Browsing by Author "Prashant Khare"

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Comparative analysis of cellular immune responses in treated Leishmania patients and hamsters against recombinant Th1 stimulatory proteins of Leishmania donovani
(Frontiers Media S.A., 2016) Sumit Joshi; Narendra K. Yadav; Keerti Rawat; Chandra Dev P. Tripathi; Anil K. Jaiswal; Prashant Khare; Rati Tandon; Rajendra K. Baharia; Sanchita Das; Reema Gupta; Pramod K. Kushawaha; Shyam Sundar; Amogh A. Sahasrabuddhe; Anuradha Dube
Our prior studies demonstrated that cellular response of T helper 1 (Th1) type was generated by a soluble antigenic fraction (ranging from 89.9 to 97.1 kDa) of Leishmania donovani promastigote, in treated Leishmania patients as well as hamsters and showed significant prophylactic potential against experimental visceral leishmaniasis (VL). Eighteen Th1 stimulatory proteins were identified through proteomic analysis of this subfraction, out of which 15 were developed as recombinant proteins. In the present work, we have evaluated these 15 recombinant proteins simultaneously for their comparative cellular responses in treated Leishmania patients and hamsters. Six proteins viz. elongation factor-2, enolase, aldolase, triose phosphate isomerase, protein disulfide isomerase, and p45 emerged as most immunogenic as they produced a significant lymphoproliferative response, nitric oxide generation and Th1 cytokine response in PBMCs and lymphocytes of treated Leishmania patients and hamsters respectively. The results suggested that these proteins may be exploited for developing a successful poly-protein and/or poly-epitope vaccine against VL. © 2016 Joshi, Yadav, Rawat, Tripathi, Jaiswal, Khare, Tandon, Baharia, Das, Gupta, Kushawaha, Sundar, Sahasrabuddhe and Dube.
Detector concepts
(Stanford Linear Accelerator Center (SLAC), 2005) Gerald Aarons; Toshinori Abe; Jason Abernathy; Medina Ablikim; Halina Abramowicz; David Adey; Catherine Adloff; Chris Adolphsen; Konstantin Afanaciev; Ilya Agapov; Jung-Keun Ahn; Hiroaki Aihara; Mitsuo Akemoto; Maria Del CarmenAlabau; Justin Albert; Hartwig Albrecht; Michael Albrecht; David Alesini; Gideon Alexander; Jim Alexander; Wade Allison; John Amann; Ramila Amirikas; Qi An; Shozo Anami; B. Ananthanarayan; Terry Anderson; Ladislav Andricek; Marc Anduze; Michael Anerella; Nikolai Anfimov; Deepa Angal-Kalinin; Sergei Antipov; Claire Antoine; Mayumi Aoki; Atsushi Aoza; Steve Aplin; Rob Appleby; Yasuo Arai; Sakae Araki; Tug Arkan; Ned Arnold; Ray Arnold; Richard Arnowitt; Xavier Artru; Kunal Arya; Alexander Aryshev; Eri Asakawa; Fred Asiri; David Asner; Muzaffer Atac; Grigor Atoian; David Attié; Jean-Eudes Augustin; David B. Augustine; Bradley Ayres; Tariq Aziz; Derek Baars; Frederique Badaud; Nigel Baddams; Jonathan Bagger; Sha Bai; David Bailey; Ian R. Bailey; David Baker; Nikolai I. Balalykin; Juan Pablo Balbuena; Jean-Luc Baldy; Markus Ball; Maurice Ball; Alessandro Ballestrero; Jamie Ballin; Charles Baltay; Philip Bambade; Syuichi Ban; Henry Band; Karl Bane; Bakul Banerjee; Serena Barbanotti; Daniele Barbareschi; Angela Barbaro-Galtieri; Desmond P. Barber; Mauricio Barbi; Dmitri Y. Bardin; Barry Barish; Timothy L. Barklow; Roger Barlow; Virgil E. Barnes; Maura Barone; Christoph Bartels; Valeria Bartsch; Rahul Basu; Marco Battaglia; Yuri Batygin; Jerome Baudot; Ulrich Baur; D. Elwyn Baynham; Carl Beard; Chris Bebek; Philip Bechtle; Ulrich J. Becker; Franco Bedeschi; Marc Bedjidian; Prafulla Behera; Ties Behnke; Leo Bellantoni; Alain Bellerive; Paul Bellomo; Lynn D. Bentson; Mustapha Benyamna; Thomas Bergauer; Edmond Berger; Matthias Bergholz; Suman Beri; Martin Berndt; Werner Bernreuther; Alessandro Bertolini; Marc Besancon; Auguste Besson; Andre Beteille; Simona Bettoni; Michael Beyer; R.K. Bhandari; Vinod Bharadwaj; Vipin Bhatnagar; Satyaki Bhattacharya; Gautam Bhattacharyya; Biplob Bhattacherjee; Ruchika Bhuyan; Xiao-Jun Bi; Marica Biagini; Wilhelm Bialowons; Otmar Biebel; Thomas Bieler; John Bierwagen; Alison Birch; Mike Bisset; S.S. Biswal; Victoria Blackmore; Grahame Blair; Guillaume Blanchard; Gerald Blazey; Andrew Blue; Johannes Blümlein; Christian Boffo; Courtlandt Bohn; V.I. Boiko; Veronique Boisvert; Eduard N. Bondarchuk; Roberto Boni; Giovanni Bonvicini; Stewart Boogert; Maarten Boonekamp; Gary Boorman; Kerstin Borras; Daniela Bortoletto; Alessio Bosco; Carlo Bosio; Pierre Bosland; Angelo Bosotti; Vincent Boudry; Djamel-Eddine Boumediene; Bernard Bouquet; Serguei Bourov; Gordon Bowden; Gary Bower; Adam Boyarski; Ivanka Bozovic-Jelisavcic; Concezio Bozzi; Axel Brachmann; Tom W. Bradshaw; Andrew Brandt; Hans Peter Brasser; Benjamin Brau; James E. Brau; Martin Breidenbach; Steve Bricker; Jean-Claude Brient; Ian Brock; Stanley Brodsky; Craig Brooksby; Timothy A. Broome; David Brown; James H. Brownell; Mélanie Bruchon; Heiner Brueck; Amanda J. Brummitt; Nicole Brun; Peter Buchholz; Yulian A. 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[No abstract available]
