Morphological and Compositional Analysis of Neutrophil Extracellular Traps Induced by Microbial and Chemical Stimuli

Almaraz-Arreortua, Alexia; Sosa-Luis, Sorely Adelina; Ríos-Ríos, William de Jesús; Romero-Tlalolini, María de los Ángeles; Aguilar-Ruiz, Sergio Roberto; Baltiérrez-Hoyos, Rafael; Aguilar, Honorio Torres

doi:10.3791/64522-v

Cited by 1 publication

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“…15250061, Thermo Fisher Scientific, Inc.) and counted on a hemocytometer slide before adjusting the neutrophil concentration to 2x10 6 cells/ml with RPMI 1640 medium. Purity was determined using Wright's staining and morphological staining (14) and was routinely >95% neutrophils. ROS measurement.…”

Section: Methodsmentioning

confidence: 99%

Helicobacter pylori extract induces purified neutrophils to produce reactive oxygen species only in the presence of plasma

Thai,

Chuenchom,

Donsa

et al. 2023

Biomed Rep

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H. pylori is a bacterial pathogen infecting over half of the world's population and induces several gastric and extra-gastric diseases through its various virulence factors, especially cagA. These factors may be released from the bacteria during interactions with host immune cells. Neutrophils play key roles in innate immunity, and their activity is regulated by plasma factors, which can alter how these cells may interact with pathogens. The aim of the present study was to determine whether purified neutrophils could produce reactive oxygen species (ROS), one of the key functions of their anti-microbial functions, in response to extracts of cagA+ and cagA-H. pylori. Extracts from either cagA+ or cagA-H. pylori were co-cultured with human neutrophils in the presence or absence of plasma, and the neutrophil ROS production was measured. In the absence of plasma, extracts from cagA+ and cagA-H. pylori did not induce neutrophil ROS production, whereas in the presence of plasma, extracts from both cagA+ and cagA-H. pylori-induced ROS production. Furthermore, when peripheral blood mononuclear cells (PBMCs) were added to the purified neutrophils in the absence of plasma, there was no neutrophil ROS production after challenging with extracts from either cagA+ or cagA-H. pylori. Thus, it is suggested that plasma contains immunological components that change the responsiveness of neutrophils, such that when neutrophils encounter the bacterial antigens in H. pylori extracts, they become activated and produce ROS. This study also revealed a potential novel immunopathogenic pathway by which cagA activation of neutrophils contributed to inflammatory damage.

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Section: Methodsmentioning

confidence: 99%