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The analysis of VH and VL genes repertoires of Fab library built from peripheral B cells of human rabies virus vaccinated donors.

Identifieur interne : 000235 ( Main/Exploration ); précédent : 000234; suivant : 000236

The analysis of VH and VL genes repertoires of Fab library built from peripheral B cells of human rabies virus vaccinated donors.

Auteurs : Mehdi Houimel [Tunisie]

Source :

RBID : pubmed:24862931

Descripteurs français

English descriptors

Abstract

A human combinatorial Fab antibody library was generated from immune repertoire based on peripheral B cells of ten rabies virus vaccinated donors. The analysis of random Fab fragments from the unselected library presented some bias of V gene usage towards IGHV-genes and IGLV-gen families. The screening of the Fab library on rabies virus allowed specific human Fab antibody fragments characterized for their gene encoding sequences, binding and specificities to RV. Genetic analysis of selected Fabs indicated that the IGHV and IGLV differ from the germ-line sequence. At the level of nucleotide sequences, the IGHV and IGLV domains were found to share 74-92% and 90-96% homology with sequences encoded by the corresponding human germ-line genes respectively. IGHV domains are characterized most frequently by IGHV3 genes, and large proportions of the anti-RV heavy chain IGHV domains are obtained following a VDJ recombination process that uses IGHD3, IGHD2, IGHD1 and IGHD6 genes. IGHJ3 and IGHJ4 genes are predominantly used in RV-Fab. The IGLV domains are dominated by IGKV1, IGLV1 and IGLV3 genes. Numerous somatic hypermutations in the RV-specific IGHV are detected, but only limited amino acid replacement in most of the RV-specific IGLV particularly in those encoded by J proximal IGLV or IGKV genes are found. Furthermore, IGHV3-IGKV1, IGHV3-IGVL1, and IGHV3-IGLV3 germ-line family pairings are preferentially enriched after the screening on rabies virus.

DOI: 10.1016/j.humimm.2014.05.005
PubMed: 24862931


Affiliations:


