http://dkr.cdri.res.in:8080/xmlui/handle/123456789/575 2026-04-19T13:41:20Z 2026-04-19T13:41:20Z Jaiswal, A K Khare, Prashant Joshi, Sumit Kushawaha, P K Sundar, Shyam Dube, Anuradha http://dkr.cdri.res.in:8080/xmlui/handle/123456789/1696 2017-05-31T11:22:01Z 2014-01-01T00:00:00Z

Th1 stimulatory proteins of Leishmania donovani: Comparative cellular and protective responses of recombinant-triose phosphate isomerase, protein disulfide isomerase, and elongation factor-2, in combination with recombinant HSP70 against visceral leishmaniasis Jaiswal, A K; Khare, Prashant; Joshi, Sumit; Kushawaha, P K; Sundar, Shyam; Dube, Anuradha 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 abovesaid 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, rLdHSP70+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 IgG2 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.

2014-01-01T00:00:00Z Prakash, Kirtika Goyal, Manish Soni, Awakash Siddiqui, A J Bhardwaj, Jyoti Puri, S K http://dkr.cdri.res.in:8080/xmlui/handle/123456789/1687 2017-05-24T11:04:47Z 2014-01-01T00:00:00Z

Molecular cloning and biochemical characterization of iron superoxide dismutase from the rodent malaria parasite Plasmodium vinckei Prakash, Kirtika; Goyal, Manish; Soni, Awakash; Siddiqui, A J; Bhardwaj, Jyoti; Puri, S K Plasmodium parasite utilizes superoxide dismutase family proteins to limit the toxicity of reactive oxygen species, such as produced through hemoglobin degradation. These proteins play an important role in parasite survival during intra-erytrocytic phase. We have identified, and biochemically characterized a putative iron dependent superoxide dismutase from rodent malaria parasite P. vinckei (PvSOD1). The recombinant PvSOD1 protein was purified to homogeneity through a combination of affinity and gel filtration chromatography. Crosslinking, Native-PAGE and FPLC gel filtration analyses documented that PvSOD1 exists as a dimer in solution, a common feature shared by other Fe-SODs. PvSOD1 is cytosolic in localization and its expression is comparatively higher during trophozoite as compared to that of ring and schizont stages. Enzymatic activity of recombinant PvSOD1 was validated using conventional zymogram analyses and xanthine-xanthine oxidase system. Under optimal conditions, PvSOD1 was highly active and catalyzed the dismutation of superoxide radicals. Furthermore, PvSOD1 showed activity over a broad range of pH and temperature. Inhibition studies suggested that PvSOD1 was inactivated by hydrogen peroxide, and peroxynitrite, but not by cyanide and azide. Since, PvSOD1 plays a central role in oxidative defence mechanism, therefore, characterization of PvSOD1 will be exploited in the screening of new superoxide dismutase inhibitors for their antimalarial activity.

2014-01-01T00:00:00Z DWIVEDI, HEMLATA SINGH, SK CHAUHAN, BS GUNJAN, S TRIPATHI, R http://dkr.cdri.res.in:8080/xmlui/handle/123456789/1680 2017-04-24T09:48:26Z 2016-01-01T00:00:00Z

Potential Cerebral malaria therapy: Intramuscular arteether and vitamin D co-administration DWIVEDI, HEMLATA; SINGH, SK; CHAUHAN, BS; GUNJAN, S; TRIPATHI, R Cerebral malaria (CM) shows lethality rate of 15-25% despite effective antimalarial chemotherapy. The effective adjunct treatment to counteract the CM pathogenesis is urgently required. In murine cerebral malaria model, most interventions studied till date are administered before the onset of CM symptoms which belittle its translational value to human. We studied intramuscular arteether – vitamin D (ART-VD) combination treatment for CM outcome improvement after the onset of neurological symptoms. The intramuscular dose of 50µg/kg VD for 3 days combined with a loading dose of 25mg/kg α/β arteether followed by 12.5mg/kg dose for two consecutive days led to significant improvement in survival (73% in combination group vs 29% and 0% in arteether and VD monotherapy respectively) and clinical recovery. The treatment in all the groups partially restored the blood brain barrier integrity and reduced the level of serum proinflammatory cytokines TNF-α (Tumor necrosis factor-α) and IFN-γ (Interferon-γ). The brain transcripts of inflammatory chemokines viz. CXCL10, CXCL9, CCL4 and CCL5 and T cell migration in the brain microvasculature were significantly diminished in all the treatment groups. ART-VD treatment significantly reduced ICAM-1(Intercellular cell adhesion molecule-1) expression. Taken together, our findings show that coordinated actions of ART-VD improve the outcome of experimental cerebral malaria.

2016-01-01T00:00:00Z Pathak, Manisha Verma, Meenakshi Srivastava, Mrigank Bhattacharya-Misra, Shailja http://dkr.cdri.res.in:8080/xmlui/handle/123456789/1670 2017-04-19T06:24:55Z 2014-01-01T00:00:00Z

Wolbachia endosymbiont of Brugia malayi elicits Th-17 mediated proinflammatory immune response through surface protein (WSP) Pathak, Manisha; Verma, Meenakshi; Srivastava, Mrigank; Bhattacharya-Misra, Shailja Wolbachia is an endosymbiotic bacterium of filarial nematode Brugia malayi. The symbiotic relationship between Wolbachia and its filarial host is dependent on some interactions between the proteins of both organisms. However, very little is known about Wolbachia proteins that are involved in inflammatory pathology of the host during lymphatic filariasis. In the present study, we cloned, expressed and purified Wolbachia surface protein (r-wsp) from Wolbachia and administered it into mice, either alone or in combination with infective larvae of B. malayi (Bm-L3) and monitored the developing immune response in infected animals. Our results show that spleens and mesenteric lymph nodes (mLNs) of mice immunized with either r-wsp or infected with Bm-L3 show increased percentages of CD4+ Th17 cells and Th1 cytokines like IFN-γ and IL-2 along with decreased percentages of Tregs, Th2 cytokines like IL-4 and IL-10 and Transforming growth factor beta (TGF-β) levels in culture supernatants of splenocytes. These observations were more prominent in mice immunized with r-wsp alone. Interestingly, when mice were first immunized with r-wsp and subsequently infected with Bm-L3, percentages of CD4+ Th17 cells and Th1 cytokines increased even further while that of Tregs, Th2 cytokines and TGF-β levels decreased. These results for the first time show that r-wsp acts synergistically with Bm-L3 in promoting pro-inflammatory response by increasing Th17 cells and at the same time diminishes host immunological tolerance by decreasing Tregs cells and TGF-β secretion.

2014-01-01T00:00:00Z