Histamine receptors have been demonstrated on lymphocyte membranes by a variety of techniques. We now report a method that allows for the flow cytometric analysis of histamine receptors on human peripheral T cells. Histamine is conjugated to fluoresceinated human albumin by the coupling agent ECDI. This conjugated histamine compound (FHA-his) binds to approximately 45% of T cells. Fluoresceinated human albumin alone (FHA), not conjugated to histamine, does not bind to T cells. In addition, unconjugated histamine can inhibit completely the binding seen with FHA-his. We conclude that this technique demonstrates specific FHA-his binding to histamine receptors on T cells and can be used to determine the number of cells bearing such receptors. In addition, the reagent could be used with a cell sorter to isolate distinct histamine-receptor-bearing (HR+) cells for further immunologic study.
It is increasingly clear that histamine mediates a variety of lymphocyte functions. Further understanding of these mechanisms requires a method for the analysis of histamine membrane receptors on the lymphocyte surface. We report now a biochemical technique for the identification and quantitation of specific histamine H1 and H2 receptors of lymphocytes. The method can be performed on small numbers of formaldehyde-fixed cells. The data this assay yields, together with that resulting from the flow cytometric analysis of histamine receptor distribution (a technique we have previously described), will be a powerful tool in the study of histamine mediation of lymphocyte function.
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