Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia
ABSTRACT: Insect saliva induces significant antibody responses associated with the intensity of exposure to bites and the risk of disease in humans. Several salivary biomarkers have been characterized to determine exposure intensity to Old World Anopheles mosquito species. However, new tools are nee...
- Autores:
-
Vásquez Cardona, Ana María
Tobón Castaño, Alberto
Londoño Rentería, Berlín Luxelly
Montiel Ramos, Jehidys
Taylor, Marissa
Lenhart, Audrey
Drame, Papa M.
Vizcaíno, Lucrecia
- Tipo de recurso:
- Article of investigation
- Fecha de publicación:
- 2020
- Institución:
- Universidad de Antioquia
- Repositorio:
- Repositorio UdeA
- Idioma:
- eng
- OAI Identifier:
- oai:bibliotecadigital.udea.edu.co:10495/42068
- Acceso en línea:
- https://hdl.handle.net/10495/42068
- Palabra clave:
- Anopheles
Formación de Anticuerpos
Antibody Formation
Antígenos
Antígens
Inmunoglobulina G
Immunoglobulin G
Mosquitos Vectores
Mosquito Vectors
Malaria
Proteínas y Péptidos Salivales
Salivary Proteins and Peptides
https://id.nlm.nih.gov/mesh/D000852
https://id.nlm.nih.gov/mesh/D000917
https://id.nlm.nih.gov/mesh/D000941
https://id.nlm.nih.gov/mesh/D007074
https://id.nlm.nih.gov/mesh/D000072138
https://id.nlm.nih.gov/mesh/D008288
https://id.nlm.nih.gov/mesh/D012471
- Rights
- openAccess
- License
- https://creativecommons.org/licenses/by/4.0/
| id |
UDEA2_0905d2e98770ec4d22e97beaab014069 |
|---|---|
| oai_identifier_str |
oai:bibliotecadigital.udea.edu.co:10495/42068 |
| network_acronym_str |
UDEA2 |
| network_name_str |
Repositorio UdeA |
| repository_id_str |
|
| dc.title.spa.fl_str_mv |
Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia |
| title |
Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia |
| spellingShingle |
Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia Anopheles Formación de Anticuerpos Antibody Formation Antígenos Antígens Inmunoglobulina G Immunoglobulin G Mosquitos Vectores Mosquito Vectors Malaria Proteínas y Péptidos Salivales Salivary Proteins and Peptides https://id.nlm.nih.gov/mesh/D000852 https://id.nlm.nih.gov/mesh/D000917 https://id.nlm.nih.gov/mesh/D000941 https://id.nlm.nih.gov/mesh/D007074 https://id.nlm.nih.gov/mesh/D000072138 https://id.nlm.nih.gov/mesh/D008288 https://id.nlm.nih.gov/mesh/D012471 |
| title_short |
Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia |
| title_full |
Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia |
| title_fullStr |
Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia |
| title_full_unstemmed |
Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia |
| title_sort |
Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia |
| dc.creator.fl_str_mv |
Vásquez Cardona, Ana María Tobón Castaño, Alberto Londoño Rentería, Berlín Luxelly Montiel Ramos, Jehidys Taylor, Marissa Lenhart, Audrey Drame, Papa M. Vizcaíno, Lucrecia |
| dc.contributor.author.none.fl_str_mv |
Vásquez Cardona, Ana María Tobón Castaño, Alberto Londoño Rentería, Berlín Luxelly Montiel Ramos, Jehidys Taylor, Marissa Lenhart, Audrey Drame, Papa M. Vizcaíno, Lucrecia |
| dc.contributor.researchgroup.spa.fl_str_mv |
Grupo Malaria |
| dc.subject.decs.none.fl_str_mv |
Anopheles Formación de Anticuerpos Antibody Formation Antígenos Antígens Inmunoglobulina G Immunoglobulin G Mosquitos Vectores Mosquito Vectors Malaria Proteínas y Péptidos Salivales Salivary Proteins and Peptides |
| topic |
Anopheles Formación de Anticuerpos Antibody Formation Antígenos Antígens Inmunoglobulina G Immunoglobulin G Mosquitos Vectores Mosquito Vectors Malaria Proteínas y Péptidos Salivales Salivary Proteins and Peptides https://id.nlm.nih.gov/mesh/D000852 https://id.nlm.nih.gov/mesh/D000917 https://id.nlm.nih.gov/mesh/D000941 https://id.nlm.nih.gov/mesh/D007074 https://id.nlm.nih.gov/mesh/D000072138 https://id.nlm.nih.gov/mesh/D008288 https://id.nlm.nih.gov/mesh/D012471 |
