Salgaller M L, Marincola F M, Cormier J N, Rosenberg S A
Surgery Branch, National Cancer Institute, NIH, Bethesda, Maryland 20892, USA.
Cancer Res. 1996 Oct 15;56(20):4749-57.
gp1OO is a melanocytic lineage-specific antigen recognized by tumor-infiltrating lymphocytes, the adoptive transfer of which is associated with tumor regression in melanoma patients. In this study, peripheral blood mononuclear cells (PBMCs) were harvested from HLA-A2+ melanoma patients before and after immunization with G9-209 (ITDQVPFSY), G9-280 (YLEPGPVTA), or G9-154 (KTWGQYWQV) peptides in Incomplete Freund's Adjuvant and were tested for the ability to be sensitized in vitro using PBMCs pulsed with the native peptides. In addition, PBMCs from patients receiving the G9-209 or G9-280 peptide were stimulated in vitro with peptides modified at anchor residues to enhance binding to HLA-A2: G9-209/2M (IMDQVPFSY) or G9-280-9V (YLEPGPVTV). In patients immunized with G9-209, a single in vitro restimulation with G9-209/2M resulted in the generation of specific antipeptide lymphocytes from seven of seven postimmune PBMCs and only three of seven preimmune PBMCs. In patients immunized with G9-280, a single in vitro restimulation with G9-280/9V resulted in the generation of specific antipeptide lymphocytes from five of six postimmune PBMCs and four of six preimmune PBMCs. In almost all cases, CTLs raised against modified epitopes were capable of recognizing targets displaying the native nonamers. Several anti-G9-209 and anti-G9-209/2M CTLs also demonstrated specific lysis of, and specific IFN-gamma release in response to, gp1OO+-established cell lines. Thus, using peptides modified to enhance immunogenicity for in vitro stimulation improved the sensitivity of immune monitoring of patients immunized with synthetic peptides. These results demonstrate that immunization with a peptide derived from a tumor-associated protein such as gp100 can provoke a measurable antitumor immune response in cancer patients.
gp100是一种黑色素细胞系特异性抗原，可被肿瘤浸润淋巴细胞识别，其过继性转移与黑色素瘤患者的肿瘤消退相关。在本研究中，从HLA - A2 +黑色素瘤患者中采集外周血单个核细胞（PBMC），这些患者在用不完全弗氏佐剂中的G9 - 209（ITDQVPFSY）、G9 - 280（YLEPGPVTA）或G9 - 154（KTWGQYWQV）肽免疫之前和之后进行采集，并使用用天然肽脉冲处理的PBMC测试其在体外被致敏的能力。此外，来自接受G9 - 209或G9 - 280肽的患者的PBMC在体外被锚定残基修饰的肽刺激，以增强与HLA - A2的结合：G9 - 209/2M（IMDQVPFSY）或G9 - 280 - 9V（YLEPGPVTV）。在用G9 - 209免疫的患者中，用G9 - 209/2M进行单次体外再刺激导致7个免疫后PBMC中有7个产生特异性抗肽淋巴细胞，而7个免疫前PBMC中只有3个产生。在用G9 - 280免疫的患者中，用G9 - 280/9V进行单次体外再刺激导致6个免疫后PBMC中有5个产生特异性抗肽淋巴细胞，6个免疫前PBMC中有4个产生。在几乎所有情况下，针对修饰表位产生的细胞毒性T淋巴细胞（CTL）能够识别展示天然九聚体的靶标。几种抗G9 - 209和抗G9 - 209/2M CTL还表现出对gp100 +建立的细胞系的特异性裂解以及对其产生特异性γ干扰素释放。因此，使用修饰以增强体外刺激免疫原性的肽提高了用合成肽免疫的患者免疫监测的敏感性。这些结果表明，用源自肿瘤相关蛋白（如gp100）的肽进行免疫可在癌症患者中引发可测量的抗肿瘤免疫反应。
