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Functional characterization and evaluation of protective efficacy of EA752–862 monoclonal antibody against B. anthracis vegetative cell and spores

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Abstract

The most promising means of controlling anthrax, a lethal zoonotic disease during the early infection stages, entail restricting the resilient infectious form, i.e., the spores from proliferating to replicating bacilli in the host. The extractible antigen (EA1), a major S-layer protein present on the vegetative cells and spores of Bacillus anthracis, is highly immunogenic and protects mice against lethal challenge upon immunization. In the present study, mice were immunized with r-EA1C, the C terminal crystallization domain of EA1, to generate a neutralizing monoclonal antibody EA752–862, that was evaluated for its anti-spore and anti-bacterial properties. The monoclonal antibody EA752–862 had a minimum inhibitory concentration of 0.08 mg/ml, was bactericidal at a concentration of 0.1 mg/ml and resulted in 100% survival of mice against challenge with B. anthracis vegetative cells. Bacterial cell lysis as observed by scanning electron microscopy and nucleic acid leakage assay could be attributed as a possible mechanism for the bactericidal property. The association of mAb EA752–862 with spores inhibits their subsequent germination to vegetative cells in vitro, enhances phagocytosis of the spores and killing of the vegetative cells within the macrophage, and subsequently resulted in 90% survival of mice upon B. anthracis Ames spore challenge. Therefore, owing to its anti-spore and bactericidal properties, the present study demonstrates mAb EA752–862 as an efficient neutralizing antibody that hinders the establishment of early infection before massive multiplication and toxin release takes place.

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Acknowledgements

S.M. is supported by Senior Research Scholarship of Lady Tata Memorial Trust, India. S.D. is funded by Senior Research Fellowship from Defence Research and Development Organization, India. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

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Authors and Affiliations

  1. Microbiology Division, Defence Food Research Laboratory, Mysore, 570011, India

    Saugata Majumder, Shreya Das, Joseph Kingston, M. S. Shivakiran & H. V. Batra

  2. School of Biotechnology, Jawaharlal Nehru University, New Delhi, 110067, India

    Vikas Kumar Somani & Rakesh Bhatnagar

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  1. Saugata Majumder
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  2. Shreya Das
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Experiments carried out in this study were performed by SM, SD and SSM. SM, SD and JK were involved in designing of experiments, data interpretation and manuscript preparation. RB and VS was involved in challenge studies data interpretation and manuscript preparation. All authors read and approved the final manuscript.

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Correspondence to Joseph Kingston.

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Majumder, S., Das, S., Kingston, J. et al. Functional characterization and evaluation of protective efficacy of EA752–862 monoclonal antibody against B. anthracis vegetative cell and spores. Med Microbiol Immunol 209, 125–137 (2020). https://doi.org/10.1007/s00430-019-00650-5

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