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Journal of Immunology Research
Volume 2016 (2016), Article ID 2908034, 17 pages
Research Article

Differential Use of Human Neutrophil Fcγ Receptors for Inducing Neutrophil Extracellular Trap Formation

1Departamento de Inmunología, Instituto de Investigaciones Biomédicas, Universidad Nacional Autónoma de México, 04510 México, DF, Mexico
2División de Estudios de Posgrado e Investigación, Facultad de Odontología, Universidad Nacional Autónoma de México, 04510 México, DF, Mexico

Received 15 September 2015; Revised 11 December 2015; Accepted 13 December 2015

Academic Editor: Kurt Blaser

Copyright © 2016 Omar Rafael Alemán et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

The Fcgamma Receptor and Integrin expression on human neutrophils was confirmed with use of the corresponding monoclonal antibodies (Fig. 1S). Neutrophils and NETs are shown both in bright field and fluorescence images (Fig. 2S). NETs are decorated with histones (Fig. 3S) and with neutrophil elastase (Fig. 4S) as shown in these higher magnification (400 X) microphotographs. Neutrophils were not in apoptosis after FcgammaR crosslinking since they did not bind Annexin V (Fig. 5S). NETs induced by Fc gamma RIIIb crosslinking was not inhibited by the Syk inhibitor iSyk (Fig. 6S). Reactive Oxygen Species (ROS) formation was inhibited by the NADPH-oxidase inhibitor diphenyleneiodinium (DPI) (Fig. 7S). Fluorescent beads used in phagocytosis experiments were efficiently opsonized by anti FcgammaR-specific antibodies (Fig. 8S), and could be easily separated from neutrophils by flow cytometry (Fig. 9S). Phagocytosis was assessed the the appearance of cells with high fluorescence (Fig. 9S). Phagocytosis was also assessed by microscopy, calculating the Phagocytic Index, the number of beads ingested by 100 neutrophils (Fig. 10S).

  1. Supplementary Material