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Interleukin-18 Regulations of Oral Epithelial Cells by Candida Albicans

Author: XuLiLi
Tutor: ZhangHong
School: Dalian Medical University
Course: Clinical Stomatology
Keywords: Candida albicans KB cells IL-18
CLC: R519
Type: Master's thesis
Year: 2010
Downloads: 56
Quote: 0
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Abstract


Background: Candida albicans is a parasite in the normal population mouth, skin, gastrointestinal, respiratory and reproductive tract and other parts of opportunistic pathogenic fungi. Recent years due to broad-spectrum antibiotics, immunosuppressants, corticosteroids wide range of applications, as well as organ transplantation, diabetes and the increasing number of AIDS patients, patients with decreased immunity or normal flora balance is upset, Candida albicans blooms, making Oral Candida albicans infection into a clear upward trend. Mucosa as a natural barrier against Candida albicans infection in the process played an important role. In addition, local mucosal immune response is also involved in the defense of Candida albicans, including present antigen to resist microbial and epithelial cells secrete a variety of cytokines, such as interleukin -18 (Interleukin-18, IL-18). Study found that IL-18 can activate the host response to Cryptococcus neoformans, S. typhimurium, Mycobacterium tuberculosis and other microorganisms defensive immune response. But on Candida albicans and IL-18 does not have much related research, and IL-18 in the local oral epithelial cells against Candida albicans in the mechanism of action is still unclear. Objective: an in vitro Candida albicans adhesion on KB cells, and to explore the role of adhesion factors and laws. Reverse transcription - polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA) detection of different forms and different concentrations of Candida albicans at different time points after infection KB cells express IL-18 situation, to further explore IL-18 in the oral mucosal epithelial cells during infection Candida albicans defense possible role. Method: 1. Selection of oral epithelial cells and Candida albicans KB cells as experimental subjects. In vitro with different concentrations of Candida albicans KB cells co-cultured at different time points after adhesion of cells seeded were fixed after Gram staining, were counted under an inverted microscope 100 KB cell adhesion on the number of Candida albicans , trypsinized cells were collected and Candida albicans, trypan blue exclusion assay the total number of Candida albicans, energy and mycelium conversion rates. 2 phases with different concentrations of spores and hyphal Candida albicans in vitro infection KB cells to KB cells cultured alone as a control group, respectively, at 3h, 6h, 12h, 24h extraction supernatants, ELISA assay on supernatant protein content of IL-18, RT-PCR detected KB cells express IL-18mRNA level. Differences between the experimental data t test analysis to determine their statistical significance. Results: 1 in 1 × 105ml to 1 × 107ml range, number of adhesion of Candida albicans with the increase of the concentration of C. albicans adhesion on KB cells in a dose-dependent manner. 2 within 24 hours, the number of C. albicans adhesion increases with time, the KB cells of Candida albicans have time-dependent adhesion. 3 albicans adhesion KB cells, its shape facing mycelium from spores in a time-dependent phase shift, over time, the mycelium conversion rate was gradually increased, and the total number of Candida albicans is gradually decreased. 4 KB cells induced by Candida albicans expression of IL-18 protein content increases with time, 12h peak, followed by a downward trend. 5 Candida albicans KB cells produced IL-18 protein content increased with concentration. 6.RT-PCR imaging analysis system showed spore adhesion and hyphal Candida albicans KB cells can make increased expression of IL-18mRNA but also significantly higher, and with the increase of concentration. Conclusions: 1. Candida albicans adhesion effect on KB cells and the concentration and time dependent relationship. 2 Candida albicans infection oral epithelial cells increased expression of IL-18, oral epithelial cells may be involved in a defensive immune response to infection Candida albicans. 3 Candida albicans oral epithelial cells express IL-18 in a time-dependent content. 4.IL-18 may be associated with oral Candida albicans infection, defense closely related.

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