Rhinovirus Infection Study Model Using Organ Culture of Turbinate Mucosa

Kim, Seong Hak; Kwon, Hyon Ja; Chung, You Sam; Lee, Bong Jae; Jang, Yong Ju

Abstract

BACKGROUND AND OBJECTIVES
To have better understanding on the pathophysiology of rhinovirus infection, an availability of an ideal experimental model is of utmost importance. We aimed to develop a new study model using the organ culture of turbinate mucosa to overcome the limitations of the conventional study methods. MATERIALS AND METHOD: The inferior turbinate mucosae harvested during the septoturbinoplasty were cultured in air-liquid interface methods, placed on the support of gelfoam soaked in the culture media. Human rhinovirus -16 was applied on the top of the mucosal surface. The success of rhinovirus infection was determined by semi-nested RT-PCR of the mucosal surface fluid taken 48 hours after incubation. Intracellular rhinovirus was visualized by in situ hybridization. Elaboration of cytokine IL-6 and IL-8 into the culture media was quantitated using the ELISA method. RESULTS: PCR product of 292 bp on semi-nested RT-PCR, representing successful rhinovirus infection, was detected in 5 tissues out of 10 mucosal tissues. In the in situ hybridization method, positively stained cells were found in epithelial layer in scattered fashion. In the analysis of cytokine production by continuous exposure to rhinovirus according to the time course, IL-6 and IL-8 secretions in the infected mucosae were significantly greater than in the control mucosa. The increase in the cytokine production was evident from 24 hours after the infection. CONCLUSION: The results of this study indicate that the organ culture of turbinate mucosa could serve as an acceptable in vitro model for studying pathophysiology of rhinovirus infection.