Efficacy of Leishmania donovani trypanothione reductase, identified as a potent Th1 stimulatory protein, for its immunogenicity and prophylactic potential against experimental visceral leishmaniasis
(Springer Verlag, 2014) Prashant Khare; Anil Kumar Jaiswal; Chandra Dev Pati Tripathi; Sumit Joshi; Shyam Sundar; Anuradha Dube
In visceral leishmaniasis (VL), Th1-type of immune responses play an important role which correlates with recovery from and resistance to disease resulting in lifelong immunity. Based on this rationale, the soluble leishmanial antigens that elicit cellular responses in peripheral blood mononuclear cells (PBMCs) from cured Leishmania patients were characterized through immunoproteomic approach which led to the identification of trypanothione reductase (TPR) (a cytosolic enzyme explored as a drug target), as one of the potent Th1 stimulatory protein. In this study, the immunogenicity of recombinant Leishmania donovani TPR (rLdTPR) was assessed in PBMCs of cured Leishmania-infected patients/hamsters and further evaluated its prophylactic efficacy against L. donovani challenges in hamsters. Substantial proliferative responses to rLdTPR, as compared to soluble L. donovani antigen, were observed in Leishmania-infected cured patients as well as in hamsters. Moreover, rLdTPR reasonably stimulated PBMCs of cured Leishmania patients to produce IFNγ, IL-12, and TNF-α but not IL-4 or IL-10. On the other hand, the protein downregulated LPS-induced IL-10 as well as soluble L. donovani antigen-induced IL-4 production in PBMCs of Leishmania patients. In case of cured hamsters, rLdTPR generates mixed Th1 and Th2 immune response. Vaccination with rLdTPR along with Bacillus Calmette-Guerin (BCG) was able to provide considerably good prophylactic efficacy (~60 %) against L. donovani challenge in hamsters. The efficacy was supported by the increased inducible NO synthase mRNA transcript and Th1-type cytokines IFNγ, IL-12, and TNF-α and downregulation of IL-4, IL-10, and TGF-β. Since rLdTPR protein is an important target, further attempts towards determination of immunodominant regions for designing fusion peptides may be taken up to optimize its prophylactic efficacy. © 2013 Springer-Verlag Berlin Heidelberg.
Immunological consequences of stress-related proteins-cytosolic tryparedoxin peroxidase and chaperonin TCP20-identified in splenic amastigotes of Leishmania donovani as Th1 stimulatory, in experimental visceral leishmaniasis
(Cambridge University Press, 2015) Anil Kumar Jaiswal; Prashant Khare; Sumit Joshi; Keerti Rawat; Narendra Yadav; Shyam Sundar; Anuradha Dube
In earlier studies, proteomic characterization of splenic amastigote fractions from clinical isolates of Leishmania donovani, exhibiting significant cellular responses in cured Leishmania subjects, led to the identification of cytosolic tryparedoxin peroxidase (LdcTryP) and chaperonin-TCP20 (LdTCP20) as Th1-stimulatory proteins. Both the proteins, particularly LdTCP20 for the first time, were successfully cloned, overexpressed, purified and were found to be localized in the cytosol of purified splenic amastigotes. When evaluated against lymphocytes of cured Leishmania-infected hamsters, the purified recombinant proteins (rLdcTryP and rLdTCP20) induced their proliferations as well as nitric oxide production. Similarly, these proteins also generated Th1-type cytokines (IFN-γ/IL-12) from stimulated PBMCs of cured/endemic Leishmania patients. Further, vaccination with rLdcTryP elicited noticeable delayed-type hypersensitivity response and offered considerably good prophylactic efficacy (~78% inhibition) against L. donovani challenge in hamsters, which was well supported by the increased mRNA expression of Th1 and Th2 cytokines. However, animals vaccinated with rLdTCP20 exhibited comparatively lesser prophylactic efficacy (~55%) with inferior immunological response. The results indicate the potentiality of rLdcTryP protein, between the two, as a suitable anti-leishmanial vaccine. Since, rLdTCP20 is also an important target, for optimization, further attempts towards determination of immunodominant regions for designing fusion peptides may be taken up. © Cambridge University Press 2014.
