Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine

Autores
de Oliveira, Bianca; Goes, Wanessa M.; Nascimento, Frederico C.; Carnielli, Juliana B. T.; Ferreira, Eden R.; de Carvalho, Alex Fiorini; dos Reis, Pablo Victor Mendes; Pereira, Milton; Ricotta, Tiago Queiroga Nery; dos Santos, Liliane Martins; de Souza, Renan Pedra; Cargnelutti, Diego Esteban; Mottram, Jeremy C.; Teixeira, Santuza R.; Fernandes, Ana Paula; Gazzinelli, Ricardo T.
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Human visceral leishmaniasis (HVL) is the second most lethal tropical parasitic disease. Currently, no prophylactic or therapeutic vaccines exist for HVL. Thus, the development of an efficacious vaccine is still needed. We previously performed an immunoproteomics analysis on Leishmania amazonensis parasite extracts to identify immunodominant antigens recognized by the sera of vaccinated and protected mice. Among the identified antigens, we discovered a novel, previously unstudied repetitive protein, initially annotated in Leishmania genomes as a kinetoplast-associated protein-like protein from Leishmania infantum (LinKAP), containing conserved domains (trichohyalin-plectin-homology [TPH] and TolA) that are associated with other mitochondrial proteins. LinKAP sequences are conserved across trypanosomatids, including Endotrypanum, Leishmania, and Trypanosoma species. Using differential centrifugation of Leishmania subcellular structures, we showed that LinKAP was enriched in fractions colocalizing with other mitochondrial proteins. mNeonGreen labeling at the endogenous locus using CRISPR-Cas9 and confocal microscopy confirmed that LinKAP is a mitochondrial-associated protein in Leishmania but not specifically colocalized with kDNA. We cloned and expressed a truncated version of LinKAP (rLinKAP), containing part (15) of the several LinKAP amino acid repeats, demonstrating over 85% homology across L. infantum, L. amazonensis, L. braziliensis, and L. mexicana species. An adjuvanted formulation of LinKAP with Poly ICLC, a polyinosinic-polycytidylic acid (Poly I:C) stabilized with carboxymethylcellulose and polylysine, was used to vaccinate mice and hamsters as a prophylactic vaccine for visceral leishmaniasis. Animals immunized with rLinKAP showed a potent cellular and humoral response and a significant decrease in tissue parasitism when challenged with L. infantum. We also tested rLinKAP as a therapeutic vaccine in mice. Following therapeutic vaccination, antibody responses were enhanced, and cellular responses became apparent. Our treatment protocol inhibited splenic parasite burden by 75% in treated mice. In conclusion, our antigen discovery strategy and the observed protective effect highlight rLinKAP as a promising vaccine candidate for leishmaniasis.
Fil: de Oliveira, Bianca. Universidade Federal de Minas Gerais; Brasil. Fundación Oswaldo Cruz; Brasil
Fil: Goes, Wanessa M.. Universidade Federal de Minas Gerais; Brasil
Fil: Nascimento, Frederico C.. Universidade Federal de Minas Gerais; Brasil
Fil: Carnielli, Juliana B. T.. University of York; Reino Unido
Fil: Ferreira, Eden R.. University of York; Reino Unido
Fil: de Carvalho, Alex Fiorini. Universidade Federal de Minas Gerais; Brasil
Fil: dos Reis, Pablo Victor Mendes. Universidade Federal de Minas Gerais; Brasil
Fil: Pereira, Milton. Universidade Federal de Minas Gerais; Brasil
Fil: Ricotta, Tiago Queiroga Nery. Universidade Federal de Minas Gerais; Brasil
Fil: dos Santos, Liliane Martins. Universidade Federal de Minas Gerais; Brasil
Fil: de Souza, Renan Pedra. Universidade Federal de Minas Gerais; Brasil
Fil: Cargnelutti, Diego Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina
Fil: Mottram, Jeremy C.. University of York; Reino Unido
Fil: Teixeira, Santuza R.. Universidade Federal de Minas Gerais; Brasil
Fil: Fernandes, Ana Paula. Universidade Federal de Minas Gerais; Brasil
Fil: Gazzinelli, Ricardo T.. Universidade Federal de Minas Gerais; Brasil
Materia
Adjuvants
Immune response
Leishmaniasis
Vaccines
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/291529
Acceder
id CONICETDig_7615067131dcd93f2e5b66aaf05f7911
oai_identifier_str oai:ri.conicet.gov.ar:11336/291529