| dc.subject.meshuri.none.fl_str_mv |
https://id.nlm.nih.gov/mesh/D000852 https://id.nlm.nih.gov/mesh/D000917 https://id.nlm.nih.gov/mesh/D000941 https://id.nlm.nih.gov/mesh/D007074 https://id.nlm.nih.gov/mesh/D000072138 https://id.nlm.nih.gov/mesh/D008288 https://id.nlm.nih.gov/mesh/D012471 |
| description |
ABSTRACT: Insect saliva induces significant antibody responses associated with the intensity of exposure to bites and the risk of disease in humans. Several salivary biomarkers have been characterized to determine exposure intensity to Old World Anopheles mosquito species. However, new tools are needed to quantify the intensity of human exposure to Anopheles bites and understand the risk of malaria in low-transmission areas in the Americas. To address this need, we conducted proteomic and bioinformatic analyses of immunogenic candidate proteins present in the saliva of uninfected Anopheles albimanus from two separate colonies-one originating from Central America (STECLA strain) and one originating from South America (Cartagena strain). A ~65 kDa band was identified by IgG antibodies in serum samples from healthy volunteers living in a malaria endemic area in Colombia, and a total of five peptides were designed from the sequences of two immunogenic candidate proteins that were shared by both strains. ELISA-based testing of human IgG antibody levels against the peptides revealed that the transferrin-derived peptides, TRANS-P1, TRANS-P2 and a salivary peroxidase peptide (PEROX-P3) were able to distinguish between malaria-infected and uninfected groups. Interestingly, IgG antibody levels against PEROX-P3 were significantly lower in people that have never experienced malaria, suggesting that it may be a good marker for mosquito bite exposure in naïve populations such as travelers and deployed military personnel. In addition, the strength of the differences in the IgG levels against the peptides varied according to location, suggesting that the peptides may able to detect differences in intensities of bite exposure according to the mosquito population density. Thus, the An. albimanus salivary peptides TRANS-P1, TRANS-P2, and PEROX-P3 are promising biomarkers that could be exploited in a quantitative immunoassay for determination of human-vector contact and calculation of disease risk. Keywords: Anopheles albimanus; antibodies; salivary gland proteins. |
| publishDate |
2020 |
| dc.date.issued.none.fl_str_mv |
2020 |
| dc.date.accessioned.none.fl_str_mv |
2024-09-12T17:58:59Z |
| dc.date.available.none.fl_str_mv |
2024-09-12T17:58:59Z |
| dc.type.spa.fl_str_mv |
Artículo de investigación |
| dc.type.coar.spa.fl_str_mv |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| dc.type.redcol.spa.fl_str_mv |
https://purl.org/redcol/resource_type/ART |
| dc.type.coarversion.spa.fl_str_mv |
http://purl.org/coar/version/c_970fb48d4fbd8a85 |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/article |
| dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
| format |
http://purl.org/coar/resource_type/c_2df8fbb1 |
| status_str |
publishedVersion |
| dc.identifier.citation.spa.fl_str_mv |
Londoño-Renteria B, Drame PM, Montiel J, Vasquez AM, Tobón-Castaño A, Taylor M, Vizcaino L, Lenhart AAE. Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia. Int J Mol Sci. 2020 Jan 21;21(3):691. doi: 10.3390/ijms21030691. |
| dc.identifier.issn.none.fl_str_mv |
1661-6596 |
| dc.identifier.uri.none.fl_str_mv |
https://hdl.handle.net/10495/42068 |
| dc.identifier.doi.none.fl_str_mv |
10.3390/ijms21030691 |
| dc.identifier.eissn.none.fl_str_mv |
1422-0067 |
| identifier_str_mv |
Londoño-Renteria B, Drame PM, Montiel J, Vasquez AM, Tobón-Castaño A, Taylor M, Vizcaino L, Lenhart AAE. Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia. Int J Mol Sci. 2020 Jan 21;21(3):691. doi: 10.3390/ijms21030691. 1661-6596 10.3390/ijms21030691 1422-0067 |
