Abstract
Leishmania martiniquensis is a neglected cause of an emerging leishmaniasis in many countries, including France, Germany, Switzerland, the United States of America, Myanmar, and Thailand, with different clinical manifestations ranging from asymptomatic, cutaneous (CL), visceral (VL), and atypically disseminated CL and VL. The persistence of parasites and the recurrence of the disease after treatment are challenges in controlling the disease. To explore efficient prophylaxis and therapy, this study aimed to investigate infection outcome and organ-specific immune responses after inoculation with L. martiniquensis (MHOM/TH/2011/PG; 5 x 106 promastigotes) in BALB/c mice via intravenous and intraperitoneal routes. A quantitative PCR technique, targeting L. martiniquensis ITS1, was primarily established to estimate the parasite burden. We found that the infection in the liver resolved; however, persistent infection was observed in the spleen. Histopathology with Leishmania-specific immunostaining revealed efficient hepatic granuloma formation, while splenic disorganization with parasitized macrophages at different locations was demonstrated. The mRNA expression of Th1 cytokines (IFN-γ, TNF-α, IL-12p40) and iNOS in the liver and spleen was upregulated. In addition, high expression of IL-10 was observed in the spleen in the chronic phase, revealing a significant moderate correlation with the parasite persistence [r(12) = 0.72, P = 0.009]. Further clarification of the mechanisms of persistent infection and experimental infection in immunosuppressed murine models are warranted.
Highlights
Leishmaniasis is a vector-borne disease caused by several species of intracellular protozoa belonging to the Leishmania genus
The clinical characteristics of VL caused by L. martiniquensis are comparable with typical VL reported in L. donovani and L. infantum infections, including prolonged fever, anemia, hepatosplenomegaly, and cachexia
A Developed Quantitative real-time PCR (qPCR) Assay for the Quantification of L. martiniquensis In this study, the newly designed primers, L.internal transcribed ribosomal RNA spacer 1 (ITS1).PCM2.4.6-qF and L.ITS1.PCM2.4.6-qR, were able to amplify L. martiniquensis Genomic DNA (gDNA), which showed a high amplification efficiency of 1.940, with correlation coefficients (r2) of 0.991 and slopes of −3.47
Summary
Leishmaniasis is a vector-borne disease caused by several species of intracellular protozoa belonging to the Leishmania genus. The main clinical manifestations, including cutaneous leishmaniasis (CL), mucocutaneous leishmaniasis (MCL), and visceral leishmaniasis (VL), are generally associated with the Leishmania species. Leishmania martiniquensis is the major species causing autochthonous leishmaniasis in Thailand in immunocompetent and immunosuppressed patients, and has a wide geographical distribution that includes France, Germany, Switzerland, the United States of America, and Myanmar [1]. Various clinical manifestations, including asymptomatic, CL alone, VL alone, and atypically disseminated cutaneous and visceral (DCL/VL) concomitant forms, have been reported in cases of L. martiniquensis infection, especially in patients with a human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) coinfection [1, 4, 5]. The clinical characteristics of VL caused by L. martiniquensis are comparable with typical VL reported in L. donovani and L. infantum infections, including prolonged fever, anemia, hepatosplenomegaly, and cachexia. All documented VL cases caused by L. martiniquensis infection present a chronic infection and are occasionally asymptomatic in human and animal reservoirs [1]
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