Induction of Th1-type cellular responses in cured/exposed Leishmania-infected patients and hamsters against polyproteins of soluble Leishmania donovani promastigotes ranging from 89.9 to 97.1 kDa
(2008) Shraddha Kumari; Mukesh Samant; Prashant Khare; Shyam Sundar; Sudhir Sinha; Anuradha Dube
Earlier, we have identified soluble antigenic fraction ranging from 68 to 97.4 kDa (F2-fraction) of Leishmania donovani promastigote, which induced Th1-immunostimulatory cellular responses in both cured Leishmania patients/hamsters and exhibited significant prophylactic potential against experimental visceral leishmaniasis (VL). In the present study, we have further fractionated F2-fraction by continuous elution SDS-PAGE and subjected them to re-evaluation for their ability to induce cellular responses. Out of seven sub-fractions: F2.1-F2.7, only four F2.4-F2.7 (89.9-97.1 kDa), individually or in pooled sub-fractions (P4-7), stimulated Th1-type remarkable lymphoproliferative, NO, IFN-γ and IL-12 responses in Leishmania-infected cured/exposed patients and hamsters. Interestingly, optimum Th1-responses were obtained with P4-7 suggesting the presence of immunostimulatory molecules in it and may be exploited for developing a successful subunit vaccine against VL. © 2008 Elsevier Ltd. All rights reserved.
Leishmania donovani Triose Phosphate Isomerase: A Potential Vaccine Target against Visceral Leishmaniasis
(2012) Pramod K. Kushawaha; Reema Gupta; Chandra Dev Pati Tripathi; Prashant Khare; Anil Kumar Jaiswal; Shyam Sundar; Anuradha Dube
Visceral leishmaniasis (VL) is one of the most important parasitic diseases with approximately 350 million people at risk. Due to the non availability of an ideal drug, development of a safe, effective, and affordable vaccine could be a solution for control and prevention of this disease. In this study, a potential Th1 stimulatory protein- Triose phosphate isomerase (TPI), a glycolytic enzyme, identified through proteomics from a fraction of Leishmania donovani soluble antigen ranging from 89.9-97.1 kDa, was assessed for its potential as a suitable vaccine candidate. The protein- L. donovani TPI (LdTPI) was cloned, expressed and purified which exhibited the homology of 99% with L. infantum TPI. The rLdTPI was further evaluated for its immunogenicity by lymphoproliferative response (LTT), nitric oxide (NO) production and estimation of cytokines in cured Leishmania patients/hamster. It elicited strong LTT response in cured patients as well as NO production in cured hamsters and stimulated remarkable Th1-type cellular responses including IFN-ã and IL-12 with extremely lower level of IL-10 in Leishmania-infected cured/exposed patients PBMCs in vitro. Vaccination with LdTPI-DNA construct protected naive golden hamsters from virulent L. donovani challenge unambiguously (~90%). The vaccinated hamsters demonstrated a surge in IFN-ã, TNF-á and IL-12 levels but extreme down-regulation of IL-10 and IL-4 along with profound delayed type hypersensitivity and increased levels of Leishmania-specific IgG2 antibody. Thus, the results are suggestive of the protein having the potential of a strong candidate vaccine. © 2012 Kushawaha et al.
