Approaches for the identification of potential excreted/secreted proteins of Leishmania major parasites.
Identifieur interne : 000025 ( Main/Exploration ); précédent : 000024; suivant : 000026Approaches for the identification of potential excreted/secreted proteins of Leishmania major parasites.
Auteurs : M. Chenik [Tunisie] ; S. Lakhal ; N. Ben Khalef ; L. Zribi ; H. Louzir ; K. DellagiSource :
- Parasitology [ 0031-1820 ] ; 2006.
Descripteurs français
- KwdFr :
- Amorces ADN (composition chimique), Analyse de profil d'expression de gènes (méthodes), Animaux (MeSH), Anticorps antiprotozoaires (immunologie), Antigènes de protozoaire (analyse), Banque de gènes (MeSH), Cellules cultivées (MeSH), Données de séquences moléculaires (MeSH), Humains (MeSH), Lapins (MeSH), Leishmania major (physiologie), Marquage isotopique (MeSH), Protein Disulfide-Isomerases (génétique), Protein Disulfide-Isomerases (métabolisme), Protéines de protozoaire (classification), Protéines de protozoaire (isolement et purification), Protéines de protozoaire (métabolisme), RT-PCR (méthodes), Radio-isotopes (MeSH), Technique de Western (méthodes), Techniques immunologiques (MeSH).
- MESH :
- analyse : Antigènes de protozoaire.
- classification : Protéines de protozoaire.
- composition chimique : Amorces ADN.
- génétique : Protein Disulfide-Isomerases.
- immunologie : Anticorps antiprotozoaires.
- isolement et purification : Protéines de protozoaire.
- métabolisme : Protein Disulfide-Isomerases, Protéines de protozoaire.
- méthodes : Analyse de profil d'expression de gènes, RT-PCR, Technique de Western.
- physiologie : Leishmania major.
- Animaux, Banque de gènes, Cellules cultivées, Données de séquences moléculaires, Humains, Lapins, Marquage isotopique, Radio-isotopes, Techniques immunologiques.
English descriptors
- KwdEn :
- Animals (MeSH), Antibodies, Protozoan (immunology), Antigens, Protozoan (analysis), Blotting, Western (methods), Cells, Cultured (MeSH), DNA Primers (chemistry), Gene Expression Profiling (methods), Gene Library (MeSH), Humans (MeSH), Immunologic Techniques (MeSH), Isotope Labeling (MeSH), Leishmania major (physiology), Molecular Sequence Data (MeSH), Protein Disulfide-Isomerases (genetics), Protein Disulfide-Isomerases (metabolism), Protozoan Proteins (classification), Protozoan Proteins (isolation & purification), Protozoan Proteins (metabolism), Rabbits (MeSH), Radioisotopes (MeSH), Reverse Transcriptase Polymerase Chain Reaction (methods).
- MESH :
- chemical , analysis : Antigens, Protozoan.
- chemical , chemistry : DNA Primers.
- chemical , classification : Protozoan Proteins.
- chemical , genetics : Protein Disulfide-Isomerases.
- chemical , immunology : Antibodies, Protozoan.
- chemical , isolation & purification : Protozoan Proteins.
- chemical , metabolism : Protein Disulfide-Isomerases, Protozoan Proteins.
- methods : Blotting, Western, Gene Expression Profiling, Reverse Transcriptase Polymerase Chain Reaction.
- physiology : Leishmania major.
- Animals, Cells, Cultured, Gene Library, Humans, Immunologic Techniques, Isotope Labeling, Molecular Sequence Data, Rabbits, Radioisotopes.
Abstract
Leishmania parasites are able to survive in host macrophages despite the harsh phagolysosomal vacuoles conditions. This could reflect, in part, their capacity to secrete proteins that may play an essential role in the establishment of infection and serve as targets for cellular immune responses. To characterize Leishmania major proteins excreted/secreted early after promastigote entry into the host macrophage, we have generated antibodies against culture supernatants of stationary-phase promastigotes collected 6 h after incubation in conditions that partially reproduce those prevailing in the parasitophorous vacuole. The screening of an L. major cDNA library with these antibodies led us to isolate 33 different cDNA clones that we report here. Sequence analysis revealed that the corresponding proteins could be classified in 3 groups: 9 proteins have been previously described as excreted/secreted in Leishmania and/or other species; 11 correspond to known proteins already characterized in Leishmania and/or other species although it is unknown whether they are excreted/secreted and 13 code for unknown proteins. Interestingly, the latter are transcribed as shown by RT-PCR and some of them are stage regulated. The L. major excreted/secreted proteins may constitute putative virulence factors, vaccine candidates and/or new drug targets.
