Messenger RNA-electroporated dendritic cells presenting MAGE-A3 simultaneously in HLA class I and class II molecules
Messenger RNA-electroporated dendritic cells presenting MAGE-A3 simultaneously in HLA class I and class II molecules
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DOI：
10.4049/jimmunol.172.11.6649
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发表时间：
2004-06-01
影响因子：
4.4
通讯作者：
Thielemans, K
中科院分区：
文献类型：
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作者：
Bonehill, A;Heirman, C;Thielemans, K
最佳的抗癌疫苗可能需要CD 4（+）Th细胞和CD 8（+）CTL的共同作用。因此，癌症免疫治疗中一种有前途的工具是通过引入肿瘤Ag的编码区对树突状细胞（DC）进行遗传修饰，其中抗原肽将在HLA I类和II类分子中呈递。这可以通过将肿瘤Ag连接到内体或溶酶体蛋白的HLA II类靶向序列来实现。在这项研究中，我们比较了恒定链，溶酶体相关膜蛋白1（LAMP 1）和DC-LAMP的靶向信号的效率。在人DC成熟之前或之后，用编码未修饰的增强型绿色荧光蛋白（eGFP）或与各种靶向信号连接的eGFP的mRNA电穿孔人DC。加入溶酶体降解抑制剂氯喹，并在电穿孔后的不同时间点评价eGFP表达。还用未修饰的MAGE-A3或与靶向信号连接的MAGE-A3对DC进行电穿孔，并研究了在HLA I类和II类分子背景下MAGE-A3衍生表位的提呈。我们的数据表明，与不同的靶向信号连接的蛋白质靶向溶酶体，并且确实在HLA I类和II类分子的背景下呈现，但具有不同的效率。与LAMP 1或DC-LAMP信号连接的蛋白质比与不变链靶向信号连接的蛋白质更有效地呈递。此外，成熟后电穿孔的DC比成熟前电穿孔的DC在Ag呈递方面更有效。
An optimal anticancer vaccine probably requires the cooperation of both CD4(+) Th cells and CD8(+) CTLs. A promising tool in cancer immunotherapy is, therefore, the genetic modification of dendritic cells (DCs) by introducing the coding region of a tumor Ag, of which the antigenic peptides will be presented in both HLA class I and class II molecules. This can be achieved by linking the tumor Ag to the HLA class II-targeting sequence of an endosomal or lysosomal protein. In this study we compared the efficiency of the targeting signals of invariant chain, lysosome-associated membrane protein-1 (LAMP1) and DC-LAMP. Human DCs were electroporated before or after maturation with mRNA encoding unmodified enhanced green fluorescent protein (eGFP) or eGFP linked to various targeting signals. The lysosomal degradation inhibitor chloroquine was added, and eGFP expression was evaluated at different time points after electroporation. DCs were also electroporated with unmodified MAGE-A3 or MAGE-A3 linked to the targeting signals, and the presentation of MAGE-A3-derived epitopes in the context of HLA class I and class II molecules was investigated. Our data suggest that proteins linked to the different targeting signals are targeted to the lysosomes and are indeed presented in the context of HLA class I and class II molecules, but with different efficiencies. Proteins linked to the LAMP1 or DC-LAMP signal are more efficiently presented than proteins linked to the invariant chain-targeting signal. Furthermore, DCs electroporated after maturation are more efficient in Ag presentation than DCs electroporated before maturation.