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Dolmatova L. S. Eliseykina M. G. Timchenko N. F. Kovaleva A. L. Shitkova O. A. 《中国海洋湖沼学报》2003,21(4):293-304
Pure fraction (92%–95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38%)-FPMC- of the holothurianEupentacta fraudatrix' (Holothuroidea, Dendrochirota) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified
by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin ofYersinia pseudotuberculosis (TST) at different concentrations (0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation.
In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction
after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively.
Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared
to that under toxin treatment alone. TST stimulated superoxide dismutase activity in concentration-dependent manner (maximum
at 0.5 μg/ml concentration in FP) after 24 h treatment, and this stimulation was prevented by a commercial catalase. Plant
lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin
stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously,
ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in
holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production
by these cells may be one of the mechanisms of antibacterial protection of holothurians.
This work was financially supported by the Russian Foundation for Fundamental Research Grant (No. 00-04-48949). 相似文献
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In view of increasing lead pollution (Pb2+) of coastal waters, the compensatory abilities of holothurians need to be assessed. The goal of the work is to clarify the functional and phenotypical differences between two types of phagocytes (P1 and P2) in Eupentacta fraudatrix exposed to Pb(NO3)2. It has been shown that 2 mg L?1 lead exposure for 48 h increases the number of P2 phagocytes as compared to P1 cells, does not significantly affect cell viability in both P1 and P2 phagocyte fractions, and significantly enhances chromatin condensation in P2 but not in P1 phagocytes. A lead concentration of 4 mg L?1 increases the number of P1 phagocytes compared to that of P2 type, and does not change cell viability and chromatin condensation in P1 phagocytes. In the P2 type, it decreases cell viability and does not influence the level of apoptosis. The protection against lead-induced apoptosis is apparently mediated by the activities of antioxidant enzymes, especially glutathione S-transferase. The differences in labeling cell surface receptors of P1 and P2 phagocytes by plant lectins also indicate the specific phenotypic properties of these cells. The results clarify the potential and GSH-dependent mechanisms of immune adaptation in holothurians that have been shortly exposed to lead at concentrations close to the maximum environmentally relevant level in coastal waters. Additionally, P1 and P2 phagocytes are first shown to have different functions and phenotypes during the response to lead, which indicates the complexity of the phagocytic system in holothurians and contributes to understanding the immunity evolution. 相似文献
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Dolmatova L.S. Eliseykina M.G. Timchenko N.F. Kovaleva A.L. Shitkova O.A. 《中国海洋湖沼学报》2003,21(4):293-304
Pure fraction (92% - 95%) of phagocytes (FP) and a mixture of amoebocytes (62%) and morula cells (38 % )-FPMC- of the holothurian Eupentacta fraudatrix ( Holothuroidea,Dendrochirota ) were obtained by using ficoll-verographine step gradient. Basal production of reactive oxygen species (ROS) in FP quantified by using reduction of nitroblue tetrazolium (NBT) was more than twice that in FPMC. Thermostable toxin of Yersinia pseudotuberculosis (TST) at differentconcentrations ( 0.2; 0.5; 2.5 μg/ml, but not 0.1 μg/ml) stimulated NBT reduction in FPMC after 24 h incubation. In FP, TST at concentrations of 0.1 and 0.2 μg/ml inhibited and at concentrations of 0.5 and 2.5 μg/ml stimulated NBT reduction after 24 h incubation. Maximal effect was observed in FP and FPMC at TST concentrations of 0.5 and 0.2 μg/ml, respectively. Addition of catalase (0.7 μg/ml) to the cells treated with TST (2.5 μg/ml) was followed by a decrease in NBT reduction compared to that under toxin treatment alone. TST stimulated superoxide dismutase activity in ccncentration-dependent manner (maximum at 0.5 μg/ml concentration in FP) after 24 htreatment, and this stimulation was prevented by a commercial catalase. Plant lectin concanavalin A stimulated NBT reduction more than 5-fold in FPMC compared to the control. With addition of TST, lectin stimulated ROS to lesser extent than that with lectin alone. When catalase, TST, and lectin were added into the FPMC simultaneously, ROS increase was similar to that under lectin treatment alone. On the whole, data obtained indicated that ROS generation in holothurian coelomocytes especially occurs in both stimulated and not stimulated phagocytes, and that changes in ROS production by these cells may be one of the mechanisms of antibacterial protection of holothurians. 相似文献
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