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31 October 2012

The influence of uremic high cystatin C concentration on neutrophil apoptosis and selected neutrophil functions isolated from healthy subjects

Ewa MajewskaABCDEFG, Natalia WittekABD, Jacek RyszADF, Zbigniew BajADEG

DOI: 10.12659/MSM.883545

Med Sci Monit 2012; 18(11): CR667-673

Abstract

Background: Cystatin C (cC) is a cysteine protease inhibitor that may influence immune response. Our aim was to test the effect of a high concentration of cC, characteristic for uremic patients, on neutrophil (PMN) apoptosis and respiratory burst, as well as the cC secretion from PMNs stimulated with proinflammatory cytokines.
Material/Methods: PMNs from 35 healthy volunteers aged 27–61 years were cultured in presence of cC, IL-1beta or TNF-alpha. The percentage of apoptotic cells based on DNA depletion, Fas, FasL and caspase -3 expression were assessed. CC concentrations were determined by ELISA test. The influence of cC on spontaneous, fMLP-, PMA- or OZ-induced burst response of PMNs was tested using chemiluminescence.
Results: PMN cultured in the presence of cC resulted in a significant drop in apoptotic cell percentage (38% [11%; 65%]) compared both to control (70% [29%; 92%], and to the cells cultured with TNF-alpha (58% [24%; 85%]). These differences were not accompanied by Fas, FasL and caspase-3 expression changes. Spontaneous, fMLP- and PMA-stimulated oxidative burst of PMNs preincubated with cC were significantly downregulated. IL-1beta markedly diminished and TNF-alpha significantly increased cC concentration in culture supernatants.
Conclusions: The presented results suggest that antiapoptotic activity of cC results from its inhibitory effect on ROS production. Thus, the higher concentration of cC characteristic for uremic patients may modulate acute inflammation through maintaining PMN longevity and inhibiting their respiratory burst and proinflammatory cytokine-related changes in cC release from PMNs.

Keywords: N-Formylmethionine Leucyl-Phenylalanine - pharmacology, Interleukin-1beta - pharmacology, Health, Fas Ligand Protein - metabolism, Cystatin C - metabolism, Cells, Cultured, Cell Separation, Cell Respiration - drug effects, Neutrophils - pathology, Tetradecanoylphorbol Acetate - pharmacology, Tumor Necrosis Factor-alpha - pharmacology, Uremia - metabolism

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750