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Le document en format XML

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<name sortKey="Houimel, Mehdi" sort="Houimel, Mehdi" uniqKey="Houimel M" first="Mehdi" last="Houimel">Mehdi Houimel</name>
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<nlm:affiliation>Laboratoire d'Epidémiologie et de Microbiologie Vétérinaire, Institut Pasteur de Tunis, Tunisia; Université Tunis El Manar, Tunis, Tunisia. Electronic address: mehdi.houimel@pasteur.rns.tn.</nlm:affiliation>
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<term>Amino Acid Sequence (MeSH)</term>
<term>Antibody Specificity (MeSH)</term>
<term>B-Lymphocytes (chemistry)</term>
<term>B-Lymphocytes (immunology)</term>
<term>Base Sequence (MeSH)</term>
<term>Gene Expression (MeSH)</term>
<term>Humans (MeSH)</term>
<term>Molecular Sequence Data (MeSH)</term>
<term>Peptide Library (MeSH)</term>
<term>Protein Binding (MeSH)</term>
<term>Rabies (immunology)</term>
<term>Rabies (prevention & control)</term>
<term>Rabies (virology)</term>
<term>Rabies Vaccines (administration & dosage)</term>
<term>Rabies virus (immunology)</term>
<term>Sequence Homology, Nucleic Acid (MeSH)</term>
<term>Single-Domain Antibodies (genetics)</term>
<term>Single-Domain Antibodies (immunology)</term>
<term>Tissue Donors (MeSH)</term>
<term>V(D)J Recombination (MeSH)</term>
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<term>Anticorps à domaine unique (génétique)</term>
<term>Anticorps à domaine unique (immunologie)</term>
<term>Banque de peptides (MeSH)</term>
<term>Donneurs de tissus (MeSH)</term>
<term>Données de séquences moléculaires (MeSH)</term>
<term>Expression des gènes (MeSH)</term>
<term>Humains (MeSH)</term>
<term>Liaison aux protéines (MeSH)</term>
<term>Lymphocytes B (composition chimique)</term>
<term>Lymphocytes B (immunologie)</term>
<term>Rage (maladie) (immunologie)</term>
<term>Rage (maladie) (prévention et contrôle)</term>
<term>Rage (maladie) (virologie)</term>
<term>Recombinaison V(D)J (MeSH)</term>
<term>Similitude de séquences d'acides nucléiques (MeSH)</term>
<term>Spécificité des anticorps (MeSH)</term>
<term>Séquence d'acides aminés (MeSH)</term>
<term>Séquence nucléotidique (MeSH)</term>
<term>Vaccins antirabiques (administration et posologie)</term>
<term>Virus de la rage (immunologie)</term>
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<keywords scheme="MESH" type="chemical" qualifier="administration & dosage" xml:lang="en">
<term>Rabies Vaccines</term>
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<keywords scheme="MESH" type="chemical" qualifier="genetics" xml:lang="en">
<term>Single-Domain Antibodies</term>
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<keywords scheme="MESH" type="chemical" qualifier="immunology" xml:lang="en">
<term>Single-Domain Antibodies</term>
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<keywords scheme="MESH" type="chemical" xml:lang="en">
<term>Peptide Library</term>
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<keywords scheme="MESH" qualifier="administration et posologie" xml:lang="fr">
<term>Vaccins antirabiques</term>
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<term>B-Lymphocytes</term>
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<term>Lymphocytes B</term>
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<keywords scheme="MESH" qualifier="génétique" xml:lang="fr">
<term>Anticorps à domaine unique</term>
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<term>Anticorps à domaine unique</term>
<term>Lymphocytes B</term>
<term>Rage (maladie)</term>
<term>Virus de la rage</term>
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<keywords scheme="MESH" qualifier="immunology" xml:lang="en">
<term>B-Lymphocytes</term>
<term>Rabies</term>
<term>Rabies virus</term>
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<term>Rabies</term>
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<term>Rage (maladie)</term>
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<term>Rabies</term>
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<term>Amino Acid Sequence</term>
<term>Antibody Specificity</term>
<term>Base Sequence</term>
<term>Gene Expression</term>
<term>Humans</term>
<term>Molecular Sequence Data</term>
<term>Protein Binding</term>
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<term>Tissue Donors</term>
<term>V(D)J Recombination</term>
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<term>Données de séquences moléculaires</term>
<term>Expression des gènes</term>
<term>Humains</term>
<term>Liaison aux protéines</term>
<term>Recombinaison V(D)J</term>
<term>Similitude de séquences d'acides nucléiques</term>
<term>Spécificité des anticorps</term>
<term>Séquence d'acides aminés</term>
<term>Séquence nucléotidique</term>
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<div type="abstract" xml:lang="en">A human combinatorial Fab antibody library was generated from immune repertoire based on peripheral B cells of ten rabies virus vaccinated donors. The analysis of random Fab fragments from the unselected library presented some bias of V gene usage towards IGHV-genes and IGLV-gen families. The screening of the Fab library on rabies virus allowed specific human Fab antibody fragments characterized for their gene encoding sequences, binding and specificities to RV. Genetic analysis of selected Fabs indicated that the IGHV and IGLV differ from the germ-line sequence. At the level of nucleotide sequences, the IGHV and IGLV domains were found to share 74-92% and 90-96% homology with sequences encoded by the corresponding human germ-line genes respectively. IGHV domains are characterized most frequently by IGHV3 genes, and large proportions of the anti-RV heavy chain IGHV domains are obtained following a VDJ recombination process that uses IGHD3, IGHD2, IGHD1 and IGHD6 genes. IGHJ3 and IGHJ4 genes are predominantly used in RV-Fab. The IGLV domains are dominated by IGKV1, IGLV1 and IGLV3 genes. Numerous somatic hypermutations in the RV-specific IGHV are detected, but only limited amino acid replacement in most of the RV-specific IGLV particularly in those encoded by J proximal IGLV or IGKV genes are found. Furthermore, IGHV3-IGKV1, IGHV3-IGVL1, and IGHV3-IGLV3 germ-line family pairings are preferentially enriched after the screening on rabies virus. </div>
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