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Cloning and characterization of a novel immunogenic protein 3 (NIP3) from Brugia malayi by immuno screening of a phage-display cDNA expression library

Identifieur interne : 008020 ( Main/Exploration ); précédent : 008019; suivant : 008021

Cloning and characterization of a novel immunogenic protein 3 (NIP3) from Brugia malayi by immuno screening of a phage-display cDNA expression library

Auteurs : Munirathinam Gnanasekar ; Balumuri Padmavathi ; Kalyanasundaram Ramaswamy

Source :

Parasitology research [ 0932-0113 ] ; 2005.

RBID : PMC:1307516

Abstract

Iterative screening of a phage display cDNA expression library of the third-stage larvae (L3) of Brugia malayi with sera from putatively immune individuals (endemic normal, EN) identified a novel clone with insert showing significant homology to Onchocerca volvulus novel immunogenic protein-3 (Ov-NIP3) gene and Caenorhabditis elegans NIP3-like protein and hence the gene was designated Brugia malayi NIP3-like protein (BmNIP3). EST database analysis showed that ESTs from several gastrointestinal nematodes such as Ancylostoma caninum, Teladorsagia circumcincta, Haemonchus contortus and Strongyloides stercoralis has BmNIP3 homologues, but the gene has not been described from these parasites. Sequence analyses showed that BmNIP3 has three potential mucin-type O-glycosylation sites and several serine/threonine phosphorylation sites. As expected, BmNIP3 protein isolated from the parasite was serine/threonine phosphorylated. Further analyses showed that BmNIP3 is differentially transcribed, with highest level of expression present in the larval (L3 and L4) stages. Mice immunized with rBmNIP3 developed strong antibody responses predominantly of the IgG1 and IgG2a subtype. A similar analyses of the sera samples from EN individuals showed that they also carry high levels of IgG1 and IgG2 antibodies against BmNIP3, whereas, chronically infected patients carry largely IgG3 antibodies and MF individuals carry high levels of IgG1 antibodies against BmNIP3. This study thus describes a novel protein from B. malayi that appears to be highly immunogenic in both humans and mice.

Url:

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1307516

DOI: 10.1007/s00436-005-1375-x
PubMed: 15952042
PubMed Central: 1307516

Affiliations:

Le document en format XML

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 by immuno screening of a phage-display cDNA expression library</title>
<author><name sortKey="Gnanasekar, Munirathinam" sort="Gnanasekar, Munirathinam" uniqKey="Gnanasekar M" first="Munirathinam" last="Gnanasekar">Munirathinam Gnanasekar</name>
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<author><name sortKey="Padmavathi, Balumuri" sort="Padmavathi, Balumuri" uniqKey="Padmavathi B" first="Balumuri" last="Padmavathi">Balumuri Padmavathi</name>
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<author><name sortKey="Ramaswamy, Kalyanasundaram" sort="Ramaswamy, Kalyanasundaram" uniqKey="Ramaswamy K" first="Kalyanasundaram" last="Ramaswamy">Kalyanasundaram Ramaswamy</name>
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<front><div type="abstract" xml:lang="en"><p id="P1">Iterative screening of a phage display cDNA expression library of the third-stage larvae (L3) of <italic>Brugia malayi</italic>
 with sera from putatively immune individuals (endemic normal, EN) identified a novel clone with insert showing significant homology to <italic>Onchocerca volvulus</italic>
 novel immunogenic protein-3 (Ov-NIP3) gene and <italic>Caenorhabditis elegans</italic>
 NIP3-like protein and hence the gene was designated <italic>Brugia malayi</italic>
 NIP3-like protein (BmNIP3). EST database analysis showed that ESTs from several gastrointestinal nematodes such as <italic>Ancylostoma caninum</italic>
, <italic>Teladorsagia circumcincta</italic>
, <italic>Haemonchus contortus</italic>
 and <italic>Strongyloides stercoralis</italic>
 has BmNIP3 homologues, but the gene has not been described from these parasites. Sequence analyses showed that BmNIP3 has three potential mucin-type <italic>O</italic>
-glycosylation sites and several serine/threonine phosphorylation sites. As expected, BmNIP3 protein isolated from the parasite was serine/threonine phosphorylated. Further analyses showed that BmNIP3 is differentially transcribed, with highest level of expression present in the larval (L3 and L4) stages. Mice immunized with rBmNIP3 developed strong antibody responses predominantly of the IgG1 and IgG2a subtype. A similar analyses of the sera samples from EN individuals showed that they also carry high levels of IgG1 and IgG2 antibodies against BmNIP3, whereas, chronically infected patients carry largely IgG3 antibodies and MF individuals carry high levels of IgG1 antibodies against BmNIP3. This study thus describes a novel protein from <italic>B. malayi</italic>
 that appears to be highly immunogenic in both humans and mice.</p>
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<name sortKey="Ramaswamy, Kalyanasundaram" sort="Ramaswamy, Kalyanasundaram" uniqKey="Ramaswamy K" first="Kalyanasundaram" last="Ramaswamy">Kalyanasundaram Ramaswamy</name>
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Cloning and characterization of a novel immunogenic protein 3 (NIP3) from Brugia malayi by immuno screening of a phage-display cDNA expression library

Cloning and characterization of a novel immunogenic protein 3 (NIP3) from Brugia malayi by immuno screening of a phage-display cDNA expression library

Source :

Abstract

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