DOI: 10.1017/S0031182005009546
PubMed: 16388694
Affiliations:
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Chenik</name><affiliation wicri:level="1"><nlm:affiliation>WHO Collaborating Center for Research and Training in Leishmaniasis, Laboratoire d'Immunopathologie, Vaccinologie et Génétique Moléculaire, Institut Pasteur de Tunis, 13, Place Pasteur 1002 Tunis-Belvédére, Tunisia. mehdi.chenik@pasteur.rns.tn</nlm:affiliation><country xml:lang="fr">Tunisie</country><wicri:regionArea>WHO Collaborating Center for Research and Training in Leishmaniasis, Laboratoire d'Immunopathologie, Vaccinologie et Génétique Moléculaire, Institut Pasteur de Tunis, 13, Place Pasteur 1002 Tunis-Belvédére</wicri:regionArea><wicri:noRegion>Place Pasteur 1002 Tunis-Belvédére</wicri:noRegion></affiliation></author><author><name sortKey="Lakhal, S" sort="Lakhal, S" uniqKey="Lakhal S" first="S" last="Lakhal">S. Lakhal</name></author><author><name sortKey="Ben Khalef, N" sort="Ben Khalef, N" uniqKey="Ben Khalef N" first="N" last="Ben Khalef">N. Ben Khalef</name></author><author><name sortKey="Zribi, L" sort="Zribi, L" uniqKey="Zribi L" first="L" last="Zribi">L. Zribi</name></author><author><name sortKey="Louzir, H" sort="Louzir, H" uniqKey="Louzir H" first="H" last="Louzir">H. Louzir</name></author><author><name sortKey="Dellagi, K" sort="Dellagi, K" uniqKey="Dellagi K" first="K" last="Dellagi">K. Dellagi</name></author></titleStmt><publicationStmt><idno type="wicri:source">PubMed</idno><date when="2006">2006</date><idno type="RBID">pubmed:16388694</idno><idno type="pmid">16388694</idno><idno type="doi">10.1017/S0031182005009546</idno><idno type="wicri:Area/PubMed/Corpus">000800</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Corpus" wicri:corpus="PubMed">000800</idno><idno type="wicri:Area/PubMed/Curation">000796</idno><idno type="wicri:explorRef" wicri:stream="PubMed" wicri:step="Curation">000796</idno><idno type="wicri:Area/PubMed/Checkpoint">000796</idno><idno type="wicri:explorRef" wicri:stream="Checkpoint" wicri:step="PubMed">000796</idno><idno type="wicri:Area/Main/Merge">000025</idno><idno type="wicri:Area/Main/Curation">000025</idno><idno type="wicri:Area/Main/Exploration">000025</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en">Approaches for the identification of potential excreted/secreted proteins of Leishmania major parasites.</title><author><name sortKey="Chenik, M" sort="Chenik, M" uniqKey="Chenik M" first="M" last="Chenik">M. Chenik</name><affiliation wicri:level="1"><nlm:affiliation>WHO Collaborating Center for Research and Training in Leishmaniasis, Laboratoire d'Immunopathologie, Vaccinologie et Génétique Moléculaire, Institut Pasteur de Tunis, 13, Place Pasteur 1002 Tunis-Belvédére, Tunisia. mehdi.chenik@pasteur.rns.tn</nlm:affiliation><country xml:lang="fr">Tunisie</country><wicri:regionArea>WHO Collaborating Center for Research and Training in Leishmaniasis, Laboratoire d'Immunopathologie, Vaccinologie et Génétique Moléculaire, Institut Pasteur de Tunis, 13, Place Pasteur 1002 Tunis-Belvédére</wicri:regionArea><wicri:noRegion>Place Pasteur 1002 Tunis-Belvédére</wicri:noRegion></affiliation></author><author><name sortKey="Lakhal, S" sort="Lakhal, S" uniqKey="Lakhal S" first="S" last="Lakhal">S. Lakhal</name></author><author><name sortKey="Ben Khalef, N" sort="Ben Khalef, N" uniqKey="Ben Khalef N" first="N" last="Ben Khalef">N. Ben Khalef</name></author><author><name sortKey="Zribi, L" sort="Zribi, L" uniqKey="Zribi L" first="L" last="Zribi">L. Zribi</name></author><author><name sortKey="Louzir, H" sort="Louzir, H" uniqKey="Louzir H" first="H" last="Louzir">H. Louzir</name></author><author><name