Effect of adenylate cyclase activators on C5a-induced human neutrophil aggregation, enzyme release and superoxide production.

The effect of adenylate cyclase activators on C5a- and f-Met-Leu-Phe-induced human neutrophil aggregation, enzyme release and superoxide production was investigated. C5a-stimulated superoxide production was markedly inhibited by adenylate cyclase activators, and the order of potency was PGE1 greater than isoproterenol greater than epinephrine greater than PGF2 alpha, which correlated with intracellular cAMP levels. However, neutrophil aggregation was inhibited by PGE1, PGE2, isoproterenol and epinephrine only at concentrations greater than 10(-6) M.

Blocking of passive sensitization of human mast cells and basophil granulocytes with IgE antibodies by a recombinant human epsilon-chain fragment of 76 amino acids.

The recombinant peptide corresponding to residues 301-376 at the junction of constant regions 2 and 3 of the human IgE epsilon chain blocked the in vivo passive sensitization of human skin mast cells and in vitro sensitization of human basophil granulocytes with human IgE antibodies. An injection of the recombinant peptide or E myeloma protein into normal skin sites 1 hr before sensitization with an allergic serum blocked passive sensitization. In this system, approximately 10-fold higher molar concentration of the recombinant peptide than E myeloma protein was required for 50% inhibition of Prausnitz-Küstner reactions.

Effect of monoclonal antihuman IgE on recombinant IgE(301-376) inhibition of specific IgE histamine release.

To explore the binding domains of rCIgE(301-376) necessary for inhibition of passive transfer of rye grass and Chinese elm IgE to human basophils, we employed monoclonal antibodies known to bind to IgE(301-336), M-272, and to IgE(367-376), M-27. By preincubating M-272, but not M-27, with rCIgE(301-376), passive transfer of specific IgE to basophils was partially inhibited. This implies that M-272 recognizes a binding site on rCIgE(301-376) or sterically interferes with the rCIgE(301-376) high-affinity binding domains on human basophils.

The use of fibronectin-coated polyvinyl chloride microtest plates to detect monoclonal antibody-binding to adherent tumor cells.

This report describes the use of fibronectin-coated polyvinyl chloride (PVC) plates as a time-saving modification in an assay measuring antibody-binding to live adherent tumor cells. Three cell lines (A-431, Colo 16 and UCLA-SO-P3) derived from human squamous cell carcinomas (SCC) and forming monolayers in cultures were plated onto flexible PVC microtest plates rather than the commonly used rigid polystyrene plates. In PVC plates, two of the three cell lines (A-431 and Colo 16) grew as foci of clumped cells instead of monolayers.