首页> 外文期刊>Acta biomaterialia >Quantitative determination of the composition of multi-shell calcium phosphate-oligonucleotide nanoparticles and their application for the activation of dendritic cells.
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Quantitative determination of the composition of multi-shell calcium phosphate-oligonucleotide nanoparticles and their application for the activation of dendritic cells.

机译:多壳磷酸钙寡核苷酸纳米颗粒组成的定量测定及其在树突状细胞激活中的应用。

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Biodegradable calcium phosphate nanoparticles as carriers for the immunoactive toll-like receptor ligands CpG and polyinosinic-polycytidylic acid for the activation of dendritic cells (DC) combined with the viral antigen hemagglutinin (HA) were prepared. A purification method based on ultracentrifugation and ultrasonication was developed to separate the nanoparticles from dissolved biomolecules. The number of biomolecules, i.e., oligonucleotides and peptide, incorporated into the nanoparticles was quantitatively determined by UV-spectroscopy, using fluorescent derivatives of the biomolecules. The immunostimulatory effects of purified calcium phosphate nanoparticles on DC were studied, i.e., cytokine production and activation of the cells in terms of the upregulation of surface molecules. Purified calcium phosphate nanoparticles, i.e., without dissolved biomolecules, are capable of inducing adaptive immunity by activation of DC. Immunostimulatory effects of purified calcium phosphate nanoparticles on DC were demonstrated by increased expression of co-stimulatory molecules and MHC II and by cytokine secretion. In addition, DC treated with purified functionalized calcium phosphate nanoparticles induced an antigen-specific T-cell response in vitro.
机译:制备了可生物降解的磷酸钙纳米粒子作为免疫活性通行费样受体配体CpG的载体,以及用于激活树突状细胞(DC)与病毒抗原血凝素(HA)结合的多肌苷-聚胞苷酸。开发了一种基于超速离心和超声处理的纯化方法,将纳米颗粒与溶解的生物分子分离。使用生物分子的荧光衍生物,通过UV光谱法定量地确定结合到纳米颗粒中的生物分子的数量,即寡核苷酸和肽。研究了纯化的磷酸钙纳米颗粒对DC的免疫刺激作用,即就表面分子的上调而言,细胞因子的产生和细胞的活化。纯化的磷酸钙纳米粒子,即没有溶解的生物分子,能够通过激活DC来诱导适应性免疫。纯化的磷酸钙纳米颗粒对DC的免疫刺激作用通过共刺激分子和MHC II的表达增加以及细胞因子的分泌得到证明。此外,用纯化的功能化磷酸钙纳米颗粒处理的DC在体外诱导了抗原特异性T细胞反应。

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