Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E

Abstract Botulinum neurotoxins (BoNTs) represent a family of bacterial toxins responsible for neuroparalytic disease ‘botulism’ in human and animals. Their potential use as biological weapon led to their classification in category ‘A’ biowarfare agent by Centers for Disease Control and Prevention (C...

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Autores principales:	Priyanka Sonkar, Vinita Chauhan, Ritika Chauhan, Nandita Saxena, Ram Kumar Dhaked
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2020
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a77fcbcca8e64d5d85a856dbbf4c6918
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spelling	oai:doaj.org-article:a77fcbcca8e64d5d85a856dbbf4c69182021-12-02T16:46:33ZCharacterization of immune response induced against catalytic domain of botulinum neurotoxin type E10.1038/s41598-020-70929-82045-2322https://doaj.org/article/a77fcbcca8e64d5d85a856dbbf4c69182020-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-020-70929-8https://doaj.org/toc/2045-2322Abstract Botulinum neurotoxins (BoNTs) represent a family of bacterial toxins responsible for neuroparalytic disease ‘botulism’ in human and animals. Their potential use as biological weapon led to their classification in category ‘A’ biowarfare agent by Centers for Disease Control and Prevention (CDC), USA. In present study, gene encoding full length catalytic domain of BoNT/E-LC was cloned, expressed and protein was purified using Ni–NTA chromatography. Humoral immune response was confirmed by Ig isotyping and cell-mediated immunity by cytokine profiling and intracellular staining for enumeration of IFN-γ secreting CD4+ and CD8+ T cells. Increased antibody titer with the predominance of IgG subtype was observed. An interaction between antibodies produced against rBoNT/E-LC was established that showed the specificity against BoNT/E in SPR assay. Animal protection with rBoNT/E-LC was conferred through both humoral and cellular immune responses. These findings were supported by cytokine profiling and flow cytometric analysis. Splenocytes stimulated with rBoNT/E-LC showed a 3.27 and 2.8 times increase in the IFN-γ secreting CD4+ and CD8+ T cells, respectively; in immunized group (P < 0.05). Protection against BoNT/E challenge tended to relate with increase in the percentage of rBoNT/E-LC specific IL-2 in the splenocytes supernatant (P = 0.034) and with IFN-γ-producing CD4+ T cell responses (P = 0.045). We have immunologically evaluated catalytically active rBoNT/E-LC. Our results provide valuable investigational report for immunoprophylactic role of catalytic domain of BoNT/E.Priyanka SonkarVinita ChauhanRitika ChauhanNandita SaxenaRam Kumar DhakedNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 10, Iss 1, Pp 1-14 (2020)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Priyanka Sonkar Vinita Chauhan Ritika Chauhan Nandita Saxena Ram Kumar Dhaked Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E
description	Abstract Botulinum neurotoxins (BoNTs) represent a family of bacterial toxins responsible for neuroparalytic disease ‘botulism’ in human and animals. Their potential use as biological weapon led to their classification in category ‘A’ biowarfare agent by Centers for Disease Control and Prevention (CDC), USA. In present study, gene encoding full length catalytic domain of BoNT/E-LC was cloned, expressed and protein was purified using Ni–NTA chromatography. Humoral immune response was confirmed by Ig isotyping and cell-mediated immunity by cytokine profiling and intracellular staining for enumeration of IFN-γ secreting CD4+ and CD8+ T cells. Increased antibody titer with the predominance of IgG subtype was observed. An interaction between antibodies produced against rBoNT/E-LC was established that showed the specificity against BoNT/E in SPR assay. Animal protection with rBoNT/E-LC was conferred through both humoral and cellular immune responses. These findings were supported by cytokine profiling and flow cytometric analysis. Splenocytes stimulated with rBoNT/E-LC showed a 3.27 and 2.8 times increase in the IFN-γ secreting CD4+ and CD8+ T cells, respectively; in immunized group (P < 0.05). Protection against BoNT/E challenge tended to relate with increase in the percentage of rBoNT/E-LC specific IL-2 in the splenocytes supernatant (P = 0.034) and with IFN-γ-producing CD4+ T cell responses (P = 0.045). We have immunologically evaluated catalytically active rBoNT/E-LC. Our results provide valuable investigational report for immunoprophylactic role of catalytic domain of BoNT/E.
format	article
author	Priyanka Sonkar Vinita Chauhan Ritika Chauhan Nandita Saxena Ram Kumar Dhaked
author_facet	Priyanka Sonkar Vinita Chauhan Ritika Chauhan Nandita Saxena Ram Kumar Dhaked
author_sort	Priyanka Sonkar
title	Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E
title_short	Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E
title_full	Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E
title_fullStr	Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E
title_full_unstemmed	Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E
title_sort	characterization of immune response induced against catalytic domain of botulinum neurotoxin type e
publisher	Nature Portfolio
publishDate	2020
url	https://doaj.org/article/a77fcbcca8e64d5d85a856dbbf4c6918
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_version_	1718383414717972480

Characterization of immune response induced against catalytic domain of botulinum neurotoxin type E

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