Separation of neutrophils from blood in human and laboratory animals and comparison of the chemotaxis

Sugawara, Takafumi; Miyamoto, Manabu; Takayama, Satoshi; Kato, Michiyuki

doi:10.1016/1056-8719(94)00062-9

Cited by 64 publications

(45 citation statements)

References 16 publications

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“…These factors are known to efficiently activate chemotaxis of human blood neutrophils. 39 Freshly isolated neutrophils responded to these chemoattractants with characteristic biphasic curves that showed maximal neutrophil responses at 10 À7 M for fMLP ( Figure 5a) and LTB 4 (data not shown) and 10 À8 M for IL-8 (data not shown). In contrast, long-lived neutrophils migrated significantly less than freshly isolated neutrophils toward chemotactic factors.…”

Section: Functions Of Long-lived Neutrophils a Chakravarti Et Almentioning

confidence: 99%

Reprogramming of a subpopulation of human blood neutrophils by prolonged exposure to cytokines

Chakravarti

Rusu

Flamand

et al. 2009

Laboratory Investigation

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Essential cells of innate immunity, neutrophils are often considered to be a homogenous population of terminally differentiated cells. During inflammation, neutrophils are extravasated cells exposed to local factors that prolong their survival and activate their production of mediators implicated in disease progression. In this study, a phenotypically distinct subset of human neutrophils that appear after prolonged exposure to cytokines was characterized. Freshly isolated neutrophils from healthy donors were incubated with granulocyte-macrophage colony-stimulating factor, tumor necrosis factor-a and interleukin (IL)-4, three cytokines that are locally present in various inflammatory conditions. Eight to 17% of neutrophils survived beyond 72 h. This subset of non-apoptotic neutrophils, as evaluated by three different markers, was enriched by discontinuous Percoll gradient centrifugation before studying their phenotype. These viable neutrophils showed neoexpression of HLA-DR, CD80 and CD49d. Compared with freshly isolated neutrophils, they responded differentially to second signals similar to formyl-methionyl-leucyl-phenylalanine with three-to four-fold increases in production of superoxide anions and leukotrienes. These cells augmented their phagocytic index by 141%, increased their adhesion to human primary fibroblasts, but reduced their migration in response to chemotactic stimuli and decreased exocytosis of primary and secondary granules. In addition, they produced substantial amounts of IL-8, IL-1Ra and IL-1b. This neutrophil subset had a unique profile of phosphorylation of intracellular signaling molecules. In conclusion, the present identification of a novel neutrophil phenotype highlights the reprogammable character of the neutrophil. This aspect is crucial for our understanding of its contribution to disease pathogenesis and host defense.

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Section: Functions Of Long-lived Neutrophils a Chakravarti Et Almentioning

confidence: 99%

Reprogramming of a subpopulation of human blood neutrophils by prolonged exposure to cytokines

Chakravarti

Rusu

Flamand

et al. 2009

Laboratory Investigation

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“…Peripheral PMNs were isolated as previously described with minor modifications (33,34). Briefly, rats were anesthetized (initial experiments were performed with halothane/metofane, and later experiments yielding similar results were performed with i.m.…”

Section: Cell Isolationmentioning

confidence: 99%

Surfactant Protein A Enhances Alveolar Macrophage Phagocytosis of Apoptotic Neutrophils

Schagat

Wofford

Wright

2001

The Journal of Immunology

191

128

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Surfactant protein A (SP-A) is an innate immune molecule that binds foreign organisms that invade the lungs and targets them for phagocytic clearance by the resident pulmonary phagocyte, the alveolar macrophage (AM). We hypothesized that SP-A binds to and enhances macrophage uptake of other nonself particles, specifically apoptotic polymorphonuclear neutrophils (PMNs). PMNs are recruited into the lungs during inflammation, but as inflammation is resolved, PMNs undergo apoptosis and are phagocytosed by AMs. We determined that SP-A increases AM phagocytosis of apoptotic PMNs 280 ± 62% above the no protein control value. The increase is dose dependent, and heat-treated SP-A still enhanced uptake, whereas deglycosylated SP-A had significantly diminished ability to enhance phagocytosis. Surfactant protein D also increased phagocytosis of apoptotic PMNs by ∼125%. However, other proteins that are structurally homologous to SP-A, mannose-binding lectin and complement protein 1q, did not. SP-A enhances phagocytosis via an opsonization-dependent mechanism and binds apoptotic PMNs ∼4-fold more than viable PMNs. Also, binding of SP-A to apoptotic PMNs does not appear to involve SP-A’s lectin domain. These data suggest that the pulmonary collectins SP-A and SP-D facilitate the resolution of inflammation by accelerating apoptotic PMN clearance.

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“…First, circulating rat PMNs are few in number compared to human (absolute PMN counts of approximately 1000/µL in the rat and 5000/µL in the human). Second, the distribution of leukocyte densities is disparate in the rat and human [6,7]. Third, and perhaps most significant, the amount of superoxide anion (O 2 Ϫ ) released from activated rat PMNs appears to be a small fraction of that released by human PMNs [8,9].…”

Section: Introductionmentioning

confidence: 99%

Disparities in the respiratory burst between human and rat neutrophils

Johnson

Moore

Hiester³

et al. 1999

Journal of Leukocyte Biology

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The importance of reactive oxygen species (ROS) in neutrophil (PMN)-mediated injury to host tissues has been strongly implicated in a number of animal models. Peculiarities of the laboratory rat PMN, including an apparent paucity of superoxide release, prompted us to examine disparities in the respiratory burst between human and rat PMNs. Using isolated PMNs, we examined oxygen consumption, superoxide release, nitrate/ nitrite release, and dihydrorhodamine (DHR) oxidation in response to an array of soluble stimuli. Our findings confirm that intact rat PMNs release little superoxide in comparison to human PMNs when primed and activated by soluble stimuli. For example, PMA-activated human PMNs released superoxide at 10.1 ؎ 2.7 times the rate of rat PMNs (P F 0.01). However, measurements of oxygen consumption, cell-associated oxidant production (by DHR oxidation) and release of superoxide from electroporated cells suggests that rat PMNs generate oxidants at rates equivalent to human PMNs but preferentially release them in an intracellular compartment. Implications for the study of PMNmediated oxidant injury in animal models are discussed. J. Leukoc. Biol. 65: 211-216; 1999.

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Separation of neutrophils from blood in human and laboratory animals and comparison of the chemotaxis

Cited by 64 publications

References 16 publications

Reprogramming of a subpopulation of human blood neutrophils by prolonged exposure to cytokines

Reprogramming of a subpopulation of human blood neutrophils by prolonged exposure to cytokines

Surfactant Protein A Enhances Alveolar Macrophage Phagocytosis of Apoptotic Neutrophils

Disparities in the respiratory burst between human and rat neutrophils

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