Montico B, Lapenta C, Ravo M, Martorelli D, Muraro E, Zeng B, Comaro E, Spada M, Donati S, Santini S M, Tarallo R, Giurato G, Rizzo F, Weisz A, Belardelli F, Dolcetti R, Dal Col J
Centro di Riferimento Oncologico, Department of Translational Research, Immunopathology and Cancer biomarkers, Aviano (PN), Italy.
Istituto Superiore di Sanità, Department of Hematology, Oncology and Molecular Medicine, Rome, Italy.
Oncoimmunology. 2017 Jul 31;6(11):e1356964. doi: 10.1080/2162402X.2017.1356964. eCollection 2017.
Although promising, the clinical benefit provided by dendritic cell (DC)-based vaccines is still limited and the choice of the optimal antigen formulation is still an unresolved issue. We have developed a new DC-based vaccination protocol for aggressive and/or refractory lymphomas which combines the unique features of interferon-conditioned DC (IFN-DC) with highly immunogenic tumor cell lysates (TCL) obtained from lymphoma cells undergoing immunogenic cell death. We show that treatment of mantle cell lymphoma (MCL) and diffuse large B-cell lymphoma (DLBCL) cell lines with 9--retinoic acid and IFNα (RA/IFNα) induces early membrane exposure of Calreticulin, HSP70 and 90 together with CD47 down-regulation and enhanced HMGB1 secretion. Consistently, RA/IFNα-treated apoptotic cells and -TCLs were more efficiently phagocytosed by DCs compared to controls. Notably, cytotoxic T cells (CTLs) generated with autologous DCs pulsed with RA/IFNα-TCLs more efficiently recognized and specifically lysed MCL or DLBCL cells or targets loaded with several HLA-A0201 cyclin D1 or HLA-B0801 survivin epitopes. These cultures also showed an expansion of Th1 and Th17 cells and an increased Th17/Treg ratio. Moreover, DCs loaded with RA/IFNα-TCLs showed enhanced functional maturation and activation. NOD/SCID mice reconstituted with human peripheral blood lymphocytes and vaccinated with autologous RA/IFNα-TCL loaded-IFN-DCs showed lymphoma-specific T-cell responses and a significant decrease in tumor growth with respect to mice treated with IFN-DC unpulsed or loaded with untreated TCLs. This study demonstrates the feasibility and efficacy of the use of RA/IFNα to generate a highly immunogenic TCL as a suitable tumor antigen formulation for the development of effective anticancer DC-based vaccines.
尽管基于树突状细胞（DC）的疫苗前景广阔，但其提供的临床益处仍然有限，最佳抗原制剂的选择仍是一个未解决的问题。我们开发了一种针对侵袭性和/或难治性淋巴瘤的新型基于DC的疫苗接种方案，该方案将干扰素预处理的DC（IFN-DC）的独特特性与从经历免疫原性细胞死亡的淋巴瘤细胞获得的高度免疫原性肿瘤细胞裂解物（TCL）相结合。我们发现，用9-维甲酸和IFNα（RA/IFNα）处理套细胞淋巴瘤（MCL）和弥漫性大B细胞淋巴瘤（DLBCL）细胞系会诱导钙网蛋白、HSP70和90的早期膜暴露，同时CD47下调并增强HMGB1分泌。一致地，与对照相比，RA/IFNα处理的凋亡细胞和-TCLs被DC更有效地吞噬。值得注意的是，用RA/IFNα-TCLs脉冲的自体DC产生的细胞毒性T细胞（CTL）更有效地识别并特异性裂解MCL或DLBCL细胞或负载有几种HLA-A0201细胞周期蛋白D1或HLA-B0801生存素表位的靶标。这些培养物还显示Th1和Th17细胞扩增以及Th17/Treg比率增加。此外，负载RA/IFNα-TCLs的DC显示出功能成熟和激活增强。用人外周血淋巴细胞重建并用自体负载RA/IFNα-TCL的IFN-DC接种的NOD/SCID小鼠显示出淋巴瘤特异性T细胞反应，并且相对于用未脉冲的IFN-DC或负载未处理TCLs的小鼠，肿瘤生长显著减少。这项研究证明了使用RA/IFNα产生高度免疫原性TCL作为开发有效的基于DC的抗癌疫苗的合适肿瘤抗原制剂的可行性和有效性。
