Evaluation of adherence of vibrio cholera Non-O1/Non-O139 to HT-29 cell line
Evaluation of adherence of vibrio cholera Non-O1/Non-O139 to HT-29 cell line
Jafar Alinezhad,1Mohammad Ali Haghighi,2,*Somayyeh Gharibi,3Omid Rasoolzadeh,4
1. Student Research Committee, Bushehr University Of Medical Sciences, Bushehr, Iran 2. Department of Microbiology and Parasitology, School of Medicine, Bushehr University of Medical Sciences, Bushehr, Iran 3. Department of Microbiology, School of Sciences, Kherad Institute of Higher Education, Bushehr, Iran 4. Student Research Committee, Bushehr University Of Medical Sciences, Bushehr, Iran
Introduction: Successful establishment of infection by vibrio cholerae requires adhesion to the intestinal mucus barrier. Human adenocarcinoma-derived intestinal cell lines offer easy and elegant ways to study bacterial adherence to the intestinal. This study was aimed to understand the adherence rate of Vibrio cholerae to HT-29 cell line.
Methods: The adhesion assay was performed on HT-29 cells and cultured in 12-well dishes .HT-29 cells were grown to confluence in DMEM containing 10% FBS and washed with PBS, and DMEM without FBS was added. V. cholerae cultures grown to the midexponential phase in LB were suspended in DMEM added to the HT-29 monolayer at different multiplicity of infection (MOI), and incubated for 45 min at 37°C in a 5% CO2 atmosphere. After 45 min the number of cfu of adhered bacteria was calculated.
Results: The adhesion strongly increased after 45 min of V. cholerae exposure to the HT-29 cell at 37 ° C in MOI 35. In the case of the highest MOI, the number of adhered bacteria was lower for adherence analyzed after the count compared to the MOI 35.
Conclusion: Bacterial attachment and host colonization are key events in V. cholerae infection and result in the ability of the infecting bacterial cells to persist and multiply. In this study, we sought to determine inoculation of which number of V. cholerae provides the best adhesion. Decrease amount of adherence was in highest MOI shows that adherence could be in a concentration-dependent manner. The results of adherence analysis were indicative of this.