EFFECTS OF IN-VITRO NICKEL EXPOSURE ON THE MACROPHAGE-MEDIATED IMMUNE FUNCTIONS OF RAINBOW-TROUT (ONCORHYNCHUS-MYKISS)
EFFECTS OF IN-VITRO NICKEL EXPOSURE ON THE MACROPHAGE-MEDIATED IMMUNE FUNCTIONS OF RAINBOW-TROUT (ONCORHYNCHUS-MYKISS)
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DOI：
10.1007/bf00197823
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发表时间：
1994-03-01
影响因子：
2.7
通讯作者：
ZELIKOFF, JT
中科院分区：
文献类型：
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作者：
BOWSER, DH;FRENKEL, K;ZELIKOFF, JT
材料和方法虹鳟（Oncorhynchus mykiss），重350-500 g，购自Muskey Trout Hatchery（阿斯伯里，NJ）。将鱼饲养在通风良好的水族箱中，温度为10- 4 ℃，每天给予鳟鱼饲料，并在喂食后5小时内将任何未消耗的食物移走。在处死前14天和7天，通过两次腹膜内注射0.4mg在弗氏不完全佐剂中的杀鲑气单胞菌来引发腹膜M~+。在用MS-222（3-氨基苯甲酸乙酯，pH 7.0）麻醉后，通过灌洗收集大分子化合物。将灌洗液以400 x g离心，并将细胞离心。将沉淀重悬于含Earle盐的Eagle最小必需培养基（EMEM-E）中。取细胞等分试样以分别通过血细胞计数器计数和台盼蓝拒染法评估M-数目和活力，并且剩余的细胞用于功能测定。
MATERIALS AND METHODSRainbow trout (Oncorhynchus mykiss), weighing 350-500 g, were purchased from Muskey Trout Hatchery (Asbury, NJ). The fish were maintained in well-aerated aquaria at 10-~+ 4 Trout chow was given daily and any unconsumed food was removed within five hours-after feeding. Trout were acclimated in the laboratory for at least ten days before use.Peritoneal M~ were elicited by two intraperitoneal injections of 0.4 mg Aeromonas salmonicida in Freund's incomplete adjuvant, fourteen and seven days prior to sacrifice. Macrophages were collected by lavage after anaesthetization with MS-222 (3-amino benzoic acid ethyl ester, pH 7.0). The lavage fluid was centrifuged at 400x g and the cel! pellet resuspended in Eagle s Minimal Essential Medium with Earle s salts (EMEM-E). Cell aliquots were taken to assess M~ number and viability by hemocytometer counting and trypan blue exclusion, respectively, and the remaining cells were used for the functional assays.