Lack of immunological cross-reactivity between parasite-derived and recombinant forms of es-62, a secreted protein of acanthocheilonema viteae

Egan, C.A. and Houston, K.M. and Alcocer, M.J.C. and Solovyova, A. and Tate, R. and Lochnit, G. and McInnes, I.B. and Harnett, M.M. (2006) Lack of immunological cross-reactivity between parasite-derived and recombinant forms of es-62, a secreted protein of acanthocheilonema viteae. Parasitology, 132 (2). pp. 263-274. ISSN 0031-1820

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Abstract

The longevity of filarial nematodes is dependent on secreted immunomodulatory products. Previous investigation of one such product, ES-62, has suggested a critical role for post-translationally attached phosphorylcholine (PC) moieties. In order to further investigate this, ES-62 lacking PC was produced, using the Pichia pastoris recombinant gene expression system. Unlike parasite-derived ES-62, which is tetrameric the recombinant material was found to consist of a mixture of apparently stable tetramers, dimers and monomers. Nevertheless, the recombinant protein was considered to be an adequate PC-free ES-62 as it was recognized by existing antisera against the parasite-derived protein. However, subsequent to this, recognition of parasite-derived ES-62 by antibodies produced against the recombinant protein was found to be absent. In an attempt to explain this, recombinant ES-62 was subjected to structural analysis and was found to (i) contain 3 changes in amino acid composition; (ii) demonstrate significant alterations in glycosylation; (iii) show major differences in protein secondary structure. The effects of these alterations in relation to the observed change in immunogenicity were investigated and are discussed. The data presented clearly show that recognition by existing antibodies is insufficient proof that recombinant proteins can be used to mimic parasite-derived material in studies on nematode immunology and vaccination.

Creators(s): Egan, C.A., Houston, K.M., Alcocer, M.J.C., Solovyova, A., Tate, R., Lochnit, G., McInnes, I.B. and Harnett, M.M.;
Item type:Article
ID code:21607
Notes:Strathprints' policy is to record up to 8 authors per publication, plus any additional authors based at the University of Strathclyde. More authors may be listed on the official publication than appear in the Strathprints' record.
Keywords:antigenic cross-reactivity, ES-62, filarial nematode, Pichia pastoris, protein folding, recombinant protein expression, Pharmacy and materia medica, Infectious Diseases, Animal Science and Zoology, Parasitology
Subjects:Medicine > Pharmacy and materia medica
Department:Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences
Faculty of Science > Strathclyde Institute of Pharmacy and Biomedical Sciences > Centre for Biophotonics
Depositing user: Strathprints Administrator
Date deposited:01 Oct 2010 19:30
Last modified:23 May 2020 00:23
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URI:https://strathprints.strath.ac.uk/id/eprint/21607
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