Leishmania infantum mimotopes and a phage–ELISA assay as tools for a sensitive and specific serodiagnosis of human visceral leishmaniasis

Salles, Beatriz C.S.; Costa, Lourena E.; Alves, Patrícia Terra; Dias, Ana C.S.; Vaz, Emília Rezende; Menezes‐Souza, Daniel; Ramos, Fernanda F.; Duarte, Mariana C.; Roatt, Bruno Mendes; Chávez-Fumagalli, Miguel Á.; Tavares, Carlos Alberto Pereira; Gonçalves, Denise Utsch; Rocha, Regina Lunardi; Goulart, Luíz Ricardo; Coelho, Eduardo Antônio Ferraz

doi:10.1016/j.diagmicrobio.2016.11.012

Cited by 26 publications

(19 citation statements)

References 57 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immunological methods have been also employed; however, problems related with the sensitivity and/or specificity of the antigens used in the tests are registered [11,12]. Thus, it is necessary to search for new candidates that will serve to design diagnostic systems with higher degree of sensitivity and specificity than current methods, such as the refinement of use of new antigens, such as recombinant proteins [13,14,15], synthetic peptides [16,17], polypeptide-based chimera [18,19] and phage clones [8,20].…”

Section: Introductionmentioning

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

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Introductionmentioning

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

Self Cite

View full text Add to dashboard Cite

show abstract

“…46 Peptide mimotopes have been shown to be able to mimic carbohydrate antigens, for example, by Umair et al 47 who used phage display to discover a peptide that mimics a glycan epitope on the surface of parasitic nematode larvae. Recently, by screening a phage display library with antibodies from VL patients, Salles et al 48 identified phages and showed their high diagnostic accuracy in ELISA with sera from VL patients and healthy controls from L. infantum –endemic regions. Following a similar approach but with antibodies from VL patients’ sera that specifically bind to the DAT antigen(s), one could potentially identify phages that mimic the DAT epitope(s), irrespective of their carbohydrate, lipid, or protein nature.…”

Section: Discussionmentioning

confidence: 99%

The Unknown Nature of the Antigen in the Direct Agglutination Test for Visceral Leishmaniasis Hampers Development of Serodiagnostic Tests

Kühne

Büscher

2019

The American Journal of Tropical Medicine and Hygiene

View full text Add to dashboard Cite

Abstract.Current diagnostic tests for visceral leishmaniasis (VL) are either not adapted for use in resource-poor settings or are insufficiently accurate in Eastern Africa. Only the direct agglutination test (DAT), based on whole Leishmania promastigotes, is highly reliable in all endemic regions, but its implementation is hampered by the need for a cold chain, minimal laboratory conditions, and long incubation times. Integrating the DAT antigen(s) in an immunochromatographic rapid diagnostic test (RDT) would overcome these disadvantages. Unfortunately, the identity of the DAT antigen(s) involved in the agglutination reaction is unknown. For this study, we reviewed all publications that might shed some light on this issue. We conclude that the DAT antigen is a mixture of Leishmania-specific epitopes of protein, carbohydrate, and lipid nature. To develop an accurate RDT for VL diagnosis in Eastern Africa, we suggest to complement the classical protein antigen discovery with approaches to identify carbohydrate and lipid epitopes.

show abstract

“…The control group (CT, n = 70) consisted of healthy individuals without clinical signs, suspicious or positive laboratory tests for VL. Samples from patients with Chagas’ disease (CD, n = 68) were confirmed by clinical evaluation associated with hemoculture in combination with specific ELISA or indirect hemagglutination assay (IHA) assays [34].…”

Section: Methodsmentioning

confidence: 99%

New antigens for the serological diagnosis of human visceral leishmaniasis identified by immunogenomic screening

Carvalho¹,

Mendes²,

Coelho³

et al. 2018

PLoS ONE

Self Cite

View full text Add to dashboard Cite

Visceral leishmaniasis (VL) still represents a serious public health problem in Brazil due to the inefficiency of the control measures currently employed, that included early diagnosis and treatment of human cases, vector control, euthanasia of infected dogs and, recently approved in Brazil, treatment with Milteforam drug. Effective clinical management depend largely on early and unequivocal diagnosis, however, cross-reactivity have also been described in serological tests, especially when it refers to individuals from areas where Chagas’ disease is also present. Thus, to discover new antigens to improve the current serological tests for VL diagnosis is urgently needed. Here, we performed an immunogenomic screen strategy to identify conserved linear B-cell epitopes in the predicted L. infantum proteome using the following criteria: i) proteins expressed in the stages found in the vertebrate host, amastigote stage, and secreted/excreted, to guarantee greater exposure to the immune system; ii) divergent from proteins present in other infectious disease pathogens with incidence in endemic areas for VL, as T. cruzi; iii) highly antigenic to humans with different genetic backgrounds, independently of the clinical stage of the disease; iv) stable and adaptable to quality-control tests to guarantee reproducibility; v) using statistical analysis to determine a suitable sample size to evaluate accuracy of diagnostic tests established by receiver operating characteristic strategy. We selected six predicted linear B-cell epitopes from three proteins of L. infantum parasite. The results demonstrated that a mixture of peptides (Mix IV: peptides 3+6) were able to identify VL cases and simultaneously able to discriminate infections caused by T. cruzi parasite with high accuracy (100.00%) and perfect agreement (Kappa index = 1.000) with direct methods performed by laboratories in Brazil. The results also demonstrated that peptide-6, Mix III (peptides 2+6) and I (peptides 2+3+6) are potential antigens able to used in VL diagnosis, represented by high accuracy (Ac = 99.52%, 99.52% and 98.56%, respectively). This study represents an interesting strategy for discovery new antigens applied to serologic diagnosis which will contribute to the improvement of the diagnosis of VL and, consequently, may help in the prevention, control and treatment of the disease in endemic areas of Brazil.

show abstract

Leishmania infantum mimotopes and a phage–ELISA assay as tools for a sensitive and specific serodiagnosis of human visceral leishmaniasis

Cited by 26 publications

References 57 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

The Unknown Nature of the Antigen in the Direct Agglutination Test for Visceral Leishmaniasis Hampers Development of Serodiagnostic Tests

New antigens for the serological diagnosis of human visceral leishmaniasis identified by immunogenomic screening

Contact Info

Product

Resources

About