FUNCTIONAL ANTIGENS OF TRICHINELLA SPIRALIS

护士细胞-寄生虫复合体是感染霍乱的独特特征 旋毛虫。它起源于一部分心肌细胞，并被诱导 由入侵的蠕虫在其作为细胞内生物的头14天内 寄生虫。护士细胞的特征是存在丰富的 细胞质内有光滑的膜，功能失调的线粒体，40 胞核增大，多见于细胞质的两端，外壁 主要由胶原蛋白组成的脱细胞胶囊。此外，《护士》 细胞被一种循环网包围，这种循环网也是在 感染。目前还不知道任何分子信号来自 一种寄生生物，它协调所有必要的变化，形成 这种不同寻常的宿主细胞。我们已经描述了一种分泌的蛋白质，这种蛋白质特定于 传染性L1幼虫，43 kDa(表观分子量)，位于ALL 护士细胞的细胞核增大。我们已经克隆并测序了该基因 编码这种蛋白，并使用针对融合蛋白的抗体 由最后95个氨基酸组成，在核质中鉴定出它，因此 弄清楚哪种蛋白质在护士细胞的细胞核中。我们现在 建议进一步表征该蛋白质与 护士细胞通过确定其差异表达模式利用 Northern分析和原位杂交。它的精确原子核 还将使用共聚焦显微镜和 免疫电子显微镜。我们将把这些发现与另外两个联系起来 哺育细胞特有的过程--即，在 包膜的形成和循环网的获取，通过 使用相似方法确定它们的差异表达模式 方法：研究方法。所有这些研究都将利用小鼠的同步感染 肌肉。计算机识别43 kDa分子的功能基序-- 辅助结构分析(例如Hlh、Cxxx盒和ras外显子1 磷酸结合位点)将通过结构域交换、删除 和突变。使用成肌细胞和成纤维细胞的体外系统将 被发掘其在协助调查职能部门方面的潜力 多个版本的43 kDa蛋白将被转染的基序 变成了他们。

The Nurse cell-parasite complex is a unique feature of infection with Trichinella spiralis. It arises from a portion of myocyte and is induced by the invading worm during its first 14 days of life as an intracellular parasite. The Nurse cell is characterized by the presence of abundant smooth membranes throughout its cytoplasm, dysfunctional mitochondria, 40+ enlarged nuclei, mostly found at either end of the cytoplasm, and an outer acellular capsule composed mainly of collagen. In addition, the Nurse cell is surrounded by a circulatory rete that is also induced during infection. Nothing is known as to any of the molecular signals from the parasite that coordinate all the changes necessary for the formation of this unusual host cell. We have described a secreted protein specific to the infectious L1 larva, 43 kDa (apparent mw), that is located in all enlarged nuclei of the Nurse cell. We have cloned and sequenced the cDNA encoding this protein and, using antibodies against a fusion protein consisting of the last 95 aa have identified it in the nucleoplasm, thus clarifying which protein is in the nuclei of the Nurse cell. We now propose to further characterize the association of this protein with the Nurse cell by determining its pattern of differential expression employing Northern analysis and in situ hybridization. Its precise nuclear localization will also be determined using co-focal microscopy and immunoelectron microscopy. We will relate these findings to two other Nurse cell-specific processes-namely, over-expression of collagen during capsule formation and the acquisition of the circulatory rete, by determining their patterns of differential expression using similar methods. All of these studies will employ synchronized infections in mouse muscle. Functional motifs of the 43 kDa molecule identified by computer- assisted structural analysis (eg. HLH, Cxxx box, and the ras exon 1 phosphate binding site) will be investigated by domain exchange, deletion and mutation. An in vitro system employing myoblasts and fibroblasts will be explored for their potential in assisting with investigating functional motifs in which several versions of the 43 kDa protein will be transfected into them.