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Effect of rhIL-17 on the expression of TLR2 and apoptotic proteins in human neutrophils

100%
Kakareko M., Jabłońska E., Jabłoński J., Ratajczak-Wrona W.
Medical Review
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2013
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issue 1
95-105
EN
Introduction. Neutrophils (PMNs) apoptosis occurs involving proteins of the Bcl-2 family, which control the permeability of the outer mitochondrial membrane. Proteins of this family are regulated by various factors, among others cytokine. Early observations of their own show the relationship between increased apoptosis and expression of Toll-like receptor (TLR2) in these cells stimulated by IL-17. The aim of the planned study was to investigate the effect of rhIL-17 on selected proteins of the Bcl-2 family, such as: the proapoptotic Bax protein and antiapoptotic Mcl-1 protein in relation to the expression of TLR2 receptor. Reference to the analysed results was to investigate the influence of fMLP on estimated parameters. Material and method. The study was conducted on human PMN isolated from peripheral blood. Apoptosis was assessed by cytometry, and the expression of TLR2, Bax, and Mcl-1 was estimated by Western blot. The results and conclusions. The results have shown that the enhancement of neutrophils intense apoptosis depends directly from the effect of rhIL-17 and probably indirectly on TLR2 receptor. The observed changes in the expression of Bax both under the influence of fMLP and rhIL-17 stress the important role of this protein in regulating neutrophils apoptosis.
PL
Wstęp. Apoptoza neutrofilów (PMN) zachodzi z udziałem białek rodziny Bcl-2, które kontrolują przepuszczalność zewnętrznej błony mitochondrialnej. Białka tej rodziny podlegają regulacji przez różne czynniki, m.in. cytokiny. Wcześniejsze obserwacje własne wskazują na relację między nasileniem apoptozy a ekspresją receptora TLR2 w komórkach stymulowanych IL-17. Celem zaplanowanych badań było zbadanie wpływu rhIL-17 na wybrane białka rodziny Bcl-2 szlaku wewnątrzkomórkowego, takich jak: proapoptotyczne białko Bax i antyapoptotyczne białko Mcl-1 w relacji do ekspresji receptora TLR2. Odniesieniem do analizowanych wyników było zbadanie wpływu fMLP na oceniane parametry. Materiał i metoda. Badania przeprowadzono na izolowanych z krwi obwodowej ludzkich PMN. Apoptozę oceniano metodą cytometrii przepływowej, natomiast ekspresję TLR2, Bax oraz Mcl-1 oznaczano metodą Western blot. Wyniki i wnioski. Uzyskane wyniki wykazały, że nasilona apoptoza neutrofilów zależy w sposób bezpośredni od wpływu rhIL-17 na badane białka szklaku wewnętrznego i prawdopodobnie w sposób zależny od receptora TLR2. Obserwowane zmiany ekspresji Bax zarówno pod wpływem fMLP, jak i rhIL-17 podkreślają
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Immunoaging – the effect of age on serum levels of NET biomarkers in men: a pilot study

76%
Garley M., Omeljaniuk W. J., Motkowski R., Ratajczak-Wrona W., Jabłońska E., Filipkowski D., Charkiewicz A. E.
International Journal of Occupational Medicine and Environmental Health
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2023
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vol. 36
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issue 3
333-348
EN
Objectives The study aimed to evaluate the impact of aging on the formation of neutrophil extracellular traps (NETs). The impaired formation of NETs is the cause of an abnormal innate immune response. Material and Methods The study included a total of 45 healthy male subjects of different age groups. Whole blood was collected from the subjects, and the concentration of myeloperoxidase (MPO), the main biocidal protein in NETs, was determined in serum using ELISA. The serum levels of circulating free DNA (cfDNA), which are the structural basis of NETs, were also measured by fluorescence. In addition, the white blood cell count was determined, whole blood smear was evaluated, and the neutrophillymphocyte ratio was calculated. The variations in the levels of NET biomarkers were analyzed in different age groups. Results The low levels of MPO (243.70 ng/ml) and cfDNA (6.24 ng/100 μl) in boys indicated neutrophil insufficiency for NETosis in children. A progressive increase in the levels of MPO and cfDNA with age was observed among adolescents (420.91, p = 0.04; 13.55, p = 0.03, respectively), with the highest level noted in the healthy adult group (466.58, p = 0.01; 14.07, p = 0.01, respectively). The levels of the studied parameters were comparable in adolescents and young adults, which proved that the NETosis process was appropriate and suggested the attainment of neutrophil maturity for the release of NETs in adolescence. The levels of MPO and cfDNA were low in older men (225.46, p < 0.01; 5.19, p < 0.01, respectively) indicating impaired NET formation. Conclusions Data on the generation of NETs in different age groups obtained in this study can allow a better understanding of the ontogenesis of the immune system in terms of the course of NETosis, and also indicate the need to support nonspecific responses in children and adults. Further research should be performed to determine the possibility of regulating the NETosis process.
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