Polymorphonuclear leukocytes cap a derivative of wheat germ agglutinin upon stimulation with formyl peptide and C5a but not leukotriene B4.
Previously, we reported that a derivative of wheat germ agglutinin (termed WGA-D) specifically inhibits human polymorphonuclear leukocyte (PMN) chemotaxis to FMLP by blocking reexpression (or recycling) of formyl peptide receptors. WGA-D (? formyl peptide receptor probe) binds to a protein on the PMN membrane that exhibits the same m.w. as the formyl peptide receptor. Since clustering (i.e., capping) of ligand-receptor complexes most likely precedes their internalization, we examined the ability of normal and stimulated PMN to cap fluoresceinated WGA-D. We found that, in contrast to capping of fluoresceinated Con A, PMN cap WGA-D in a chemotactic factor-specific fashion. Fluoresceinated WGA-D (5.0 to 20 micrograms/ml) alone did not induce either PMN shape changes (i.e., activation) or capping. Both FMLP (1 to 1000 nM) and human C5a (0.1 to 1.0 nM) induced PMN to polarize and to cap bound WGA-D, in a concentration-dependent fashion. Interestingly, leukotriene B4 (LTB4) (5.0 nM), while inducing the same degree of PMN polarization as FMLP (100 nM) and C5a (0.5 nM), failed to induce PMN to cap bound WGA-D. In contrast, FMLP (100 nM), C5a (0.5 nM), and LTB4 (5.0 nM) induced PMN to cap bound fluoresceinated Con A (10 micrograms/ml) to the same extent. The effect of suboptimal concentrations of FMLP and C5a on capping of WGA-D by PMN was additive. LTB4 did not enhance either FMLP or C5a-induced capping of WGA-D by PMN. Also, FMLP and C5a (but not LTB4) were capable of inducing both desensitization and cross-desensitization of WGA-D capping by PMN. Studies using rhodamine-labeled WGA-D and a fluoresceinated analog of FMLP revealed that both capped to the same place on the PMN membrane. Thus, the data suggest that WGA-D binds to a site on the PMN membrane that is either the FMLP receptor or very closely associated with it.
Kurrasch
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Rosalind Russell Arthritis Research Laboratory, Department of Medicine, University of California, San Francisco.
Elfman
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Perez
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D000328:Adult; D002462:Cell Membrane / Q000502:physiology; D002634:Chemotaxis, Leukocyte; D003182:Complement C5 / Q000502:physiology*; D015936:Complement C5a; D003208:Concanavalin A; D003888:Desensitization, Immunologic; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006801:Humans; D007152:Immunologic Capping; D007975:Leukotriene B4 / Q000502:physiology*; D008564:Membrane Potentials; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000378:metabolism; D009504:Neutrophils / Q000276:immunology* / Q000378:metabolism / Q000502:physiology; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic / Q000032:analysis; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins / Q000031:analogs & derivatives / Q000276:immunology* / Q000378:metabolism
D003182:Complement C5; D005452:Fluoresceins; D044042:Receptors, Formyl Peptide; D011971:Receptors, Immunologic; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins; D003208:Concanavalin A; D007975:Leukotriene B4; D009240:N-Formylmethionine Leucyl-Phenylalanine; D015936:Complement C5a; D016650:Fluorescein-5-isothiocyanate