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

Published online by Cambridge University Press:  11 October 2005

C. A. EGAN ,
K. M. HOUSTON ,
M. J. C. ALCOCER ,
A. SOLOVYOVA ,
R. TATE ,
G. LOCHNIT ,
I. B. McINNES ,
M. M. HARNETT ,
R. GEYER  and
O. BYRON
...Show all authors
C. A. EGAN
Affiliation:
Department of Immunology, University of Strathclyde, Glasgow G4 0NR, UK
K. M. HOUSTON
Affiliation:
Department of Immunology, University of Strathclyde, Glasgow G4 0NR, UK
M. J. C. ALCOCER
Affiliation:
Division of Nutritional Sciences, University of Nottingham, Loughborough LE12 5RD, UK
A. SOLOVYOVA
Affiliation:
Division of Infection and Immunity, Institute of Biomedical and Life Sciences, University of Glasgow G12 8QQ, UK
R. TATE
Affiliation:
Department of Physiology and Pharmacology, University of Strathclyde, Glasgow G4 0NR, UK
G. LOCHNIT
Affiliation:
Institute of Biochemistry, University of Giessen, Giessen D35392, Germany
I. B. McINNES
Affiliation:
Division of Immunology, Infection and Inflammation, University of Glasgow, Glasgow G11 6NT, UK
M. M. HARNETT
Affiliation:
Division of Immunology, Infection and Inflammation, University of Glasgow, Glasgow G11 6NT, UK
R. GEYER
Affiliation:
Institute of Biochemistry, University of Giessen, Giessen D35392, Germany
O. BYRON
Affiliation:
Division of Infection and Immunity, Institute of Biomedical and Life Sciences, University of Glasgow G12 8QQ, UK
W. HARNETT
Affiliation:
Department of Immunology, University of Strathclyde, Glasgow G4 0NR, UK
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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.

Keywords

antigenic cross-reactivityES-62filarial nematodePichia pastorisprotein foldingrecombinant protein expression
Type
Research Article
Information
Parasitology , Volume 132 , Issue 2 , February 2006 , pp. 263 - 274
Copyright
2005 Cambridge University Press

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