Brando Clara, Mukhopadhyay Sunil, Kovacs Eniko, Medina Rosa, Patel Pritesh, Catina Tracey L, Campbell Kerry S, Santoli Daniela
The Wistar Institute, Philadelphia, PA, USA.
J Leukoc Biol. 2005 Aug;78(2):359-71. doi: 10.1189/jlb.0604360. Epub 2005 Jun 3.
The major histocompatibility complex nonrestricted cytotoxic leukemic T cell line T acute lymphoblastic leukemia (TALL)-104 is being pursued as a therapeutic agent for cancer. However, the receptors and effector mechanisms responsible for its broad tumoricidal function remain undefined. Here, we examined the roles played by natural cytotoxicity receptors (NCR), killer cell immunoglobulin-like receptors, cytolytic granule components, and tumor necrosis factor (TNF) family members in tumor recognition and lysis by TALL-104 cells. The perforin-granzyme pathway, TNF-related apoptosis-inducing ligand (TRAIL), and Fas were each involved in the lysis of particular tumor targets by TALL-104. Furthermore, phorbol 12-myristate 13-acetate/ionomycin treatment induced surface expression of Fas-L and TRAIL. In addition, supernatants from CD3-stimulated TALL-104 cultures exhibited antiproliferative activity, which was blocked 50-90% by anti-TNF-alpha monoclonal antibody (mAb). Although negative for the NCR natural killer (NK)p44, this cell line was found to express NKp46. An anti-NKp46 antibody strongly blocked TALL-104-mediated lysis of certain targets and directly induced cytokine production, granule release, and redirected lysis responses. Anti-NKG2D and anti-2B4 also stimulated redirected cytotoxicity by TALL-104. By contrast, anti-NKG2A mAb did not stain the cells or inhibit killing responses. Alternatively, KIR3DL2 was detected on TALL-104, and expression of its reported ligand, human leukocyte antigen (HLA)-A, on target cells provided protection from cytotoxicity. Thus, NKp46, NKG2D, and 2B4 are activating receptors, and KIR3DL2 is an inhibitory receptor on TALL-104. The data demonstrate the ability of TALL-104 cells to recognize a wide variety of tumors with NK cell receptors and kill them with a broad arsenal of cytolytic effector mechanisms, including cytolytic granules and TNF family ligands.
主要组织相容性复合体非限制性细胞毒性白血病T细胞系T急性淋巴细胞白血病（TALL）-104正被作为一种癌症治疗药物进行研究。然而，其广泛的杀瘤功能所涉及的受体和效应机制仍不明确。在此，我们研究了自然细胞毒性受体（NCR）、杀伤细胞免疫球蛋白样受体、溶细胞颗粒成分和肿瘤坏死因子（TNF）家族成员在TALL-104细胞识别和裂解肿瘤中的作用。穿孔素-颗粒酶途径、TNF相关凋亡诱导配体（TRAIL）和Fas均参与了TALL-104对特定肿瘤靶标的裂解。此外，佛波酯12-肉豆蔻酸酯13-乙酸盐/离子霉素处理可诱导Fas-L和TRAIL的表面表达。另外，CD3刺激的TALL-104培养上清液表现出抗增殖活性，抗TNF-α单克隆抗体（mAb）可阻断50%-90%的该活性。尽管该细胞系的NCR自然杀伤（NK）p44呈阴性，但发现其表达NKp46。抗NKp46抗体强烈阻断TALL-104介导的对某些靶标的裂解，并直接诱导细胞因子产生、颗粒释放和重定向裂解反应。抗NKG2D和抗2B4也刺激TALL-104产生重定向细胞毒性。相比之下，抗NKG2A mAb未对细胞进行染色或抑制杀伤反应。另外，在TALL-104上检测到了KIR3DL2，其报道的配体人类白细胞抗原（HLA）-A在靶细胞上的表达提供了对细胞毒性的保护。因此，NKp46、NKG2D和2B4是激活受体，而KIR3DL2是TALL-104上的抑制性受体。数据表明TALL-104细胞能够利用NK细胞受体识别多种肿瘤，并通过包括溶细胞颗粒和TNF家族配体在内的广泛溶细胞效应机制将其杀死。
