Butterfield L H, Jilani S M, Chakraborty N G, Bui L A, Ribas A, Dissette V B, Lau R, Gamradt S C, Glaspy J A, McBride W H, Mukherji B, Economou J S
Division of Surgical Oncology, University of California Los Angeles Medical Center 90095-1782, USA.
J Immunol. 1998 Nov 15;161(10):5607-13.
Dendritic cells (DC) are potent stimulators of primary T cell responses. In this study, we demonstrate that DC, genetically engineered to express the MART-1/Melan-A (MART-1) tumor-associated Ag, express MART-1 mRNA and protein, correctly process and present the HLA-A2.1-restricted immunodominant MART-1 peptide (MART-1(27-35)), and serve as potent stimulators of MART-1-specific CTL in vitro. A replication-defective E1-deleted adenovirus (AdV) was constructed that expresses MART-1 (AdVMART1). Transduced DC produce full length MART-1 mRNA as well as MART-1 protein. AdVMART1 does not significantly down-regulate cell surface class I expression despite having an intact E3 region. Transduction of an HLA-A2-positive/MART-1-negative cell line with AdVMART1 renders these cells sensitive to lysis by CTL specific for the MART-1(27-35) immunodominant peptide. In addition, DC transduced with AdVMART1 stimulated MART-1(27-35)-specific tumor-infiltrating lymphocytes to synthesize IFN-gamma. Finally, AdVMART1-transduced DC were able to generate MART-1(27-35) peptide-specific, class I-restricted CTL in PBL cultures from normal donors. This study supports the use of tumor Ag-engineered DC in genetic immunotherapy.
树突状细胞（DC）是原发性T细胞反应的有效刺激物。在本研究中，我们证明，经基因工程改造以表达MART-1/Melan-A（MART-1）肿瘤相关抗原的DC，表达MART-1 mRNA和蛋白，能正确加工并呈递HLA-A2.1限制性免疫显性MART-1肽（MART-1(27-35)），并在体外作为MART-1特异性CTL的有效刺激物。构建了一种表达MART-1的复制缺陷型E1缺失腺病毒（AdV）（AdVMART1）。转导的DC产生全长MART-1 mRNA以及MART-1蛋白。尽管AdVMART1具有完整的E3区域，但它不会显著下调细胞表面I类分子的表达。用AdVMART1转导HLA-A2阳性/MART-1阴性细胞系，使这些细胞对针对MART-1(27-35)免疫显性肽的CTL裂解敏感。此外，用AdVMART1转导的DC刺激MART-1(27-35)特异性肿瘤浸润淋巴细胞合成IFN-γ。最后，AdVMART1转导的DC能够在来自正常供体的外周血淋巴细胞培养物中产生MART-1(27-35)肽特异性、I类限制性CTL。本研究支持在基因免疫治疗中使用肿瘤抗原工程化的DC。
