Abstract
Background: Gnathostoma spinigerum is a harmful parasitic nematode that causes severe morbidity and mortality in humans and animals. Effective drugs and vaccines and reliable diagnostic methods are needed to prevent and control the associated diseases; however, the lack of genome, transcriptome, and proteome databases remains a major limitation. In this study, transcriptomic and secretomic analyses of advanced third-stage larvae of G. spinigerum (aL3Gs) were performed using next-generation sequencing, bioinformatics, and proteomics. Results: An analysis that incorporated transcriptome and bioinformatics data to predict excretory–secretory proteins (ESPs) classified 171 and 292 proteins into classical and non-classical secretory groups, respectively. Proteins with proteolytic (metalloprotease), cell signaling regulatory (i.e., kinases and phosphatase), and metabolic regulatory function (i.e., glucose and lipid metabolism) were significantly upregulated in the transcriptome and secretome. A two-dimensional (2D) immunomic analysis of aL3Gs-ESPs with G. spinigerum-infected human sera and related helminthiases suggested that the serine protease inhibitor (serpin) was a promising antigenic target for the further development of gnathostomiasis immunodiagnostic methods. Conclusions: The transcriptome and excretory–secretory proteome of aL3Gs can facilitate an understanding of the basic molecular biology of the parasite and identifying multiple associated factors, possibly promoting the discovery of novel drugs and vaccines. The 2D-immunomic analysis identified serpin, a protein secreted from aL3Gs, as an interesting candidate for immunodiagnosis that warrants immediate evaluation and validation.
Highlights
Gnathostomiasis is an important foodborne parasitic zoonosis caused by nematodes of the genus Gnathostoma Owen, 1836
Outbreaks of human gnathostomiasis have frequently occurred in Asia and Central America [1, 17], and sporadic cases reported in several countries have been associated with travelers who returned home after visiting endemic areas [31, 43]
The baseand raw read-qualities from next-generation sequencing (NGS) sequencing are shown in Supplementary Figure S3 and Supplementary Table S1, respectively
Summary
Gnathostomiasis is an important foodborne parasitic zoonosis caused by nematodes of the genus Gnathostoma Owen, 1836. There are six species including G. spinigerum, G. binucleatum, G. doloresi, G. hispidum, G. malaysiae, and G. nipponicum that have been reported to infect humans [10]. Outbreaks of human gnathostomiasis have frequently occurred in Asia and Central America [1, 17], and sporadic cases reported in several countries have been associated with travelers who returned home after visiting endemic areas [31, 43]. In Southeast Asia, G. spinigerum is considered a major gnathostomiasis-causing species in humans. G. spinigerum is classified in Phylum Nematoda, Order Rhabditida, Family Gnathostomatidae, and Genus Gnathostoma. Humans are infected by consumption of raw or improperly cooked fishes, amphibians, and eels that harbor infective (advanced third-stage) larvae. Swamp eels (Monopterus albus) are a common source of human gnathostomiasis [16]
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