Development of rhinovirus study model using organ culture of turbinate mucosa.
To better understand the pathophysiology of rhinovirus (RV) infection, a development of a study model using organ culture of turbinate mucosa was sought. Inferior turbinate mucosal tissues were cultured using air-liquid interface methods, on a support of gelfoam soaked in culture media. RV-16 was applied to the mucosal surface and washed off, and histological changes were evaluated. The success of RV infection was assayed by semi-nested RT-PCR of the mucosal surface fluid taken 48h after incubation. Intracellular RVs were visualized by in situ hybridization (ISH). Secretion of the cytokines, IL-6 and IL-8, into the culture media was quantitated by ELISA. After 7 days of culture, the turbinate mucosae did not show significant damage. A PCR product indicating successful RV infection was detected in 5 out of 10 mucosal tissues. ISH showed a very small number of positively stained cells focally located in the epithelial layer. In the beginning 24h after infection, secretion of IL-6 and IL-8 into the culture media of infected mucosae was significantly greater than into the media of control mucosae. Our results indicate that the air-liquid interface organ culture of turbinate mucosa could serve as an acceptable in vitro model for studying RV infection.
Jang
Yong Ju
YJ
Department of Otolaryngolgy, Asan Medical Center, University of Ulsan College of Medicine, 388-1 Pungnap-2dong, Songpa-gu, Seoul 138-736, South Korea. jangyj@amc.seoul.kr
Lee
Si Hyeong
SH
Kwon
Hyon-Ja
HJ
Chung
Yoo-Sam
YS
Lee
Bong-Jae
BJ
D000293:Adolescent; D000328:Adult; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D017403:In Situ Hybridization; D015850:Interleukin-6 / Q000032:analysis; D016209:Interleukin-8 / Q000032:analysis; D008875:Middle Aged; D009297:Nasal Mucosa / Q000254:growth & development / Q000473:pathology / Q000821:virology*; D009924:Organ Culture Techniques / Q000379:methods*; D010850:Picornaviridae Infections / Q000821:virology*; D012367:RNA, Viral / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012229:Rhinovirus / Q000254:growth & development*; D014420:Turbinates / Q000473:pathology / Q000821:virology*
D015850:Interleukin-6; D016209:Interleukin-8; D012367:RNA, Viral
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