奈米金與免疫系統間的生物奈米相互作用

Abstract

奈米金粒子已被用來做為抗原或標靶藥物傳遞的載體，根據我們之前的實驗結果顯示奈米金會依尺寸大小進入脾臟內。脾臟是人體重要的免疫器官。不過，奈米金在體液免疫的相關功能機制目前仍不清楚。 本文的目的主要是在研究奈米金與免疫系統間的相互關係。 全文分為兩個部分。第一個部分: 我們將不同粒徑的奈米金 (2, 5, 8, 12, 17, 37 and 50nm)分別與小鼠antibody-generating cell一起培養6, 12, 18 和24小時。結果顯示細胞的存活率不受不同粒徑的奈米金影響。2, 5, 8 和12奈米的奈米金與細胞培養12小時後會增強細胞分泌出更多的免疫球蛋白G。而且我們發現2, 5, 8 和12奈米的奈米金刺激細胞分泌出更多的抗體，這機制是透過上調blimp1及下調pax5的基因表現。 因此，2到12奈米的奈米金可被用來當佐劑去刺激體液免疫的B 細胞產生更多的抗體。但如果是運用在標靶藥物運送的話，2到12奈米的奈米金可能會使體內分泌出辨認藥物或辨認配體的抗體，使藥物失效，無法達到治療的效果。第二部分:製備出辨認奈米金的單株抗體。將2奈米和5奈米的奈米金分別注入老鼠體內，接著犧牲老鼠，取出脾臟和FO 細胞進行細胞融合形成融合瘤，再利用HAT培養液篩選出融合成功的融合瘤，然後將細胞單株化，最後進行大量的ELISA 篩選出會分泌辨認奈米金抗體的融合瘤。以利實驗室之後可用來研究生物與奈米間的相互關係或探討到底這個抗體到底是如何被產生出來的。

Gold nanoparticles (GNPs) have been used to as antigen and targeted drug delivery carrier that showed significant size-dependent biodistribution in the spleen. The spleen is the largest and the most important immune organ in the body. However mechanistic details about the function of GNPs in humoral immunity system is still unclear at present time. The aim of this study was to explore the interaction of GNPs and immune system. This study is divided into two sections. The first section of this study was to identify GNPs associated with antibody secretion in B cell. The different size of GNPs (2, 5, 8, 12, 17, 37, 50 nm) were used to treat murine antibody-generating cells for 6, 12, 18 and 24 hours. Cell viability was not affected by the GNP treatment. Secretion of immunoglobulin G was enhanced 12 hrs after the treatment of 2, 5, 8, and 12 nm GNPs. GNPs treatment up-regulated blimp1 and down-regulated pax5 gene expressions. Here we show that GNPs ranging from 2-12nm stimulate secretion of antibody of B cells through blimp1 pathway. Thus, the 2-12nm GNPs can be used as an adjuvant to stimulate B cell producing a lot of antibodies, in which is enhance humoral immunity. If use in targeted drug delivery, 2-12 nm GNPs may cause the body generate against drugs or against ligands antibody, resulting in drug failure, so that drugs eventually lead to lack of therapeutic efficacy. The second section of this study was to generation of monoclonal antibody against GNP. Construction of cell lines producing a desired monoclonal antibody requires the following four steps: immunizing mouse, obtaining immune cells from its spleen, fusing the spleen cells with myeloma cells to make them immortal, and selecting the hybridoma cell lines producing the desired monoclonal antibody. In the future, we can explore bio-nano interaction and how anti-GNP antibody to generate in vivo .