Cancer Bio-immunotherapy Unit, Centro di Riferimento Oncologico, National Cancer Institute, Aviano, Italy.
Cancer Immunol Immunother. 2012 Jun;61(6):881-92. doi: 10.1007/s00262-011-1157-5. Epub 2011 Nov 17.
The generation of efficacious vaccines against self-antigens expressed in tumor cells requires breakage of tolerance, and the refocusing of immune responses toward epitopes for which tolerance may not be established. While the presentation of tumor antigens by mature dendritic cells (mDC) may surpass tolerance, broadening of the antigenic repertoire remains an issue. We report that fusion of the candidate idiotype vaccine IGKV3-20 to the Gly-Ala repeat (GAr) of the Epstein-Barr virus nuclear antigen (EBNA)-1 inhibits degradation by the proteasome and redirects processing to the lysosome. mDCs transduced with a recombinant lentivirus encoding the chimeric idiotype efficiently primed CD4+ and CD8+ cytotoxic T-cell (CTL) responses that lysed autologous blasts expressing IGKV3-20 or pulsed with IGKV3-20 synthetic peptides, and HLA-matched IGKV3-20-positive tumor cell lines. Comparison of the cytotoxic response of CD4+ and CD8+ T lymphocytes activated by mDCs expressing the wild-type or chimeric IGKV3-20 reveled largely non-overlapping epitope repertoires in both CD4+ and CD8+ effectors. Thus, fusion to the GAr may provide an effective means to broaden the immune response to an endogenous protein by promoting the presentation of antigenic epitopes that require a lysosome-dependent processing step.
针对肿瘤细胞中表达的自身抗原生成有效疫苗需要打破耐受,并且重新聚焦免疫反应以针对可能未建立耐受的表位。虽然成熟树突状细胞(mDC)呈现肿瘤抗原可能超过耐受,但抗原谱的拓宽仍然是一个问题。我们报告说,候选独特型疫苗 IGKV3-20 与 Epstein-Barr 病毒核抗原(EBNA)-1 的甘氨酸-丙氨酸重复序列(GAr)融合可抑制蛋白酶体降解,并将加工重新定向到溶酶体。用编码嵌合独特型的重组慢病毒转导的 mDC 可有效引发 CD4+和 CD8+细胞毒性 T 细胞(CTL)反应,这些反应可裂解表达 IGKV3-20 的自体母细胞或脉冲 IGKV3-20 合成肽,并裂解 HLA 匹配的 IGKV3-20 阳性肿瘤细胞系。比较表达野生型或嵌合 IGKV3-20 的 mDC 激活的 CD4+和 CD8+T 淋巴细胞的细胞毒性反应,揭示了 CD4+和 CD8+效应物中抗原表位谱的大部分不重叠。因此,与 GAr 的融合可能通过促进需要溶酶体依赖性加工步骤的抗原表位的呈现,为内源性蛋白的免疫反应提供一种有效手段。