Kershaw Michael H, Wang Gang, Westwood Jennifer A, Pachynski Russell K, Tiffany H Lee, Marincola Francesco M, Wang Ena, Young Howard A, Murphy Philip M, Hwu Patrick
Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892, USA.
Hum Gene Ther. 2002 Nov 1;13(16):1971-80. doi: 10.1089/10430340260355374.
T-cell-based immunotherapies provide a promising means of cancer treatment although durable antitumor responses are infrequent. A potential reason for these shortcomings may lie in the observed lack of trafficking of specific T cells to tumor. Our increasing knowledge of the process of trafficking involving adhesion molecules and chemokines affords us the opportunity to intervene and correct deficiencies in this process. Chemokines can be expressed by a range of tumors and may serve as suitable targets for directing specific T cells toward tumor. We initially sought to identify which chemokines were produced by a range of human tumor cell lines, and which chemokines and chemokine receptors were expressed by cultured T cells. We identified two chemokines: Growth-Regulated Oncogene-alpha (Gro-alpha; CXCL1) and Regulated on Activation Normal T Cell-Expressed and Secreted (RANTES; CCL5), to be secreted by several human tumor cell lines. Expression was also detected in fine-needle aspirates of melanoma from patients. In addition, we determined the expression of several chemokine receptors on cultured human T cells including CCR1, CCR2, CCR4, CCR5, CXCR3, and CXCR4. Cultured, activated human T cells expressed the chemokines lymphotactin (XCL1), RANTES, macrophage inflammatory protein-1 alpha (MIP-1 alpha; CCL3) and MIP-1 beta (CCL4), but no appreciable Gro-alpha. In a strategy to direct T cells toward chemokines expressed by tumors we chose Gro-alpha as the target chemokine because it was produced by tumor and not by T cells themselves. However, T cells did not express the receptor for Gro-alpha, CXCR2, and therefore, T cells were transduced with a retroviral vector encoding CXCR2. Calcium ion mobilization, an important first step in chemokine receptor signaling, was subsequently demonstrated in transduced T cells in response to Gro-alpha. In addition, Gro-alpha was chemotactic for T cells expressing CXCR2 in vitro toward both recombinant protein and tumor-derived chemokine. Interestingly we demonstrate, for the first time, that Gro-alpha was able to induce interferon-gamma (IFN-gamma) secretion from transduced T cells, thereby extending our knowledge of other potential functions of CXCR2. This study demonstrates the feasibility of redirecting the migration properties of T cells toward chemokines secreted by tumors.
基于T细胞的免疫疗法为癌症治疗提供了一种很有前景的手段，尽管持久的抗肿瘤反应并不常见。这些缺点的一个潜在原因可能在于观察到特定T细胞向肿瘤的迁移不足。我们对涉及黏附分子和趋化因子的迁移过程的了解日益增加，这为我们提供了干预和纠正这一过程中缺陷的机会。趋化因子可由多种肿瘤表达，并可作为将特定T细胞导向肿瘤的合适靶点。我们最初试图确定一系列人类肿瘤细胞系产生哪些趋化因子，以及培养的T细胞表达哪些趋化因子和趋化因子受体。我们鉴定出两种趋化因子：生长调节致癌基因α（Gro-α；CXCL1）和活化正常T细胞表达和分泌调节因子（RANTES；CCL5），它们由几种人类肿瘤细胞系分泌。在患者黑色素瘤的细针穿刺抽吸物中也检测到了表达。此外，我们确定了培养的人类T细胞上几种趋化因子受体的表达，包括CCR1、CCR2、CCR4、CCR5、CXCR3和CXCR4。培养的活化人类T细胞表达趋化因子淋巴细胞趋化因子（XCL1）、RANTES、巨噬细胞炎性蛋白-1α（MIP-1α；CCL3）和MIP-1β（CCL4），但没有明显的Gro-α。在将T细胞导向肿瘤表达的趋化因子的策略中，我们选择Gro-α作为目标趋化因子，因为它是由肿瘤产生的，而不是由T细胞自身产生的。然而，T细胞不表达Gro-α的受体CXCR2，因此，用编码CXCR2的逆转录病毒载体转导T细胞。随后在转导的T细胞中证明了钙离子动员，这是趋化因子受体信号传导的重要第一步，以响应Gro-α。此外，Gro-α在体外对表达CXCR2的T细胞具有趋化作用，使其向重组蛋白和肿瘤来源的趋化因子迁移。有趣的是，我们首次证明Gro-α能够诱导转导的T细胞分泌干扰素-γ（IFN-γ），从而扩展了我们对CXCR2其他潜在功能的认识。这项研究证明了将T细胞的迁移特性重定向到肿瘤分泌的趋化因子的可行性。
