Comparative Proteomic Analysis of Murine Cutaneous Lesions Induced by Leishmania amazonensis or Leishmania major

Abstract

Cutaneous leishmaniasisis is the most common clinical form of leishmaniasis and one of the most relevant neglected diseases. It is known that the progress of the disease is species specific and the host's immune response plays an important role in its outcome. However, the pathways that lead to parasite clearance or survival remain unknown. In this work, skin tissue from mice experimentally infected with L. amazonensis, one of the causative agents of cutaneous leishmaniasis in the Amazon region, L. major, another causative agent of cutaneous leishmaniasis in Africa, the Middle East, China, and India, or lipopolysaccharides from Escherichia coli as an inflammation model were investigated using label-free proteomics to unveil Leishmaniaspecific protein alterations. Proteomics is a powerful tool to investigate host−pathogen relationships to address biological questions. In this work, proteins from mice skin biopsies were identified and quantified using nano-LC coupled with tandem mass spectrometry analyses. Integrated Proteomics Pipeline was used for peptide/protein identification and quantification. Western blot was used for validation of protein quantification by mass spectrometry, and protein pathways were predicted using Ingenuity Pathway Analysis. In this proteomics study, several proteins were pointed out as hypothetical targets to guide future studies on Leishmania-specific modulation of proteins in the host. We identified hundreds of exclusively modulated proteins after Leishmania spp. infection and 17 proteins that were differentially modulated in the host after L. amazonensis or L. major infection.

Differentially modulated proteins associated with Leishmaniasis—a systematic review of in-vivo and in-vitro studies

“…Despite the different experimental conditions and setup, heat-shock and Tryparedoxin related proteins were differentially regulated in 17 studies, indicating that those proteins are often modulated in the parasite. Moreover, several proteins were consistently identifiable and also to be similarly modulating in studies with similar experimental conditions [ 42 , 40 , 41 ]. Only three studies explored the host response (human subjects) during Leishmania infection.…”

“…Also, the study reported U6 snRNA-associated Sm-like protein and heat repeat-containing protein 3 to be precisely up-regulated in infection with L. major , while protein tweety homolog 3, phosphatidate cytidylyltransferase, and macrophage metalloelastase to be precisely up-regulated in infection with L. amazonensis . Overall, the studies by Negrão et al [ 40 , 42 , 41 ] demonstrated the species-specific protein modulation in Leishmania infection as well as differentiated the host response through LPS inoculation and Leishmania infection. To observe the immune response effects of malnutrition in Leishmania infection, Losada-Barragán et al [ 43 , 44 ] conducted the differential proteomic studies involving malnourished mice.…”

“…amazonensis and L . major Western blot; Nano LC–MS/MS; LTQ-OrbitrapVelos MS Infected vs healthy 69 51 2019 [ 41 ] 7 L . infantum Flow Cytometry; ITRAQ; Nano LC–MS/MS Protein Malnourished infected vs well nutritioned un-infected 38 71 2019 [ 43 ] 8 L .…”

“…The RAS-related protein was up-regulated in this study as well as in the Menezes et al [ 39 ] study. The same group [ 41 ] performed a similar experiment with skin biopsies and reported 17 DMP in infected mice. Further, they [ 42 ] adopted the labeled (tandem mass tag) proteomic approach and confirmed their earlier findings.…”

How can proteomics overhaul our understanding of Leishmania biology?

“…Proteomic MS has enabled detection of changes in the host protein expression profiles in macrophage in vitro infection models [15,16], murine and human cutaneous lesions [17,18] and in the serum of infected individuals [19]. Proteomic MS studies have demonstrated that the Leishmania infection causes a species-specific reprogramming of major metabolic pathways in the host cell [16].…”