Identification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach

Summary: Background: Bartonella bacilliformis is the aetiological agent of Carrión's disease, a biphasic and highly lethal illness formerly restricted to the South American Andes that is now spreading to adjacent areas. Reliable serodiagnostic approaches and vaccines are urgently needed. In th...

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Autores principales:	Alexander A Dichter, MSc, Tilman G Schultze, PhD, Anne Wenigmann, BSc, Wibke Ballhorn, Andreas Latz, PhD, Elif Schlüfter, BSc, Palmira Ventosilla, MSc, Humberto Guerra Allison, PhD, Cesar Ugarte-Gil, PhD, Pablo Tsukayama, ProfPhD, Volkhard A J Kempf, ProfMD
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	Medicine (General) R5-920 Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c1bf22ff51f446bdbe6a00e3c60c79d6
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spelling	oai:doaj.org-article:c1bf22ff51f446bdbe6a00e3c60c79d62021-12-02T05:03:51ZIdentification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach2666-524710.1016/S2666-5247(21)00184-1https://doaj.org/article/c1bf22ff51f446bdbe6a00e3c60c79d62021-12-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2666524721001841https://doaj.org/toc/2666-5247Summary: Background: Bartonella bacilliformis is the aetiological agent of Carrión's disease, a biphasic and highly lethal illness formerly restricted to the South American Andes that is now spreading to adjacent areas. Reliable serodiagnostic approaches and vaccines are urgently needed. In this study, we aimed to identify immunodominant proteins of B bacilliformis and to establish novel and reliable serodiagnostic tools. Methods: We used a reverse vaccinology approach in combination with an analysis of heterologous genomic expression libraries to identify immunodominant proteins, on the basis of the genome sequences of B bacilliformis strains KC583 and KC584. Antigens were screened with serum samples collected from Peruvian patients with B bacilliformis infections and from German healthy blood donors without history of travel to South America. We further analysed immunoreactive proteins of B bacilliformis with immunoblotting and line blots. We used selected target proteins to develop a diagnostic ELISA. To assess the performance of this ELISA, we did receiver operating characteristic analyses to assess the area under the curve, cutoff values, sensitivities, and specificities with 95% CIs. Findings: We used serum samples obtained between Dec 23, 1990, and May 5, 2018, from 26 Peruvian patients with B bacilliformis infections and serum samples taken between Aug 28 and Aug 31, 2020, from 96 healthy German blood donors. 21 potentially immunodominant proteins were identified and recombinantly expressed, and their reactivity was assessed with immunoblotting and line blots. Of these 21 antigens, 14 were found to be immunoreactive. By using serum samples of Peruvian patients with Carrión's disease and of healthy German blood donors, we identified three antigens (porin B, autotransporter E, and hypothetical protein B) as suitable immunodominant antigens, and we applied them in a diagnostic ELISA using two different antigen combinations (porin B plus autotransporter E and porin B plus autotransporter E plus hypothetical protein B). For the combination of porin B and autotransporter E, with optical density measured at 450 nm (OD450) cutoff value of 0·29, sensitivity was 80·8% (95% CI 60·7–93·5) and specificity was 94·8% (88·3–98·3) for all Peruvian patient samples. For a combination of porin B, autotransporter E, and hypothetical protein B, with an OD450 cutoff of 0·34, sensitivity was 76·9% (56·4–91·0) and specificity was 93·8% (86·9–97·7) for all Peruvian patient samples. Interpretation: This novel ELISA could represent a useful serodiagnostic tool for future epidemiological studies of B bacilliformis in endemic areas. Additionally, the immunodominant antigens we have identified could provide a first basis for future vaccine development to prevent the highly lethal Carrión's disease. Funding: DRUID (Novel Drug Targets against Poverty-Related and Neglected Tropical Infectious Diseases) Initiative and Robert Koch Institute. Translations: For the Spanish and Quechua translations of the abstract see Supplementary Materials section.Alexander A Dichter, MScTilman G Schultze, PhDAnne Wenigmann, BScWibke BallhornAndreas Latz, PhDElif Schlüfter, BScPalmira Ventosilla, MScHumberto Guerra Allison, PhDCesar Ugarte-Gil, PhDPablo Tsukayama, ProfPhDVolkhard A J Kempf, ProfMDElsevierarticleMedicine (General)R5-920MicrobiologyQR1-502ENThe Lancet Microbe, Vol 2, Iss 12, Pp e685-e694 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine (General) R5-920 Microbiology QR1-502
