Molecular characterization, expression profile, and preliminary evaluation of diagnostic potential of CD63 in Schistosoma japonicum

Parasitol Res. 2018 Nov;117(11):3625-3631. doi: 10.1007/s00436-018-6063-8. Epub 2018 Sep 3.

Abstract

Schistosomes are the causative agents of human schistosomiasis, which is endemic in tropical and subtropical zones. CD63 is a member of the tetraspanin protein family widely expressed among eukaryotes. Previously, we identified a CD63 homolog from extracellular vesicles isolated from Schistosoma japonicum. In this study, we characterized this CD63 homolog using a molecular approach and evaluated the potential of its recombinant protein for the diagnosis of schistosomiasis. A sequence alignment indicated that S. japonicum CD63 (SjCD63) has sequence identities of 76 and 28% with S. mansoni and human CD63, respectively. A phylogenetic analysis displayed that S. japonicum CD63 is related to S. mansoni and Opisthorchis viverrini CD63. The cDNA of SjCD63 was 740 bp long with an expected protein size of 23.58 kDa. A RT-qPCR analysis revealed significantly higher expression of SjCD63 mRNA in adult worms on days 21, 28, and 35 than in 7-day schistosomula, cercariae, and eggs. In addition, recombinant SjCD63 protein detected by ELISA revealed significantly higher optical density values compared to that of the negative control in both S. japonicum-infected mouse and rabbit sera, providing preliminary evidence for its diagnostic potential. Overall, these results provide insight into the molecular properties of SjCD63, its expression profiles, and its preliminary diagnostic potential.

Keywords: Diagnosis; Expression profile; Recombinant protein; Schistosoma japonicum; Tetraspanins.

MeSH terms

  • Amino Acid Sequence / genetics
  • Animals
  • Cloning, Molecular
  • DNA, Complementary / genetics
  • Enzyme-Linked Immunosorbent Assay
  • Female
  • Humans
  • Mice
  • Phylogeny
  • RNA, Messenger / genetics
  • Rabbits
  • Real-Time Polymerase Chain Reaction
  • Recombinant Proteins / genetics*
  • Schistosoma japonicum / genetics*
  • Schistosoma japonicum / metabolism
  • Schistosomiasis japonica / diagnosis*
  • Schistosomiasis japonica / parasitology
  • Schistosomiasis japonica / veterinary*
  • Sequence Alignment
  • Tetraspanin 30 / genetics*

Substances

  • DNA, Complementary
  • RNA, Messenger
  • Recombinant Proteins
  • Tetraspanin 30