title: Proteomic analysis identification of antigenic proteins in Gnathostoma spinigerum larvae.
abstract: Gnathostoma spinigerum is the causative agent of human gnathostomiasis. The advanced third stage larva (AL3) of this nematode can migrate into the subcutaneous tissues, including vital organs, often producing severe pathological effects. This study performed immuno-proteomic analysis of antigenic spots, derived from G. spinigerum advanced third stage larva (GSAL3) and recognized by human gnathostomiasis sera, using two-dimensional (2-DE) gel electrophoresis based-liquid chromatography/tandem mass spectrometry (LC/MS-MS), and followed by the aid of a database search. The crude GSAL3 extract was fractionated using IPG strips (pH 3-11NL) and followed by SDS-PAGE in the second dimension. Each gel was stained with colloidal Coomassie blue or was electro-transferred onto a nitrocellulose membrane and probed with gnathostomiasis human sera by immunoblotting. Individual Coomassie-stained protein spots corresponding to the antigenic spots recognized by immunoblotting were excised and processed using LC/MS-MS. Of the 93 antigenic spots excised, 87 were identified by LC/MS-MS. Twenty-seven protein types were found, the most abundant being Ascaris suum37. Six spots showed good quality spectra, but could not be identified. This appears to be the first attempt to characterize antigenic proteins from GSAL3 using a proteomic approach. Immuno-proteomics shows promise to assist the search for candidate proteins for diagnosis and vaccine/drug design and may provide better understand of the host-parasite relationship in human gnathostomiasis.
authors: Department of Medical Technology, School of Allied Health Sciences and Public Health, Walailak University, Nakhon Si Thammarat 80161, Thailand; Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Penchom
authors: P
authors: Janwan
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Pewpan M
authors: PM
authors: Intapan
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Porntip
authors: P
authors: Laummaunwai
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Rutchanee
authors: R
authors: Rodpai
authors: Department of Biochemistry, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Chaisiri
authors: C
authors: Wongkham
authors: Khon Kaen University Research Instrument Center, Research Affairs, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand.
authors: Tonkla
authors: T
authors: Insawang
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand; Faculty of Medicine, Mahasarakham University, Maha Sarakham 44000, Thailand.
authors: Tongjit
authors: T
authors: Thanchomnang
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand; Faculty of Medicine, Mahasarakham University, Maha Sarakham 44000, Thailand.
authors: Oranuch
authors: O
authors: Sanpool
authors: Research and Diagnostic Center for Emerging Infectious Diseases, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand; Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand. Electronic address: wanch_ma@kku.ac.th.
authors: Wanchai
authors: W
authors: Maleewong
mesh_terms: D000818:Animals; D000947:Antigens, Helminth / Q000032:analysis*; D006039:Gnathostoma / Q000276:immunology* / Q000502:physiology; D058429:Gnathostomiasis / Q000276:immunology* / Q000469:parasitology; D006790:Host-Parasite Interactions; D006801:Humans; D007106:Immune Sera / Q000276:immunology; D007814:Larva / Q000276:immunology / Q000502:physiology; D040901:Proteomics; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D000947:Antigens, Helminth; D007106:Immune Sera
keywords: Antigenic protein spots; Gnathostoma spinigerum; Proteomic analysis