Subproject (IV)---Establishment of an ex vivo Culture Platform of Human Respiratory Tract Epithelial Cells---Association of Replication and Drug Susceptibility of Avian Influenza A (H7N9) Virus with Donors’ Anthropological Characteristics (I)

Background: Although novel avian influenza A(H7N9) virus infection has not been epidemic in Taiwan, the presence of a natural reservoir and the disease severity highlight the need to evaluate its risk on human public health and to understand the possible pathogenesis mechanism. According to epidemiologic studies in China, it seemed to frequently infect older people and to cause more severe complications in men of patients with coexisting medical conditions. Therefore, we hypothesize that replication and drug susceptibility of the H7N9 virus are different in ex vivo culture of the human respiratory tract epithelia obtained from different anthropological characteristics. Purposes: We aim to establish an complete ex vivo culture platform of human respiratory tract epithelial cells including the upper and lower airways and to compare the differences in virus replication and drug susceptibility of avian influenza A(H7N9) virus in different age, sex, or medical comorbidity groups. Material and Methods: This is a prospective, case-control, 2-year study. We will perform ex vivo cultures of respiratory tract epithelia of residual surgical specimens from 60 patients undergoing upper airway or lung surgery between January 1, 2014 and December 31, 2015. The patients will be recruited according to their age, gender, and medical comorbidity. We will inoculate the H7N9 into the ex vivo cultures and the 50% infectious doses (TCID50) in these supernatants will be determined by either cytopathic effects or real-time reverse transcriptase PCR (rRT-PCR) assay for replication measurement. Drug (oseltamivir) susceptibility will be determined by TCID50 reduction assay or rRT-PCR assay. We will further compare the differences of replication and oseltamivir susceptibility of the H7N9 virus between different anatomic site, age, gender, or medical comorbidity groups. Anticipating Outcome: This study can establish a complete ex vivo culture platform of the human respiratory tract epithelial cells in order to rapidly study emerging infectious diseases and understand the tropisms, replications, and drug susceptibility of avian influenza A(H7N9) virus infections in different patient groups using this platform.

Project ID:PC10304-0017
External Project ID:MOST103-2321-B182-012