Diagnostic application of recombinant Leishmania proteins and evaluation of their in vitro immunogenicity after stimulation of immune cells collected from tegumentary leishmaniasis patients and healthy individuals

Lima, Mariana P.; Costa, Lourena E.; Lage, Daniela P.; Dias, Daniel Lucas Santos; Ribeiro, Patrícia A.F.; Machado, Amanda S.; Ramos, Fernanda F.; Salles, Beatriz C.S.; Fagundes, Mírian Ívens; Carvalho, Gerusa B.; Franklin, Michelle L.; Chávez-Fumagalli, Miguel Á.; Machado-de-Ávila, Ricardo Andrez; Menezes‐Souza, Daniel; Duarte, Mariana C.; Teixeira, Antônio Lúcio; Gonçalves, Denise Utsch; Coelho, Eduardo Antônio Ferraz

doi:10.1016/j.cellimm.2018.09.006

Cited by 13 publications

(9 citation statements)

References 47 publications

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“…They represent housekeeping proteins found in the cytoplasm and nucleus of parasites, being abundantly expressed in both promastigote and amastigote forms, and considered as important components of leishmanial flagella [24]. β-tubulins have been previously described as Th1 cell-stimulating agents; then demonstrating their potential to be evaluated as vaccine candidates against leishmaniasis [25,26].…”

Section: Discussionmentioning

confidence: 99%

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Costa

Alves

Carneiro

et al. 2019

IJMS

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Two Leishmania infantum mimotopes (B10 and C01) identified by phage display showed to be antigenic and immunogenic for visceral (VL) and tegumentary (TL) leishmaniasis; however, their biological targets in the parasites have not been identified. The aim of the present study was to investigate the native antigens expressing both mimotopes, and to use them in distinct immunological assays. For this, a subtractive phage display technology was used, where a combinatorial library of single-chain variable fragments (scFv) was employed and the most reactive monoclonal antibodies for each target were captured, being the target antigens identified by mass spectrometry. Results in immunoblotting and immunoprecipitation assays showed that both monoclonal scFvs antibodies identified the β-tubulin protein as the target antigen in L. infantum. To validate these findings, the recombinant protein was cloned, purified and tested for the serodiagnosis of human leishmaniasis, and its immunogenicity was evaluated in PBMC derived from healthy subjects and treated or untreated VL patients. Results showed high diagnostic efficacy, as well as the development of a specific Th1 immune response in the cell cultures, since higher IFN-γ and lower IL-10 production was found.

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Section: Discussionmentioning

confidence: 99%

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Costa

Alves

Carneiro

et al. 2019

IJMS

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“…This protein was shown also to be antigenic in previous immunoproteomics studies developed by our group, such as when L. infantum extracts were reacted against canine (Coelho et al ., 2012) and human (Lage et al ., 2019) VL sera, as well as when L. braziliensis antigenic preparations were reacted against TL patients sera (Duarte et al ., 2015). In addition, when β -tubulin was applied in a recombinant version in ELISA assays, it showed satisfactory diagnostic value to detect TL (Duarte et al ., 2015; Lima et al ., 2018) and VL (Costa et al ., 2019) cases.…”

Section: Discussionmentioning

confidence: 99%

An immunoproteomics approach to identifyLeishmania infantumproteins to be applied for the diagnosis of visceral leishmaniasis and human immunodeficiency virus co-infection

et al. 2020

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The co-infection between visceral leishmaniasis (VL) and human immunodeficiency virus (HIV) has increased in several countries in the world. The current serological tests are not suitable since they present low sensitivity to detect the most of VL/HIV cases, and a more precise diagnosis should be performed. In this context, in the present study, an immunoproteomics approach was performed using Leishmania infantum antigenic extracts and VL, HIV and VL/HIV patients sera, besides healthy subjects samples; aiming to identify antigenic markers for these clinical conditions. Results showed that 43 spots were recognized by antibodies in VL and VL/HIV sera, and 26 proteins were identified by mass spectrometry. Between them, β-tubulin was expressed, purified and tested in ELISA experiments as a proof of concept for validation of our immunoproteomics findings and results showed high sensitivity and specificity values to detect VL and VL/HIV patients. In conclusion, the identified proteins in the present work could be considered as candidates for future studies aiming to improvement of the diagnosis of VL and VL/HIV co-infection.

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“…This protein is a multifunctional metalloenzyme with the Enzyme Commission number E.C. code 4.2.1.11, which catalyzes the reversible dehydration of D-2-phosphoglycerate to phosphoenolpyruvate [ 24 ]; enolase as an immunogen has been capable of generating a Th1 immune response (considered fundamental for intracellular parasite elimination), Th2, or a mixture of both in response to the kind of microorganism being studied (intracellular or extracellular); therefore, there is no predominance of one of these two types of the immune response, both cell populations can be expressed [ 21 , 22 , 25 , 26 ]. Subsequent studies detected the presence of antibodies against the recombinant enolase from T. cruzi (anti-rTcENO antibodies) in sera from mice experimentally immunized with rTcENO, demonstrating that the purified recombinant enolase was recognized by mouse sera.…”

Section: Introductionmentioning

confidence: 99%

Consensus Enolase of Trypanosoma Cruzi: Evaluation of Their Immunogenic Properties Using a Bioinformatics Approach

Díaz-Hernández

González-Vázquez

Arce-Fonseca

et al. 2022

Life

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There is currently no vaccine against American trypanosomiasis, caused by the parasite Trypanosoma cruzi. This is due to the genomic variation observed in the six DTUs of T. cruzi. This work aims to propose a consensus sequence of the enolase protein from different strains of T. cruzi and mainly evaluate its immunogenic properties at the bioinformatic level. From specialized databases, 15 sequences of the enolase gene were aligned to obtain a consensus sequence, where this sequence was modeled and then evaluated and validated through different bioinformatic programs to learn their immunogenic potential. Finally, chimeric peptides were designed with the most representative epitopes. The results showed high immunogenic potential with six epitopes for MHC-I, and seven epitopes for MHC-II, all of which were highly representative of the enolase present in strains from the American continent as well as five epitopes for B cells. Regarding the computational modeling, molecular docking with Toll-like receptors showed a high affinity and low constant of dissociation, which could lead to an innate-type immune response that helps to eliminate the parasite. In conclusion, the consensus sequence proposed for enolase is capable of providing an ideal immune response; however, the experimental evaluation of this enolase consensus and their chimeric peptides should be a high priority to develop a vaccine against Chagas disease.

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Diagnostic application of recombinant Leishmania proteins and evaluation of their in vitro immunogenicity after stimulation of immune cells collected from tegumentary leishmaniasis patients and healthy individuals

Cited by 13 publications

References 47 publications

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

Leishmania infantum β-Tubulin Identified by Reverse Engineering Technology through Phage Display Applied as Theranostic Marker for Human Visceral Leishmaniasis

An immunoproteomics approach to identifyLeishmania infantumproteins to be applied for the diagnosis of visceral leishmaniasis and human immunodeficiency virus co-infection

Consensus Enolase of Trypanosoma Cruzi: Evaluation of Their Immunogenic Properties Using a Bioinformatics Approach

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