Characterization of glycolytic enzymes--rAldolase and rEnolase of Leishmania donovani, identified as Th1 stimulatory proteins, for their immunogenicity and immunoprophylactic efficacies against experimental visceral leishmaniasis

Th1 immune responses play an important role in controlling Visceral Leishmaniasis (VL) hence, Leishmania proteins stimulating T-cell responses in host, are thought to be good vaccine targets. Search of such antigens eliciting cellular responses in Peripheral blood mononuclear cells (PBMCs) from cure...

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Bibliographic Details
Published in:PloS one 2014-01, Vol.9 (1), p.e86073-e86073
Main Authors: Gupta, Reema, Kumar, Vikash, Kushawaha, Pramod Kumar, Tripathi, Chandradev Pati, Joshi, Sumit, Sahasrabuddhe, Amogh Anant, Mitra, Kalyan, Sundar, Shyam, Siddiqi, Mohammad Imran, Dube, Anuradha
Format: Article
Language:English
Subjects:
Aldolase
Amino Acid Sequence
Animals
Antigens
Antigens, Protozoan - immunology
Biology
Committees
Computer applications
Cricetinae
Cytokines
Cytokines - biosynthesis
Dihydrofolate reductase
Disease Models, Animal
Enzymes
Epitopes
Epitopes, T-Lymphocyte - chemistry
Epitopes, T-Lymphocyte - immunology
Ethics
Fructose-Bisphosphate Aldolase - chemistry
Fructose-Bisphosphate Aldolase - genetics
Fructose-Bisphosphate Aldolase - metabolism
Gene expression
Glycolysis
Hamsters
Hypersensitivity, Delayed - immunology
Immune response
Immunogenicity
Immunoglobulin G - immunology
Immunology
Infections
Interferon
Interleukin 10
Interleukin 12
Interleukin 4
Laboratory animals
Leishmania
Leishmania donovani
Leishmania donovani - enzymology
Leishmania donovani - genetics
Leishmania donovani - immunology
Leishmania major
Leishmaniasis
Leishmaniasis Vaccines - immunology
Leishmaniasis, Visceral - immunology
Leishmaniasis, Visceral - prevention & control
Leukocytes (mononuclear)
Leukocytes, Mononuclear - immunology
Leukocytes, Mononuclear - metabolism
Lymphocyte Activation - immunology
Lymphocytes
Lymphocytes T
Male
Medicine
Models, Molecular
Mycobacterium bovis - immunology
Nitric oxide
Nitric Oxide - metabolism
Nitric-oxide synthase
Parasites
Parasitic diseases
Parasitology
Patients
Peripheral blood mononuclear cells
Phosphopyruvate hydratase
Phosphopyruvate Hydratase - chemistry
Phosphopyruvate Hydratase - genetics
Phosphopyruvate Hydratase - metabolism
Pneumonia
Protein Conformation
Proteins
Recombinant proteins
T cells
Th1 Cells - immunology
Th1 Cells - metabolism
Three dimensional models
Tropical diseases
Tumor necrosis factor-α
Vaccination
Vaccines
Vector-borne diseases
Visceral leishmaniasis
γ-Interferon
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Summary:Th1 immune responses play an important role in controlling Visceral Leishmaniasis (VL) hence, Leishmania proteins stimulating T-cell responses in host, are thought to be good vaccine targets. Search of such antigens eliciting cellular responses in Peripheral blood mononuclear cells (PBMCs) from cured/exposed/Leishmania patients and hamsters led to the identification of two enzymes of glycolytic pathway in the soluble lysate of a clinical isolate of Leishmania donovani--Enolase (LdEno) and aldolase (LdAld) as potential Th1 stimulatory proteins. The present study deals with the molecular and immunological characterizations of LdEno and LdAld. The successfully cloned and purified recombinant proteins displayed strong ability to proliferate lymphocytes of cured hamsters' along with significant nitric-oxide production and generation of Th1-type cytokines (IFN-γ and IL-12) from stimulated PBMCs of cured/endemic VL patients. Assessment of their prophylactic potentials revealed ∼ 90% decrease in parasitic burden in rLdEno vaccinated hamsters against Leishmania challenge, strongly supported by an increase in mRNA expression levels of iNOS, IFN-γ, TNF-α and IL-12 transcripts along with extreme down-regulation of TGF-β, IL-4 and IL-10. However, animals vaccinated with rLdAld showed comparatively lesser prophylactic efficacy (∼ 65%) with inferior immunological response. Further, with a possible implication in vaccine design against VL, identification of potential T-cell epitopes of both the proteins was done using computational approach. Additionally, in-silico 3-D modelling of the proteins was done in order to explore the possibility of exploiting them as potential drug targets. The comparative molecular and immunological characterizations strongly suggest rLdEno as potential vaccine candidate against VL and supports the notion of its being effective T-cell stimulatory protein.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0086073