Evaluation of the transcription level of the protein disulfide isomerase in different stages from Ancylostoma caninum with a real-time PCR assay

Abstract

The protein disulfide isomerase (PDI) is a ubiquitous protein, which contributes in building disulfide bridges. In the work presented here, the expression of the PDI in different stages of the canine hookworm Ancylostoma caninum was investigated. Third-stage larvae (L3), adults, as well as serum-stimulated and hypobiotic L3 were used. For quantification of the PDI gene transcription, a real-time PCR was used establishing a hybridization probe (TaqMantrade mark probes) for detection of PDI copy numbers in different populations. 18S ribosomal ribonucleic acid (rRNA) was used as a housekeeping gene for normalization. The results show differences in the transcription level of the investigated A. caninum populations: The serum-stimulated larvae representing the switch to parasitism showed the highest PDI expression. The hypobiotic larvae representing a resting stage showed the lowest expression level. Male adults showed an elevated expression compared to female adult worms. The L3 expression level was just below the serum-stimulated population. This work confirms the upregulated gene expression of PDI during host penetration and invasion.

Evaluation of reference genes for quantitative real-time PCR to investigate protein disulfide isomerase transcription pattern in the bovine lungworm Dictyocaulus viviparus

“…As expected, there is no difference in the mixed gender L5 transcription level of the HannoverDv2000 and the Intervet vaccine strain, respectively. The imbalance between males and females continues in adult parasites and is in accordance with observations in A. caninum (Epe et al, 2007), perhaps indicating an additional male-specific role of the pdi-2. The rather low pdi-2 transcription in eggs might be the consequence of the egg content, representing a compilation of ova, blastomeres, embryos, and L1, of which developing L1 need collagen for cuticule synthesis only.…”

Molecular characterization and immunolocalization of a protein disulfide isomerase from Angiostrongylus cantonensis

“…Our results showed that active enzyme can be obtained through heterologous expression and AcPDI was tested to behave oxidation-reduction activities as in other parasites (Mouray et al 2007;Hong and Soong 2008). These enzymatic activities were essential for the correct folding of nascent proteins and validate PDI as potential strategic targets for parasite diseases control (Epe et al 2007;Khalaf et al 2011). The co-expression of PDI and ancrod in Pichia pastoris can increase the production of ancrod, and the enzymatic activities of rAncrod were similar to that of the native protein (Zhang et al 2011).…”

Screening trematodes for novel intervention targets: a proteomic and immunological comparison of Schistosoma haematobium, Schistosoma bovis and Echinostoma caproni

“…PDI has been identified in the E/S products of adult E. caproni, E. friedi and F. hepatica worms, suggesting that it may be important in host-parasite interactions (Salazar-Calderon et al 2003;Bernal et al 2006;Sotillo et al 2010). Moreover, PDI is immunogenic in human S. haematobium infections (Mutapi et al 2005) and experimental F. hepatica (Moxon et al 2010) and it has been shown to be immunologically protective against the hookworm, Ancylostoma (Epe et al 2007). Differences in PDI kDa Fig.…”

“…Moreover, PDI is immunogenic in human S. haematobium infections (Mutapi et al 2005) and experimental F. hepatica (Moxon et al . 2010) and it has been shown to be immunologically protective against the hookworm, Ancylostoma (Epe et al 2007). Differences in PDI molecular weight between S. haematobium and E. caproni could be due to post-translational modifications, akin to the PDI glycosylation reported in Trypansoma brucei where it is related to parasite defence (Rubotham et al 2005).…”