Abstract

BackgroundNeutrophils are the most abundant leukocytes in peripheral blood and represent one of the most important elements of innate immunity. Recent subcellular proteomic studies have focused on the identification of human neutrophil proteins in various subcellular membrane and granular fractions. Although there are relatively few studies dealing with the analysis of the total extract of human neutrophils, many biological problems such as the role of chemokines, adhesion molecules, and other activating inputs involved in neutrophil responses and signaling can be approached on the basis of the identification of the total cellular proteins.ResultsUsing gel-LC-MS/MS, 251 total cellular proteins were identified from resting human neutrophils. This is more than ten times the number of proteins identified by an initial proteome analysis of human neutrophils and almost five times the number of proteins identified by the first 2-DE map of extracts of rat polymorphonuclear leukocytes. Most of the proteins identified in the present study are well-known, but some of them, such as neutrophil-secreted proteins and centaurin beta-1, a cytoplasmic protein involved in the regulation of NF-κB activity, are described here for the first-time.ConclusionThe present report provides new information about the protein content of human neutrophils. Importantly, our study resulted in the discovery of a series of proteins not previously reported to be associated with human neutrophils. These data are relevant to the investigation of comparative pathological states and models for novel classes of pharmaceutical drugs that could be useful in the treatment of inflammatory disorders in which neutrophils participate.

Highlights

  • Neutrophils are the most abundant leukocytes in peripheral blood and represent one of the most important elements of innate immunity

  • In order to eliminate residual contaminants, we introduced an additional step to lyses the contaminating red blood cells (RBCs) and 3 washes with Hanks' solution to eliminate contaminating monocytes/lymphocytes

  • Since our main objective was to identify the largest number of proteins, the present report describes a catalog of human neutrophil proteins

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Summary

Introduction

Neutrophils are the most abundant leukocytes in peripheral blood and represent one of the most important elements of innate immunity. Neutrophils (polymorphonuclear leukocytes, PMNs) are highly specialized blood cells which play an important role in innate immunity [1]. They constitute the first line of defense of the organism and are the most abundant leukocytes in human peripheral blood [2]. PMNs are able to instantly kill pathogens in innate immunity, neutrophils have the potential to orchestrate adaptive immune responses. They release proinflammatory chemokines and cytokines, which attract and stimulate T cells and dendritic cells (DCs) [6]. DCs can present PMN-derived antigens to T cells, instructing the responses of Th1 and Th2, and acquire antigen-presenting functions themselves [8] or transfer antigens directly to DCs [9]

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