Terlikowski S, Sulkowski S, Sulkowska M, Nowak H F
Department of Gynecology and Septic Obstetrics, Medical University of Bialystok, Poland.
J Submicrosc Cytol Pathol. 1999 Apr;31(2):265-72.
The aim of the present study was the comparative analysis of morphological changes found in the lungs of Buffalo rats in the course of Morris hepatoma 5123 after i.t. treatment with recombinant human TNF-alpha (rhTNF-alpha) and its muteins. Modification of the native TNF-alpha molecule and synthesis of mutagenized analogues can prevent undesirable symptoms observed in the case of therapeutic administration of rhTNF-alpha. TNF-alpha has been shown to interact with two distinct membrane receptors (TNF-R): p55R and p75R. Mutagenized mutein V binds selectively with p55R. Mutein VI fails to recognize either TNF-R. The cytokines were applied in a dose of 10 micrograms protein in a cycle of 8 days. The control group consisted of tumor-bearing animals which were given PBS. Ultrastructural examinations were based on transmission electron microscope (TEM). Mutein VI-receiving animals showed enhanced changes of cytotoxic nature. Severe damage to endothelial cells (necrosis inclusive) was observed. Blood vascular lumen showed accumulation of neutrophils and monocytes. Features of enhanced activity of endothelial cells were noted. Focally, within pulmonary alveoli conglomerates of fibrin and fragments of damaged cells were found, with erythrocytes, neutrophils and macrophages in their vicinity. The epithelium of pulmonary alveoli showed signs of considerable damage, including necrosis. The lumen of pulmonary capillaries in rhTNF-alpha-treated animals showed a predominance of eosinophils and monocytic cells. Features of endothelial stimulation were observed, although without a tendency to form microthrombi. Much less pronounced changes both in the lung capillary bed and in the alveolar epithelial cells were noted in the mutein V-given animals. Our findings confirm the possibility of peripheral activation of cells involved in the cytokine-induced antitumor response. Mutein V with the smallest effect on the lung tissue rebuilding seems to be a rhTNF-alpha derivative which can delimit the undesirable symptoms in the course of antitumor therapy reduced to i.t. injections.
本研究的目的是对经腹腔注射重组人肿瘤坏死因子-α（rhTNF-α）及其突变体后，布法罗大鼠在莫里斯肝癌5123病程中肺部发现的形态学变化进行比较分析。天然TNF-α分子的修饰和诱变类似物的合成可以预防rhTNF-α治疗给药时出现的不良症状。TNF-α已被证明可与两种不同的膜受体（TNF-R）相互作用：p55R和p75R。诱变突变体V与p55R选择性结合。突变体VI不能识别任何一种TNF-R。细胞因子以10微克蛋白质的剂量在8天的周期内给药。对照组由给予磷酸盐缓冲盐水（PBS）的荷瘤动物组成。超微结构检查基于透射电子显微镜（TEM）。接受突变体VI的动物表现出细胞毒性性质的增强变化。观察到内皮细胞严重受损（包括坏死）。血管腔内可见中性粒细胞和单核细胞聚集。注意到内皮细胞活性增强的特征。在肺泡内局部发现了纤维蛋白团块和受损细胞碎片，其附近有红细胞、中性粒细胞和巨噬细胞。肺泡上皮显示出明显受损的迹象，包括坏死。rhTNF-α处理动物的肺毛细血管腔内以嗜酸性粒细胞和单核细胞为主。观察到内皮细胞刺激的特征，尽管没有形成微血栓的倾向。在给予突变体V的动物中，肺毛细血管床和肺泡上皮细胞的变化要少得多。我们的研究结果证实了参与细胞因子诱导的抗肿瘤反应的细胞发生外周激活的可能性。对肺组织重建影响最小的突变体V似乎是一种rhTNF-α衍生物，它可以在抗肿瘤治疗（简化为腹腔注射）过程中限制不良症状。
