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    Determination of extracellular traps structures from sheep polymorphonuclear leukocytes to Echinococcus granulosus protoscoleces
    (Academic Press Inc Elsevier Science, 2022) Yildiz, Kader; Simsek, Neslihan Sursal; Gurcan, I. Safa
    It was aimed to detect extracellular traps structures from sheep polymorphonuclear leukocytes (PMN) after being confronted with Echinococcus granulosus protoscoleces in vitro. Also, the effect of cyst fluid was examined on the development of extracellular traps. At the end of the incubation for 1 h, the extracellular traps augmented with neutrophil elastase, histone (H3) and myeloperoxidase were visualized in the protoscoleces-PMN co-culture microscopically. Some protoscoleces lysed and the chitinous hooks released were surrounded by the extracellular traps. The other protoscoleces were still intact and the extracellular trap structures were observed around them. The relationship between the extracellular DNA contents and the protoscoleces concentration was not found statistically significant (P > 0.05). The extracellular DNA amount in the co-cultures diluted in RPMI-1640 increased with the incubation time (P < 0.05). However, the time-dependent relationship was not found in the co-cultures diluted in the cyst fluid (P > 0.05). The difference in the extracellular DNA amount was detected as statistically significant (P < 0.05) between the two co-culture groups (diluted in RPMI-1640 or the cyst fluid), except for 30 min incubation. To the Author's knowledge, NETosis reaction was firstly observed in sheep PMN after being confronted with protoscoleces in vitro. The cyst fluid had some negative effects on the development of extracellular traps from sheep PMNs at the 1-h incubation time. It should be investigated which molecules are responsible for NETosis inhibition in hydatid cyst fluid. Future studies may clarify whether neutrophils fight with protoscoleces by using their different mechanisms.
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    Öğe
    In vitro investigation on extracellular traps formation of cat polymorphonuclear leucocytes against Toxoplasma gondii
    (Tubitak Scientific & Technological Research Council Turkey, 2021) Simsek, Neslihan Sursal; Cakmak, Ayse; Yildiz, Kader
    Neutrophil granulocytes are one of the most important defenders of the innate immune system in the host. Moreover, neutrophils are able to reach the inflammation area and kill the pathogens using various defense strategies including neutrophil extracellular traps (NETs). However, there is still not enough information available regarding the innate immunity against Toxoplasma gondii in cats that are both definitive and intermediate hosts of this parasite. Therefore, we investigated the in vitro NETs formation which is induced by cat polymorphonuclear leucocytes (PMNs) against T. gondii tachyzoites. Firstly, PMNs were isolated from cat venous blood samples by using discontinues Percoll dilutions (72%, 63%, 54%, and 45%). Afterward, MN-tachyzoites cocultures were stained against histone (H3), neutrophil elastase (NE), and myeloperoxidase (MPO) by using monoclonal antibodies and were examined under a fluorescence microscope. The effect of different parasite doses (1:1, 1:3, and 1:5) and incubation times (30, 60, 90, and 120 min) on NETs formation was also evaluated. The presence of the extracellular DNA content was measured using a fluorometer. Confluent Vero cell cultures were used to assess the effect of NETs on the tachyzoites viability. The classical structures of NETs, such as extracellular DNA, NE, H3, and MPO were microscopically observed in the NETs formation released from cat PMNs. The amount of extracellular DNA increased in parallel with the incubation time (p < 0.001). The influence of the tachyzoites dose on the NETs formation was not statistically significant (p > 0.05). Zymosan was used as a positive control in the experiments and it was shown to be an important inducer for the NETs formation. In conclusion, as mentioned previous studies and considering our results, the NETs may be a conserved strategy to control T. gondii infection in hosts because of the immobilization and lethal effects.