Wang M, Chen P W, Bronte V, Rosenberg S A, Restifo N P
Howard Hughes Medical Institute-National Institutes of Health Research Scholars Program, National Institutes of Health, Bethesda, Maryland, USA.
J Immunother Emphasis Tumor Immunol. 1995 Oct;18(3):139-46. doi: 10.1097/00002371-199510000-00001.
The recent cloning of tumor-associated antigens (TAAs) recognized by CD8+ T lymphocytes (TCD8+) has made it possible to use recombinant and synthetic forms of TAAs to generate TCD8+ with anti-tumor activity. To explore new therapeutic strategies in a mouse model, we retrovirally transduced the experimental murine tumor CT26(H-2d), with the lacZ gene encoding our model TAA, beta-galactosidase (beta-gal). The transduced cell line, CT26.CL25, grew as rapidly and as lethally as the parental cell line in normal, immunocompetent animals. In an attempt to elicit TCD8+ directed against our model TAA by using purely recombinant and synthetic forms of our model TAA, we synthesized a nine-amino-acid long immunodominant peptide of beta-gal (TPH-PARIGL), corresponding to amino acid residues 876-884, which was known to be presented by the Ld major histocompatibility complex (MHC) class I molecule, and a recombinant vaccinia virus encoding the full-length beta-gal protein (VJS6). Splenocytes obtained from naïve mice and co-cultured with beta-gal peptide could not be expanded in primary ex vivo cultures. However, mice immunized with VJS6, but not with a control recombinant vaccinia virus, yielded splenocytes that were capable of specifically lysing CT26.CL25 in vitro after co-culture with beta-gal peptide. Most significantly, adoptive transfer of these cells could effectively treat mice bearing 3-day-old established pulmonary metastases. These observations show that therapeutic TCD8+ directed against a model TAA could be generated by using purely recombinant and synthetic forms of this antigen. These findings point the way to a potentially useful immunotherapeutic strategy, which has been made possible by the recent cloning of immunogenic TAAs that are expressed by human malignancies.
最近克隆出了可被CD8⁺ T淋巴细胞（TCD8⁺）识别的肿瘤相关抗原（TAA），这使得利用重组和合成形式的TAA来产生具有抗肿瘤活性的TCD8⁺成为可能。为了在小鼠模型中探索新的治疗策略，我们用逆转录病毒转导了实验性鼠肿瘤CT26（H - 2d），使其带有编码我们的模型TAAβ - 半乳糖苷酶（β - gal）的lacZ基因。在正常的、具有免疫活性的动物中，转导后的细胞系CT26.CL25的生长速度和致死性与亲代细胞系一样快。为了尝试通过使用我们模型TAA的纯重组和合成形式来引发针对我们模型TAA的TCD8⁺，我们合成了一段由九个氨基酸组成的β - gal免疫显性肽（TPH - PARIGL），对应于氨基酸残基876 - 884，已知该肽段由Ld主要组织相容性复合体（MHC）I类分子呈递，还构建了一种编码全长β - gal蛋白的重组痘苗病毒（VJS6）。从未接触过抗原的小鼠获得的脾细胞与β - gal肽共培养后，在原代体外培养中无法扩增。然而，用VJS6免疫的小鼠（而非用对照重组痘苗病毒免疫的小鼠）产生的脾细胞，在与β - gal肽共培养后，能够在体外特异性裂解CT26.CL25。最显著的是，这些细胞的过继转移能够有效治疗患有3日龄已形成肺转移瘤的小鼠。这些观察结果表明，通过使用该抗原的纯重组和合成形式，可以产生针对模型TAA的治疗性TCD8⁺。这些发现为一种潜在有用的免疫治疗策略指明了方向，而最近克隆出的由人类恶性肿瘤表达的免疫原性TAA使得这种策略成为可能。
