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小鼠血吸虫病肉芽肿内血管活性肠肽的检测及其mRNA的定位

Detection of vasoactive intestinal peptide and localization of its mRNA within granulomas of murine schistosomiasis.

作者信息

Weinstock J V, Blum A M

机构信息

Department of Internal Medicine, University of Iowa, Iowa City 52242.

出版信息

Cell Immunol. 1990 Feb;125(2):291-300. doi: 10.1016/0008-8749(90)90085-6.

Abstract

Schistosomiasis mansoni is a parasitic disease resulting in the deposition of ova predominantly in the liver and intestines. These ova secrete antigens which induce host sensitization and evoke focal granulomas. The granulomas are intricate delayed-hypersensitivity reactions governed by numerous cellular and humoral interactions. They displace or destroy normal tissue. Vasoactive intestinal peptide (VIP) is one of several neuropeptides which exert a broad range of biologic actions that may include modulation of immune responses. In this study, VIP was sought within liver granulomas isolated from Schistosoma mansoni-infected, CBA/J mice. Granuloma extracts contained appreciable amounts of immunoreactive VIP as detected by radioimmunoassay. Immunoreactive VIP was shown, by each of two chromatographic methods, to elute as a single peak coinciding with that of synthetic VIP. In situ hybridization was performed with an oligonucleotide probe complementary to a portion of the nucleotide sequence encoding VIP on preproVIP mRNA (antisense probe). Radiolabeled VIP probe adhered exclusively to granuloma eosinophils and to eosinophils within a peritonitis induced in normal mice by proteose peptone. Hybridization of radiolabeled VIP probe in the presence of unlabeled probe substantially attenuated binding. A sense probe failed to bind. These data suggest that the granulomas contain authentic VIP and that eosinophils express the gene for this molecule.

摘要

曼氏血吸虫病是一种寄生虫病,主要导致虫卵沉积在肝脏和肠道。这些虫卵分泌抗原,诱导宿主致敏并引发局灶性肉芽肿。肉芽肿是由众多细胞和体液相互作用控制的复杂迟发型超敏反应。它们会取代或破坏正常组织。血管活性肠肽(VIP)是几种神经肽之一,具有广泛的生物学作用,可能包括调节免疫反应。在本研究中,在从感染曼氏血吸虫的CBA/J小鼠分离的肝脏肉芽肿中寻找VIP。通过放射免疫测定法检测到肉芽肿提取物中含有相当数量的免疫反应性VIP。通过两种色谱方法中的每一种都表明,免疫反应性VIP作为单个峰洗脱,与合成VIP的峰一致。用与前体VIP mRNA上编码VIP的核苷酸序列的一部分互补的寡核苷酸探针(反义探针)进行原位杂交。放射性标记的VIP探针仅附着于肉芽肿嗜酸性粒细胞以及在蛋白胨诱导的正常小鼠腹膜炎中的嗜酸性粒细胞。在未标记探针存在下,放射性标记的VIP探针的杂交大大减弱了结合。正义探针未能结合。这些数据表明肉芽肿含有真正的VIP,并且嗜酸性粒细胞表达该分子的基因。

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