Over-Expression of 60s Ribosomal L23a Is Associated with Cellular Proliferation in SAG Resistant Clinical Isolates of Leishmania donovani
(Public Library of Science, 2013) Sanchita Das; Priyanka Shah; Rajendra K. Baharia; Rati Tandon; Prashant Khare; Shyam Sundar; Amogh A. Sahasrabuddhe; M.I. Siddiqi; Anuradha Dube
Background:Sodium antimony gluconate (SAG) unresponsiveness of Leishmania donovani (Ld) had effectively compromised the chemotherapeutic potential of SAG. 60s ribosomal L23a (60sRL23a), identified as one of the over-expressed protein in different resistant strains of L.donovani as observed with differential proteomics studies indicates towards its possible involvement in SAG resistance in L.donovani. In the present study 60sRL23a has been characterized for its probable association with SAG resistance mechanism.Methodology and principal findings:The expression profile of 60s ribosomal L23a (60sRL23a) was checked in different SAG resistant as well as sensitive strains of L.donovani clinical isolates by real-time PCR and western blotting and was found to be up-regulated in resistant strains. Ld60sRL23a was cloned, expressed in E.coli system and purified for raising antibody in swiss mice and was observed to have cytosolic localization in L.donovani. 60sRL23a was further over-expressed in sensitive strain of L.donovani to check its sensitivity profile against SAG (Sb V and III) and was found to be altered towards the resistant mode.Conclusion/Significance:This study reports for the first time that the over expression of 60sRL23a in SAG sensitive parasite decreases the sensitivity of the parasite towards SAG, miltefosine and paramomycin. Growth curve of the tranfectants further indicated the proliferative potential of 60sRL23a assisting the parasite survival and reaffirming the extra ribosomal role of 60sRL23a. The study thus indicates towards the role of the protein in lowering and redistributing the drug pressure by increased proliferation of parasites and warrants further longitudinal study to understand the underlying mechanism. © 2013 Das et al.
Recent insights on pattern recognition receptors and the interplay of innate immune responses against West Nile Virus infection
(Academic Press Inc., 2024) Jatin Behari; Kajal Yadav; Prashant Khare; Brijesh Kumar; Ambuj Kumar Kushwaha
The recent outbreaks of neurotropic West Nile Virus (WNV) in humans are of grave public health concern, requiring a thorough understanding of the host immune response to develop effective therapeutic interventions. Innate immunity contributes to the primary immune response against WNV infection aimed at controlling and eliminating the virus from the body. As soon as WNV infects the body, pattern recognition receptors (PRRs) recognize viral pathogen-associated molecular patterns, particularly viral RNA, and initiate innate immune responses. This review explores the diverse PRRs in sensing WNV infection and orchestrating immune defenses. Specifically, this paper reviews the role of PRRs in WNV infection, encompassing both findings from mouse models and current clinical studies. Activation of PRRs triggers signaling pathways that induce the expression of antiviral proteins to inhibit viral replication. Understanding the intricacies of the immune response is crucial for developing effective vaccines and therapeutic interventions against WNV infection. © 2024 Elsevier Inc.
Th1 stimulatory proteins of Leishmania donovani: Comparative cellular and protective responses of rTriose phosphate isomerase, rProtein disulfide isomerase and relongation factor-2 in combination with rHSP70 against visceral leishmaniasis
(Public Library of Science, 2014) Anil Kumar Jaiswal; Prashant Khare; Sumit Joshi; Pramod Kumar Kushawaha; Shyam Sundar; Anuradha Dube
In visceral leishmaniasis, the recovery from the disease is always associated with the generation of Th1-type of cellular responses. Based on this, we have previously identified several Th1-stimulatory proteins of Leishmania donovani -triose phosphate isomerase (TPI), protein disulfide isomerase (PDI) and elongation factor-2 (EL-2) etc. including heat shock protein 70 (HSP70) which induced Th1-type of cellular responses in both cured Leishmania patients/hamsters. Since, HSPs, being the logical targets for vaccines aimed at augmenting cellular immunity and can be early targets in the immune response against intracellular pathogens; they could be exploited as vaccine/adjuvant to induce long-term immunity more effectively. Therefore, in this study, we checked whether HSP70 can further enhance the immunogenicity and protective responses of the above said Th1-stimulatory proteins. Since, in most of the studies, immunogenicity of HSP70 of L donovani was assessed in native condition, herein we generated recombinant HSP70 and tested its potential to stimulate immune responses in lymphocytes of cured Leishmania infected hamsters as well as in the peripheral blood mononuclear cells (PBMCs) of cured patients of VL either individually or in combination with above mentioned recombinant proteins. rLdHSP70 alone elicited strong cellular responses along with remarkable up-regulation of IFN-γ and IL-12 cytokines and extremely lower level of IL-4 and IL-10. Among the various combinations, rLdFISP70 + rLdPDI emerged as superior one augmenting improved cellular responses followed by rLdHSP70 + rLdEL-2. These combinations were further evaluated for its protective potential wherein rLdHSP70 + rLdPDI again conferred utmost protection ( 80%) followed by rLdHSP70 + rLdEL-2 (∼75%) and generated a strong cellular immune response with significant increase in the levels of iNOS transcript as well as IFN-γ and IL-12 cytokines which was further supported by the high level of lgG2 antibody in vaccinated animals. These observations indicated that vaccine(s) based on combination of HSP70 with Th1-stimulatory protein(s) may be a viable proposition against intracellular pathogens. © 2014 Jaiswal et al.