利用銀耳多醣塗覆於微脂粒包覆新城病毒（NDV）做為死毒疫苗之黏膜佐劑之研究

新城病(Newcastle Disease Virus，NDV)是一種高度傳染性的家禽疾病，主要感染呼吸、消化、神經與生殖系統，屬於副粘液病毒科（Paramyxoviridae）。因此，本研究研發一種新型態黏膜佐劑 — 多醣體黏性微脂粒，由DPPC組成之微脂粒包裹不活化新城病毒，掺和於銀耳多醣( Tremella )或三仙膠( Xanthan Gum )溶液。動物實驗部分：於四週齡來亨雞分為五組每組10隻兩次點鼻接種後，取得鼻腔/氣管沖洗液與血清以酵素免疫連結分析法（ELASA）分析s-IgA，結果發現，Tremella微脂粒其s-IgA相較於其餘四組，皆有顯著的差異(P<0.05)。血清抗體IgG中在Tremella組相較於其餘四組也有顯著的差異(P<0.05)。血球抑制凝集試驗(HI test)，Tremella微脂粒的titer值為256。在攻毒試驗以Sa to 強毒株1000MLD點鼻攻毒後，Tremella組可達到100％的存活率，control組為30％存活率。多醣體黏性微脂粒佐劑之免疫機制部分：由西方點墨法（Western Blot）顯示Tremella微脂粒是唯一在ERK pathway有被磷酸化；最後為瞭解多醣體黏性微脂粒完整的保護能力，將攻毒後存活雞隻的臟器如：氣管、肝、肺、脾臟，然後以RT-PCR偵測病毒殘存量，結果顯示Tremella微脂粒在各個臟器幾乎沒有病毒殘留。由此結果可證實，Tremella微脂粒有效的提高動物體內血清的IgG和鼻腔/氣管的s-IgA，且能在第一道防線鼻黏膜免疫上阻擋病毒的入侵，免於病毒的感染，使存活率達到100％

關鍵字

微脂粒；銀耳多醣；新城病

並列摘要

Newcastle disease virus (NDV) is a high contagious disease, it infects respiration、digestion、nervous system and procreation, it is an an avian paramyxovirus。We propose a new adjuvant of mucosal─ polysaccharide viscosity liposome, inactive NDV encapsulated in liposomes and coated with Tremella or Xanthan Gum. In animal test，four-week-old specific-pathogen-free (SPF) white leghorn chickens, it was divided into five group of every group fetches 10 animals, after twice intranasal deliver, we collect its nasal wash liquid and serum to analyse the formulation amount of its specific sIgA and IgG with ELASA. The result verify, The titers of tremella-coated liposome-encapsulated NDV (T-lip) mucosal secretary immunoglobulin A (s-IgA) and serum IgG was the highest than the others（p＜0.05）. It display the highest ability of hemagglutination inhibition in HI TEST.（titer＝256）. After challenge with strong poisonous Sato of 1000MLD respectively, Chickens receiving virus alone had a 30% survival rate then NDV only、NDV-Liposome and Xanthan Gum coated liposome-encapsulated NDV had 60% survival rate. In contrast, groups that received T-lip had a higher survival rate of 100%. In immune system of polysaccharide viscosity liposome, T-lip phosphorylated in ERK pathway. To understand protection ability of polysaccharide viscosity liposome, we collected internal organs of alive chicken after challenge, example: throat、liver、lung and spleen, then we detected virus residue with RT-PCR. T-lip did not have virus residue in all internal organs almost. The result suggest that T-lip could enhance sIgA and IgG, then protected virus to invade in nasal mucosa immune system, it made survival rate up to 100%.

並列關鍵字

Liposome ； Tremella ； NDV

參考文獻

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【5】Chang, R., S. Nir, et al. (1998). "Analysis of binding and membrane destabilization of phospholipid membranes by surfactant apoprotein B." Biochim Biophys Acta 1371(2): 254-64.

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利用銀耳多醣塗覆於微脂粒包覆新城病毒（NDV）做為死毒疫苗之黏膜佐劑之研究

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