Isolation and characterization of gelatinase granules from human neutrophils

Kjeldsen, Lars; Sengeløv, Henrik; Lollike, Karsten; Nielsen, Maria Haahr; Borregaard, Niels

doi:10.1182/blood.v83.6.1640.bloodjournal8361640

Cited by 47 publications

(61 citation statements)

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“…The failure of cycloheximide to modulate the release of MMP‐9, combined with the inability of PMA and fMLP to upregulate the MMP‐9 mRNA levels, indicates that MMP‐9 was released from preformed stocks located in a specific granules subtype called “gelatinases granules” (2, 22). Interestingly, we found not only the pro‐form (92kDa) but also the activated form (88 kDa) of MMP‐9 in the supernatant of cultured granulocytes.…”

Section: Discussionmentioning

confidence: 99%

“…First, neutrophils are potent secretors of serine proteases (mainly elastase) that might, in vitro , activate the MMP‐9 by proteolytic cleavage of the aminoterminal prodomain. These enzymes are located in azurophil granules of mature neutrophils, which are partially released after PMA stimulation (2). In vitro , it has been shown that serine protease inhibitors strongly inhibit the activation of neutrophil‐derived MMP‐9, suggesting an important contribution of the neutrophil elastase in this process (25).…”

Section: Discussionmentioning

confidence: 99%

“…Among the MMPs, MMP‐9 (gelatinase B) is thought to play a key role in tissue remodeling and repair through the degradation of type IV collagen, the major component of basal membranes. This enzyme is expressed, produced and secreted by a large array of human cells including eosinophils and neutrophils (2, 3) In neutrophils, MMP‐9 is thought to be synthesized during the maturation of the precursor in the bone marrow and stored in specific granules until needed (4). MMPs are selectively inhibited by tissue inhibitors of metalloproteinases (TIMPs).…”

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confidence: 99%

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Matrix metalloproteinases and TIMP‐1 production by peripheral blood granulocytes from COPD patients and asthmatics

Cataldo

Munaut

Noël

et al. 2001

Allergy

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Both asthmatic and COPD patients were found to have increased amounts of granulocytes and matrix metalloproteinase-9 (MMP-9) in their sputum. The present study was conducted to investigate whether the elevated amounts of MMP-9 and TIMP-1 found in such patients' airways may be linked to an enhanced secretion by granulocytes. Blood granulocytes from asthmatics (n = 10), COPD patients (n = 11), and healthy controls (n = 11) were isolated and cultured under basal conditions or after stimulation by phorbol 12-myristate 13-. MMP-9 activity was detected by zymography while MMP-8 and TIMP-1 levels were measured by ELISA. In zymography, pro-and activated forms of MMP-9 were present in each group (healthy subjects, asthmatics, and COPD patients). Spontaneous release was not different between the three groups. Stimulation by fMLP and PMA increased to a similar extent the release of MMP-9 by granulocytes in all the three groups. TIMP-1 levels were also increased after stimulation by PMA and fMLP only in healthy subjects and COPD patients. MMP-8 levels were barely detectable. We conclude that circulating granulocytes from COPD patients and asthmatics do not display an abnormal secretion of MMP-9, and that granulocytes from asthmatics have an impaired ability to release TIMP-1 upon stimulation.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Matrix metalloproteinases and TIMP‐1 production by peripheral blood granulocytes from COPD patients and asthmatics

Cataldo

Munaut

Noël

et al. 2001

Allergy

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“…Specific and gelatinase granules contain the membrane component of the NADPH oxidase, cytochrome b 558, composed by the subunits gp91 phox and p22 phox . They also include modulators of the immune and inflammatory responses, such as gelatinase B (MMP‐9) (Chakrabarti and Patel, ), and lactoferrin (Kjeldsen et al ., ). Secretory vesicles also play an important role in the innate immune response by providing the β2‐integrin family member macrophage antigen 1 (CD11b/CD18) (Sengelov et al ., ; Uriarte et al ., ), which are translocated to the plasma membrane upon neutrophil activation by pathogen‐derived molecules.…”

Section: Regulation Of Neutrophil Exocytosismentioning

confidence: 97%

The role of Rab27a in the regulation of neutrophil function

Catz

2014

Cell Microbiol

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Summary Neutrophils are central regulators of the innate immune response and help shape the adaptive immune response. Malfunction and unregulated neutrophil activation leads to disease and inflammation. During the host response to infection, neutrophils display several mechanisms of defense mediated by their arsenal of granular proteins. Regulation of granular trafficking, docking and fusion is at the core of the neutrophil defense response to pathogens. The small GTPase Rab27a has emerged as a central regulator of the neutrophil response through its tight control of vesicular trafficking and degranulation. This review focuses on the latest research that has led to the characterization of Rab27a as an essential regulator of neutrophil function.

