Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.

BACKGROUND: The development of a vaccine conferring long-lasting immunity remains a challenge against visceral leishmaniasis (VL). Immunoproteomic characterization of Leishmania donovani proteins led to the identification of a novel protein NAD+-dependent Silent Information regulatory-2 (SIR2 family...

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Published in:PLOS Neglected Tropical Diseases
Main Authors: Rajendra K Baharia, Rati Tandon, Tanuj Sharma, Manish K Suthar, Sanchita Das, Mohammad Imran Siddiqi, Jitendra Kumar Saxena, Shaym Sundar, Anuradha Dube
Format: Article in Journal/Newspaper
Language:English
Published: Public Library of Science (PLoS) 2015
Subjects:
Online Access:https://doi.org/10.1371/journal.pntd.0003557
https://doaj.org/article/f632526349984a9391a36ce6a5687ed6
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spelling ftdoajarticles:oai:doaj.org/article:f632526349984a9391a36ce6a5687ed6 2023-05-15T15:15:00+02:00 Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis. Rajendra K Baharia Rati Tandon Tanuj Sharma Manish K Suthar Sanchita Das Mohammad Imran Siddiqi Jitendra Kumar Saxena Shaym Sundar Anuradha Dube 2015-03-01T00:00:00Z https://doi.org/10.1371/journal.pntd.0003557 https://doaj.org/article/f632526349984a9391a36ce6a5687ed6 EN eng Public Library of Science (PLoS) http://europepmc.org/articles/PMC4351947?pdf=render https://doaj.org/toc/1935-2727 https://doaj.org/toc/1935-2735 1935-2727 1935-2735 doi:10.1371/journal.pntd.0003557 https://doaj.org/article/f632526349984a9391a36ce6a5687ed6 PLoS Neglected Tropical Diseases, Vol 9, Iss 3, p e0003557 (2015) Arctic medicine. Tropical medicine RC955-962 Public aspects of medicine RA1-1270 article 2015 ftdoajarticles https://doi.org/10.1371/journal.pntd.0003557 2022-12-31T10:24:58Z BACKGROUND: The development of a vaccine conferring long-lasting immunity remains a challenge against visceral leishmaniasis (VL). Immunoproteomic characterization of Leishmania donovani proteins led to the identification of a novel protein NAD+-dependent Silent Information regulatory-2 (SIR2 family or sirtuin) protein (LdSir2RP) as one of the potent immunostimulatory proteins. Proteins of the SIR2 family are characterized by a conserved catalytic domain that exerts unique NAD-dependent deacetylase activity. In the present study, an immunobiochemical characterization of LdSir2RP and further evaluation of its immunogenicity and prophylactic potential was done to assess for its possible involvement as a vaccine candidate against leishmaniasis. METHODOLOGY/PRINCIPAL FINDINGS: LdSir2RP was successfully cloned, expressed and purified. The gene was present as a monomeric protein of ~45 kDa and further established by the crosslinking experiment. rLdSir2RP shown cytosolic localization in L. donovani and demonstrating NAD+-dependent deacetylase activity. Bioinformatic analysis also confirmed that LdSir2RP protein has NAD binding domain. The rLdSir2RP was further assessed for its cellular response by lymphoproliferative assay and cytokine ELISA in cured Leishmania patients and hamsters (Mesocricetus auratus) in comparison to soluble Leishmania antigen and it was observed to stimulate the production of IFN-γ, IL-12 and TNF-α significantly but not the IL-4 and IL-10. The naïve hamsters when vaccinated with rLdSir2RP alongwith BCG resisted the L. donovani challenge to the tune of ~75% and generated strong IL-12 and IFN-γ mediated Th1 type immune response thereof. The efficacy was further supported by remarkable increase in IgG2 antibody level which is indicative of Th1 type of protective response. Further, with a possible implication in vaccine design against VL, identification of potential T-cell epitopes of rLdSir2RP was done using computational approach. CONCLUSION/SIGNIFICANCE: The immunobiochemical characterization ... Article in Journal/Newspaper Arctic Directory of Open Access Journals: DOAJ Articles Arctic PLOS Neglected Tropical Diseases 9 3 e0003557
institution Open Polar
collection Directory of Open Access Journals: DOAJ Articles
op_collection_id ftdoajarticles
language English
topic Arctic medicine. Tropical medicine
RC955-962
Public aspects of medicine
RA1-1270
spellingShingle Arctic medicine. Tropical medicine
RC955-962
Public aspects of medicine
RA1-1270
Rajendra K Baharia
Rati Tandon
Tanuj Sharma
Manish K Suthar
Sanchita Das
Mohammad Imran Siddiqi
Jitendra Kumar Saxena
Shaym Sundar
Anuradha Dube
Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.