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccinede Oliveira, BiancaGoes, Wanessa M.Nascimento, Frederico C.Carnielli, Juliana B. T.Ferreira, Eden R.de Carvalho, Alex Fiorinidos Reis, Pablo Victor MendesPereira, MiltonRicotta, Tiago Queiroga Nerydos Santos, Liliane Martinsde Souza, Renan PedraCargnelutti, Diego EstebanMottram, Jeremy C.Teixeira, Santuza R.Fernandes, Ana PaulaGazzinelli, Ricardo T.AdjuvantsImmune responseLeishmaniasisVaccineshttps://purl.org/becyt/ford/3.3https://purl.org/becyt/ford/3Human visceral leishmaniasis (HVL) is the second most lethal tropical parasitic disease. Currently, no prophylactic or therapeutic vaccines exist for HVL. Thus, the development of an efficacious vaccine is still needed. We previously performed an immunoproteomics analysis on Leishmania amazonensis parasite extracts to identify immunodominant antigens recognized by the sera of vaccinated and protected mice. Among the identified antigens, we discovered a novel, previously unstudied repetitive protein, initially annotated in Leishmania genomes as a kinetoplast-associated protein-like protein from Leishmania infantum (LinKAP), containing conserved domains (trichohyalin-plectin-homology [TPH] and TolA) that are associated with other mitochondrial proteins. LinKAP sequences are conserved across trypanosomatids, including Endotrypanum, Leishmania, and Trypanosoma species. Using differential centrifugation of Leishmania subcellular structures, we showed that LinKAP was enriched in fractions colocalizing with other mitochondrial proteins. mNeonGreen labeling at the endogenous locus using CRISPR-Cas9 and confocal microscopy confirmed that LinKAP is a mitochondrial-associated protein in Leishmania but not specifically colocalized with kDNA. We cloned and expressed a truncated version of LinKAP (rLinKAP), containing part (15) of the several LinKAP amino acid repeats, demonstrating over 85% homology across L. infantum, L. amazonensis, L. braziliensis, and L. mexicana species. An adjuvanted formulation of LinKAP with Poly ICLC, a polyinosinic-polycytidylic acid (Poly I:C) stabilized with carboxymethylcellulose and polylysine, was used to vaccinate mice and hamsters as a prophylactic vaccine for visceral leishmaniasis. Animals immunized with rLinKAP showed a potent cellular and humoral response and a significant decrease in tissue parasitism when challenged with L. infantum. We also tested rLinKAP as a therapeutic vaccine in mice. Following therapeutic vaccination, antibody responses were enhanced, and cellular responses became apparent. Our treatment protocol inhibited splenic parasite burden by 75% in treated mice. In conclusion, our antigen discovery strategy and the observed protective effect highlight rLinKAP as a promising vaccine candidate for leishmaniasis.Fil: de Oliveira, Bianca. Universidade Federal de Minas Gerais; Brasil. Fundación Oswaldo Cruz; BrasilFil: Goes, Wanessa M.. Universidade Federal de Minas Gerais; BrasilFil: Nascimento, Frederico C.. Universidade Federal de Minas Gerais; BrasilFil: Carnielli, Juliana B. T.. University of York; Reino UnidoFil: Ferreira, Eden R.. University of York; Reino UnidoFil: de Carvalho, Alex Fiorini. Universidade Federal de Minas Gerais; BrasilFil: dos Reis, Pablo Victor Mendes. Universidade Federal de Minas Gerais; BrasilFil: Pereira, Milton. Universidade Federal de Minas Gerais; BrasilFil: Ricotta, Tiago Queiroga Nery. Universidade Federal de Minas Gerais; BrasilFil: dos Santos, Liliane Martins. Universidade Federal de Minas Gerais; BrasilFil: de Souza, Renan Pedra. Universidade Federal de Minas Gerais; BrasilFil: Cargnelutti, Diego Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; ArgentinaFil: Mottram, Jeremy C.. University of York; Reino UnidoFil: Teixeira, Santuza R.. Universidade Federal de Minas Gerais; BrasilFil: Fernandes, Ana Paula. Universidade Federal de Minas Gerais; BrasilFil: Gazzinelli, Ricardo T.. Universidade Federal de Minas Gerais; BrasilAmerican Society for Microbiology2025-05info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/291529de Oliveira, Bianca; Goes, Wanessa M.; Nascimento, Frederico C.; Carnielli, Juliana B. T.; Ferreira, Eden R.; et al.; Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine; American Society for Microbiology; mSphere; 10; 5; 5-2025; 1-222379-5042CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://journals.asm.org/doi/10.1128/msphere.00097-25info:eu-repo/semantics/altIdentifier/doi/10.1128/msphere.00097-25info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-08-25T14:49:25Zoai:ri.conicet.gov.ar:11336/291529instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982026-08-25 14:49:25.829CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine
title Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine
spellingShingle Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine
de Oliveira, Bianca
Adjuvants
Immune response
Leishmaniasis
Vaccines
title_short Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine
title_full Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine
title_fullStr Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine
title_full_unstemmed Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine
title_sort Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine
dc.creator.none.fl_str_mv de Oliveira, Bianca
Goes, Wanessa M.