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“…Essentially the method described by Kjeldsen et al [] was used. The PMNs were lysed by nitrogen cavitation for 20 min, and the lysate of 5 × 10 8 PMNs was placed on top of a two‐layer Percoll gradient (density 1050/1120) and centrifuged for 30 min at 20,000 rpm (SS34 rotor, Sorvall, Fisher Scientific, UK).…”

Section: Methodsmentioning

confidence: 99%

Human polymorphonuclear neutrophils express RANK and are activated by its ligand, RANKL

et al. 2012

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The receptor activator of NF-κB (RANK) is especially well studied in the context of bone remodeling, and RANK and its ligand, RANKL, are key molecules in the induction of bone resorbing osteoclasts. We now report that polymorphonuclear neutrophils (PMNs) contain preformed RANK, stored in secretory vesicles and in specific granules. Upon stimulation of PMNs in vitro, RANK was translocated to the cell membrane. In patients with persistent bacterial infections, RANK surface expression was enhanced compared with that of healthy individuals. The functional activity of RANK was assessed by determining migration of PMNs toward RANKL. A time-and dose-dependent migration was seen, leading to the conclusion that RANK on PMNs is functional. We presume that regulated RANK expression contributes to the fine tuning of PMN migration, for example, on and through inflamed endothelium that is known to express RANKL. Keywords: Chemotaxis · Neutrophils · polymorphonuclear neutrophil (PMN) · RANK IntroductionThe receptor activator of NF-κB (RANK) is especially well studied in the context of bone remodeling, and RANK and its ligand, RANKL, are key molecules in the induction of bone resorbing osteoclasts (reviewed in [1,2]). Expression of RANK, however, is not limited to osteoclast precursor cells; peripheral blood monocytes and dendritic cells also express RANK. More recently, the expression of RANK on polymorphonuclear neutrophils (PMNs) derived from the synovial fluid of patients with rheumatoid arthritis was described [3], which suggests that RANK can be upregulated on PMNs under inflammatory conditions.PMNs contain a multitude of preformed receptors and enzymes that are stored in specialized granules or vesicles and are mobilized in a controlled fashion in response to selected stimuli [4,5]. While cytotoxic and bactericidal entities are predominantly stored in granules, receptors such as CD14 or CD16 and the adhesion molecules CD11b/CD18 are associated with the membrane of secretory vesicles [4,6,7]. Mobilization of the latter generates a Correspondence: Prof. G. Maria Hänsch e-mail: n50@ix.urz.uni-heidelberg.de PMN phenotype that attaches to endothelial cells and allows transmigration, emphasizing that controlled mobilization of granules and vesicles is central to the PMN function. A limited number of receptors are also de novo synthesized by PMNs. Interferon-γ (IFN-γ) or lipopolysaccharides (LPS), for example, induce synthesis of CD14, major histocompatibility complex (MHC) class II antigens, and of the costimulatory antigens CD80 and CD86 [8].In this study, we analyzed the expression of RANK by PMNs and found that it is preformed and stored in secretory vesicles and in specific granules as well, and that it is translocated to the surface upon stimulation in vitro and in vivo. Moreover, we found that RANK-positive PMNs can be activated via RANKL. Results Neutrophils of healthy donors express RANKPMNs of healthy donors express RANK. By cytofluorometry, RANK was detected intracellularly and on the cell surface (Fig. 1A) less tha...

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Isolation and characterization of gelatinase granules from human neutrophils

Cited by 47 publications

References 1 publication

Matrix metalloproteinases and TIMP‐1 production by peripheral blood granulocytes from COPD patients and asthmatics

Matrix metalloproteinases and TIMP‐1 production by peripheral blood granulocytes from COPD patients and asthmatics

The role of Rab27a in the regulation of neutrophil function

Human polymorphonuclear neutrophils express RANK and are activated by its ligand, RANKL

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