Neutral polymer micelle carriers with pH-responsive, endosome-releasing activity modulate antigen trafficking to enhance CD8(+) T cell responses.
Neutral polymer micelle carriers with pH-responsive, endosome-releasing activity modulate antigen trafficking to enhance CD8(+) T cell responses.
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DOI：
10.1016/j.jconrel.2014.03.041
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发表时间：
2014-10-10
期刊：
影响因子：
--
通讯作者：
Stayton PS
中科院分区：
文献类型：
--
作者：
Keller S;Wilson JT;Patilea GI;Kern HB;Convertine AJ;Stayton PS
合成亚单位疫苗需要诱导CD 8+细胞毒性T细胞（CTL）应答以有效接种对抗细胞内病原体。大多数亚单位疫苗主要产生体液免疫应答，具有弱于期望的CD 8+细胞毒性T细胞应答。在这里，中性的，pH响应性聚合物胶束载体，改变细胞内抗原运输显示，以增强CD 8 + T细胞的反应与相关的增加细胞溶质的交付和减少胞吐。聚合物二嵌段载体由N-（2-羟丙基）甲基丙烯酰胺冠状嵌段和用于硫醇化卵清蛋白可逆缀合的侧基吡啶基二硫化物基团组成，以及由丙基丙烯酸（PAA）、甲基丙烯酸二甲氨基乙酯（DMAEMA）和甲基丙烯酸丁酯（BMA）组成的三元共聚物两性核形成嵌段。二嵌段共聚物自组装成25-30 nm直径的胶束纳米粒子。相对于游离卵清蛋白、卵清蛋白和聚合物的未缀合的物理混合物以及非pH响应性胶束卵清蛋白对照，卵清蛋白与胶束的缀合显著增强了体外抗原交叉呈递。在小鼠树突状细胞系（DC2.4）的机制研究表明，胶束介导的增强细胞内抗原保留和胞质抗原积累。大约90%的初始内化的卵清蛋白缀合的胶束保留在细胞1.5小时后，相比之下，只有约40%的对照。此外，与缀合物给药的细胞显示67倍高的细胞溶质抗原水平相对于可溶性卵清蛋白4小时后摄取。与用可溶性蛋白、卵白蛋白和聚合物混合物以及没有内体释放活性的对照胶束免疫相比，用卵白蛋白-聚合物缀合物皮下免疫小鼠显著增强了抗原特异性CD 8 + T细胞应答（CD 8+的0.4%IFN-γ+）。此外，pH响应性载体促进抗原递送至引流淋巴结中的抗原呈递细胞。早在注射后90分钟，卵-胶束缀合物分别与28%和55%的树突状细胞和巨噬细胞相关。24小时后，偶联物优先与树突状细胞结合，相对于游离蛋白、物理混合物和非pH响应性偶联物对照，摄取分别增强30倍、3倍和3倍。这些结果证明了pH响应性聚合物胶束用于依赖于CD 8 + T细胞活化的疫苗应用的潜力。
Synthetic subunit vaccines need to induce CD8+ cytotoxic T-cell (CTL) responses for effective vaccination against intracellular pathogens. Most subunit vaccines primarily generate humoral immune responses, with a weaker than desired CD8+ cytotoxic T-cell response. Here, a neutral, pH-responsive polymer micelle carrier that alters intracellular antigen trafficking was shown to enhance CD8+ T-cell responses with a correlated increase in cytosolic delivery and a decrease in exocytosis. Polymer diblock carriers consisted of a N-(2-hydroxypropyl) methacrylamide corona block with pendant pyridyl disulfide groups for reversible conjugation of thiolated ovalbumin, and a tercopolymer ampholytic core-forming block composed of propylacrylic acid (PAA), dimethylaminoethyl methacrylate (DMAEMA), and butyl methacrylate (BMA). The diblock copolymers self-assembled into 25–30 nm diameter micellar nanoparticles. Conjugation of ovalbumin to the micelles significantly enhanced antigen cross-presentation in vitro relative to free ovalbumin, an unconjugated physical mixture of ovalbumin and polymer, and a non pH-responsive micelle-ovalbumin control. Mechanistic studies in a murine dendritic cell line (DC2.4) demonstrated micelle-mediated enhancements in intracellular antigen retention and cytosolic antigen accumulation. Approximately 90% of initially internalized ovalbumin-conjugated micelles were retained in cells after 1.5 h, compared to only ~40% for controls. Furthermore, cells dosed with conjugates displayed 67-fold higher cytosolic antigen levels relative to soluble ovalbumin 4 h post uptake. Subcutaneous immunization of mice with ovalbumin-polymer conjugates significantly enhanced antigen-specific CD8+ T cell responses (0.4 % IFN-γ+ of CD8+) compared to immunization with soluble protein, ovalbumin and polymer mixture, and the control micelle without endosome-releasing activity. Additionally, pH-responsive carrier facilitated antigen delivery to antigen presenting cells in the draining lymph nodes. As early as 90 min post injection ova-micelle conjugates were associated with 28% and 55% of dendritic cells and macrophages, respectively. After 24 h, conjugates preferentially associated with dendritic cells, affording 30-, 3-, and 3-fold enhancements in uptake relative to free protein, physical mixture, and the non pH-responsive conjugate controls, respectively. These results demonstrate the potential of pH-responsive polymeric micelles for use in vaccine applications that rely on CD8+ T cell activation.
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影响因子：
4.7
作者：
Lundy, Brittany B.;Convertine, Anthony;Miteva, Martina;Stayton, Patrick S.
通讯作者：
Stayton, Patrick S.
DOI：
10.1016/j.jconrel.2008.10.004
发表时间：
2009-02-10
期刊：
Journal of controlled release : official journal of the Controlled Release Society
影响因子：
--
作者：
Convertine AJ;Benoit DS;Duvall CL;Hoffman AS;Stayton PS
通讯作者：
Stayton PS
DOI：
10.1084/jem.166.6.1654
发表时间：
1987-12-01
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Macatonia SE;Knight SC;Edwards AJ;Griffiths S;Fryer P
通讯作者：
Fryer P
影响因子：
10.8
作者：
Murthy, N;Robichaud, JR;Hoffman, AS
通讯作者：
Hoffman, AS
影响因子：
4.7
作者：
Foster, Suzanne;Duvall, Craig L.;Crownover, Emily F.;Hoffman, Allan S.;Stayton, Patrick S.
通讯作者：
Stayton, Patrick S.