Seternes T, Dalmo R A, Hoffman J, Bøgwald J, Zykova S, Smedsrød B
Department of Experimental Pathology, Institute of Medical Biology, University of Tromsø, N-9037 Tromsø, Norway.
J Exp Biol. 2001 Dec;204(Pt 23):4055-64. doi: 10.1242/jeb.204.23.4055.
The mechanism of elimination of blood-borne Vibrio salmonicida lipopolysaccharide (LPS) from Atlantic cod (Gadus morhua L.) was studied. The anatomical distribution of LPS was determined using both morphological and radiotracing methods. Immunohistochemistry performed on tissue specimens after injection of LPS disclosed that the endocardial endothelial cells (EECs) represented the cellular site of uptake in heart. Co-injection of trace amounts of [(125)I]LPS together with excess amounts of formaldehyde-treated albumin (FSA), a ligand for the scavenger receptor, significantly inhibited the accumulation of the radiotracer in heart only. Studies on purified monolayer cultures of atrial EECs showed that fluorescein-labelled LPS was taken up in structures reminiscent of endosomal/lysosomal vesicles. Incubation of cultures with [(125)I]LPS together with excess amounts of FSA, fucoidan and dextran sulphate, molecules known to compete for endocytosis via the scavenger receptor, reduced uptake of the probe by 80 %. Mannan, a ligand for the mannose receptor, did not compete for uptake. Kinetic studies on the uptake and degradation of [(125)I]LPS in cultured atrial endocardial cells revealed no degradation after 48 h of culture. In conclusion, we have shown that the EECs of cod remove V. salmonicida LPS from the circulation by scavenger-receptor-mediated endocytosis.
研究了大西洋鳕鱼(Gadus morhua L.)清除血源性病原体杀鲑气单胞菌脂多糖(LPS)的机制。使用形态学和放射性示踪方法确定了LPS的解剖分布。在注射LPS后对组织标本进行免疫组织化学分析,结果显示心内膜内皮细胞(EECs)是心脏中摄取LPS的细胞部位。将微量的[(125)I]LPS与过量的经甲醛处理的白蛋白(FSA,一种清道夫受体的配体)共同注射,仅显著抑制了放射性示踪剂在心脏中的积累。对心房EECs纯化单层培养物的研究表明,荧光素标记的LPS被摄取到类似于内体/溶酶体囊泡的结构中。将培养物与[(125)I]LPS以及过量的FSA、岩藻多糖和硫酸葡聚糖一起孵育,这些分子已知可通过清道夫受体竞争内吞作用,使探针的摄取减少了80%。甘露糖受体的配体甘露聚糖不竞争摄取。对培养的心房内膜细胞中[(125)I]LPS摄取和降解的动力学研究表明,培养48小时后没有降解。总之,我们已经表明,鳕鱼的EECs通过清道夫受体介导的内吞作用从循环中清除杀鲑气单胞菌LPS。