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Holzhausen, J.* ; Haake, C.* ; Schicht, S.* ; Hinse, P.* ; Jordan, D.* ; Kremmer, E. ; Strube, C.*

Biological function of Dictyocaulus viviparus asparaginyl peptidase legumain-1 and its suitability as a vaccine target.

Parasitology 145, 378-392 (2018)
Postprint DOI PMC
Open Access Green
The present study characterized the biological function of the asparaginyl peptidase legumain-1 (LEG-1) of the bovine lungworm Dictyocaulus viviparus and its suitability as a recombinant vaccine against dictyocaulosis. Quantitative real-time PCR and immunoblot analysis revealed LEG-1 to be almost exclusively transcribed and expressed in parasitic lungworm stages. Immunohistochemistry localized the enzyme in the parasite's gut, which was confirmed by immunoblots detecting LEG-1 in the gut as well as male testes. LEG-1 was recombinantly (rLEG-1) expressed in the yeast Pichia pastoris and subsequently analysed in activity assays for its enzyme functions and substrate specificity. For sufficient functionality, rLEG-1 needed trans-activation through D. viviparus cathepsin L-2, indicating a novel mechanism of legumain activation. After trans-activation, rLEG-1 worked best at pH 5.5 and 35-39 degrees C and cleaved a legumain-specific artificial substrate as well as the natural substrates bovine collagen types I and II. In a clinical vaccination trial, rLEG-1 did not protect against challenge infection. Results of in vitro characterization, transcription pattern and localization enhance the presumption that LEG-1 participates in digestion processes of D. viviparus. Since rLEG-1 needs trans-activation through a cathepsin, it is probably involved in an enzyme cascade and therefore remains interesting as a candidate in a multi-component vaccine.
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Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Korrespondenzautor
Schlagwörter Legumain ; Bovine Lungworm ; Nematode ; Enzyme Activity ; Immunohistochemistry ; Vaccination ; Immunization ; Hidden Antigen; Haemonchus-contortus; Schistosoma-mansoni; Gastrointestinal Nematode; Hemoglobin Digestion; Endopeptidase Sm32; Bovine; Calves; Parasites; Genes; Identification
ISSN (print) / ISBN 0031-1820
e-ISSN 1469-8161
Zeitschrift Parasitology
Quellenangaben Band: 145, Heft: 3, Seiten: 378-392 Artikelnummer: , Supplement: ,
Verlag Cambridge Univ. Press
Verlagsort London [u.a.]
Nichtpatentliteratur Publikationen
Institut(e) CF Monoclonal Antibodies (CF-MAB)