sortKey="Dellagi, K" sort="Dellagi, K" uniqKey="Dellagi K" first="K" last="Dellagi">K. Dellagi</name></author></analytic><series><title level="j">Parasitology</title><idno type="ISSN">0031-1820</idno><imprint><date when="2006" type="published">2006</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass><keywords scheme="KwdEn" xml:lang="en"><term>Animals (MeSH)</term><term>Antibodies, Protozoan (immunology)</term><term>Antigens, Protozoan (analysis)</term><term>Blotting, Western (methods)</term><term>Cells, Cultured (MeSH)</term><term>DNA Primers (chemistry)</term><term>Gene Expression Profiling (methods)</term><term>Gene Library (MeSH)</term><term>Humans (MeSH)</term><term>Immunologic Techniques (MeSH)</term><term>Isotope Labeling (MeSH)</term><term>Leishmania major (physiology)</term><term>Molecular Sequence Data (MeSH)</term><term>Protein Disulfide-Isomerases (genetics)</term><term>Protein Disulfide-Isomerases (metabolism)</term><term>Protozoan Proteins (classification)</term><term>Protozoan Proteins (isolation & purification)</term><term>Protozoan Proteins (metabolism)</term><term>Rabbits (MeSH)</term><term>Radioisotopes (MeSH)</term><term>Reverse Transcriptase Polymerase Chain Reaction (methods)</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>Amorces ADN (composition chimique)</term><term>Analyse de profil d'expression de gènes (méthodes)</term><term>Animaux (MeSH)</term><term>Anticorps antiprotozoaires (immunologie)</term><term>Antigènes de protozoaire (analyse)</term><term>Banque de gènes (MeSH)</term><term>Cellules cultivées (MeSH)</term><term>Données de séquences moléculaires (MeSH)</term><term>Humains (MeSH)</term><term>Lapins (MeSH)</term><term>Leishmania major (physiologie)</term><term>Marquage isotopique (MeSH)</term><term>Protein Disulfide-Isomerases (génétique)</term><term>Protein Disulfide-Isomerases (métabolisme)</term><term>Protéines de protozoaire (classification)</term><term>Protéines de protozoaire (isolement et purification)</term><term>Protéines de protozoaire (métabolisme)</term><term>RT-PCR (méthodes)</term><term>Radio-isotopes (MeSH)</term><term>Technique de Western (méthodes)</term><term>Techniques immunologiques (MeSH)</term></keywords><keywords scheme="MESH" type="chemical" qualifier="analysis" xml:lang="en"><term>Antigens, Protozoan</term></keywords><keywords scheme="MESH" type="chemical" qualifier="chemistry" xml:lang="en"><term>DNA Primers</term></keywords><keywords scheme="MESH" type="chemical" qualifier="classification" xml:lang="en"><term>Protozoan Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en"><term>Protein Disulfide-Isomerases</term></keywords><keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en"><term>Antibodies, Protozoan</term></keywords><keywords scheme="MESH" type="chemical" qualifier="isolation & purification" xml:lang="en"><term>Protozoan Proteins</term></keywords><keywords scheme="MESH" type="chemical" qualifier="metabolism" xml:lang="en"><term>Protein Disulfide-Isomerases</term><term>Protozoan Proteins</term></keywords><keywords scheme="MESH" qualifier="analyse" xml:lang="fr"><term>Antigènes de protozoaire</term></keywords><keywords scheme="MESH" qualifier="classification" xml:lang="fr"><term>Protéines de protozoaire</term></keywords><keywords scheme="MESH" qualifier="composition chimique" xml:lang="fr"><term>Amorces ADN</term></keywords><keywords scheme="MESH" qualifier="génétique" xml:lang="fr"><term>Protein Disulfide-Isomerases</term></keywords><keywords scheme="MESH" qualifier="immunologie" xml:lang="fr"><term>Anticorps antiprotozoaires</term></keywords><keywords scheme="MESH" qualifier="isolement et