Abstract

A xenogeneic anti-human Fc-receptor serum was prepared by injecting rabbits with Fc-receptor (FcR) material isolated from the NP-40 lysate of an IgM-secreting human lymphoblastoid cell line. By indirect immunofluorescence, the immune IgG (or F(ab')₂ fragment) from such animals reacted with a fraction of normal human peripheral blood leukocytes (PBL), corresponding closely to the proportion of FcR-bearing cells (assessed by immune complex binding), in the same cell population. The anti-FcR serum stained almost all the cells from five surface Ig-bearing lymphoblastoid cell lines, including the original cell line from which the FcR material was obtained, but not the FcR^- T cell-like lymphoma, MOLT-4, phagocytic cells, or a granulocyte-like human cell line, K562. When human PBL were incubated with anti-FcR serum, the proportion of rosettes subsequently formed with antibody-sensitized ox erythrocytes (EA) was greatly reduced. The inhibitory activity of the antiserum was removed when it was absorbed with human PBL enriched for EA rosette-forming cells (RFC), but not human PBL depleted of such cells (T cells). The antiserum reacted with a fraction of Ig-bearing lymphocytes in normal mouse spleen and PBL, and with allogeneic activated T cells (30 to 40% FcR⁺) from irradiated F₁ hybrid mice transplanted with parental T cells, but not with normal thymocytes. The determinants recognized by the anti-FcR serum are distinct from serologically detectable ₂ microglobulin, HLA, H-2 and Ia-like determinants: a) The anti-FcR serum precipitated from an NP-40 lysate of a human lymphoblastoid cell line (WT-18), material with an apparent m.w. of 46,000 daltons and that appears distinct from Ia molecules or ₂ microglobulin. b) The anti-FcR serum did not react with MOLT-4 cells nor with T cells in normal human PBL, mouse spleen, and thymus, although these cells express serologically detectable HLA or H-2 specificities. c) The anti-FcR serum, but not an anti-human Ia-like serum, reacted with normal mouse spleen cells. d) The anti-FcR serum reacted with an FcR⁺ T cell tumor, but not with an FcR^- T cell tumor, both derived from the same AKR lymphoma.

Footnotes

¹ Research Scholar of the National Cancer Institute of Canada.

² Send correspondence to Bruce E. Elliott, Ph.D., Cancer Research Division, Department of Pathology, Botterell Hall Queen's University, Kingston, Ontario, Canada K7L 3N6.