Characterization of two ELISAs for NGAL, a newly described lipocalin in human neutrophils

Kjeldsen, Lars; Koch, Claus; Arnljots, Kristina; Borregaard, Niels

doi:10.1016/s0022-1759(96)00153-6

Cited by 64 publications

(63 citation statements)

References 23 publications

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“…The plasma concentration of NGAL and gelatinase has previously been shown to closely mirror the granulocyte count in healthy adults. 24,25 Thus, the interindividual fluctuation in ficolin-1 concentrations in normal individuals may not within certain limits be dependent on the actual granulocyte count.…”

Section: Discussionmentioning

confidence: 99%

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Variation in FCN1 affects biosynthesis of ficolin-1 and is associated with outcome of systemic inflammation

et al. 2012

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Ficolin-1 is a recognition molecule of the lectin complement pathway. The ficolin-1 gene FCN1 is polymorphic, but the functional and clinical consequences are unknown.The concentration of ficolin-1 in plasma and FCN1 polymorphisms in positions À 1981 (rs2989727), À 791 (rs28909068), À 542 (rs10120023), À 271 (rs28909976), À 144 (rs10117466) and þ 7918 (rs1071583) were determined in 100 healthy individuals. FCN1 expression by isolated monocytes and granulocytes and ficolin-1 levels in monocyte culture supernatants were assessed in 21 FCN1-genotyped individuals. FCN1 polymorphisms were determined in a cohort of 251 patients with systemic inflammation. High ficolin-1 plasma levels were significantly associated with the minor alleles in position À 542 and À 144. These alleles were also significantly associated with high FCN1 mRNA expression. The level of ficolin-1 in culture supernatants was significantly higher in individuals homozygous for the minor alleles at positions À 542 and À 144. Homozygosity for these alleles was significantly associated with fatal outcome in patients with systemic inflammation. None of the other investigated polymorphisms were associated with FCN1 and ficolin-1 expression, concentration or disease outcome. Functional polymorphic sites in the promoter region of FCN1 regulate both the expression and synthesis of ficolin-1 and are associated with outcome in severe inflammation.

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Section: Discussionmentioning

confidence: 99%

“…It has been shown that neutrophils might be the primary source of ficolin-1 in the blood, 21 thus we determined the level of neutrophil gelatinase-associated lipocalin (NGAL) and gelatinase, which reflects the pool of circulating neutrophils. 24,25 No correlation was observed with either NGAL or gelatinase (data not shown).…”

Section: Plasma Levels Of Ficolin-1mentioning

confidence: 90%

Variation in FCN1 affects biosynthesis of ficolin-1 and is associated with outcome of systemic inflammation

et al. 2012

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“…After 15 min of incubation, cells were pelleted by centrifugation at 200 g for 6 min, the supernatant decanted, and the pellet resuspended to the initial volume in KRP with glucose. The content of myeloperoxidase (azurophil granule marker), lysozyme (azurophil, and specific granule marker), lactoferrin, and NGAL (specific granule markers) and gelatinase (marker for gelatinase granules) was measured in supernatant and pellet by enzyme-linked immunosorbent assays (ELISAs) [18][19][20]. Release of granule marker protein was expressed as content in supernantant in percent of total content (supernatant ϩ pellet).…”

Section: Exocytosis Experimentsmentioning

confidence: 99%

Giant granules of neutrophils in Chediak-Higashi syndrome are derived from azurophil granules but not from specific and gelatinase granules

Kjeldsen

Calafat

Borregaard

1998

Journal of Leukocyte Biology

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The abnormal giant granules of ChediakHigashi syndrome (CHS) neutrophils in humans are thought to be derived from both azurophil and specific granules, whereas the presence of gelatinase granules and their contribution to giant granule formation has not been investigated previously. We have examined the ultrastructure and mobilization of neutrophil granules from a patient with CHS by immunogold electron microscopy and exocytosis experiments of isolated leukocytes. The giant granules contained the azurophil granule components myeloperoxidase and CD63. We found no evidence of involvement of specific or gelatinase granules in the formation of giant granules because lactoferrin and gelatinase were contained in normalappearing peroxidase-negative granules. On stimulation of leukocytes with N-formyl-methionyl-leucylphenylalanine and the calcium ionophore, ionomycin, there was a diminished exocytosis of myeloperoxidase in CHS compared with a healthy control, indicating a lack of mobilization of the giant granules. On the other hand, there was a normal or augmented release of lactoferrin and gelatinase in CHS neutrophils, with gelatinase granules being the most easily mobilized, as known from normal neutrophils. In conclusion, giant granules from CHS neutrophils originate from azurophil granules but not from the specific and gelatinase granules. J. Leukoc. Biol. 64: 72-77; 1998.

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“…Several immunoassays have been developed for the measurement of HNL/NGAL. The assays are based on different formats and include RIA (20), Western blotting (21), ELISA (22,23), Triage device (24), and the Architect platform (16). Several research groups used one of these assays to determine the levels of HNL/NGAL in urine and drew the conclusion that HNL/NGAL is a biomarker of AKI (12,13,16 -18).…”

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confidence: 99%

The Origin of Multiple Molecular Forms in Urine of HNL/NGAL

Cai

Rubin

Wang

et al. 2010

Clinical Journal of the American Society of Nephrology

251

213

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Results: The major molecular form of HNL/NGAL secreted by neutrophils is dimeric, whereas the major form secreted by HK-2 cells is monomeric. This was reflected by a predominance of the monomeric form in urine from patients with AKI and the dimeric form in patients with UTIs. The epitope specificities of the antibody used in the ELISAs had a profound effect on assay performance and paralleled differences of the antibodies to identify the different forms of urine HNL/NGAL.Conclusions: The monomeric form is the predominant form secreted by tubular epithelial cells, and the dimeric form is the predominant form secreted by neutrophils. The development of molecular form-specific assays for HNL/NGAL may be a means to identify the origin of HNL/NGAL in urine and construct more specific tools for the diagnosis of AKI.

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Characterization of two ELISAs for NGAL, a newly described lipocalin in human neutrophils

Cited by 64 publications

References 23 publications

Variation in FCN1 affects biosynthesis of ficolin-1 and is associated with outcome of systemic inflammation

Variation in FCN1 affects biosynthesis of ficolin-1 and is associated with outcome of systemic inflammation

Giant granules of neutrophils in Chediak-Higashi syndrome are derived from azurophil granules but not from specific and gelatinase granules

The Origin of Multiple Molecular Forms in Urine of HNL/NGAL

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