Nascimento, Frederico C.
Carnielli, Juliana B. T.
Ferreira, Eden R.
de Carvalho, Alex Fiorini
dos Reis, Pablo Victor Mendes
Pereira, Milton
Ricotta, Tiago Queiroga Nery
dos Santos, Liliane Martins
de Souza, Renan Pedra
Cargnelutti, Diego Esteban
Mottram, Jeremy C.
Teixeira, Santuza R.
Fernandes, Ana Paula
Gazzinelli, Ricardo T.
author de Oliveira, Bianca
author_facet de Oliveira, Bianca
Goes, Wanessa M.
Nascimento, Frederico C.
Carnielli, Juliana B. T.
Ferreira, Eden R.
de Carvalho, Alex Fiorini
dos Reis, Pablo Victor Mendes
Pereira, Milton
Ricotta, Tiago Queiroga Nery
dos Santos, Liliane Martins
de Souza, Renan Pedra
Cargnelutti, Diego Esteban
Mottram, Jeremy C.
Teixeira, Santuza R.
Fernandes, Ana Paula
Gazzinelli, Ricardo T.
author_role author
author2 Goes, Wanessa M.
Nascimento, Frederico C.
Carnielli, Juliana B. T.
Ferreira, Eden R.
de Carvalho, Alex Fiorini
dos Reis, Pablo Victor Mendes
Pereira, Milton
Ricotta, Tiago Queiroga Nery
dos Santos, Liliane Martins
de Souza, Renan Pedra
Cargnelutti, Diego Esteban
Mottram, Jeremy C.
Teixeira, Santuza R.
Fernandes, Ana Paula
Gazzinelli, Ricardo T.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Adjuvants
Immune response
Leishmaniasis
Vaccines
topic Adjuvants
Immune response
Leishmaniasis
Vaccines
purl_subject.fl_str_mv https://purl.org/becyt/ford/3.3
https://purl.org/becyt/ford/3
dc.description.none.fl_txt_mv Human visceral leishmaniasis (HVL) is the second most lethal tropical parasitic disease. Currently, no prophylactic or therapeutic vaccines exist for HVL. Thus, the development of an efficacious vaccine is still needed. We previously performed an immunoproteomics analysis on Leishmania amazonensis parasite extracts to identify immunodominant antigens recognized by the sera of vaccinated and protected mice. Among the identified antigens, we discovered a novel, previously unstudied repetitive protein, initially annotated in Leishmania genomes as a kinetoplast-associated protein-like protein from Leishmania infantum (LinKAP), containing conserved domains (trichohyalin-plectin-homology [TPH] and TolA) that are associated with other mitochondrial proteins. LinKAP sequences are conserved across trypanosomatids, including Endotrypanum, Leishmania, and Trypanosoma species. Using differential centrifugation of Leishmania subcellular structures, we showed that LinKAP was enriched in fractions colocalizing with other mitochondrial proteins. mNeonGreen labeling at the endogenous locus using CRISPR-Cas9 and confocal microscopy confirmed that LinKAP is a mitochondrial-associated protein in Leishmania but not specifically colocalized with kDNA. We cloned and expressed a truncated version of LinKAP (rLinKAP), containing part (15) of the several LinKAP amino acid repeats, demonstrating over 85% homology across L. infantum, L. amazonensis, L. braziliensis, and L. mexicana species. An adjuvanted formulation of LinKAP with Poly ICLC, a polyinosinic-polycytidylic acid (Poly I:C) stabilized with carboxymethylcellulose and polylysine, was used to vaccinate mice and hamsters as a prophylactic vaccine for visceral leishmaniasis. Animals immunized with rLinKAP showed a potent cellular and humoral response and a significant decrease in tissue parasitism when challenged with L. infantum. We also tested rLinKAP as a therapeutic vaccine in mice. Following therapeutic vaccination, antibody responses were enhanced, and cellular responses became apparent. Our treatment protocol inhibited splenic parasite burden by 75% in treated mice. In conclusion, our antigen discovery strategy and the observed protective effect highlight rLinKAP as a promising vaccine candidate for leishmaniasis.
