Human single-chain antibodies that neutralize Pseudomonas aeruginosa-exotoxin A-mediated cellular apoptosis
| Autor: | Nitat Sookrung, Wanpen Chaicumpa, Sumate Ampawong, Sirijan Santajit, Kodchakorn Mahasongkram, Watee Seesuay, Pornphan Diraphat, Onrapak Reamtong, Nitaya Indrawattana |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Phage display Bacterial toxins Virulence Factors 030106 microbiology Virulence lcsh:Medicine Exotoxins Apoptosis Biology medicine.disease_cause Article Microbiology 03 medical and health sciences Medical research Peptide Library Catalytic Domain medicine Pseudomonas exotoxin Humans Pseudomonas Infections Peptide library lcsh:Science ADP Ribose Transferases Multidisciplinary Pseudomonas aeruginosa lcsh:R respiratory system Complementarity Determining Regions Recombinant Proteins Anti-Bacterial Agents Elongation factor Molecular Docking Simulation 030104 developmental biology lcsh:Q Single-Chain Antibodies Exotoxin HeLa Cells |
| Zdroj: | Scientific Reports Scientific Reports, Vol 9, Iss 1, Pp 1-15 (2019) |
| ISSN: | 2045-2322 |
| Popis: | Targeting bacterial virulence factors directly provides a new paradigm for the intervention and treatment of bacterial diseases. Pseudomonas aeruginosa produces a myriad of virulence factors to cause fatal diseases in humans. In this study, human single-chain antibodies (HuscFvs) that bound to P. aeruginosa exotoxin A (ETA) were generated by phage display technology using recombinant ETA, ETA-subdomains and the synthetic peptide of the ETA-catalytic site as baits for selecting ETA-bound-phages from the human-scFv phage display library. ETA-bound HuscFvs derived from three phage-transfected E. coli clones neutralized the ETA-induced mammalian cell apoptosis. Computerized simulation demonstrated that these HuscFvs used several residues in their complementarity-determining regions (CDRs) to form contact interfaces with the critical residues in ETA-catalytic domain essential for ADP-ribosylation of eukaryotic elongation factor 2, which should consequently rescue ETA-exposed-cells from apoptosis. The HuscFv-treated ETA-exposed cells also showed decremented apoptosis-related genes, i.e., cas3 and p53. The effective HuscFvs have high potential for future evaluation in animal models and clinical trials as a safe, novel remedy for the amelioration of exotoxin A-mediated pathogenesis. HuscFvs may be used either singly or in combination with the HuscFv cognates that target other P. aeruginosa virulence factors as an alternative therapeutic regime for difficult-to-treat infections. |
| Databáze: | OpenAIRE |
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