title: Leptin augments protective immune responses in murine macrophages and enhances potential of miltefosine against experimental visceral leishmaniasis.
abstract: Adverse side effects and drug resistance issues are the two most important drawbacks which influence the widespread use of existing antileishmanial drugs. Use of immune stimulating agent with standard antileishmanial might be helpful to minimize the toxic effect of drug, shorten the dose regimen and delay the emergence of resistance. In the present study, we explored the in vitro immunomodulatory potential of an immunomodulator, leptin with lower concentration of standard drug, miltefosine. The level of Th1/Th2 cytokines, production of nitric oxide and reactive oxygen species and phagocytic activity was assessed by ELISA, Griess reaction and flow cytometric analysis, respectively. Leptin at a concentration of 15μg/mL showed heightened level of Th1 cytokines and nitric oxide generation from murine macrophages (J-774A.1 cells). Leptin (15μg/mL) also reduces the effective concentration of miltefosine by 2-folds from 7.5μM to 3.7μM. When given in conjunction with lower concentration of miltefosine (4μM), leptin (15μg/mL) significantly (***p<0.001) elevated the level of IL-12 (7.7 fold), TNF-α (8.1 fold) and nitric oxide (6.6 fold) along with markedly (***p<0.001) suppressed level of IL-10 and TGF-β when compared with untreated infected macrophages. Leptin plus miltefosine also induces the phagocytic ability (**p<0.01) of macrophages in comparison to leptin alone and miltefosine alone treated groups. These finding illustrate that leptin activates host macrophages to generate protective immune response for the successful elimination of Leishmania parasite at lower concentration of miltefosine and has potential for further exploration in experimental animal model of visceral leishmaniasis (VL).
authors.affiliation: Department of Pathology, King George's Medical University, Lucknow 226 003, India. Electronic address: rahul_biotech08@yahoo.com.
authors.forename: Rahul
authors.initials: R
authors.lastname: Shivahare
authors.affiliation: Department of Pathology, King George's Medical University, Lucknow 226 003, India.
authors.forename: Wahid
authors.initials: W
authors.lastname: Ali
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow 226 031, India.
authors.forename: Preeti
authors.initials: P
authors.lastname: Vishwakarma
authors.affiliation: Department of Pathology, King George's Medical University, Lucknow 226 003, India.
authors.forename: S M
authors.initials: SM
authors.lastname: Natu
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow 226 031, India.
authors.forename: Sunil K
authors.initials: SK
authors.lastname: Puri
authors.affiliation: Division of Parasitology, CSIR-Central Drug Research Institute, Lucknow 226 031, India.
authors.forename: Suman
authors.initials: S
authors.lastname: Gupta
mesh_terms: D000818:Animals; D000981:Antiprotozoal Agents / Q000494:pharmacology*; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D007898:Leishmaniasis, Visceral / Q000188:drug therapy* / Q000276:immunology; D020738:Leptin / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects* / Q000276:immunology; D051379:Mice; D009569:Nitric Oxide / Q000096:biosynthesis; D010767:Phosphorylcholine / Q000031:analogs & derivatives* / Q000494:pharmacology
chemical_list: D000981:Antiprotozoal Agents; D016207:Cytokines; D020738:Leptin; D010767:Phosphorylcholine; D009569:Nitric Oxide; C039128:miltefosine
keywords: Combination therapy; Immune response; J-774.A1 cells; Leishmania donovani; Leptin; Miltefosine