Generation and application of a monoclonal antibody against the 18-kDa oncosphere antigen of Taenia pisiformis

Exp Parasitol. 2021 May:224:108096. doi: 10.1016/j.exppara.2021.108096. Epub 2021 Mar 17.

Abstract

Taenia pisiformis is a parasite that causes cysticercosis pisiformis, which has acquired economic relevance because of its effects on animal welfare and production. A useful assay for the detection of T. pisiformis is needed for the prevention of cysticercosis pisiformis and control of the parasite. The 18-kDa oncosphere antigen is expressed in the oncosphere of several cysticerci in species of the genus Taenia, including T. pisiformis. This protein plays an important role in tissue invasion and has extensive applications in diagnosis. In this study, the T. pisiformis 18-kDa oncosphere antigen (TPO18) was expressed in soluble form and successfully purified for use in the production of monoclonal antibodies (MAbs) against TPO18. Twenty hybridomas were obtained using ELISA, and the subcloning process identified three positive hybridoma cell lines, which were designated as 4E8, 5G5, and 7E8. MAb 7E8 exhibited the highest titer and had an IgG2b heavy chain and a kappa light chain. Western blot analysis demonstrated that MAb 7E8 reacted with GST-TPO18. Immunohistochemistry showed that TPO18 was widely distributed in the drape and wall of uteri in adults of T. pisiformis adults and in the fibrous layer of the sucker and cyst cavity of T. pisiformis cysticerci. This research will provide a foundation for the development of diagnostic tools and will contribute to a better understanding of the functions of TPO18.

Keywords: 18-kDa oncosphere antigen; Cysticerci; Immunohistochemistry; Monoclonal antibody; Taenia pisiformis.

MeSH terms

  • Animals
  • Antibodies, Monoclonal / immunology
  • Antibodies, Monoclonal / metabolism*
  • Antigens, Helminth / immunology*
  • Antigens, Helminth / isolation & purification
  • Blotting, Western
  • Cloning, Molecular
  • Cysticercus / immunology
  • Dogs
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme-Linked Immunosorbent Assay
  • Hybridomas
  • Immunohistochemistry
  • Mice
  • Mice, Inbred BALB C
  • Rabbits
  • Taenia / immunology*

Substances

  • Antibodies, Monoclonal
  • Antigens, Helminth