A recombinant cysteine proteinase from Leishmania (Leishmania) chagasi as an antigen for delayed-type hypersensitivity assays and serodiagnosis of canine visceral leishmaniasis

Pinheiro, Paulo Henrique da Costa; Pinheiro, Adriana Nunes; Ferreira, Josie Haydée Lima; Costa, Francisco Assis Lima; Katz, Simone; Barbiéri, Clara Lúcia

doi:10.1016/j.vetpar.2009.02.011

Cited by 23 publications

(20 citation statements)

References 34 publications

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“…In canine epidemiological screening, a test with high sensitivity and specificity is desirable. The performance observed for the synthetic antigens in this work is consistent with the performance of many antigens (mainly recombinant proteins) that have been developed in recent years for CVL diagnosis by ELISA (23,24), with the advantages of being cheaper, simpler, reproducible, useful for large-scale testing and, in most cases, more specific and sensitive (26). Our results show an improvement in the sensitivity of the synthetic antigen ELISA, compared to the indirect immunofluorescence assay, in asymptomatic dogs, due to the fact that the IFAT shows a lack of sensitivity for the detection of specific antibodies in asymptomatic dogs (20) and in healthy dogs, due to the large number of false-positive IFAT reactions (18).…”

Section: Discussionsupporting

confidence: 85%

“…and Trypanosoma cruzi parasites are superposed and, due to the phylogenetic similarity between those parasites, serological cross-reactions and/or false-positive results are quite common (21,22). As a strategy to develop a more sensitive and specific method for serodiagnosis of CVL, some individual Leishmania proteins were used as recombinant antigens (23,24). However, due to the high variability observed in the humoral responses of infected dogs, efficient diagnosis based on purified antigens might require a mixture of antigens or the use of chimeric antigens con-taining several leishmanial proteins (25).…”

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confidence: 99%

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Sensitive and Specific Serodiagnosis of Leishmania infantum Infection in Dogs by Using Peptides Selected from Hypothetical Proteins Identified by an Immunoproteomic Approach

Chávez‐Fumagalli

Martins

Testasicca

et al. 2013

Clin Vaccine Immunol

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In Brazil, the percentage of infected dogs living in areas where canine visceral leishmaniasis (CVL) is endemic ranges from 10 to 62%; however, the prevalence of infection in dogs is probably higher than figures reported from serological studies. In addition, problems with the occurrence of false-positive or false-negative results in the serodiagnosis of CVL have been reported. The present work analyzed the potential of synthetic peptides mapped from hypothetical proteins for improvement of the serodiagnosis of Leishmania infantum infection in dogs. From 26 identified leishmanial proteins, eight were selected, considering that no homologies between these proteins and others from trypanosomatide sequence databases were encountered. The sequences of these proteins were mapped to identify linear B-cell epitopes, and 17 peptides were synthesized and tested in enzyme-linked immunosorbent assays (ELISAs) for the serodiagnosis of L. infantum infection in dogs. Of these, three exhibited sensitivity and specificity values higher than 75% and 90%, respectively, to differentiate L. infantum-infected animals from Trypanosoma cruziinfected animals and healthy animals. Soluble Leishmania antigen (SLA) showed poor sensitivity (4%) and specificity (36%) to differentiate L. infantum-infected dogs from healthy and T. cruzi-infected dogs. Lastly, the three selected peptides were combined in different mixtures and higher sensitivity and specificity values were obtained, even when sera from T. cruzi-infected dogs were used. The study's findings suggest that these three peptides can constitute a potential tool for more sensitive and specific serodiagnosis of L. infantum infection in dogs.

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Section: Discussionsupporting

confidence: 85%

mentioning

confidence: 99%

Sensitive and Specific Serodiagnosis of Leishmania infantum Infection in Dogs by Using Peptides Selected from Hypothetical Proteins Identified by an Immunoproteomic Approach

Chávez‐Fumagalli

Martins

Testasicca

et al. 2013

Clin Vaccine Immunol

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“…To our knowledge, only one recently published study has used a recombinant Leishmania protein as a DTH test reagent (the recombinant cysteine proteinase rLdccys1) to detect CMI in vivo in naturally infected dogs, with very promising results. 39 Our study is the first to use recombinant proteins expressed in baculovirus-infected larvae for DTH testing. One limitation of other expression platforms is the high cost associated with large-scale production.…”

Section: Discussionmentioning

confidence: 99%

Humoral and In Vivo Cellular Immunity against the Raw Insect-Derived Recombinant Leishmania infantum Antigens KMPII, TRYP, LACK, and papLe22 in Dogs from an Endemic Area

