不同血清型腺病毒感染肺癌細胞株A549所分泌之前發炎細胞激素研究

Characterization of Proinflammatory Cytokine Profiles Produced in Different Serotypes of Adenoviruses-infected Human Lung Alveolar A549 Cells

Approximately 5%-10% of children with acute airway infections can be attributed to adenovirus infection. Adenovirus (AdV) infection usually causes fever and further severe acute respiratory and conjunctival infection, which may 1ead to death in vu1nerab1e subjects such children younger than 6 years old. In c1inics, infectious AdVs are divided into 6 species and 51 serotypes. Among the identified infectious AdVs, species B (serotypes AdV3 and AdV7) and species C (serotypes AdVl, AdV2 and AdV5) are highly associated with respiratory tract infections and may persist in children without causing symptoms for years. Intriguingly, infection of AdV3 and AdV7 usually causes severe syndromes compared to infections by other serotypes AdV. The immune responses induced by the different serotypes AdV are rarely investigated. In the present study, we collected AdV samples isolated from patients with respiratory infection in middle Taiwan during 2004-2005 and measured the different proinflammatory cytokines produced in the AdV-infected lung alveolar A549 cells. Production of cytokines was quantitated using sandwich ELISA. Our results revealed that AdV3 induced the highest level ofinterleukin-l (IL-1), IL-6, IL-8 and tumor necrosis factor alpha (TNF-α) compared to the other AdVl, AdV2, AdV5 and AdV6. These findings provide evidence that AdV3, compared with other AdVs, robustly induces the production of various inflammatory cytokine, which may further contribute to severe airway inflammation found c1inically. 5%-10%的兒童急性呼吸道感染皆由腺病毒感染所引起，並進而造成嚴重的急性呼吸道感染、結膜感染甚至死亡。人類腺病毒目前已知有6個亞屬包含51種血清型。血清型第1、2 、3 、5 、6型為常見造成呼吸道感染的腺病毒，其中又以血清型第3型與第7型為最容易導致嚴重病症。腺病毒呼吸道感染及進一步誘發的呼吸道發炎與損傷機制至今尚未完全明瞭，但一般認為前發炎細胞激素的誘發扮演相當重要的角色。因此，本研究擬由2004-2005年間感染腺病毒的病人檢體中，分離與鑑定不同血清型的腺病毒，並進一步探討其對於誘發肺癌細胞株A549所分泌的前發炎細胞激素之類型與濃度進行分析探討。我們利用分離所得之血清型第1、2 、3 、5 、6型腺病毒進行對A549細胞的感染，培養8 、24 、48小時後收集培養基進行前發炎細胞激素IL-1ß 、IL-6 、IL-8及TNF-α的定量分析。結果顯示，腺病毒感染後確實顯著提升前發炎細胞激素的分泌，這些與先前的臨床檢驗相符。此外，相較於本實驗的其他腺病毒，血清型第3型的腺病毒所誘發的前發炎激素最高，推測可能與血清型第3型腺病毒在臨床上最容易導致呼吸道重症有關。綜合本研究的結果，發現不同血清型腺病毒會誘發不同程度的前發炎細胞激素之分泌，而且這些差異與其在臨床上所引起之症狀嚴重情形相關。另外，基於本研究結果，我們建議監控腺病毒感染患者的IL-1ß 、IL-8及TNF-α的濃度，可以在完成病毒血清型鑑定完成之前，或許可以幫助醫師評估病人的病情發展與監測藥物療效。