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

C.A. Egan, K.M. Houston, M.J.C. Alcocer, A. Solovyova, R. Tate, G. Lochnit, I.B. McInnes, M.M. Harnett

Research output: Contribution to journalArticle

8 Citations (Scopus)

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.
LanguageEnglish
Pages263-274
Number of pages11
JournalParasitology
Volume132
Issue number2
DOIs
Publication statusPublished - Feb 2006

Fingerprint

Acanthocheilonema
Acanthocheilonema viteae
cross reaction
Phosphorylcholine
Parasites
Recombinant Proteins
recombinant proteins
parasites
Proteins
proteins
Nematoda
protein secondary structure
Secondary Protein Structure
Pichia pastoris
antibodies
Pichia
Antibodies
glycosylation
Allergy and Immunology
immunology

Keywords

  • antigenic cross-reactivity
  • ES-62
  • filarial nematode
  • Pichia pastoris
  • protein folding
  • recombinant protein expression

Cite this

Egan, C.A. ; Houston, K.M. ; Alcocer, M.J.C. ; Solovyova, A. ; Tate, R. ; Lochnit, G. ; McInnes, I.B. ; Harnett, M.M. / Lack of immunological cross-reactivity between parasite-derived and recombinant forms of es-62, a secreted protein of acanthocheilonema viteae. In: Parasitology. 2006 ; Vol. 132, No. 2. pp. 263-274.
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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.",
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Lack of immunological cross-reactivity between parasite-derived and recombinant forms of es-62, a secreted protein of acanthocheilonema viteae. / Egan, C.A.; Houston, K.M.; Alcocer, M.J.C.; Solovyova, A.; Tate, R.; Lochnit, G.; McInnes, I.B.; Harnett, M.M.

In: Parasitology, Vol. 132, No. 2, 02.2006, p. 263-274.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Lack of immunological cross-reactivity between parasite-derived and recombinant forms of es-62, a secreted protein of acanthocheilonema viteae

AU - Egan, C.A.

AU - Houston, K.M.

AU - Alcocer, M.J.C.

AU - Solovyova, A.

AU - Tate, R.

AU - Lochnit, G.

AU - McInnes, I.B.

AU - Harnett, M.M.

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N2 - 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.

AB - 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.

KW - antigenic cross-reactivity

KW - ES-62

KW - filarial nematode

KW - Pichia pastoris

KW - protein folding

KW - recombinant protein expression

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DO - 10.1017/S0031182005009005

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SP - 263

EP - 274

JO - Parasitology

T2 - Parasitology

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SN - 0031-1820

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ER -