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Ex vivo evolution of kidney leukocytes immune parameters in European bullhead (Cottus sp.)


In all living organisms, immunity is essential for body protection against biological pathogens. This physiological system is particularly sensitive to chemical contamination. Immune biomarker measurements can be used in laboratory experiments to assess the impacts of xenobiotics on cellular and humoral functions in an immunotoxicological concern. Moreover, these immune indicators can be also applied under field context to characterize the health of wild fish. In environmental immunotoxicology, analysis of immune parameters in native fish populations may be particularly difficult as immune cellular or biochemical responses have to be studied in fresh tissues with living cells. The aim of the present study was to determine in laboratory, required conditions to analyze cellular immune responses from kidney tissue samples in European bull head (Cottus sp.). For this purpose, eight bullheads were sacrificed and kidneys were immediately dissected out in order to study some immune markers. Each immune response was analyzed either immediately after tissue homogenization (T0) or 12 and 24 hours after tissue kidney treatment. Oxidative burst activity was measured by a chemiluminescence technique. Cellularity, cell mortality (apoptosis versus necrosis and leukocyte phagocytosis activity were performed by flow cytometry. Results obtained from analysis realized 12 or 24 hours after fish dissection were compared with those obtained when analysis were done immediately after kidney dissection and homogenization. In this context, no significant variation in kidney leukocyte composition was observed at 12 or 24 hours. Cellular mortality and phagocytosis activity were reduced 12 hours after kidney dissection whereas the oxidative burst activity decreased only 24 hours after tissue sampling. Simultaneously, leukocyte viability was increased 12 hours after kidney treatment. The sampling procedure and tissue treatment may be responsible for stressful conditions for isolated leukocytes which must be taken into account for the following cytometric procedure. Under field conditions, kidneys must be homogenized immediately after dissection but cellular analysis and immune marker measurements may be done only 12 hours after kidney sampling procedure. So, in the field context of environmental immunotoxicology, it is not necessary to analyze immune cellular responses directly in field and a time of 12 hours is useful to obtain low stressed-isolated leukocytes suspensions.
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ineris-00970786 , version 1 (02-04-2014)



Sabrina Jolly, Anne Bado-Nilles, Alain Geffard, Jean-Marc Porcher, Stéphane Betoulle, et al.. Ex vivo evolution of kidney leukocytes immune parameters in European bullhead (Cottus sp.). 21. SETAC Europe annual meeting "Ecosystem protection in a sustainable world : a challenge for science and regulation", May 2011, Milan, Italy. ⟨ineris-00970786⟩
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