Shigella Pathogenesis Modeling with Tissue Culture Assays

Area of Science:

• Microbiology
• Pathogenesis
• Infectious Diseases

Background:

• Shigella causes Shigellosis, a diarrheal disease, infecting the colonic epithelium.
• Lack of vaccines and increasing antibiotic resistance necessitate understanding Shigella pathogenesis.
• Small animal models fail to accurately replicate Shigellosis.

Purpose of the Study:

• To describe protocols for enumerating Shigella invasion, cell-cell spread, and plaque formation.
• To introduce a novel method for studying Shigella invasion in primary intestinal enteroids.
• To provide tools for examining Shigella virulence factors.

Main Methods:

• Utilized tissue-cultured cell lines (Henle-407 and CoN-841) to study Shigella.
• Developed and described protocols for quantifying bacterial invasion and spread.
• Established a new method using primary intestinal enteroids for invasion studies.

Main Results:

• Established protocols for assessing Shigella invasion, cell-cell spread, and plaque formation in cell lines.
• Successfully developed a new method to study Shigella invasion in primary intestinal enteroids.
• Validated methods for examining Shigella virulence in relevant in vitro models.

Conclusions:

• The described protocols offer valuable tools for Shigella pathogenesis research.
• These methods can advance the understanding of Shigella virulence mechanisms.
• The developed techniques support the search for novel antimicrobial therapies against Shigella infections.