Clin Cancer Res 2011 Nov;17 (22): 7047-57. [IF:7.338]
Tumor-derived autophagosome vaccine: mechanism of cross-presentation and therapeutic efficacy.
Li Y , Wang LX , Pang P , Cui Z , Aung S , Haley D , Fox BA , Urba WJ , Hu HM .
Authors' Affiliations: Laboratories of Cancer Immunobiology, Molecular Tumor Immunology, and Immunological Monitoring, Robert W. Franz Cancer Research Center, Earle A. Chiles Research Institute, Providence Portland Medical Center; UbiVac, LLC, Portland, Oregon; Cancer Research and Biotherapy Center, The Second Nanjing Hospital; and Department of Microbiology and Immunology, School of Medicine, Southeast University, Nanjing, Jiangsu, People's Republic of China.
东南大学医学院病原生物学与免疫学系 ，南京第二医院，美国俄勒冈州波特兰医学中心癌症研究
Abstract
We previously reported that autophagy in tumor cells plays a critical role in cross-presentation of tumor antigens and that autophagosomes are efficient antigen carriers for cross-priming of tumor-reactive CD8(+) T cells. Here, we sought to characterize further the autophagosome-enriched vaccine named DRibble (DRiPs-containing blebs), which is derived from tumor cells after inhibition of protein degradation, and to provide insights into the mechanisms responsible for their efficacy as a novel cancer immunotherapy. DRibbles were characterized by Western blot and light or transmission electron microscopy. The efficiency of cross-presentation mediated by DRibbles was first compared with that of whole-tumor cells and pure proteins. The mechanisms of antigen cross-presentation by DRibbles were analyzed, and the antitumor efficacy of the DRibble vaccine was tested in 3LL Lewis lung tumors and B16F10 melanoma. The DRibbles sequester both long-lived and short-lived proteins, including defective ribosomal products (DRiP), and damage-associated molecular pattern molecules exemplified by HSP90, HSP94, calreticulin, and HMGB1. DRibbles express ligands for CLEC9A, a newly described C-type lectin receptor expressed by a subset of conventional and plasmacytoid dendritic cells (DC), and cross-presentation was partially CLEC9A dependent. Furthermore, this autophagy-assisted antigen cross-presentation pathway involved both caveolae- and clathrin-mediated endocytosis and endoplasmic reticulum-associated degradation machinery. It depends on proteasome and TAP1, but not lysosome functions of antigen-presenting cells. Importantly, DCs loaded with autophagosome-enriched DRibbles can eradicate 3LL Lewis lung tumors and significantly delay the growth of B16F10 melanoma. These data documented the unique characteristics and potent antitumor efficacy of the autophagosome-based DRibble vaccine. The efficacy of DRibble cancer vaccine will be further tested in clinical trials. Clin Cancer Res; 17(22); 7047-57. 2011 AACR.
摘要：
之前我们对自体吞噬在肿瘤抗原交叉提呈方面的重要作用以及自噬体作为有效抗原载体可以广泛刺激肿瘤特异性CD8+T细胞进行了报道。
现在我们准备对自噬体修饰疫苗Dribble（含有DRiPs的疱疹）进行进一步的表征，为其将来可以作为一种新的癌症免疫疗法提供一次那个的机制理论。
Dribbles用Western blot和光电子显微镜来表征。
Dribbles介导的交叉抗原提呈效率首先和整个肿瘤细胞以及纯蛋白的交叉抗原提成效率进行了比较。
我们分析了Dribbles抗原交叉提呈的机理，其抗肿瘤的效应在3LL Lewis 肺肿瘤细胞和B16F10黑色素瘤细胞中进行了测试。
DRibbles隔离了长寿命和短寿命的蛋白，包括一些收到损伤的核糖体产物(DRiP)和受损相关分子标识分子例如HSP90，HSP94，Calreticulin和HMGB1。
Dribbles可以表达CLEC9A的配体，这是正常组织的和浆细胞瘤组织中的树突状细胞（DC）的亚单位表达的一种C型凝集素受体，可以实现部分CLEC9A依赖性的交叉提呈。
并且，这种自噬体协助的抗原交叉提呈的过程同时包括细胞膜内穴样凹陷和网格蛋白介导的内吞以及内质网相关降解机制。
它依靠的是蛋白酶体和TAP1，并不是溶酶体介导的抗原提呈细胞。
更重要的是，负载了自噬体修饰Dribbles的树突状细胞可以杀死3LL Lewis 肺癌细胞并且可以显著抑制B16F10黑色素瘤细胞的生长。
这些数据证明了自噬体修饰的Dribble疫苗独特的性质和潜在的抗肿瘤效应。Dribble癌症疫苗的效果将会在临床试验中得到更加全面的测试。
