Protein profile analysis from advanced third-stage larvae (AdvL3) and adult worms of Gnathostoma binucleatum (Nematoda: Spirurida)

Abstract

Proteins from crude extracts of advanced third-stage larvae and adult Gnathostoma binucleatum nematode worms showed protein profiles in SDS-PAGE analysis similar to Echinococcus granulosus, Trichinella spiralis, Dipylidium caninum, Ancylostoma caninum, Ascaris lumbricoides and Toxocara canis. The immunoblot analysis of the human serum infected or suspected to be infected with G. binucleatum using the total larvae extract recognized the 40, 60, 80 and 115 kDa proteins and using the total adult worm extract recognized only the 80 and 115 kDa proteins. However, the 115 kDa protein showed cross-reactions with A. caninum, A. lumbricoides, T. canis and D. caninum with human serum positive to gnathostomosis, while the 40 kDa protein was only recognized with the G. binucleatum total larvae extract. The results obtained suggest that the use of antigens from the advanced third-stage larvae of the parasite were best recognized for immunodiagnosis of gnathostomosis.

Identification of Immunodominant Peptides from Gnathostoma binucleatum

“…11 This antigen was not found in L3A from G. binucleatum obtained from experimentally infected mice. 17 However, in the present study, using larvae recovered from infected fish, we detected an antigenic band of 24 kDa (Figure 2).…”

“…These results are consistent with reported SDS-PAGE patterns of antigens of G. binucleatum larvae and adults in which the protein molecular weight range was 20-208 kDa. 17 We did not find bands with high molecular weights. One possible reason for such differences could be the source of the larvae.…”

“…Previous IET analysis using a total G. binucleatum extract 17 showed only four antigenic proteins (40, 60, 80, and 115 kDa), and a total adult worm extract showed that only the 80-kDa and the 115-kDa two proteins were recognized, but the 115 kDa protein was cross-reactive with A. lumbricoides, Toxocara canis, and Ancylostoma caninum. In the same study, it was also reported that the 40-kDa protein did not cross-react with serum samples of patients with other parasitic diseases.…”

“…One possible reason for such differences could be the source of the larvae. In the present study, we used larvae from intermediate host fishes, but larvae used by Caballero-Garcia and others 17 were from experimentally infected mice. Continuous growth of G. spinigerum L3A in mice has been reported.…”

“…Two of the peptides were metalloproteases (80 and 208 kDa). 16,17 The purpose of this study was to identify and characterize G. binucleatum immunodominant antigens derived from somatic antigens (AgS) and excretory-secretory antigens (AgES) that could be useful for immunodiagnosis of human gnathostomiasis cases.…”