鸚鵡多瘤病毒次單位疫苗之開發

鸚鵡多瘤病毒(Avian Polyomavirus；APV)感染症在鸚鵡科鳥類中為常見的病毒性疾病。受APV感染之鸚鵡科鳥類可能出現厭食、精神不濟、羽毛掉落、皮下出血等症狀，甚至死亡。所有年齡之鸚鵡科鳥類皆可受感染，而受感染之個體年齡越小死亡率越高，屬於高傳染性病毒疾病。鸚鵡多瘤病毒歸類於乳多瘤病毒科(Papovaviridae)，多瘤病毒屬(Polymavirus)之一員，為不具封套之二十面對稱體、環狀雙股DNA病毒。鸚鵡多瘤病毒具有四種結構蛋白VP1、VP2、VP3及VP4，其中VP1為病毒主要外殼蛋白，具有誘導中和抗體的抗原決定位。隨著台灣飼養鸚鵡的數量增加，加上高密度的飼養環境，病毒的擴散速度也日益提升，但市面上僅在美國地區有販售死毒APV疫苗，台灣目前仍無相關疫苗能用於預防。此外死毒APV疫苗陸續被報導出注射部位出現肌肉壞死的情形，經研究後認為此情形與死毒疫苗搭配使用之油質佐劑有關。因此本研究目的旨在製備有效之APV次單位疫苗，作為鸚鵡疾病預防之用。目前已成功轉殖VP1基因至pBacPAK8中，使用昆蟲桿狀病毒表達系統生產重組pBacPAK8_APV VP1蛋白，搭配O/W劑型之佐劑製成APV疫苗免疫於SPF小雞與虎皮鸚鵡幼鳥，透過ELISA檢測血清中之特異性抗體反應及中和試驗檢測免疫血清中和病毒之能力。結果顯示，注射APV疫苗之組別特異性抗體反應皆升高且血清具有中和病毒之能力，表示製備出的APV疫苗具有發展成商業疫苗之潛力。

關鍵字

鸚鵡多瘤病毒；鸚鵡；桿狀病毒表現系統；次單位疫苗

並列摘要

Avian polyomavirus (APV) is the common viral diseases in the Psittacidae family. Infection by APV may lead to no appetite, cutaneous hemorrhage, feather abnormalities, and even death. All age of parrots can be infected by APV and the mortality is associated with young age. APV is a highly contagious viral disease. Avian polyomavirus belongs to the family polyomavirus, which is non-enveloped, double-stranded DNA virus. The icosahedral capsid is composed of the major structural protein VP1 and minor structural proteins VP2, VP3 and VP4. Among them, VP1 has an epitope that induces neutralizing antibodies. With the increase in the number of parrots raised in Taiwan, the rate of spread of the virus also increased. However, there is no effective commercial APV vaccine available in Taiwan. Inactivated APV vaccines are only sold in the United States, but the oil-adjuvanted vaccines were however found to cause muscular necrosis. We aim to prepare an effective and safe APV vaccine for the prevention of parrot disease. So far, we have successfully cloned the VP1 gene into pBacPAK8 and expressed protein using the baculovirus expression system. Then, we used o/w adjuvant to prepare APV vaccine and immunized SPF chicks and parrot chicks. The serum antibody response was detected by ELISA and the ability of the immune serum to neutralize the virus was detected by neutralization test. The results showed that the specific antibody response of the injection groups was increased and the immune serum had the ability to neutralize the virus, indicating that the prepared APV vaccine has the potential to develop into a commercial vaccine.

並列關鍵字

avian polyomavirus ； parrot ； baculovirus expression system ； subunit vaccine

參考文獻

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