Cloning, characterization and DNA immunization of an Onchocerca volvulus glyceraldehyde-3-phosphate dehydrogenase (Ov-GAPDH)

Klaus D. Erttmann, Andre Kleensang, Erik Schneider, Sven Hammerschmidt, Dietrich W. Büttner, Michaela Gallin

Research output: Contribution to journalArticle

Abstract

In the search for Onchocerca volvulus antigens possibly involved in protection against human onchocerciasis, partial amino acid sequence analysis of one of the O. volvulus antigens of the serologically identified proteins showed a close relationship to the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein family. Subsequent adult worm cDNA library screening and cloning produced a clone of 1650 bp. An open reading frame spans over 1020 bp encoding for a protein of 340 amino acids with an apparent molecular weight of 38 000. Comparison of the complete amino acid sequence identified this protein as a member of the GAPDH protein family. The recombinantly expressed protein shows GAPDH enzymatic activity as well as plasminogen-binding capacity. DNA sequence analysis of the corresponding gene revealed the presence of two introns. Using immunohistology Ov-GAPDH was observed in microfilariae, infective larvae, and adult male and female worms. Most striking was the labelling of the musculature of the body wall. Labelling was also observed in the pseudocoeloma cavity and in a subset of cell nuclei, suggesting additional, non-glycolytic functions of the Ov-GAPDH. Gene gun immunization with the DNA-construct in cattle led to specific humoral immune responses. Thus, the protective potential of the DNA-construct of Ov-GAPDH can be evaluated in vaccination trials using animal models such as the cattle/Onchocerca ochengi model.

Original languageEnglish (US)
Pages (from-to)85-94
Number of pages10
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1741
Issue number1-2
DOIs
Publication statusPublished - Jun 30 2005
Externally publishedYes

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Keywords

  • DNA immunization
  • Filaria
  • Glyceraldehyde-3-phosphate dehydrogenase
  • Onchocerca volvulus
  • Plasminogen-binding
  • Protective antigen

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine
  • Biophysics

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