Protein transduction of dendritic cells for NY-ESO-1-based immunotherapy of myeloma
Protein transduction of dendritic cells for NY-ESO-1-based immunotherapy of myeloma
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DOI：
10.1158/0008-5472.can-05-1383
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发表时间：
2005-11-01
期刊：
影响因子：
11.2
通讯作者：
van Rhee, F
中科院分区：
文献类型：
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作者：
Batchu, RB;Moreno, AM;van Rhee, F
基于独特型或肿瘤裂解物脉冲的树突状细胞的骨髓瘤疫苗取得了有限的成功，部分原因可能是由于骨髓瘤抗原的交叉启动不足。将骨髓瘤相关抗原引入树突状细胞细胞质的一种有效方法是蛋白质转导，这是一种与蛋白质转导结构域（PTD）融合的蛋白质自由穿过膜屏障的过程。NY-ESO-1是一种免疫原性抗原，其本身在60%的高危骨髓瘤患者中高度表达，该抗原被纯化为接近均匀性，无论是单独还是作为重组融合蛋白与源自HIV-Tat的PTD。通过显微镜、Western blotting和细胞内流式细胞术证实，PTD-NY-ESO-1有效进入树突状细胞，而不影响树突状细胞表型。通过抑制内吞作用的阿米洛利和蛋白酶体抑制剂n -乙酰基-l -亮氨酸-l -去亮氨酸的实验，证实PTD-NY-ESO-1通过蛋白转导进入树突状细胞，并被蛋白酶体降解。四聚体分析显示，与NY-ESO-1对照蛋白相比，PTD-NY-ESO-1特异性HLA-A2.1, CD8(+) T淋巴细胞（157-165）的生成优于NY-ESO-1（分别为44%和2%）。PTD-NY-ESO-1产生的ny - eso -1特异性T淋巴细胞分泌ifn - γ，表明tc1型细胞因子反应。因此，PTD-NY-ESO-1通过蛋白转导进入细胞质，被蛋白酶体处理，HLA I类呈递的NY-ESO-1肽引发NY-ESO-1特异性T淋巴细胞。
Myeloma vaccines, based on dendritic cells pulsed with idiotype or tumor lysate, have been met with limited success, probably in part due to insufficient cross-priming of myeloma antigens. A powerful method to introduce myeloma-associated antigens into the cytosol of dendritic cells is protein transduction, a process by which proteins fused with a protein transduction domain (PTD) freely traverse membrane barriers. NY-ESO-1, an immunogenic antigen by itself highly expressed in 60% of high-risk myeloma patients, was purified to near homogeneity both alone and as a recombinant fusion protein with a PTD, derived from HIV-Tat. Efficient entry of PTD-NY-ESO-1 into dendritic cells, confirmed by microscopy, Western blotting, and intracellular flow cytometry, was achieved without affecting dendritic cell phenotype. Experiments with amiloride, which inhibits endocytosis, and N-acetyl-L-leucinyl-L-norleucinal, a proteasome inhibitor, confirmed that PTD-NY-ESO-1 entered dendritic cells by protein transduction and was degraded by the proteasome. Tetramer analysis indicated superior generation of HLA-A2.1, CD8(+) T lymphocytes specific for NY-ESO-1(157-165) with PTD-NY-ESO-1 compared with NY-ESO-1 control protein (44% versus 2%, respectively). NY-ESO-1-specific T lymphocytes generated with PTD-NY-ESO-1 secreted IFN-gamma indicative of a Tc1-type cytokine response. Thus, PTD-NY-ESO-1 accesses the cytoplasm by protein transduction, is processed by the proteasome, and NY-ESO-1 peptides presented by HLA class I elicit NY-ESO-1-specific T lymphocytes.