流行性感冒病毒 (influenza virus, IAV) 具高度罹病率及死亡率，因此被列為相當嚴重的呼吸道感染疾病之一。由於病毒套膜上的血球凝集素 (hemagglutinin) 和神經氨酸酶 (neuraminidase) 具有高變異特性，因此極需發展廣效性的流行性感冒病毒疫苗。目前已有許多研究致力於改進或開發新型流行性感冒病毒疫苗，但仍有許多問題需要解決。半乳糖凝集素-1 (galectin-1) 屬於S型凝集素 (lectin)，其具有序列保守的醣類辨識區域 (carbohydrate recognition domain)，透過這段區域可辨識帶有半乳糖的寡醣鏈。由實驗室先前發表過的文章發現，galectin-1可直接結合上A型流行性感冒病毒套膜上的醣蛋白，並抑制其血球凝集素的活性和病毒的感染力。此外，我們實驗室最近的研究亦顯示，galectin-1共價接合上金奈米粒子 (AuNP/Gal-1) 與單純galectin-1相比，能顯著增強與流行性感冒病毒結合的親合力，並顯著降低抗病毒的有效濃度。因此，在本研究中，我們探討以AuNP/Gal-1結合去活性流行性感冒病毒是否可發展為一種新型流行性感冒病毒的疫苗。首先，以肌肉注射方式給予小鼠AuNP/Gal-1混合去活性流行性感冒病毒 (IAV/AuNP/Gal-1)，使小鼠血清產生抗病毒的IgM和IgG，但不無IgA之產生。進而將免疫接種IAV/AuNP/Gal-1後的小鼠脾臟細胞，在體外以高劑量的去活性流行性感冒病毒刺激則有較好的增生現象。此外，給予IAV/AuNP/Gal-1的小鼠比單純給予去活性病毒的組別，會隨著AuNP/Gal-1劑量提升而增加抗流行性感冒病毒的IgG量。然而，免疫接種IAV/AuNP/Gal-1和IAV/Alum的小鼠組別，在誘導個體產生IgG、IgG1及IgG2a的效力上，並無顯著差異。進一步以AuNP/Gal-1刺激骨髓細胞分化的樹突細胞，相較於單獨以galectin-1刺激的組別，AuNP/Gal-1誘導產生甲型腫瘤壞死因子 (tumor necrosis factor-α, TNF-α)、白介素6 (interlukin-6, IL-6)、白介素10 (interlukin-10, IL-10) 的量顯著提升。值得注意的是，以IAV/AuNP/Gal-1免疫接種的小鼠，相較單獨給予去活性病毒的組別，血清可中和流行性感冒病毒感染的能力較好。總而言之，AuNP/Gal-1確實透過直接包裹病毒及刺激樹突細胞的活性來促進抗流行性感冒病毒抗體的產生。由於galectin-1可與不同亞型的流行性感冒病毒結合，因此我們認為AuNP/Gal-1可廣泛的應用於開發抵抗不同亞型的流行性感冒病毒的新型疫苗。

Influenza is an important infectious disease that causes significant morbidity and mortality. Because of the high variability of viral hemagglutinin and neuraminidase, it is highly desirable to develop broad-spectrum influenza vaccines. Much effort has been devoted to developing improved or novel vaccines for influenza; however, there is still a long way to go. Galectin-1, which belongs to S-type lectins, contains conserved carbohydrate recognition domains (CRDs) recognizing galactose-containing oligosaccharides. We have reported previously that galectin-1 can directly bind to the envelope glycoproteins of influenza A virus and inhibit its hemagglutination activity and infectivity. Moreover, our recent studies showed that galectin-1 conjugated on gold nanoparticle (AuNP/Gal-1) dramatically enhanced the binding affinity to influenza virus and reduced the EC50 of anti-influenza viral activity as compared to galectin-1 alone. In this study, we tested whether AuNP/Gal-1 combined with inactivated influenza virus could be used as a novel vaccine for influenza. We found that mice immunized intramuscularly with AuNP/Gal-1 admixed with inactivated influenza A virus (IAV/AuNP/Gal-1) produced IgM and IgG, but not IgA against influenza virus in the serum. The splenocytes from mice treated with IAV/AuNP/Gal-1 proliferated in response to high doses of inactivated influenza virus in vitro. Furthermore, mice treated with IAV/AuNP/Gal-1 induced higher anti-influenza virus IgG in a dose-dependent manner than those treated with inactivated virus alone. However, there was no significant difference in the efficacy of IgG, IgG1 and IgG2a production between mice immunized with IAV/AuNP/Gal-1 and with IAV/Alum. In addition, bone-marrow-derived dendritic cells stimulated with AuNP/Gal-1 induce higher TNF-α, IL-6 and IL-10 than those stimulated with galectin-1 alone. Notably, levels of neutralizing antibodies against influenza virus produced from mice immunized with IAV/AuNP/Gal-1 were higher than those immunized with inactivated virus alone. In conclusion, AuNP/Gal-1 indeed helped production of anti-influenza virus antibody through directly wrapping virus and stimulating dendritic cells activation. Since galectin-1 can bind to different subtypes of influenza virus, AuNP/Gal-1 may have broad applications in the development of novel vaccines against different subtypes and strains of influenza virus.

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