purification" xml:lang="fr"><term>Protéines de protozoaire</term></keywords><keywords scheme="MESH" qualifier="methods" xml:lang="en"><term>Blotting, Western</term><term>Gene Expression Profiling</term><term>Reverse Transcriptase Polymerase Chain Reaction</term></keywords><keywords scheme="MESH" qualifier="métabolisme" xml:lang="fr"><term>Protein Disulfide-Isomerases</term><term>Protéines de protozoaire</term></keywords><keywords scheme="MESH" qualifier="méthodes" xml:lang="fr"><term>Analyse de profil d'expression de gènes</term><term>RT-PCR</term><term>Technique de Western</term></keywords><keywords scheme="MESH" qualifier="physiologie" xml:lang="fr"><term>Leishmania major</term></keywords><keywords scheme="MESH" qualifier="physiology" xml:lang="en"><term>Leishmania major</term></keywords><keywords scheme="MESH" xml:lang="en"><term>Animals</term><term>Cells, Cultured</term><term>Gene Library</term><term>Humans</term><term>Immunologic Techniques</term><term>Isotope Labeling</term><term>Molecular Sequence Data</term><term>Rabbits</term><term>Radioisotopes</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>Animaux</term><term>Banque de gènes</term><term>Cellules cultivées</term><term>Données de séquences moléculaires</term><term>Humains</term><term>Lapins</term><term>Marquage isotopique</term><term>Radio-isotopes</term><term>Techniques immunologiques</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">Leishmania parasites are able to survive in host macrophages despite the harsh phagolysosomal vacuoles conditions. This could reflect, in part, their capacity to secrete proteins that may play an essential role in the establishment of infection and serve as targets for cellular immune responses. To characterize Leishmania major proteins excreted/secreted early after promastigote entry into the host macrophage, we have generated antibodies against culture supernatants of stationary-phase promastigotes collected 6 h after incubation in conditions that partially reproduce those prevailing in the parasitophorous vacuole. The screening of an L. major cDNA library with these antibodies led us to isolate 33 different cDNA clones that we report here. Sequence analysis revealed that the corresponding proteins could be classified in 3 groups: 9 proteins have been previously described as excreted/secreted in Leishmania and/or other species; 11 correspond to known proteins already characterized in Leishmania and/or other species although it is unknown whether they are excreted/secreted and 13 code for unknown proteins. Interestingly, the latter are transcribed as shown by RT-PCR and some of them are stage regulated. The L. major excreted/secreted proteins may constitute putative virulence factors, vaccine candidates and/or new drug targets.</div></front></TEI><affiliations><list><country><li>Tunisie</li></country></list><tree><noCountry><name sortKey="Ben Khalef, N" sort="Ben Khalef, N" uniqKey="Ben Khalef N" first="N" last="Ben Khalef">N. Ben Khalef</name><name sortKey="Dellagi, K" sort="Dellagi, K" uniqKey="Dellagi K" first="K" last="Dellagi">K. Dellagi</name><name sortKey="Lakhal, S" sort="Lakhal, S" uniqKey="Lakhal S" first="S" last="Lakhal">S. Lakhal</name><name sortKey="Louzir, H" sort="Louzir, H" uniqKey="Louzir H" first="H" last="Louzir">H. Louzir</name><name sortKey="Zribi, L" sort="Zribi, L" uniqKey="Zribi L" first="L" last="Zribi">L. Zribi</name></noCountry><country name="Tunisie"><noRegion><name sortKey="Chenik, M" sort="Chenik, M" uniqKey="Chenik M" first="M" last="Chenik">M. Chenik</name></noRegion></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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