PLGA nanoparticle-mediated delivery of tumor antigenic peptides elicits effective immune responses.
PLGA nanoparticle-mediated delivery of tumor antigenic peptides elicits effective immune responses.
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DOI：
10.2147/ijn.s29506
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发表时间：
2012
影响因子：
8
通讯作者：
Minev B
中科院分区：
文献类型：
--
作者：
Ma W;Chen M;Kaushal S;McElroy M;Zhang Y;Ozkan C;Bouvet M;Kruse C;Grotjahn D;Ichim T;Minev B
由于在稳定性和递送方面存在困难，导致抗原呈递效率低下，癌症患者的应答率低，肽疫苗临床试验取得的成功有限。本研究的目的是开发一种新的肿瘤抗原肽递送方法，通过使用包封肿瘤抗原肽的聚（dl -乳酸-co-glycolide）纳米颗粒（PLGA-NPs）来引发增强的免疫反应。采用双乳液-溶剂蒸发法制备PLGA-NPs。人工抗原提呈细胞是由人树突状细胞（dc）负载包封肿瘤抗原肽的PLGA-NPs生成的。抗原呈递效率采用干扰素-γ ELISpot测定（Vector Laboratories, Burlingame， CA）。产生抗原特异性细胞毒性T淋巴细胞（ctl），并通过CytoTox 96®非放射性细胞毒性试验（Promega, Fitchburg， WI）进行评估。比较了不完全弗氏佐剂乳化和PLGA-NPs包封两种方法的肽递送效率。结果表明，大多数PLGA-NPs的直径在150 ~ 500 nm之间，在pH 7.4时带负电荷，平均zeta电位为- 15.53±0.71 mV；PLGA-NPs可以在30分钟的培养时间内在人dc中进行共定位。负载PLGA-NPs包封肽的人树突状细胞诱导的CTL毒性显著高于自由肽，而负载PLGA-NPs包封三肽鸡尾酒的人树突状细胞诱导的CTL反应显著高于包封两肽鸡尾酒的人树突状细胞。最重要的是，PLGA-NPs包封的肽剂量比不完全弗氏佐剂乳化的肽剂量少63倍，但在体内诱导的CTL反应更强。这些结果表明，递送包裹在PLGA-NPs中的肽是一种在体内诱导有效的抗肿瘤CTL反应的有希望的方法。
The peptide vaccine clinical trials encountered limited success because of difficulties associated with stability and delivery, resulting in inefficient antigen presentation and low response rates in patients with cancer. The purpose of this study was to develop a novel delivery approach for tumor antigenic peptides in order to elicit enhanced immune responses using poly(DL-lactide-co-glycolide) nanoparticles (PLGA-NPs) encapsulating tumor antigenic peptides. PLGA-NPs were made using the double emulsion-solvent evaporation method. Artificial antigen-presenting cells were generated by human dendritic cells (DCs) loaded with PLGA-NPs encapsulating tumor antigenic peptide(s). The efficiency of the antigen presentation was measured by interferon-γ ELISpot assay (Vector Laboratories, Burlingame, CA). Antigen-specific cytotoxic T lymphocytes (CTLs) were generated and evaluated by CytoTox 96® Non-Radioactive Cytotoxicity Assay (Promega, Fitchburg, WI). The efficiency of the peptide delivery was compared between the methods of emulsification in incomplete Freund’s adjuvant and encapsulation in PLGA-NPs. Our results showed that most of the PLGA-NPs were from 150 nm to 500 nm in diameter, and were negatively charged at pH 7.4 with a mean zeta potential of −15.53 ± 0.71 mV; the PLGA-NPs could be colocalized in human DCs in 30 minutes of incubation. Human DCs loaded with PLGA-NPs encapsulating peptide induced significantly stronger CTL cytotoxicity than those pulsed with free peptide, while human DCs loaded with PLGA-NPs encapsulating a three-peptide cocktail induced a significantly greater CTL response than those encapsulating a two-peptide cocktail. Most importantly, the peptide dose encapsulated in PLGA-NPs was 63 times less than that emulsified in incomplete Freund’s adjuvant, but it induced a more powerful CTL response in vivo. These results demonstrate that the delivery of peptides encapsulated in PLGA-NPs is a promising approach to induce effective antitumor CTL responses in vivo.