Recombinant Leishmania (Leishmania) infantum Ecto-Nucleoside Triphosphate Diphosphohydrolase NTPDase-2 as a new antigen in canine visceral leishmaniasis diagnosis

Abstract

Canine visceral leishmaniasis is an important public health concern. In the epidemiological context of human visceral leishmaniasis, dogs are considered the main reservoir of Leishmania parasites; therefore, dogs must be epidemiologically monitored constantly in endemic areas. Furthermore, dog to human transmission has been correlated with emerging urbanization and increasing rates of leishmaniasis infection worldwide. Leishmania (Leishmania) infantum (L. chagasi) is the etiologic agent of visceral leishmaniasis in the New World. In this work, a new L. (L.) infantum (L. chagasi) recombinant antigen, named ATP diphosphohydrolase (rLic-NTPDase-2), intended for use in the immunodiagnosis of CVL was produced and validated. The extracellular domain of ATP diphosphohydrolase was cloned and expressed in the pET21b-Escherichia coli expression system. Indirect ELISA assays were used to detect the purified rLic-NTPDase-2 antigen using a standard canine sera library. This library contained CVL-positive samples, leishmaniasis-negative samples and samples from Trypanosoma cruzi-infected dogs. The results show a high sensitivity of 100% (95% CI=92.60-100.0%) and a high specificity of 100% (95% CI=86.77-100.0%), with a high degree of confidence (k=1). These findings demonstrate the potential use of this recombinant protein in immune diagnosis of canine leishmaniasis and open the possibility of its application to other diagnostic approaches, such as immunochromatography fast lateral flow assays and human leishmaniasis diagnosis.

NTPDase activities: possible roles on Leishmania spp infectivity and virulence

“…These data showed for the first time that LicNTPDases are expressed in the tissues of naturally infected dogs, which indicates the importance of E‐NTPDase in the natural infection. The results also corroborated the previous findings (de Souza et al, ), which demonstrated the potential application of rLicNTPDase‐2 in investigating the presence of amastigotes in infected dogs (Vasconcellos et al, ).…”

The Antileishmanial Potential of C-3 Functionalized Isobenzofuranones against Leishmania (Leishmania) Infantum Chagasi

“…These studies have sought new methods and targets for diagnosis, new vaccine candidates and new rationally designed drugs that can be applied not only in humans, but also in dogs because canines are considered the major reservoirs of several species of Leishmania in the home environment and outdoors [6,7].…”

The Leishmania-Macrophage Interactions: Role of E-NTPDases and Purinergic Signaling

“…In this work, anti-E-NTPDase2 antibodies were detected in 100% of the dogs with CVL from an endemic region of Brazil, regardless of the stage of the disease (asymptomatic, oligosymptomatic or polisymptomatic) [48] . This work has shown the expression of E-NTPDase2 on this parasite and highlighted its antigenicity.…”

“…The isoform called as E-NTPDase-1 (approximately 70 kDa) is also known as guanosine diphosphatase and is more similar to ENTPDase-1 from T. cruzi that was described for the first time by our research group [40] . Unfortunately, the literature on this topic is not uniform and other research groups have named these proteins differently, e.g., E-NTPDase-2 (approximately 40 kDa) is also called as ATPdiphosphohydrolase or nucleoside diphosphatase [48] . In order to stimulate a uniformity in the parasite E-NTPDase nomenclature, we have performed phylogenetic and bioinformatic analyses and proposed the following uniform nomenclature: TpNTPDase-1 and TpNTPDase-2 (Tp = trypanosomatids NTPDases; isoform 1 is approximately 70 kDa and isoform 2 is 40 kDa) [46] .…”

“…The GP63, a zinc-metalloprotease named glycoprotein 63 (GP63), is another critical virulence factor that has been fully explored [34][35][36][37][38] . However, the E-NTPDases (ecto-nucleoside triphosphate diphosphohydrolases), while less-extensively studied, have been demonstrated and believed to be important infectivity and virulence factors [39][40][41][42][43][44][45][46][47][48] .…”