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Mosquito cell line glycoproteins: an unsuitable model system for the Plasmodium ookinete-mosquito midgut interaction?

BACKGROUND: Mosquito midgut glycoproteins may act as key recognition sites for the invading malarial ookinete. Effective transmission blocking strategies require the identification of novel target molecules. We have partially characterised the surface glycoproteins of two cell lines from two mosquit...

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Published in:Parasites & vectors 2010-03, Vol.3 (1), p.22-22
Main Authors: Wilkins, Simon, Billingsley, Peter F
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description BACKGROUND: Mosquito midgut glycoproteins may act as key recognition sites for the invading malarial ookinete. Effective transmission blocking strategies require the identification of novel target molecules. We have partially characterised the surface glycoproteins of two cell lines from two mosquito species; Anopheles stephensi and Anopheles gambiae, and investigated the binding of Plasmodium berghei ookinetes to carbohydrate ligands on the cells. Cell line extracts were run on SDS-PAGE gels and carbohydrate moieties determined by blotting against a range of biotinylated lectins. In addition, specific glycosidases were used to cleave the oligosaccharides. RESULTS: An. stephensi 43 and An. gambiae 55 cell line glycoproteins expressed oligosaccharides containing oligomannose and hybrid oligosaccharides, with and without α1-6 core fucosylation; N-linked oligosaccharides with terminal Galβ1-3GalNAc or GalNAcβ1-3Gal; O-linked α/βGalNAc. An. stephensi 43 cell line glycoproteins also expressed N-linked Galβ1-4R and O-linked Galβ1-3GalNAc. Although P. berghei ookinetes bound to both mosquito cell lines, binding could not be inhibited by GlcNAc, GalNAc or Galactose. CONCLUSIONS: Anopheline cell lines displayed a limited range of oligosaccharides. Differences between the glycosylation patterns of the cell lines and mosquito midgut epithelial cells could be a factor why ookinetes did not bind in a carbohydrate inhibitable manner. Anopheline cell lines are not suitable as a potential model system for carbohydrate-mediated adhesion of Plasmodium ookinetes.
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Effective transmission blocking strategies require the identification of novel target molecules. We have partially characterised the surface glycoproteins of two cell lines from two mosquito species; Anopheles stephensi and Anopheles gambiae, and investigated the binding of Plasmodium berghei ookinetes to carbohydrate ligands on the cells. Cell line extracts were run on SDS-PAGE gels and carbohydrate moieties determined by blotting against a range of biotinylated lectins. In addition, specific glycosidases were used to cleave the oligosaccharides. RESULTS: An. stephensi 43 and An. gambiae 55 cell line glycoproteins expressed oligosaccharides containing oligomannose and hybrid oligosaccharides, with and without α1-6 core fucosylation; N-linked oligosaccharides with terminal Galβ1-3GalNAc or GalNAcβ1-3Gal; O-linked α/βGalNAc. An. stephensi 43 cell line glycoproteins also expressed N-linked Galβ1-4R and O-linked Galβ1-3GalNAc. Although P. berghei ookinetes bound to both mosquito cell lines, binding could not be inhibited by GlcNAc, GalNAc or Galactose. CONCLUSIONS: Anopheline cell lines displayed a limited range of oligosaccharides. Differences between the glycosylation patterns of the cell lines and mosquito midgut epithelial cells could be a factor why ookinetes did not bind in a carbohydrate inhibitable manner. Anopheline cell lines are not suitable as a potential model system for carbohydrate-mediated adhesion of Plasmodium ookinetes.</description><identifier>ISSN: 1756-3305</identifier><identifier>EISSN: 1756-3305</identifier><identifier>DOI: 10.1186/1756-3305-3-22</identifier><identifier>PMID: 20338056</identifier><language>eng</language><publisher>England: Springer-Verlag</publisher><subject>adhesion ; Anopheles gambiae ; Anopheles stephensi ; Carbohydrates ; Care and treatment ; epithelial cells ; Galactose ; Glycoproteins ; glycosidases ; glycosylation ; Health aspects ; hybrids ; Lectins ; Ligands ; Malaria ; midgut ; Mosquitoes ; oligosaccharides ; ookinetes ; Parasites ; Physiological aspects ; Plasmodium berghei ; polyacrylamide gel electrophoresis ; Standard deviation ; Vaccines</subject><ispartof>Parasites &amp; vectors, 2010-03, Vol.3 (1), p.22-22</ispartof><rights>COPYRIGHT 2010 BioMed Central Ltd.</rights><rights>2010 Wilkins and Billingsley; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.