Innate Immune Response to Vibrio cholerae β-Barrel Pore-Forming Hemolysin: Unraveling Cytotoxic, Oxidative, and Inflammatory Pathways

Document Type : Original Research Papers

Authors

1 Bahgat

2 Sherbin, Mansoura, Egypt

3 Professor of Botany and Microbiology Department , Faculty of Science, Benha University

4 Botany and Microbiology Department, Faculty of Science, Benha University, Egypt

10.21608/bjas.2025.369208.1626

Abstract

Abstract
Background
Vibrio cholerae β-barrel pore-forming hemolysin (Vchβ-PFH) is a key virulence factor that disrupts host cell membranes, triggering immune responses. However, the transcriptional landscape of innate immunity in response to Vchβ-PFH remains poorly understood. This study investigates the cytotoxic, oxidative, inflammasome, and immune signaling responses to Vchβ-PFH in macrophage-like THP-1.
Methods
Cells were exposed to varying concentrations of Vchβ-PFH, and cytotoxicity was assessed using MTT and LDH assays. Oxidative stress levels were quantified via intracellular ROS measurements. Inflammasome activation was evaluated by NLRP3 and IL-1β using qRT-PCR. Immune signaling responses were analyzed by measuring the transcriptional expression of TNF-α, IFN-β, IL-10, and CD200. Primer sequences used for qRT-PCR were designed and validated through Primer-BLAST.
Results
Exposure to Vchβ-PFH resulted in significant dose-dependent cytotoxicity, as indicated by reduced cell viability and increased LDH release (p < 0.01). ROS levels were markedly elevated, suggesting oxidative stress as a key mechanism of toxin-induced damage. qRT-PCR analysis revealed upregulation of NLRP3, IL-1β, indicating inflammasome activation. Pro-inflammatory cytokines (TNF-α, IFN-β, and IL-1β) were significantly upregulated, while CD200 and IL-10 were also increased, suggesting a potential immune-regulatory feedback mechanism.
Conclusion
Vchβ-PFH induces cellular cytotoxicity, oxidative stress, inflammasome activation, and immune modulation, revealing its potential role in V. cholerae pathogenesis. These findings highlight novel molecular targets for therapeutic interventions against pore-forming toxin-mediated infections. Further studies are needed to explore the mechanisms underlying host immune evasion and inflammatory resolution in response to V. cholerae hemolysins.

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Benha Journal of Applied Sciences
Volume 10, Issue 2 - Serial Number 2
Applied and Basic Sciences:
February 2025
Pages 115-119

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