IDENTIFICATION OF T-LYMPHOCYTES ON FROZEN TISSUE SECTIONS OF HUMAN LYMPHOID TISSUES BY ROSETTE-FORMING ASSAY

Abstract

Localization of cells possessing immunological cell surface markers in the human lymphoid tissues including the thymus, lymph node and spleen was analysed by the rosette forming assay on the frozen tissue sections. E receptor-bearing cells were detected by the adherence of neuraminidase-treated sheep erythrocytes (EN). ENadhered to the thymocytes in the medullary and cortical area of the thymus, the paracortex of the lymph node and the splenic periarteriolar lymphatic sheath of the spleen. Complement receptor-bearing cells were identified by the adherence of ox erythrocytes sensitized with the rabbit anti-ox erythrocyte IgM fraction and human complement (IgMEACh). IgMEACh adhered to the lymphoid follicle of the lymph node and splenic white pulp. The IgG Fc receptor-bearing cells were identified by the adherence of ox erythrocytes sensitized with the rabbit anti-ox erythrocyte IgG fraction (IgGEA). IgGEA adhered to the lymph sinus of the lymph node and splenic red pulp. From these results, it is considered that the T lymphocytes on the frozen tissue section of the human lymphoid tissues are detectable by the adherence of EN, In this rosette-forming assay on the frozen tissue sections, most of the IgMEACh adhered to the B lymphocytes and the IgGEA adhered to mainly the histiocytes or macrophages. Abbreviations used in this paper: E, sheep erythrocytes; E N, neuraminidase-treated sheep erythrocytes; IgMEACh, ox erythrocytes sensitized with rabbit anti-ox erythrocyte 19S (IgM) fraction and human complement; IgGEA, ox erythrocytes sensitized with rabbit anti-ox erythrocyte 7S (IgG) fraction; IgMEA, ox erythrocytes sensitized with rabbit anti-ox erythrocyte l9S (lgM) fraction; PBS, phosphate-buffered saline, pH 7.2; GVB++, gelatin Veronal (barbital sodium) buffer with 0.15 mM CaCl2 and 0.5 mM MgCl2; FCS, fetal calf serum.