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Toxicity, membrane binding and uptake of the Sclerotinia sclerotiorum agglutinin (SSA) in different insect cell lines

Authors
  • Shen, Ying1, 2
  • De Schutter, Kristof1, 2
  • Walski, Tomasz1, 2
  • Van Damme, Els J. M.2
  • Smagghe, Guy1
  • 1 Ghent University, Laboratory of Agrozoology, Department of Crop Protection, Faculty of Bioscience Engineering, Coupure links 653, Ghent, 9000, Belgium , Ghent (Belgium)
  • 2 Ghent University, Laboratory of Biochemistry and Glycobiology, Department of Molecular Biotechnology, Faculty of Bioscience Engineering, Coupure links 653, Ghent, 9000, Belgium , Ghent (Belgium)
Type
Published Article
Journal
In Vitro Cellular & Developmental Biology - Animal
Publisher
Springer-Verlag
Publication Date
Jul 11, 2017
Volume
53
Issue
8
Pages
691–698
Identifiers
DOI: 10.1007/s11626-017-0176-8
Source
Springer Nature
Keywords
License
Yellow

Abstract

The fungal lectin purified from Sclerotinia sclerotiorum, further referred to as Sclerotinia sclerotiorum agglutinin or SSA, possesses insecticidal activity against important pest insects such as pea aphids (Acyrthosiphon pisum). This paper aims at a better understanding of its activity at cellular level. Therefore, different insect cell lines were treated with SSA. These cell lines were derived from different tissues and represent the three major orders of insects important in agriculture: CF-203 (midgut Choristoneura fumiferana, Lepidoptera), GUTAW1 (midgut, Helicoverpa zea, Lepidoptera), High5 cells (ovary, Trichoplusia ni, Lepidoptera), Sf9 (ovary cells from Spodoptera frugiperda, Lepidoptera), S2 (hemocyte, Drosophila melanogaster, Diptera), and TcA (whole body, Tribolium castaneum, Coleoptera). Although the sensitivity to SSA differs between the cell lines, SSA clearly showed toxicity in all six cell lines with median effect concentrations (EC50) ranging between 9 and 42 μg/ml. An in-depth analysis of the mechanism of uptake in the cells revealed superior amounts of FITC-SSA at the membrane of CF-203 cells compared to Sf9 cells, while a similar small amount of SSA was internalized in both cell lines. Pre-incubation with the clathrin-mediated endocytosis inhibitor phenylarsine oxide inhibited the internalization of SSA into the CF-203 and Sf9 cells with a respective reduction of 6- and 1.7-fold. The data are discussed in relation to the importance of cellular uptake mechanism for SSA binding and cytotoxicity.

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