title: Combination of liposomal CpG oligodeoxynucleotide 2006 and miltefosine induces strong cell-mediated immunity during experimental visceral leishmaniasis.
abstract: Immuno-modulators in combination with antileishmanial drug miltefosine is a better therapeutic approach for treatment of Visceral Leishmaniasis (VL) as it not only reduces the dose of miltefosine but also shortens the treatment regimen. However, immunological mechanisms behind the perceived benefits of this combination therapy have not been investigated in detail. In the present study, we hypothesized that potential use of drugs that target the host in addition to the parasite might represent an alternative strategy for combination therapy. We investigated immune responses generated in Leishmania donovani infected animals (hamsters and mice) treated with combination of CpG-ODN-2006 and miltefosine at short dose regimen. Infected animals were administered CpG-ODN-2006 (0.4 mg/kg, single dose), as free and liposomal form, either alone or in combination with miltefosine for 5 consecutive days and parasite clearance was evaluated at day 4 and 7 post treatment. Animals that received liposomal CpG-ODN-2006 (lipo-CpG-ODN-2006) and sub-curative miltefosine (5 mg/kg) showed the best inhibition of parasite multiplication (∼97%) which was associated with a biased Th1 immune response in. Moreover, compared to all the other treated groups, we observed increased mRNA expression levels of pro-inflammatory cytokines (IFN-γ, TNF-α and IL-12) and significantly suppressed levels of Th2 cytokines (IL-10 and TGF-β) on day 4 post treatment in animals that underwent combination therapy with lipo-CpG-ODN-2006 and sub-curative miltefosine. Additionally, same therapy also induced heightened iNOS mRNA levels and NO generation, increased IgG2 antibody level and strong T-cell response in these hamsters compared with all the other treated groups. Collectively, our results suggest that combination of lipo-CpG-ODN-2006 and sub-curative miltefosine generates protective T-cell response in an animal model of visceral leishmaniasis which is characterized by strong Th1 biased immune response thereby underlining our hypothesis that combination therapy, at short dose regimen can be used as a novel way of treating visceral leishmaniasis.
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Rahul
authors.initials: R
authors.lastname: Shivahare
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Preeti
authors.initials: P
authors.lastname: Vishwakarma
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Naveen
authors.initials: N
authors.lastname: Parmar
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Pawan Kumar
authors.initials: PK
authors.lastname: Yadav
authors.affiliation: Division of Medicinal and Process Chemistry, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Wahajul
authors.initials: W
authors.lastname: Haq
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Mrigank
authors.initials: M
authors.lastname: Srivastava
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Suman
authors.initials: S
authors.lastname: Gupta
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow, India.
authors.forename: Susanta
authors.initials: S
authors.lastname: Kar
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D049109:Cell Proliferation; D006224:Cricetinae; D004797:Enzyme-Linked Immunosorbent Assay; D007111:Immunity, Cellular; D007074:Immunoglobulin G / Q000276:immunology; D007155:Immunologic Factors / Q000276:immunology; D007898:Leishmaniasis, Visceral / Q000188:drug therapy* / Q000276:immunology*; D008081:Liposomes / Q000494:pharmacology; D008647:Mesocricetus; D051379:Mice; D008807:Mice, Inbred BALB C; D009569:Nitric Oxide / Q000378:metabolism; D009838:Oligodeoxyribonucleotides / Q000494:pharmacology*; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D013601:T-Lymphocytes / Q000166:cytology / Q000469:parasitology
chemical_list: D000981:Antiprotozoal Agents; C513533:CpG ODN 2006; D007074:Immunoglobulin G; D007155:Immunologic Factors; D008081:Liposomes; D009838:Oligodeoxyribonucleotides; D012333:RNA, Messenger; D010767:Phosphorylcholine; D009569:Nitric Oxide; C039128:miltefosine
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