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犬感染性幼虫C型凝集素的系统发育分析及其与小鼠和人类免疫系统中C型凝集素家族的比较。

Phylogenetic Analysis of C Type Lectin from canis Infective Larvae and Comparison with the C Type Lectin Family in the Immune System of Mouse and Human.

作者信息

Etebar Fazeleh, Hosseini Seyed Hossein, Jalousian Fatemeh, Ranjbar Mohammad Mehdi

机构信息

Dept. of Parasitology, Faculty of Veterinary Medicine, University of Tehran, Tehran, Iran.

Dept. of Immunology, Razi Vaccine and Serum Research Institute, Karaj, Iran.

出版信息

Iran J Parasitol. 2018 Jan-Mar;13(1):49-57.

Abstract

BACKGROUND

C type lectin (CTL) family is a type of calcium-dependent proteins found in vertebrates and invertebrates. The objective of this study was to perform a comparative analysis and phylogenetic inferring for understanding the similarities and differences of carbohydrate recognition domain (CRD) domain of CTL and other nematodes, and similar C type lectin involved in the immune system of mouse and human as their host.

METHODS

The female was retrieved from the 2-6 months puppies (Department of Parasitology, Faculty of Veterinary Medicine, University of Tehran, 2015). To collect eggs, the worms were cultured for 5 d until they were embryonated. The hatching process was accelerated for collecting the stage 2 larvae, and the larvae were cultured for a week. A cDNA library was made from the total mRNA of infective larvae. The PCR amplification for C type lectin gene was performed and the amino acids were analyzed using the alignment method and the construction of phylogenetic tree.

RESULTS

The suspension sample maintained at 30 ºC for four weeks could embryonate 90%-100% of eggs. CTL gene was 657 bp in length and encoded a protein with 219 amino acids. The CTL of species of Strongylida order were closely placed in the tree, whereas the members of Ascaridida orders were located in a separate branch. High levels of similarity (36%-44%) and conservation of C type lectin from with mouse and human C type lectins. Its C type lectin showed a higher similarity with asialoglycoprotein receptor (ASGPR), macrophage lectin, dendritic cell-specific intercellular adhesion molecule 3-grabbing nonintegrin (DC-SIGN), MINCLE receptor of mouse and human.

CONCLUSION

Analysis of CRD domain of C type lectin protein could make a better understanding of their role in the interaction of nematode parasite with their hosts.

摘要

背景

C型凝集素(CTL)家族是在脊椎动物和无脊椎动物中发现的一类钙依赖性蛋白。本研究的目的是进行比较分析和系统发育推断,以了解CTL的碳水化合物识别结构域(CRD)与其他线虫的异同,以及作为宿主的小鼠和人类免疫系统中涉及的类似C型凝集素。

方法

从2 - 6个月大的幼犬(德黑兰大学兽医学院寄生虫学系,2015年)中获取雌性线虫。为了收集虫卵,将线虫培养5天直至胚胎化。加速孵化过程以收集二期幼虫,并将幼虫培养一周。从感染性幼虫的总mRNA构建cDNA文库。进行C型凝集素基因的PCR扩增,并使用比对方法和构建系统发育树来分析氨基酸。

结果

在30℃下保存4周的悬浮样本可使90% - 100%的虫卵胚胎化。CTL基因长度为657 bp,编码一个含219个氨基酸的蛋白质。圆线目物种的CTL在系统发育树中紧密排列,而蛔目成员位于一个单独的分支中。该线虫的C型凝集素与小鼠和人类的C型凝集素具有高度相似性(36% - 44%)和保守性。其C型凝集素与去唾液酸糖蛋白受体(ASGPR)、巨噬细胞凝集素、树突状细胞特异性细胞间粘附分子3结合非整合素(DC - SIGN)、小鼠和人类的MINCLE受体具有更高的相似性。

结论

对C型凝集素蛋白的CRD结构域进行分析可以更好地了解它们在寄生虫与宿主相互作用中的作用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/baa4/6019583/57e358863c81/IJPA-13-49-g001.jpg

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