| url |
https://hdl.handle.net/10495/42068 |
| dc.language.iso.spa.fl_str_mv |
eng |
| language |
eng |
| dc.relation.ispartofjournalabbrev.spa.fl_str_mv |
Int. J. Mol. Sci. |
| dc.relation.citationendpage.spa.fl_str_mv |
15 |
| dc.relation.citationissue.spa.fl_str_mv |
3 |
| dc.relation.citationstartpage.spa.fl_str_mv |
1 |
| dc.relation.citationvolume.spa.fl_str_mv |
21 |
| dc.relation.ispartofjournal.spa.fl_str_mv |
International Journal of Molecular Sciences |
| dc.rights.uri.spa.fl_str_mv |
https://creativecommons.org/licenses/by/4.0/ |
| dc.rights.uri.*.fl_str_mv |
http://creativecommons.org/licenses/by/2.5/co/ |
| dc.rights.accessrights.spa.fl_str_mv |
info:eu-repo/semantics/openAccess |
| dc.rights.coar.spa.fl_str_mv |
http://purl.org/coar/access_right/c_abf2 |
| rights_invalid_str_mv |
https://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/2.5/co/ http://purl.org/coar/access_right/c_abf2 |
| eu_rights_str_mv |
openAccess |
| dc.format.extent.spa.fl_str_mv |
15 páginas |
| dc.format.mimetype.spa.fl_str_mv |
application/pdf |
| dc.publisher.spa.fl_str_mv |
MDPI |
| dc.publisher.place.spa.fl_str_mv |
Basilea, Suiza |
| institution |
Universidad de Antioquia |
| bitstream.url.fl_str_mv |
https://bibliotecadigital.udea.edu.co/bitstreams/d4aa6f6f-b831-4214-86a7-50627141a6ba/download https://bibliotecadigital.udea.edu.co/bitstreams/e454b6ed-3b95-43ca-a066-a4aab03da4e4/download https://bibliotecadigital.udea.edu.co/bitstreams/b7010443-8027-4525-9b71-9a8185dd167a/download https://bibliotecadigital.udea.edu.co/bitstreams/40b1cc13-71b3-4c6e-bfa7-14a6ff2de1d3/download https://bibliotecadigital.udea.edu.co/bitstreams/8f6d6330-7b51-433b-ad58-9f6352034b83/download |
| bitstream.checksum.fl_str_mv |
1646d1f6b96dbbbc38035efc9239ac9c 8a4605be74aa9ea9d79846c1fba20a33 90072400077137f3caca81023d229c6b 9ab3e3eaf973e80317363ca98adb5c16 b19c2cb3925cfcef3d28623601ff1ed4 |
| bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 |
| repository.name.fl_str_mv |
Repositorio Institucional de la Universidad de Antioquia |
| repository.mail.fl_str_mv |
aplicacionbibliotecadigitalbiblioteca@udea.edu.co |
| _version_ |
1851052242184962048 |
| spelling |
Vásquez Cardona, Ana MaríaTobón Castaño, AlbertoLondoño Rentería, Berlín LuxellyMontiel Ramos, JehidysTaylor, MarissaLenhart, AudreyDrame, Papa M.Vizcaíno, LucreciaGrupo Malaria2024-09-12T17:58:59Z2024-09-12T17:58:59Z2020Londoño-Renteria B, Drame PM, Montiel J, Vasquez AM, Tobón-Castaño A, Taylor M, Vizcaino L, Lenhart AAE. Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in Colombia. Int J Mol Sci. 2020 Jan 21;21(3):691. doi: 10.3390/ijms21030691.1661-6596https://hdl.handle.net/10495/4206810.3390/ijms210306911422-0067ABSTRACT: Insect saliva induces significant antibody responses associated with the intensity of exposure to bites and the risk of disease in humans. Several salivary biomarkers have been characterized to determine exposure intensity to Old World Anopheles mosquito species. However, new tools are needed to quantify the intensity of human exposure to Anopheles bites and understand the risk of malaria in low-transmission areas in the Americas. To address this need, we conducted proteomic and bioinformatic analyses of immunogenic candidate proteins present in the saliva of uninfected Anopheles albimanus from two separate colonies-one originating from Central America (STECLA strain) and one originating from South America (Cartagena strain). A ~65 kDa band was identified by IgG antibodies in serum samples from healthy volunteers living in a malaria endemic area in Colombia, and a total of five peptides were designed from the sequences of two immunogenic candidate proteins that were shared by both strains. ELISA-based testing of human IgG antibody levels against the peptides revealed that the transferrin-derived peptides, TRANS-P1, TRANS-P2 and a salivary peroxidase