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

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....

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Bibliographic Details
Published in:Scientific reports 2020-08, Vol.10 (1), p.13932-14, Article 13932
Main Authors: Sonkar, Priyanka, Chauhan, Vinita, Chauhan, Ritika, Saxena, Nandita, Dhaked, Ram Kumar
Format: Article
Language:English
Subjects:
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Animals
Antibodies, Neutralizing - immunology
Botulinum toxin
Botulinum Toxins - chemistry
Botulinum Toxins - genetics
Botulinum Toxins - immunology
Botulinum Toxins, Type A - chemistry
Botulinum Toxins, Type A - immunology
Botulism
Botulism - metabolism
Botulism - prevention & control
Catalytic Domain - genetics
Catalytic Domain - immunology
CD4 antigen
CD8 antigen
CD8-Positive T-Lymphocytes - immunology
Cell-mediated immunity
Cloning, Molecular - methods
Clostridium botulinum - genetics
Cytokines
Enumeration
Flow cytometry
Food contamination
Humanities and Social Sciences
Humans
Immune response
Immune response (humoral)
Immunization
Immunoglobulin G
Interleukin 2
Lymphocytes T
Male
Mice
Mice, Inbred BALB C
multidisciplinary
Neurotoxins
Science
Science (multidisciplinary)
Splenocytes
Toxins
γ-Interferon
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Summary: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 
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-020-70929-8