Abstract
Leishmania infantum is the etiologic agent of zoonotic visceral leishmaniasis (VL) in countries in the Mediterranean basin, where dogs are the domestic reservoirs and represent important elements in the transmission of the disease. Since the major focal areas of human VL exhibit a high prevalence of seropositive dogs, the control of canine VL could reduce the infection rate in humans. Efforts toward this have focused on the improvement of diagnostic tools, as well as on vaccine development. The identification of parasite antigens including suitable major histocompatibility complex (MHC) class I- and/or II-restricted epitopes is very important since disease protection is characterized by strong and long-lasting CD8+ T and CD4+ Th1 cell-dominated immunity. In the present study, total protein extract from late-log phase L. infantum promastigotes was analyzed by two-dimensional western blots and probed with sera from asymptomatic and symptomatic dogs. A total of 42 protein spots were found to differentially react with IgG from asymptomatic dogs, while 17 of these identified by Coommasie stain were extracted and analyzed. Of these, 21 proteins were identified by mass spectrometry; they were mainly involved in metabolism and stress responses. An in silico analysis predicted that the chaperonin HSP60, dihydrolipoamide dehydrogenase, enolase, cyclophilin 2, cyclophilin 40, and one hypothetical protein contain promiscuous MHCI and/or MHCII epitopes. Our results suggest that the combination of immunoproteomics and bioinformatics analyses is a promising method for the identification of novel candidate antigens for vaccine development or with potential use in the development of sensitive diagnostic tests.
Highlights
Leishmania is an obligate intracellular protozoan parasite that is transmitted via phlebotomine sand flies
Drugs used to treat zoonotic Visceral leishmaniasis (VL) (ZVL) are not sufficiently effective to eradicate disease owing to the development of resistant parasitic strains and still the epidemiological risk persists [2,3]
In southern Europe and the Mediterranean region, ZVL is the most widespread form of leishmaniasis caused by a single parasite species, Leishmania (L.) infantum, with dogs representing the main domestic reservoirs of the parasite and playing significant role in disease transmission
Summary
Leishmania is an obligate intracellular protozoan parasite that is transmitted via phlebotomine sand flies. It causes a broad spectrum of human diseases ranging from self-healing cutaneous lesions to severe visceral dissemination. Visceral leishmaniasis (VL) is the most severe clinical form, giving rise to approximately 500,000 new cases each year, and is usually fatal if not treated properly [1]. In southern Europe and the Mediterranean region, ZVL is the most widespread form of leishmaniasis caused by a single parasite species, Leishmania (L.) infantum, with dogs representing the main domestic reservoirs of the parasite and playing significant role in disease transmission. The control of ZVL considerably reduces the number of reservoirs, and the infection rate in humans [8]
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