Effect of Collection and Preprocessing Methods on Neutrophil Elastase Plasma Concentrations

Fischer, Joachim E.; Janousek, Martin; Fischer, Marianne; Seifarth, Federico G.; Blau, Nenad; Fanconi, Sergio

doi:10.1016/s0009-9120(98)00008-3

Cited by 5 publications

(4 citation statements)

References 16 publications

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“…Significant differences in sampling conditions when determining the plasma elastase levels were previously demonstrated. 31 Our observation of increased plasma elastase levels after IVIg treatment is supported by the fact that previous studies showed an increase in proinflammatory cytokines after IVIg infusions, including neutrophil-activating cytokines TNF-α, IL-6, and IL-8. 32,33 The observation of an increase in plasma elastase levels following IVIg infusion is supported by other reports of in vitro elastase release 27 and general granulocyte activation 29,30 after in vitro stimulation of whole blood by IVIg.…”

Section: Discussionsupporting

confidence: 76%

Common variable immunodeficiency patients display elevated plasma levels of granulocyte activation markers elastase and myeloperoxidase

Litzman

Chovancová

Bejdák

et al. 2019

Int J Immunopathol Pharmacol

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Common variable immunodeficiency disorders (CVIDs) represent a group of primary immunodeficiency diseases characterized by hypogammaglobulinemia and dysfunctional immune response to invading pathogens. Previous studies have indicated that CVID is associated with microbial translocation and systemic myeloid cell activation. The goal of this study was to determine whether patients with CVID display elevated systemic levels of markers of granulocyte activation and whether the levels are further influenced by intravenous immunoglobulin (IVIg) infusions. The plasma levels of granulocyte activation markers elastase and myeloperoxidase were determined using enzyme-linked immunosorbent assay (ELISA) in 46 CVID patients and 44 healthy controls. All CVID patients were in a stable state with no apparent acute infection. In addition, granulocyte activation markers’ plasma levels in 24 CVID patients were determined prior to and 1 h following IVIg administration. Neutrophil elastase and myeloperoxidase plasma levels were significantly higher in CVID patients than in healthy controls. Systemic elastase levels were further increased following IVIg administration. In vitro stimulation of 13 CVID patients’ whole blood using IVIg in a therapeutically relevant dose for 2 h resulted in a significant increase in plasma elastase levels compared to unstimulated blood. The data presented here indicate that CVID is associated with chronic granulocytic activation which is further exacerbated by administering IVIg. Increased myeloperoxidase and elastase levels may contribute to associated comorbidities in CVID patients.

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Section: Discussionsupporting

confidence: 76%

Common variable immunodeficiency patients display elevated plasma levels of granulocyte activation markers elastase and myeloperoxidase

Litzman

Chovancová

Bejdák

et al. 2019

Int J Immunopathol Pharmacol

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“…Our results indicate that plasma samples may be more suitable for MS protein profiling (Table 1). During coagulation, release of cellular components, mainly from thrombocytes and leukocytes, is induced (10,11 ). Furthermore, proteases seem to be more active in serum samples (12 ).…”

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confidence: 99%

Preanalytical Impact of Sample Handling on Proteome Profiling Experiments with Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Findeisen¹,

Sismanidis

Riedl

et al. 2005

Clinical Chemistry

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Mass spectrometry (MS) offers a wide range of possibilities for analyzing biological samples containing complex mixtures of proteins and peptides, by generating proteome profiles. The aim of most profiling experiments is identification of changes in protein patterns that are related to a certain disease or clinical status and might therefore be used to improve diagnosis, staging, and monitoring (1 ). Reproducibility of spectra generated by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS is crucial (2 ), however, and early enthusiastic reports about the diagnostic power of proteome-profiling approaches for early disease classification have recently been critically assessed and reevaluated (3 ). The reproducibility of profiling experiments depends on sample quality, which in turn is greatly influenced by preanalytical storage conditions and the choice of anticoagulants (4 -6 ). In this study, we examined the impact of preanalytical sample handling and storage times on MALDI-TOF MS data for serum and plasma samples and identified a candidate marker to allow objective classification of given samples with respect to age and, hence, appropriateness for MS protein profiling and interpretation of data.We collected 10-mL venous blood samples into serum tubes, EDTA-coated tubes, and ammonium-heparincoated tubes (Sarstedt). After collection, all samples were initially kept at room temperature for 30 min, to allow clot formation, before centrifugation at 3000g for 10 min in a precooled (4°C) centrifuge (Rotina 48R; Hettich). After centrifugation, 1 aliquot of each sample was immediately frozen at Ϫ80°C (time 0); other aliquots were incubated for distinct time periods at room temperature as follows: Specimens from 1 healthy volunteer were stored at room temperature until 1, 2, and 4 h after collection and then stored at Ϫ80°C before further use. For classification analysis, serum samples from 6 healthy volunteers and from 20 patients consecutively entering the emergency unit of the university hospital at Mannheim were collected and processed as described above, with the exception that aliquots were prepared only at time 0 and from samples stored at room temperature for 4 h after collection. The study was conducted in accordance with the regulations of the local ethics committee. All samples were deidentified.Serum and plasma samples were processed with the following magnetic bead (MB)-based affinity chromatography resins, used according to optimized protocols recommended by the manufacturer (Bruker Daltonics): hydrophobic interaction chromatography C 8 (MB-HIC-C8), weak cation-exchange chromatography (MB-WCX), and immobilized metal-ion affinity chromatography (MB-IMAC-Cu). Specifically, 5 L of the sample was mixed with the appropriate MB suspension, washed 3 times with 100 L of wash buffer, and eluted with 5 L of appropriate elution solution for the MB-HIC-C8 and MB-WCX reagent sets and 10 L of appropriate elution solution for the MB-IMAC-Cu reagent set. A portion of the eluted sample was diluted 1:...

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“…When performed in addition to other analyses, this assay requires only 4 ll of plasma. Unfortunately, routine E-a1PI measurements are hampered by the sampling procedures available as these result in unpredictable post-collection artefacts [1]. Recently, we identi®ed a collection method that minimises the risk of falsepositive E-a1PI results [1].…”

Section: Introductionmentioning

confidence: 99%

“…Unfortunately, routine E-a1PI measurements are hampered by the sampling procedures available as these result in unpredictable post-collection artefacts [1]. Recently, we identi®ed a collection method that minimises the risk of falsepositive E-a1PI results [1]. Here we present data on the diagnostic performance of E-a1PI, applying the new assay and sampling procedure to a heterogeneous cohort of critically ill newborn infants.…”

Section: Introductionmentioning

confidence: 99%

Diagnostic potential of neutrophil elastase inhibitor complex in the routine care of critically ill newborn infants

et al. 2000

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Effect of Collection and Preprocessing Methods on Neutrophil Elastase Plasma Concentrations

Cited by 5 publications

References 16 publications

Common variable immunodeficiency patients display elevated plasma levels of granulocyte activation markers elastase and myeloperoxidase

Common variable immunodeficiency patients display elevated plasma levels of granulocyte activation markers elastase and myeloperoxidase

Preanalytical Impact of Sample Handling on Proteome Profiling Experiments with Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

Diagnostic potential of neutrophil elastase inhibitor complex in the routine care of critically ill newborn infants

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