Analysis of Neutrophil Responses to Biological Exposures.

Neutrophils are an important part of the innate immune system and among the first cells to respond to infections and inflammation. Responses include chemotaxis towards stimuli, extravasation from the vasculature, and antimicrobial actions such as phagocytosis, granule release, reactive oxygen species (ROS) production, and neutrophil extracellular trap (NET) formation (NETosis). Studying how neutrophils respond to a variety of stimuli, from biomaterial interactions to microbial insults, is therefore an essential undertaking to fully comprehend the immune response. While there are some immortalized cell lines available that recapitulate many neutrophil responses, ex vivo or in vivo studies are required to fully understand the complete range of neutrophil phenotypes. Here we describe two protocols for neutrophil isolation for further ex vivo study: recovery of neutrophils from human peripheral blood, and isolation of neutrophils from the oral cavity. We also discuss an in vivo model of general inflammation with the murine air pouch that can be used to assess numerous parameters of neutrophil and immune activation, including neutrophil recruitment and biological activity. In these protocols, the cells are isolated to allow for a high degree of experimental control. The protocols are relatively straightforward and can be successfully used by labs with no prior primary cell experience. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Neutrophil isolation from human blood Basic Protocol 2: Neutrophil isolation from the oral cavity Basic Protocol 3: Murine air pouch model of general inflammation.

Innate Phagocyte Polarization in the Oral Cavity.

The oral cavity is a complex environment constantly exposed to antigens from food and the oral microbiota. Innate immune cells play an essential role in maintaining health and homeostasis in the oral environment. However, these cells also play a significant role in disease progression. This review will focus on two innate phagocytes in the oral cavity: macrophages and neutrophils, and examine their roles during homeostasis and disease development, with a focus on periodontal disease and cancer. Macrophages have a well-known ability to polarize and be activated towards a variety of phenotypes. Several studies have found that macrophages' polarization changes can play an essential role in maintaining health in the oral cavity and contribute to disease. Recent data also finds that neutrophils display phenotypic heterogeneity in the oral cavity. In both cases, we focus on what is known about how these cellular changes alter these immune cells' interactions with the oral microbiota, including how such changes can lead to worsening, rather than improving, disease states.