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Studies of skin-window exudate human neutrophils: Complex patterns of adherence to serum-coated surfaces in dependence on FMLP doses

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Abstract

Human neutrophils were isolated both from peripheral blood (PB) and from aseptic inflammatory exudates obtained by the Senn's skin-window (SW) technique. The respiratory burst (O −2 release) and the adherence to serum-coated wells of culture microplates was investigated using a simultaneous assay. Unstimulated PB resting neutrophils did not produce a significant amount of O −2 and were incapable of adhering to serum-coated plastic surfaces, while unstimulated SW neutrophils showed augmented adhesion to serum-coated culture wells. SW neutrophils were primed to enhanced FMLP-dependent O −2 release in response ton-formyl-methionyl-leucylphenylalanine (FMLP). Adhesion of SW neutrophils was significantly decreased by addition of low doses (10−10−10−8M) of FMLP (from 17.1% to 8.4%,P< 0.01, N=12), while fully activating doses (>5×10−8 M) of FMLP induced a marked increase of the cell adhesion, more pronounced in SW (39.2%) than in PB cells (27.2%). Low (5×10−9 M) and high (5×10−7 M) FMLP doses induced morphological changes (polarization) and actin polymerization in the neutrophils from both sources. Biphasic dose-response curves of SW neutrophil adherence were observed using FMLP, but not using concanavalin A or phorbol myristate acetate as stimulatory agents. Therefore, the adherence of SW cells appears to be regulated in a complex fashion, nonlinearly dependent on the chemotactic peptide doses and specifically regulated according to the receptors involved.

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Authors and Affiliations

  1. Istituto di Chimica e Microscopia Clinica, University of Verona, Italy

    Paolo Bellavite & Fulvio Suttora

  2. Istituto di Patologia Medica, University of Verona, Italy

    Antonio Carletto, Domenico Biasi, Fabio Poli & Lisa M. Bambara

  3. Istituto di Clinica Medica, University of Verona, Italy

    Paola Caramaschi

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  1. Paolo Bellavite
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Bellavite, P., Carletto, A., Biasi, D. et al. Studies of skin-window exudate human neutrophils: Complex patterns of adherence to serum-coated surfaces in dependence on FMLP doses. Inflammation 18, 575–587 (1994). https://doi.org/10.1007/BF01535256

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