topic_facet Arctic medicine. Tropical medicine
RC955-962
Public aspects of medicine
RA1-1270
description BACKGROUND: The development of a vaccine conferring long-lasting immunity remains a challenge against visceral leishmaniasis (VL). Immunoproteomic characterization of Leishmania donovani proteins led to the identification of a novel protein NAD+-dependent Silent Information regulatory-2 (SIR2 family or sirtuin) protein (LdSir2RP) as one of the potent immunostimulatory proteins. Proteins of the SIR2 family are characterized by a conserved catalytic domain that exerts unique NAD-dependent deacetylase activity. In the present study, an immunobiochemical characterization of LdSir2RP and further evaluation of its immunogenicity and prophylactic potential was done to assess for its possible involvement as a vaccine candidate against leishmaniasis. METHODOLOGY/PRINCIPAL FINDINGS: LdSir2RP was successfully cloned, expressed and purified. The gene was present as a monomeric protein of ~45 kDa and further established by the crosslinking experiment. rLdSir2RP shown cytosolic localization in L. donovani and demonstrating NAD+-dependent deacetylase activity. Bioinformatic analysis also confirmed that LdSir2RP protein has NAD binding domain. The rLdSir2RP was further assessed for its cellular response by lymphoproliferative assay and cytokine ELISA in cured Leishmania patients and hamsters (Mesocricetus auratus) in comparison to soluble Leishmania antigen and it was observed to stimulate the production of IFN-γ, IL-12 and TNF-α significantly but not the IL-4 and IL-10. The naïve hamsters when vaccinated with rLdSir2RP alongwith BCG resisted the L. donovani challenge to the tune of ~75% and generated strong IL-12 and IFN-γ mediated Th1 type immune response thereof. The efficacy was further supported by remarkable increase in IgG2 antibody level which is indicative of Th1 type of protective response. Further, with a possible implication in vaccine design against VL, identification of potential T-cell epitopes of rLdSir2RP was done using computational approach. CONCLUSION/SIGNIFICANCE: The immunobiochemical characterization ...
format Article in Journal/Newspaper
author Rajendra K Baharia
Rati Tandon
Tanuj Sharma
Manish K Suthar
Sanchita Das
Mohammad Imran Siddiqi
Jitendra Kumar Saxena
Shaym Sundar
Anuradha Dube
author_facet Rajendra K Baharia
Rati Tandon
Tanuj Sharma
Manish K Suthar
Sanchita Das
Mohammad Imran Siddiqi
Jitendra Kumar Saxena
Shaym Sundar
Anuradha Dube
author_sort Rajendra K Baharia
title Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.
title_short Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.
title_full Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.
title_fullStr Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.
title_full_unstemmed Recombinant NAD-dependent SIR-2 protein of Leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.
title_sort recombinant nad-dependent sir-2 protein of leishmania donovani: immunobiochemical characterization as a potential vaccine against visceral leishmaniasis.
publisher Public Library of Science (PLoS)
publishDate 2015
url https://doi.org/10.1371/journal.pntd.0003557
https://doaj.org/article/f632526349984a9391a36ce6a5687ed6
geographic Arctic
geographic_facet Arctic
genre Arctic
genre_facet Arctic
op_source PLoS Neglected Tropical Diseases, Vol 9, Iss 3, p e0003557 (2015)
op_relation http://europepmc.org/articles/PMC4351947?pdf=render
https://doaj.org/toc/1935-2727
https://doaj.org/toc/1935-2735
1935-2727
1935-2735
doi:10.1371/journal.pntd.0003557
https://doaj.org/article/f632526349984a9391a36ce6a5687ed6
op_doi https://doi.org/10.1371/journal.pntd.0003557
container_title PLOS Neglected Tropical Diseases
container_volume 9
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