Chen S C, Adams A, Thompson K D, Richards R H
Department of Veterinary Medicine, National Pingtung University of Science and Technology, Neipu, Taiwan.
Dis Aquat Organ. 1998 Mar 5;32(2):99-110. doi: 10.3354/dao032099.
The cytological response of rainbow trout Oncorhynchus mykiss head kidney macrophages to ingested Mycobacterium spp. was examined in vitro. Mycobacterium marinum or Mycobacterium sp. TB267 isolated from snakehead fish Channa striata Bloch were opsonised with either fresh rainbow trout serum, serum which had been heat-inactivated, or rainbow trout antiserum against the extracellular products (ECP) of the 2 Mycobacterium spp. A monoclonal antibody against the ECP was also used as an opsonin. Suspensions of macrophages were prepared (1 ml of 1 x 10(7) cells ml-1), mixed with the opsonised bacteria (100 microliters of 2 x 10(9) ml-1), and incubated at 18 degrees C for 0.5, 1, 2, 4 or 6 h to allow phagocytosis to occur. A quantitative evaluation of the phagocytosis of the mycobacteria by the macrophages was carried out by electron microscopy. Macrophage phagosomes and their contents were examined and numbers of intact and partially degraded bacteria determined. Pre-labelling dense granules (secondary lysosomes) with ferritin enabled phagosome lysosome fusion to be identified and their frequency determined. Opsonisation of the mycobacteria was found to greatly enhance the phagocytic and killing activity of the rainbow trout macrophages.
在体外研究了虹鳟Oncorhynchus mykiss头肾巨噬细胞对摄入的分枝杆菌属细菌的细胞学反应。用新鲜虹鳟血清、热灭活血清或虹鳟针对这两种分枝杆菌属细胞外产物（ECP）的抗血清对从条纹鳢Channa striata Bloch分离出的海分枝杆菌或分枝杆菌TB267进行调理。一种针对ECP的单克隆抗体也用作调理素。制备巨噬细胞悬液（1 ml，1×10⁷ 个细胞/ml⁻¹），与调理过的细菌（100 μl，2×10⁹ 个/ml⁻¹）混合，并在18℃下孵育0.5、1、2、4或6小时以使吞噬作用发生。通过电子显微镜对巨噬细胞吞噬分枝杆菌的情况进行定量评估。检查巨噬细胞吞噬体及其内容物，并确定完整和部分降解细菌的数量。用铁蛋白对致密颗粒（次级溶酶体）进行预标记能够识别吞噬体 - 溶酶体融合并确定其频率。发现对分枝杆菌进行调理可大大增强虹鳟巨噬细胞的吞噬和杀伤活性。
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