rMELEISH: A Novel Recombinant Multiepitope-Based Protein Applied to the Serodiagnosis of Both Canine and Human Visceral Leishmaniasis

Abstract

Background: visceral leishmaniasis (VL) is a critical public health problem in over ninety countries. The control measures adopted in Brazil have been insufficient when it comes to preventing the spread of this overlooked disease. In this context, a precise diagnosis of VL in dogs and humans could help to reduce the number of cases of this disease. Distinct studies for the diagnosis of VL have used single recombinant proteins in serological assays; however, the results have been variable, mainly in relation to the sensitivity of the antigens. In this context, the development of multiepitope-based proteins could be relevant to solving such problem. Methods: a chimeric protein (rMELEISH) was constructed based on amino acid sequences from kinesin 39 (k39), alpha-tubulin, and heat-shock proteins HSP70 and HSP 83.1, and tested in enzyme-linked immunosorbent (ELISA) for the detection of L. infantum infection using canine (n = 140) and human (n = 145) sera samples. Results: in the trials, rMELEISH was able to discriminate between VL cases and cross-reactive diseases and healthy samples, with sensitivity and specificity values of 100%, as compared to the use of a soluble Leishmania antigenic extract (SLA). Conclusions: the preliminary data suggest that rMELEISH has the potential to be tested in future studies against a larger serological panel and in field conditions for the diagnosis of canine and human VL.

Immunotherapy Combining Mimotopes Selected by Phage Display Plus Amphotericin B Is Effective for Treatment Against Visceral Leishmaniasis

“…The treatment for VL causes toxicity in patients, entails high cost and/or leads to the emergence of resistant strains [23]. In addition, diagnosis is hampered by the variable sensitivity and specificity of the tests, and there is no human vaccine available [24,25]. New strategies have been searched to control the disease, and immunotherapy based on the association between the therapeutic agents and immunogens could represent a contribution in the field as it will be possible to reduce drug toxicity using lower concentrations and stimulate the long-lasting immune response by the use of prophylactic vaccines, protecting the host against reinfection by the parasites [26].…”

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis

“…infantum -positive serum samples/30 healthy human and 50 healthy dog serum samples/80 human and 45 dog serum samples from non- L . infantum diseases Human and canine diagnosis Sensitivity: 100% Specificity: 100% [ 4 ]/Brazil Human Chagas disease/ Trypanosoma cruzi rTC E . coli BL21(λDE3) pLysS cells ELISA 58 Chagas-positive serum samples/30 serum samples from healthy individuals/60 serum samples from non-Chagas diseases Sensitivity: 98.28% Specificity: 96.67% [ 5 ]/Brazil …”

“…Results derived from an ELISA assay demonstrated 93.1% sensitivity and 77.4% specificity. Next, Dias et al (2023) [ 4 ] conducted a study to evaluate the diagnostic efficiency of a new RMP, rMELEISH, for both human and canine visceral leishmaniasis, caused by L . infantum .…”

“…The CD technique has been applied in several studies to estimate the secondary structure profile of the RMPs in solution for application in diagnostic tests [ 2 , 4 , 19 , 107 , 114 ]. Typically, the RMPs are rationally designed containing solvent-exposed bound epitopes.…”

Design and evaluation of a novel multi-epitope antigen for evaluate the diagnostic immunity responses against Leishmania infantum infection

“…The high sensitivity and speci city of serological tests has been made possible by technological advances in recombinant antigens used as reagents for serological diagnosis of VL (Zhou, Chen et al 2022). The detection of VL has been made possible by a number of high-performance recombinant proteins, but more work is still required to nd the right antigens and achieve high-potency performance (Dias, Machado et al 2023).…”