Mark T. Quinn - Neutrophil: Methods and Protocols

Neutrophils (also known as polymorphonuclear neutrophils (PMNs) or granulocytes) are the most abundant white cell in humans. Granulocytes and/or granulocyte precursors normally comprise ~60% of the nucleated cells in bone marrow and the bloodstream. Mature neutrophils have a typical circulating half-life of 68 h in the blood and then migrate through tissues for ~23 days. Their relatively short lifespan is devoted largely to surveillance for invading microorganisms. During infection, the neutrophil lifespan is extended, granulopoiesis increases, and large numbers of neutrophils are rapidly recruited to the site(s) of infection. Following recognition (binding) and phagocytosis of microorganisms, neutrophils utilize an extraordinary array of oxygen-dependent and oxygen-independent microbicidal weapons to destroy infectious agents. Oxygen-dependent mechanisms involve the production of reactive oxygen species (ROS), while oxygen-independent mechanisms include degranulation and release of lytic enzymes and bactericidal peptides. Inasmuch as these processes are highly effective at killing most ingested microbes, neutrophils serve as the primary cellular defense against infection.

The aim ofNeutrophils: Methods and Protocols, Third Edition,is to provide (1) a set of protocols to assess basic neutrophil functions and (2) protocols for investigating specialized areas in neutrophil research. This volume is designed for the basic researcher involved in the study of neutrophil function. A wide variety of methods have been developed to assess neutrophil function, and these methods have been instrumental in advancing our understanding of the role of neutrophils in host defense and inflammatory disease. For those researchers and clinicians interested in the study of neutrophils, the availability of a comprehensive source of protocols describing the most modern methodological advances in neutrophil biology is invaluable, as many publications do not provide information on the finer details critical to success of a given method. As such, we have compiled a series of protocols written by leading researchers in the field that provide detailed guidelines for establishing and performing the most common neutrophil function assays. Hints of the best way to perform these methods as well as guidance in detecting associated problems are included, so novice investigators will also be able to effectively utilize these assays.

In the third edition ofNeutrophils: Methods and Protocols, chapters retained from previous editions have been updated and include many new approaches. In addition, theThird Editioncontains a number of new chapters that were not included in the first two editions. Part I contains overviews of neutrophil biology and function, and disorders of neutrophils. Part II describes commonly used methods to isolate neutrophils from humans and other animal species. This section also contains a chapter that describes use of a neutrophil transplant model with zebrafish larva. Part III details methods for investigating chemotaxis, transmigration, phagocytosis, and bactericidal activity. Three chapters provide methods used to assess neutrophil transmigration, chemotaxis, or swarming against live microbes. One of the chapters updated from the