A novel Thioredoxin-related protein 14 from Fasciola gigantica has an immunodiagnostic potential for fasciolosis

Narin Changklungmoa; Pornanan Kueakhai; Kant Sangpairoj; Supawadee Osotprasit; Athit Chaiwichien; Tepparit Samrit; Prasert Sobhon; Kulathida Chaithirayanon

doi:10.1016/j.actatropica.2020.105471

A novel Thioredoxin-related protein 14 from Fasciola gigantica has an immunodiagnostic potential for fasciolosis

Acta Trop. 2020 Jul:207:105471. doi: 10.1016/j.actatropica.2020.105471. Epub 2020 Apr 14.

Authors

Narin Changklungmoa¹, Pornanan Kueakhai¹, Kant Sangpairoj², Supawadee Osotprasit¹, Athit Chaiwichien¹, Tepparit Samrit¹, Prasert Sobhon³, Kulathida Chaithirayanon⁴

Affiliations

¹ Faculty of Allied Health Sciences, Burapha University, Long-Hard Bangsaen Road, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand.; Research unit of vaccine and diagnosis of parasitic diseases, Burapha University, Long-Hard Bangsaen Road, Saen Sook Sub-district, Mueang District, Chonburi 20131, Thailand.
² Division of Anatomy, Department of Preclinical Science, Faculty of Medicine, Thammasat University, Pathumthani 12120, Thailand; Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok 10400, Thailand.
³ Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok 10400, Thailand.
⁴ Department of Anatomy, Faculty of Science, Mahidol University, Rama VI Rd, Bangkok 10400, Thailand.. Electronic address: kulathida.cha@mahidol.ac.th.

PMID: 32302691
DOI: 10.1016/j.actatropica.2020.105471

Abstract

In the definitive host, a trematode parasite can survive and evade the damage by reactive oxygen species that are generated from its metabolism and the host immune cells. Several anti-oxidant proteins are found in Fasciola spp. which play essential roles in cellular redox balance. One of them is thioredoxin-related protein 14 (TRP14) that has a highly conserved WCPDC motif and serves as a disulfide reductase-like thioredoxin (Trx). In the present study, a cDNA encoding TRP14 from F. gigantica (FgTRP14) was selected and cloned by immunoscreening with a rabbit infected serum. Phylogenetic analysis was performed by MEGA X program showed that FgTRP14 was most highly related to the Fasciola hepatica. Immunoblotting analysis of the polyclonal antibody rabbit serum against recombinant FgTRP14 (rFgTRP14) revealed that the molecular weight of natural FgTRP14 was at 14 kDa from metacercariae, NEJ, 4-week old juvenile and adult stage. The native FgTRP14 was expressed in caecal epithelial cells and preferentially localized on the cells' surface lamellae of adult stage. By sandwich ELISA assay, the circulating FgTRP14 could be recognized in sera of experimentally F. gigantica metacercariae infection in mice. The native FgTRP14 in the excretory-secretory (ES) and whole body (WB) of adult F. gigantica were detected at the concentrations 6.3 ng/ml, and 45 ng/ml, respectively. Therefore, it could be considered for immunodiagnostic candidate for fasciolosis.

Keywords: Fasciola gigantica; antioxidant activity; immunodiagnosis; sandwich ELISA; thioredoxin-related protein.

MeSH terms

Animals
Enzyme-Linked Immunosorbent Assay
Fasciola / immunology*
Fascioliasis / diagnosis*
Immunologic Tests
Male
Mice
Mice, Inbred ICR
Rabbits
Thioredoxins / immunology*

Substances

Thioredoxins