peptide (PEROX-P3) were able to distinguish between malaria-infected and uninfected groups. Interestingly, IgG antibody levels against PEROX-P3 were significantly lower in people that have never experienced malaria, suggesting that it may be a good marker for mosquito bite exposure in naïve populations such as travelers and deployed military personnel. In addition, the strength of the differences in the IgG levels against the peptides varied according to location, suggesting that the peptides may able to detect differences in intensities of bite exposure according to the mosquito population density. Thus, the An. albimanus salivary peptides TRANS-P1, TRANS-P2, and PEROX-P3 are promising biomarkers that could be exploited in a quantitative immunoassay for determination of human-vector contact and calculation of disease risk. Keywords: Anopheles albimanus; antibodies; salivary gland proteins.Universidad de AntioquiaColombia. Ministerio de Ciencia, Tecnología e Innovación - MinCienciasNational Institute of Food and AgricultureCOL000752415 páginasapplication/pdfengMDPIBasilea, Suizahttps://creativecommons.org/licenses/by/4.0/http://creativecommons.org/licenses/by/2.5/co/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Identification and Pilot Evaluation of Salivary Peptides from Anopheles albimanus as Biomarkers for Bite Exposure and Malaria Infection in ColombiaArtículo de investigaciónhttp://purl.org/coar/resource_type/c_2df8fbb1https://purl.org/redcol/resource_type/ARThttp://purl.org/coar/version/c_970fb48d4fbd8a85info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionAnophelesFormación de AnticuerposAntibody FormationAntígenosAntígensInmunoglobulina GImmunoglobulin GMosquitos VectoresMosquito VectorsMalariaProteínas y Péptidos SalivalesSalivary Proteins and Peptideshttps://id.nlm.nih.gov/mesh/D000852https://id.nlm.nih.gov/mesh/D000917https://id.nlm.nih.gov/mesh/D000941https://id.nlm.nih.gov/mesh/D007074https://id.nlm.nih.gov/mesh/D000072138https://id.nlm.nih.gov/mesh/D008288https://id.nlm.nih.gov/mesh/D012471Int. J. Mol. Sci.153121International Journal of Molecular SciencesUdeA 2015-7765MinCiencias RC-075-2016, Code 1115712498201014798 CDC-USAID grant # 200-2018-M-00025RoR:03bp5hc83RoR:03fd5ne08RoR:05qx3fv49PublicationCC-LICENSElicense_rdflicense_rdfapplication/rdf+xml; charset=utf-8927https://bibliotecadigital.udea.edu.co/bitstreams/d4aa6f6f-b831-4214-86a7-50627141a6ba/download1646d1f6b96dbbbc38035efc9239ac9cMD52falseAnonymousREADLICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://bibliotecadigital.udea.edu.co/bitstreams/e454b6ed-3b95-43ca-a066-a4aab03da4e4/download8a4605be74aa9ea9d79846c1fba20a33MD53falseAnonymousREADORIGINALTobonAlberto_2020_Salivary_Peptides_Anopheles.pdfTobonAlberto_2020_Salivary_Peptides_Anopheles.pdfArtículo de investigaciónapplication/pdf1938328https://bibliotecadigital.udea.edu.co/bitstreams/b7010443-8027-4525-9b71-9a8185dd167a/download90072400077137f3caca81023d229c6bMD51trueAnonymousREADTEXTTobonAlberto_2020_Salivary_Peptides_Anopheles.pdf.txtTobonAlberto_2020_Salivary_Peptides_Anopheles.pdf.txtExtracted texttext/plain70832https://bibliotecadigital.udea.edu.co/bitstreams/40b1cc13-71b3-4c6e-bfa7-14a6ff2de1d3/download9ab3e3eaf973e80317363ca98adb5c16MD54falseAnonymousREADTHUMBNAILTobonAlberto_2020_Salivary_Peptides_Anopheles.pdf.jpgTobonAlberto_2020_Salivary_Peptides_Anopheles.pdf.jpgGenerated Thumbnailimage/jpeg15055https://bibliotecadigital.udea.edu.co/bitstreams/8f6d6330-7b51-433b-ad58-9f6352034b83/downloadb19c2cb3925cfcef3d28623601ff1ed4MD55falseAnonymousREAD10495/42068oai:bibliotecadigital.udea.edu.co:10495/420682025-03-26 19:10:28.61https://creativecommons.org/licenses/by/4.0/open.accesshttps://bibliotecadigital.udea.edu.coRepositorio Institucional de la Universidad de Antioquiaaplicacionbibliotecadigitalbiblioteca@udea.edu.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 |
