Identification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays

ABSTRACT Immunoassays are currently needed to quantify Loa loa microfilariae (mf). To address this need, we have conducted proteomic and bioinformatic analyses of proteins present in the urine of a Loa mf-infected patient and used this information to identify putative biomarkers produced by L. loa m...

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Autores principales:	Papa M. Drame, Zhaojing Meng, Sasisekhar Bennuru, Jesica A. Herrick, Timothy D. Veenstra, Thomas B. Nutman
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/85a2e55e0d4c4d7e9c2b4b8de7c856f4
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spelling	oai:doaj.org-article:85a2e55e0d4c4d7e9c2b4b8de7c856f42021-11-15T15:49:39ZIdentification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays10.1128/mBio.02132-152150-7511https://doaj.org/article/85a2e55e0d4c4d7e9c2b4b8de7c856f42016-03-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.02132-15https://doaj.org/toc/2150-7511ABSTRACT Immunoassays are currently needed to quantify Loa loa microfilariae (mf). To address this need, we have conducted proteomic and bioinformatic analyses of proteins present in the urine of a Loa mf-infected patient and used this information to identify putative biomarkers produced by L. loa mf. In total, 70 of the 15,444 described putative L. loa proteins were identified. Of these 70, 18 were L. loa mf specific, and 2 of these 18 (LOAG_16297 and LOAG_17808) were biologically immunogenic. We developed novel reverse luciferase immunoprecipitation system (LIPS) immunoassays to quantify these 2 proteins in individual plasma samples. Levels of these 2 proteins in microfilaremic L. loa-infected patients were positively correlated to mf densities in the corresponding blood samples (r = 0.71 and P < 0.0001 for LOAG_16297 and r = 0.61 and P = 0.0002 for LOAG_17808). For LOAG_16297, the levels in plasma were significantly higher in Loa-infected (geometric mean [GM], 0.045 µg/ml) than in uninfected (P < 0.0001), Wuchereria bancrofti-infected (P = 0.0005), and Onchocerca volvulus-infected (P < 0.0001) individuals, whereas for LOAG_17808 protein, they were not significantly different between Loa-infected (GM, 0.123 µg/ml) and uninfected (P = 0.06) and W. bancrofti-infected (P = 0.32) individuals. Moreover, only LOAG_16297 showed clear discriminative ability between L. loa and the other potentially coendemic filariae. Indeed, the specificity of the LOAG_16297 reverse LIPS assay was 96% (with a sensitivity of 77%). Thus, LOAG_16297 is a very promising biomarker that will be exploited in a quantitative point-of-care immunoassay for determination of L. loa mf densities. IMPORTANCE Loa loa, the causative agent of loiasis, is a parasitic nematode transmitted to humans by the tabanid Chrysops fly. Some individuals infected with L. loa microfilariae (mf) in high densities are known to experience post-ivermectin severe adverse events (SAEs [encephalopathy, coma, or death]). Thus, ivermectin-based mass drug administration (MDA) programs for onchocerciasis and for lymphatic filariasis control have been interrupted in parts of Africa where these filarial infections coexist with L. loa. To allow for implementation of MDA for onchocerciasis and lymphatic filariasis, tools that can accurately identify people at risk of developing post-ivermectin SAEs are needed. Our study, using host-based proteomics in combination with novel immunoassays, identified a single Loa-specific antigen (LOAG_16297) that can be used as a biomarker for the prediction of L. loa mf levels in the blood of infected patients. Therefore, the use of such biomarker could be important in the point-of-care assessment of L. loa mf densities.Papa M. DrameZhaojing MengSasisekhar BennuruJesica A. HerrickTimothy D. VeenstraThomas B. NutmanAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 7, Iss 1 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Papa M. Drame Zhaojing Meng Sasisekhar Bennuru Jesica A. Herrick Timothy D. Veenstra Thomas B. Nutman Identification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays
description	ABSTRACT Immunoassays are currently needed to quantify Loa loa microfilariae (mf). To address this need, we have conducted proteomic and bioinformatic analyses of proteins present in the urine of a Loa mf-infected patient and used this information to identify putative biomarkers produced by L. loa mf. In total, 70 of the 15,444 described putative L. loa proteins were identified. Of these 70, 18 were L. loa mf specific, and 2 of these 18 (LOAG_16297 and LOAG_17808) were biologically immunogenic. We developed novel reverse luciferase immunoprecipitation system (LIPS) immunoassays to quantify these 2 proteins in individual plasma samples. Levels of these 2 proteins in microfilaremic L. loa-infected patients were positively correlated to mf densities in the corresponding blood samples (r = 0.71 and P < 0.0001 for LOAG_16297 and r = 0.61 and P = 0.0002 for LOAG_17808). For LOAG_16297, the levels in plasma were significantly higher in Loa-infected (geometric mean [GM], 0.045 µg/ml) than in uninfected (P < 0.0001), Wuchereria bancrofti-infected (P = 0.0005), and Onchocerca volvulus-infected (P < 0.0001) individuals, whereas for LOAG_17808 protein, they were not significantly different between Loa-infected (GM, 0.123 µg/ml) and uninfected (P = 0.06) and W. bancrofti-infected (P = 0.32) individuals. Moreover, only LOAG_16297 showed clear discriminative ability between L. loa and the other potentially coendemic filariae. Indeed, the specificity of the LOAG_16297 reverse LIPS assay was 96% (with a sensitivity of 77%). Thus, LOAG_16297 is a very promising biomarker that will be exploited in a quantitative point-of-care immunoassay for determination of L. loa mf densities. IMPORTANCE Loa loa, the causative agent of loiasis, is a parasitic nematode transmitted to humans by the tabanid Chrysops fly. Some individuals infected with L. loa microfilariae (mf) in high densities are known to experience post-ivermectin severe adverse events (SAEs [encephalopathy, coma, or death]). Thus, ivermectin-based mass drug administration (MDA) programs for onchocerciasis and for lymphatic filariasis control have been interrupted in parts of Africa where these filarial infections coexist with L. loa. To allow for implementation of MDA for onchocerciasis and lymphatic filariasis, tools that can accurately identify people at risk of developing post-ivermectin SAEs are needed. Our study, using host-based proteomics in combination with novel immunoassays, identified a single Loa-specific antigen (LOAG_16297) that can be used as a biomarker for the prediction of L. loa mf levels in the blood of infected patients. Therefore, the use of such biomarker could be important in the point-of-care assessment of L. loa mf densities.
format	article
author	Papa M. Drame Zhaojing Meng Sasisekhar Bennuru Jesica A. Herrick Timothy D. Veenstra Thomas B. Nutman
author_facet	Papa M. Drame Zhaojing Meng Sasisekhar Bennuru Jesica A. Herrick Timothy D. Veenstra Thomas B. Nutman
author_sort	Papa M. Drame
title	Identification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays
title_short	Identification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays
title_full	Identification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays
title_fullStr	Identification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays
title_full_unstemmed	Identification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays
title_sort	identification and validation of <italic toggle="yes">loa loa</italic> microfilaria-specific biomarkers: a rational design approach using proteomics and novel immunoassays
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/85a2e55e0d4c4d7e9c2b4b8de7c856f4
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Identification and Validation of <italic toggle="yes">Loa loa</italic> Microfilaria-Specific Biomarkers: a Rational Design Approach Using Proteomics and Novel Immunoassays

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