Fil: de Oliveira, Bianca. Universidade Federal de Minas Gerais; Brasil. Fundación Oswaldo Cruz; Brasil
Fil: Goes, Wanessa M.. Universidade Federal de Minas Gerais; Brasil
Fil: Nascimento, Frederico C.. Universidade Federal de Minas Gerais; Brasil
Fil: Carnielli, Juliana B. T.. University of York; Reino Unido
Fil: Ferreira, Eden R.. University of York; Reino Unido
Fil: de Carvalho, Alex Fiorini. Universidade Federal de Minas Gerais; Brasil
Fil: dos Reis, Pablo Victor Mendes. Universidade Federal de Minas Gerais; Brasil
Fil: Pereira, Milton. Universidade Federal de Minas Gerais; Brasil
Fil: Ricotta, Tiago Queiroga Nery. Universidade Federal de Minas Gerais; Brasil
Fil: dos Santos, Liliane Martins. Universidade Federal de Minas Gerais; Brasil
Fil: de Souza, Renan Pedra. Universidade Federal de Minas Gerais; Brasil
Fil: Cargnelutti, Diego Esteban. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mendoza. Instituto de Medicina y Biología Experimental de Cuyo; Argentina
Fil: Mottram, Jeremy C.. University of York; Reino Unido
Fil: Teixeira, Santuza R.. Universidade Federal de Minas Gerais; Brasil
Fil: Fernandes, Ana Paula. Universidade Federal de Minas Gerais; Brasil
Fil: Gazzinelli, Ricardo T.. Universidade Federal de Minas Gerais; Brasil
description Human visceral leishmaniasis (HVL) is the second most lethal tropical parasitic disease. Currently, no prophylactic or therapeutic vaccines exist for HVL. Thus, the development of an efficacious vaccine is still needed. We previously performed an immunoproteomics analysis on Leishmania amazonensis parasite extracts to identify immunodominant antigens recognized by the sera of vaccinated and protected mice. Among the identified antigens, we discovered a novel, previously unstudied repetitive protein, initially annotated in Leishmania genomes as a kinetoplast-associated protein-like protein from Leishmania infantum (LinKAP), containing conserved domains (trichohyalin-plectin-homology [TPH] and TolA) that are associated with other mitochondrial proteins. LinKAP sequences are conserved across trypanosomatids, including Endotrypanum, Leishmania, and Trypanosoma species. Using differential centrifugation of Leishmania subcellular structures, we showed that LinKAP was enriched in fractions colocalizing with other mitochondrial proteins. mNeonGreen labeling at the endogenous locus using CRISPR-Cas9 and confocal microscopy confirmed that LinKAP is a mitochondrial-associated protein in Leishmania but not specifically colocalized with kDNA. We cloned and expressed a truncated version of LinKAP (rLinKAP), containing part (15) of the several LinKAP amino acid repeats, demonstrating over 85% homology across L. infantum, L. amazonensis, L. braziliensis, and L. mexicana species. An adjuvanted formulation of LinKAP with Poly ICLC, a polyinosinic-polycytidylic acid (Poly I:C) stabilized with carboxymethylcellulose and polylysine, was used to vaccinate mice and hamsters as a prophylactic vaccine for visceral leishmaniasis. Animals immunized with rLinKAP showed a potent cellular and humoral response and a significant decrease in tissue parasitism when challenged with L. infantum. We also tested rLinKAP as a therapeutic vaccine in mice. Following therapeutic vaccination, antibody responses were enhanced, and cellular responses became apparent. Our treatment protocol inhibited splenic parasite burden by 75% in treated mice. In conclusion, our antigen discovery strategy and the observed protective effect highlight rLinKAP as a promising vaccine candidate for leishmaniasis.
publishDate 2025
dc.date.none.fl_str_mv 2025-05
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/291529
de Oliveira, Bianca; Goes, Wanessa M.; Nascimento, Frederico C.; Carnielli, Juliana B. T.; Ferreira, Eden R.; et al.; Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine; American Society for Microbiology; mSphere; 10; 5; 5-2025; 1-22
2379-5042
CONICET Digital
CONICET
url http://hdl.handle.net/11336/291529
identifier_str_mv de Oliveira, Bianca; Goes, Wanessa M.; Nascimento, Frederico C.; Carnielli, Juliana B. T.; Ferreira, Eden R.; et al.; Characterization of a novel Leishmania antigen containing a repetitive domain and its potential use as a prophylactic and therapeutic vaccine; American Society for Microbiology; mSphere; 10; 5; 5-2025; 1-22
2379-5042
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://journals.asm.org/doi/10.1128/msphere.00097-25
info:eu-repo/semantics/altIdentifier/doi/10.1128/msphere.00097-25
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv American Society for Microbiology
publisher.none.fl_str_mv American Society for Microbiology
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.24418

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