title: Characterization of the immunostimulatory properties of Leishmania infantum HSP70 by fusion to the Escherichia coli maltose-binding protein in normal and nu/nu BALB/c mice.
abstract: Leishmania infantum HSP70 has been described as an immunodominant antigen in both humans and dogs suffering from visceral leishmaniasis. In this study, we used L. infantum HSP70 fused to Escherichia coli maltose-binding protein (MBP), as the reporter protein, to analyze the influence of HSP70 on the immunogenicity of MBP in BALB/c mice. Plasmids were constructed to produce the three recombinant proteins used in this study, namely, MBP, L. infantum HSP70, and MBP-HSP70, which consists of MBP fused to the L. infantum HSP70 amino terminus. Immunization of BALB/c mice with the MBP-HSP70 fusion protein elicited humoral and cellular responses against MBP that were higher by an order of magnitude than those elicited by immunization with MBP alone or with a mixture of MBP and HSP70. Covalent linkage of MBP to HSP70 was essential for eliciting a strong anti-MBP immune response. Cytokine secretion and immunoglobulin G isotype analyses indicated that immunization with the MBP-HSP70 fusion protein preferentially induces a Th1 immune response. Immunization of athymic nu/nu mice with the MBP-HSP70 fusion protein unexpectedly gave rise to an anti-MBP humoral response showing features of a T-cell-dependent response. Thus, we present evidence that L. infantum HSP70 demonstrates an adjuvant effect in the immune response against a covalently linked reporter protein.
authors.affiliation: Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas-Universidad Autónoma de Madrid, Cantoblanco, Spain.
authors.forename: A I
authors.initials: AI
authors.lastname: Rico
authors.forename: G
authors.initials: G
authors.lastname: Del Real
authors.forename: M
authors.initials: M
authors.lastname: Soto
authors.forename: L
authors.initials: L
authors.lastname: Quijada
authors.forename: C
authors.initials: C
authors.lastname: Martinez-A
authors.forename: C
authors.initials: C
authors.lastname: Alonso
authors.forename: J M
authors.initials: JM
authors.lastname: Requena
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000032:analysis / Q000276:immunology; D000918:Antibody Specificity; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology*; D029968:Escherichia coli Proteins; D005260:Female; D006360:Heat-Shock Proteins / Q000235:genetics / Q000276:immunology*; D007111:Immunity, Cellular; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D007371:Interferon-gamma / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D007376:Interleukin-2 / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D015847:Interleukin-4 / Q000032:analysis / Q000276:immunology / Q000378:metabolism; D018314:Leishmania infantum / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D033902:Periplasmic Binding Proteins; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D018417:Th1 Cells / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D016056:Immunodominant Epitopes; D007074:Immunoglobulin G; D007376:Interleukin-2; C461619:MalE protein, E coli; D033902:Periplasmic Binding Proteins; D011993:Recombinant Fusion Proteins; D015847:Interleukin-4; D007371:Interferon-gamma