Todolí

Solano‐Gallego²,

Juan³

et al. 2010

The American Society of Tropical Medicine and Hygiene

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Abstract. Leishmania infantum causes visceral leishmaniasis, a severe zoonotic and systemic disease that is fatal if left untreated. Identification of the antigens involved in Leishmania -specific protective immune response is a research priority for the development of effective control measures. For this purpose, we evaluated, in 27 dogs from an enzootic zone, specific humoral and cellular immune response by delayed-type hypersensitivity (DTH) skin test both against total L. infantum antigen and the raw Trichoplusia ni insect-derived kinetoplastid membrane protein-11 (rKMPII), tryparedoxin peroxidase (rTRYP), Leishmania homologue of receptors for activated C kinase (rLACK), and 22-kDa potentially aggravating protein of Leishmania (rpapLe22) antigens from this parasite. rTRYP induced the highest number of positive DTH responses (55% of leishmanin skin test [LST]-positive dogs), showing that TRYP antigen is an important T cell immunogen, and it could be a promising vaccine candidate against this disease. When TRYP-DTH and KMPII-DTH tests were evaluated in parallel, 82% of LST-positive dogs were detected, suggesting that both antigens could be considered as components of a standardized DTH immunodiagnostic tool for dogs.

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“…This response evolves erythematous or nodular injury resulting in a chronic inflammatory infiltrate in the dermis, composed mainly of macrophages and lymphocytes (20,21). The LST is a marker of effective cell-mediated immunity (CMI).…”

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confidence: 99%

Genome-Wide Association Study of Cell-Mediated Response in Dogs Naturally Infected by Leishmania infantum

et al. 2016

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A genome-wide association study (GWAS) could unravel the complexity of the cell-mediated immunity (CMI) to canine leishmaniasis (CanL). Therefore, we scanned 110,165 single-nucleotide polymorphisms (SNPs), aiming to identify chromosomal regions associated with the leishmanin skin test (LST), lymphocyte proliferation assay (LPA), and cytokine responses to further understand the role played by CMI in the outcome of natural Leishmania infantum infection in 189 dogs. Based on LST and LPA, four CMI profiles were identified (LST ؊ /LPA ؊ , LST ؉ /LPA ؊ , LST ؊ /LPA ؉ , and LST ؉ /LPA ؉ ), which were not associated with subclinically infected or diseased dogs. LST ؉ /LPA ؉ dogs showed increased interferon gamma (IFN-␥) and tumor necrosis factor alpha (TNF-␣) levels and mild parasitism in the lymph nodes, whereas LST ؊ /LPA ؉ dogs, in spite of increased IFN-␥, also showed increased interleukin-10 (IL-10) and transforming growth factor ␤ (TGF-␤) levels and the highest parasite load in lymph nodes. Low T cell proliferation under low parasite load suggested that L. infantum was not able to induce effective CMI in the early stage of infection. Altogether, genetic markers explained 87%, 16%, 15%, 11%, 0%, and 0% of phenotypic variance in TNF-␣, TGF-␤, LST, IL-10, IFN-␥, and LPA, respectively. GWAS showed that regions associated with TNF-␣ include the following genes: IL12RB1, JAK3, CCRL2, CCR2, CCR3, and CXCR6, involved in cytokine and chemokine signaling; regions associated with LST, including COMMD5 and SHARPIN, involved in regulation of NF-B signaling; and regions associated with IL-10, including LTBP1 and RASGRP3, involved in T regulatory lymphocytes differentiation. These findings pinpoint chromosomic regions related to the cell-mediated response that potentially affect the clinical complexity and the parasite replication in canine L. infantum infection.

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A recombinant cysteine proteinase from Leishmania (Leishmania) chagasi as an antigen for delayed-type hypersensitivity assays and serodiagnosis of canine visceral leishmaniasis

Cited by 23 publications

References 34 publications

Sensitive and Specific Serodiagnosis of Leishmania infantum Infection in Dogs by Using Peptides Selected from Hypothetical Proteins Identified by an Immunoproteomic Approach

Sensitive and Specific Serodiagnosis of Leishmania infantum Infection in Dogs by Using Peptides Selected from Hypothetical Proteins Identified by an Immunoproteomic Approach

Humoral and In Vivo Cellular Immunity against the Raw Insect-Derived Recombinant Leishmania infantum Antigens KMPII, TRYP, LACK, and papLe22 in Dogs from an Endemic Area

Genome-Wide Association Study of Cell-Mediated Response in Dogs Naturally Infected by Leishmania infantum

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