Heterologous expression of three antigenic proteins from Angiostrongylus cantonensis: ES-7, Lec-5, and 14-3-3 in mammalian cells

Publication date: Available online 16 March 2018 Source:Molecular and Biochemical Parasitology Author(s): Bianca B. Cognato, Sukwan Handali, Alessandra L. Morassutti, Alexandre Januário da Silva, Carlos Graeff-Teixeira Angiostrongylus cantonensis is a parasitic nematode and the main causative agent of human cerebral eosinophilic meningoencephalitis (EoM). A definitive diagnosis of EoM usually requires serologic or molecular analysis of the patient’s clinical sample. Currently, a 31 kDa antigen is used in immunological tests for this purpose, however as a crude antigen preparation it may present cross-reactivity with other helminthic infections, especially echinococcosis. Heterologous expression studies using prokaryotic systems failed on producing antigenic proteins. The aim of this study was to express and purify three recombinant glycoproteins representing A. cantonensis antigens: ES-7, Lec-5, and 14-3-3, in Chinese hamster ovary (CHO) cells and ES-7 in human embryonic kidney (HEK) cells to develop a source of specific antigens to be used in the diagnosis of angiostrongyliasis. The potential diagnostic value of these three proteins was subsequently characterized in one- and two-dimensional electrophoresis and Western blot to dot blot analyses, with Angiostrongylus-positive sera, normal human sera (NHS), and a pool of Echinococcus-positive sera (included as a specificity control) used for detection. In addition, recognition of these three proteins following treat...
Source: Molecular and Biochemical Parasitology - Category: Parasitology Source Type: research