title: In vitro effect of cadmium and copper on separated blood leukocytes of Dicentrarchus labrax.
abstract: The immunotoxic effects of heavy metals on blood leukocytes of sea bass (Dicentrarchus labrax) were examined. The cells, separated by a discontinuous Percoll-gradients, were exposed in vitro to various sublethal concentrations of cadmium and copper (10(-7)M, 10(-5)M, and 10(-3)M) and their immunotoxic effect was then evaluated by measuring neutral red uptake, MTT assay, DNA fragmentation and Hsp70 gene expression. First of all, we demonstrated that the cells treated in vitro could incorporate Cd and Cu. A relationship between heavy metal exposure and dose-time-dependent alterations in responses of leukocytes from blood was found for both metals, but copper was more immunotoxic than cadmium in all assays performed. A significant reduction in the cells׳ ability to uptake neutral red and viability by MTT assay was recorded, indicating that both cadmium and copper could change the membrane permeability, inducing cellular apoptosis when the concentration of metals reached 10(-3)M. The apoptotic effect may also explain the high level of cytotoxicity found when the leukocytes were exposed to higher concentration of metals. These results demonstrated that toxic effect of copper and cadmium affect on the mechanisms of cell-mediated immunity reducing the immune defences of the organism.
authors: Dipartimento STEBICEF, Università degli Studi di Palermo,Via Archirafi 18, Palermo, Italy.
authors: Mirella
authors: M
authors: Vazzana
authors: Dipartimento STEBICEF, Università degli Studi di Palermo,Via Archirafi 18, Palermo, Italy.
authors: Monica
authors: M
authors: Celi
authors: Dipartimento DISTEM, Università degli Studi di Palermo, Via Archirafi 18, Palermo, Italy.
authors: Cecilia
authors: C
authors: Tramati
authors: Istituto Zooprofilattico della Sicilia "A. Mirri", Palermo, Italy.
authors: Vincenzo
authors: V
authors: Ferrantelli
authors: Dipartimento STEBICEF, Università degli Studi di Palermo,Via Archirafi 18, Palermo, Italy. Electronic address: vincenzo.arizza@unipa.it.
authors: Vincenzo
authors: V
authors: Arizza
authors: Dipartimento STEBICEF, Università degli Studi di Palermo,Via Archirafi 18, Palermo, Italy.
authors: Nicolò
authors: N
authors: Parrinello
mesh_terms: D000818:Animals; D001492:Bass / Q000235:genetics / Q000502:physiology*; D002104:Cadmium / Q000633:toxicity*; D003300:Copper / Q000633:toxicity*; D005786:Gene Expression Regulation / Q000187:drug effects; D018840:HSP70 Heat-Shock Proteins / Q000235:genetics; D007962:Leukocytes / Q000187:drug effects*; D009499:Neutral Red / Q000378:metabolism; D014874:Water Pollutants, Chemical / Q000633:toxicity*
chemical_list: D018840:HSP70 Heat-Shock Proteins; D014874:Water Pollutants, Chemical; D002104:Cadmium; D009499:Neutral Red; D003300:Copper
keywords: Heavy metal; Hsp70 gene; Leukocytes; MTT; NR; Real-time-PCR