title: Expression, purification, immunogenicity and protective efficacy of a recombinant nucleoside hydrolase from Leishmania donovani, a vaccine candidate for preventing cutaneous leishmaniasis.
abstract: The nucleoside hydrolase gene from Leishmania donovani was cloned and expressed in Escherichia coli as a full length 36-kDa protein (LdNH36). Following lysis and extraction, the protein was purified by anion exchange and gel filtration chromatography. The purified protein had a molecular mass of approximately 36-kDa and was confirmed to be >99% pure. Using a nucleoside hydrolase assay, the protein was found to exhibit a Km of 741 ± 246 μM. Protein integrity was confirmed by lithium dodecyl sulfate polyacrylamide gel electrophoresis (LDS-PAGE), mass spectrometry (MS), and enzymatic assay. Analysis of antibody levels from immunized mice indicated that LdNH36 alone or in a stable emulsion with the Toll-like receptor-4 ligand glucopyranosyl lipid adjuvant (GLA-SE) as immunostimulant induced high levels of antigen-specific IgG antibodies. The cellular immune response indicated a Th1 response in mice immunized with LdNH36, but only when formulated with GLA-SE. Mice immunized with the LdNH36 antigen in combination with the GLA-SE adjuvant and challenged with Leishmania mexicana showed significant reductions (>20 fold) in parasite burden, confirming the protective efficacy of this vaccine candidate.
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA.
authors: C Patrick
authors: CP
authors: McAtee
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA.
authors: Christopher A
authors: CA
authors: Seid
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA.
authors: Molly
authors: M
authors: Hammond
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA.
authors: Elissa
authors: E
authors: Hudspeth
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA.
authors: Brian P
authors: BP
authors: Keegan
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA.
authors: Zhuyun
authors: Z
authors: Liu
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA.
authors: Junfei
authors: J
authors: Wei
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA.
authors: Bin
authors: B
authors: Zhan
authors: Laboratory of Parasitology, Centro de Investigaciones Regionales "Dr. Hideyo Noguchi", Universidad Autónoma de Yucatán, Mérida, Yucatán, Mexico.
authors: Raul
authors: R
authors: Arjona-Sabido
authors: Laboratory of Parasitology, Centro de Investigaciones Regionales "Dr. Hideyo Noguchi", Universidad Autónoma de Yucatán, Mérida, Yucatán, Mexico.
authors: Vladimir
authors: V
authors: Cruz-Chan
authors: Laboratory of Parasitology, Centro de Investigaciones Regionales "Dr. Hideyo Noguchi", Universidad Autónoma de Yucatán, Mérida, Yucatán, Mexico.
authors: Eric
authors: E
authors: Dumonteil
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA; James A. Baker III Institute for Public Policy, Rice University, Houston, TX, USA; Scowcroft Institute of International Affairs, Bush School of Government and Public Service at Texas A&M University, College Station, TX, USA.
authors: Peter J
authors: PJ
authors: Hotez
authors: National School of Tropical Medicine, Baylor College of Medicine, Houston, TX, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX, USA. Electronic address: bottazzi@bcm.edu.
authors: Maria Elena
authors: ME
authors: Bottazzi
mesh_terms: D000818:Animals; D005260:Female; D000071497:Immunogenicity, Vaccine; D007893:Leishmania donovani / Q000201:enzymology / Q000235:genetics / Q000276:immunology; D054332:Leishmaniasis Vaccines / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000493:pharmacokinetics; D016773:Leishmaniasis, Cutaneous / Q000276:immunology / Q000517:prevention & control; D051379:Mice; D008807:Mice, Inbred BALB C; D009699:N-Glycosyl Hydrolases / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D015800:Protozoan Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D054332:Leishmaniasis Vaccines; D015800:Protozoan Proteins; D011994:Recombinant Proteins; D009699:N-Glycosyl Hydrolases
keywords: LdNH36; Leishmania donovani; NTDs; Neglected tropical diseases; Vaccines