spellingShingle	Medicine (General) R5-920 Microbiology QR1-502 Alexander A Dichter, MSc Tilman G Schultze, PhD Anne Wenigmann, BSc Wibke Ballhorn Andreas Latz, PhD Elif Schlüfter, BSc Palmira Ventosilla, MSc Humberto Guerra Allison, PhD Cesar Ugarte-Gil, PhD Pablo Tsukayama, ProfPhD Volkhard A J Kempf, ProfMD Identification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach
description	Summary: Background: Bartonella bacilliformis is the aetiological agent of Carrión's disease, a biphasic and highly lethal illness formerly restricted to the South American Andes that is now spreading to adjacent areas. Reliable serodiagnostic approaches and vaccines are urgently needed. In this study, we aimed to identify immunodominant proteins of B bacilliformis and to establish novel and reliable serodiagnostic tools. Methods: We used a reverse vaccinology approach in combination with an analysis of heterologous genomic expression libraries to identify immunodominant proteins, on the basis of the genome sequences of B bacilliformis strains KC583 and KC584. Antigens were screened with serum samples collected from Peruvian patients with B bacilliformis infections and from German healthy blood donors without history of travel to South America. We further analysed immunoreactive proteins of B bacilliformis with immunoblotting and line blots. We used selected target proteins to develop a diagnostic ELISA. To assess the performance of this ELISA, we did receiver operating characteristic analyses to assess the area under the curve, cutoff values, sensitivities, and specificities with 95% CIs. Findings: We used serum samples obtained between Dec 23, 1990, and May 5, 2018, from 26 Peruvian patients with B bacilliformis infections and serum samples taken between Aug 28 and Aug 31, 2020, from 96 healthy German blood donors. 21 potentially immunodominant proteins were identified and recombinantly expressed, and their reactivity was assessed with immunoblotting and line blots. Of these 21 antigens, 14 were found to be immunoreactive. By using serum samples of Peruvian patients with Carrión's disease and of healthy German blood donors, we identified three antigens (porin B, autotransporter E, and hypothetical protein B) as suitable immunodominant antigens, and we applied them in a diagnostic ELISA using two different antigen combinations (porin B plus autotransporter E and porin B plus autotransporter E plus hypothetical protein B). For the combination of porin B and autotransporter E, with optical density measured at 450 nm (OD450) cutoff value of 0·29, sensitivity was 80·8% (95% CI 60·7–93·5) and specificity was 94·8% (88·3–98·3) for all Peruvian patient samples. For a combination of porin B, autotransporter E, and hypothetical protein B, with an OD450 cutoff of 0·34, sensitivity was 76·9% (56·4–91·0) and specificity was 93·8% (86·9–97·7) for all Peruvian patient samples. Interpretation: This novel ELISA could represent a useful serodiagnostic tool for future epidemiological studies of B bacilliformis in endemic areas. Additionally, the immunodominant antigens we have identified could provide a first basis for future vaccine development to prevent the highly lethal Carrión's disease. Funding: DRUID (Novel Drug Targets against Poverty-Related and Neglected Tropical Infectious Diseases) Initiative and Robert Koch Institute. Translations: For the Spanish and Quechua translations of the abstract see Supplementary Materials section.
format	article
author	Alexander A Dichter, MSc Tilman G Schultze, PhD Anne Wenigmann, BSc Wibke Ballhorn Andreas Latz, PhD Elif Schlüfter, BSc Palmira Ventosilla, MSc Humberto Guerra Allison, PhD Cesar Ugarte-Gil, PhD Pablo Tsukayama, ProfPhD Volkhard A J Kempf, ProfMD
author_facet	Alexander A Dichter, MSc Tilman G Schultze, PhD Anne Wenigmann, BSc Wibke Ballhorn Andreas Latz, PhD Elif Schlüfter, BSc Palmira Ventosilla, MSc Humberto Guerra Allison, PhD Cesar Ugarte-Gil, PhD Pablo Tsukayama, ProfPhD Volkhard A J Kempf, ProfMD
author_sort	Alexander A Dichter, MSc
title	Identification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach
title_short	Identification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach
title_full	Identification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach
title_fullStr	Identification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach
title_full_unstemmed	Identification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach
title_sort	identification of immunodominant bartonella bacilliformis proteins: a combined in-silico and serology approach
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/c1bf22ff51f446bdbe6a00e3c60c79d6
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Identification of immunodominant Bartonella bacilliformis proteins: a combined in-silico and serology approach

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