</rights><rights>Copyright ©2010 Wilkins and Billingsley; licensee BioMed Central Ltd. 2010 Wilkins and Billingsley; licensee BioMed Central Ltd.</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-b665t-853ff1072288cefe12cb3fb74cce1e6d476faa706e65a8bbe399dc757db08cc43</citedby></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC2861666/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.proquest.com/docview/902432711?pq-origsite=primo$$EHTML$$P50$$Gproquest$$Hfree_for_read</linktohtml><link.rule.ids>230,314,727,780,784,885,25753,27924,27925,37012,37013,44590,53791,53793</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20338056$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Wilkins, Simon</creatorcontrib><creatorcontrib>Billingsley, Peter F</creatorcontrib><title>Mosquito cell line glycoproteins: an unsuitable model system for the Plasmodium ookinete-mosquito midgut interaction?</title><title>Parasites &amp; vectors</title><addtitle>Parasit Vectors</addtitle><description>BACKGROUND: Mosquito midgut glycoproteins may act as key recognition sites for the invading malarial ookinete. Effective transmission blocking strategies require the identification of novel target molecules. We have partially characterised the surface glycoproteins of two cell lines from two mosquito species; Anopheles stephensi and Anopheles gambiae, and investigated the binding of Plasmodium berghei ookinetes to carbohydrate ligands on the cells. Cell line extracts were run on SDS-PAGE gels and carbohydrate moieties determined by blotting against a range of biotinylated lectins. In addition, specific glycosidases were used to cleave the oligosaccharides. RESULTS: An. stephensi 43 and An. gambiae 55 cell line glycoproteins expressed oligosaccharides containing oligomannose and hybrid oligosaccharides, with and without α1-6 core fucosylation; N-linked oligosaccharides with terminal Galβ1-3GalNAc or GalNAcβ1-3Gal; O-linked α/βGalNAc. An. stephensi 43 cell line glycoproteins also expressed N-linked Galβ1-4R and O-linked Galβ1-3GalNAc. 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vectors</jtitle><addtitle>Parasit Vectors</addtitle><date>2010-03-25</date><risdate>2010</risdate><volume>3</volume><issue>1</issue><spage>22</spage><epage>22</epage><pages>22-22</pages><issn>1756-3305</issn><eissn>1756-3305</eissn><abstract>BACKGROUND: Mosquito midgut glycoproteins may act as key recognition sites for the invading malarial ookinete. Effective transmission blocking strategies require the identification of novel target molecules. We have partially characterised the surface glycoproteins of two cell lines from two mosquito species; Anopheles stephensi and Anopheles gambiae, and investigated the binding of Plasmodium berghei ookinetes to carbohydrate ligands on the cells. Cell line extracts were run on SDS-PAGE gels and carbohydrate moieties determined by blotting against a range of biotinylated lectins. In addition, specific glycosidases were used to cleave the oligosaccharides. RESULTS: An. stephensi 43 and An. gambiae 55 cell line glycoproteins expressed oligosaccharides containing oligomannose and hybrid oligosaccharides, with and without α1-6 core fucosylation; N-linked oligosaccharides with terminal Galβ1-3GalNAc or GalNAcβ1-3Gal; O-linked α/βGalNAc. An. stephensi 43 cell line glycoproteins also expressed N-linked Galβ1-4R and O-linked Galβ1-3GalNAc. Although P. berghei ookinetes bound to both mosquito cell lines, binding could not be inhibited by GlcNAc, GalNAc or Galactose. CONCLUSIONS: Anopheline cell lines displayed a limited range of oligosaccharides. Differences between the glycosylation patterns of the cell lines and mosquito midgut epithelial cells could be a factor why ookinetes did not bind in a carbohydrate inhibitable manner. Anopheline cell lines are not suitable as a potential model system for carbohydrate-mediated adhesion of Plasmodium ookinetes.</abstract><cop>England</cop><pub>Springer-Verlag</pub><pmid>20338056</pmid><doi>10.1186/1756-3305-3-22</doi><oa>free_for_read</oa></addata></record>
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subjects adhesion
Anopheles gambiae
Anopheles stephensi
Carbohydrates
Care and treatment
epithelial cells
Galactose
Glycoproteins
glycosidases
glycosylation
Health aspects
hybrids
Lectins
Ligands
Malaria
midgut
Mosquitoes
oligosaccharides
ookinetes
Parasites
Physiological aspects
Plasmodium berghei
polyacrylamide gel electrophoresis
Standard deviation
Vaccines
title Mosquito cell line glycoproteins: an unsuitable model system for the Plasmodium ookinete-mosquito midgut interaction?
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