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Taming C-terminal peptides of Staphylococcus aureus leukotoxin M for B-cell response: Implication in improved subclinical bovine mastitis diagnosis and protective efficacy in vitro

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Relation http://ir.cftri.com/12372/
http://dx.doi.org/10.1016/j.toxicon.2016.05.018
 
Title Taming C-terminal peptides of Staphylococcus aureus leukotoxin M for
B-cell response: Implication in improved subclinical bovine mastitis
diagnosis and protective efficacy in vitro
 
Creator Padmaja, R. J.
Halami, P. M.
 
Subject 17 Toxicology
 
Description Leukotoxin M/F0-PV (LukM/F0-PV) produced by bovine mastitis causing Staphylococcus aureus structurally
comprises three domains, the b-sandwich, rim and stem domain. The rim and stem domains interacting
with target cell membrane lipid rafts contributes to the virulent trait of the toxin. In the present study,
two facts were hypothesized that neutralization of these domains will ebb LukM/F0-PV leukotoxicity.
Secondly, the neutralizing antibodies can improve the leukotoxin detection sensitivity in bovine mastitis
milk samples. The in silico mapping of S. aureus LukM C-termini comprising these domains predicted
seven linear B-cell antigenic epitopes. The immune response of C-terminal truncated recombinant
peptides rCtM19 (19 kDa; near carboxy-terminal) having four epitopes and rCtM15 (15 kDa; C-terminal)
with three epitopes were evaluated for their diagnostic and neutralization potential. Anti-rCtM19 and
anti-rCtM15 antibodies with enhanced immunogenicity had the most striking outcome in IgG-ELISA for
detecting native determinants of leukotoxin. For the obtained ELISA values, ROC curve inferred a cut-off
score of >0.102 OD405. The assay sensitivity in the range of 90e96% along with 100% specificity and AUC
of 0.93e0.98 categorized subclinical and clinical from healthy bovine milk samples. As observed through
in vitro neutralization and LDH assays, C-terminus specific antibodies (1:42 titer) deactivating leukotoxicity
abolished LukM from interacting with lipid bilayer and LukF for forming pores on bovine
neutrophil membrane. As a proof of concept, it was proved that peptide antibodies can be a more specific
serodiagnostic and passive therapeutic molecules.
 
Publisher 2016
 
Date 2016
 
Type Article
PeerReviewed
 
Format application/pdf
 
Language en
 
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Identifier http://ir.cftri.com/12372/1/Journal%20of%20Food%20Enginrering%2032%20%28%201997%29%20125-I%2031.pdf
Padmaja, R. J. and Halami, P. M. (2016) Taming C-terminal peptides of Staphylococcus aureus leukotoxin M for B-cell response: Implication in improved subclinical bovine mastitis diagnosis and protective efficacy in vitro. Toxicon, 119. pp. 99-105.