Joshi B H, Leland P, Silber J, Kreitman R J, Pastan I, Berger M, Puri R K
Laboratory of Molecular Tumour Biology, Division of Cellular and Gene Therapies, Center for Biologics, Evaluation and Research, Food and Drug Administration, NIH Building 29B, Room 2NN10, 29 Lincoln Dr., Bethesda, Maryland, MD 20892, USA.
Br J Cancer. 2002 Jan 21;86(2):285-91. doi: 10.1038/sj.bjc.6600034.
Cytotoxins directed to interleukin-4 receptors have shown to mediate relatively selective cytotoxicity against a variety of human cancer cells in vitro and in vivo. In an ongoing Phase I clinical trial, a recombinant protein comprised of circularly permuted IL-4 fused to a mutated form of Pseudomonas exotoxin (the fusion protein termed IL-4(38-37)-PE38KDEL or cpIL4-PE) has shown antitumour activity against malignant glioma. Human medulloblastomas are neuroectodermal tumours that occur in children and have a poor prognosis. The goal of this study was to determine whether human medulloblastoma derived cell lines express interleukin-4 receptor and whether interleukin-4 receptor expression is accompanied by sensitivity to cpIL4-PE. Medulloblastoma cell lines express interleukin-4 receptor at the protein and mRNA levels as determined by binding, indirect immunofluorescence and RT--PCR studies. These cells expressed IL-4Ralpha (also known as IL-4Rbeta) and IL-13Ralpha1 (also known as IL-13Ralpha') chains, however common gamma(c), a component of the interleukin-4 receptor system in immune cells was not detected. Consistent with the expression of IL-4R, cpIL4-PE was found to be highly and specifically cytotoxic to four of five medulloblastoma cell lines. Susceptibility of medulloblastoma cell lines to cpIL4-PE seemed to correlate closely to the functional IL-4 binding sites in general as demonstrated by 125I-IL-4 binding, but did not seem to correlate with mRNA or cell surface immunoreactive receptor protein expression. The sensitivity of medulloblastoma cells to cpIL4-PE could be eliminated by concurrent incubation with IL-4 or IL-13, but not with IL-2. None of these cell lines showed any change in proliferation upon treatment with exogenous IL-4. These studies establish the interleukin-4 receptor as a medulloblastoma-associated target for possible tumour-directed cancer therapy. Further studies are warranted to investigate interleukin-4 receptor expression in primary medulloblastoma tumours and sensitivity to cpIL-4PE in vitro and in vivo.
靶向白细胞介素-4受体的细胞毒素已显示在体外和体内对多种人类癌细胞介导相对选择性的细胞毒性。在一项正在进行的I期临床试验中,一种由环状排列的IL-4与突变形式的铜绿假单胞菌外毒素融合而成的重组蛋白(称为IL-4(38-37)-PE38KDEL或cpIL4-PE的融合蛋白)已显示出对恶性胶质瘤的抗肿瘤活性。人类髓母细胞瘤是发生于儿童的神经外胚层肿瘤,预后较差。本研究的目的是确定源自人类髓母细胞瘤的细胞系是否表达白细胞介素-4受体,以及白细胞介素-4受体表达是否伴随着对cpIL4-PE的敏感性。通过结合、间接免疫荧光和RT-PCR研究确定,髓母细胞瘤细胞系在蛋白质和mRNA水平表达白细胞介素-4受体。这些细胞表达IL-4Rα(也称为IL-4Rβ)和IL-13Rα1(也称为IL-13Rα')链,然而未检测到免疫细胞中白细胞介素-4受体系统的一个组成部分——共同γ链(common gamma(c))。与IL-4R的表达一致,发现cpIL4-PE对五个髓母细胞瘤细胞系中的四个具有高度特异性的细胞毒性。如125I-IL-4结合所示,髓母细胞瘤细胞系对cpIL4-PE的敏感性总体上似乎与功能性IL-4结合位点密切相关,但似乎与mRNA或细胞表面免疫反应性受体蛋白表达无关。髓母细胞瘤细胞对cpIL4-PE的敏感性可通过与IL-4或IL-13同时孵育而消除,但与IL-2同时孵育则不能消除。用外源性IL-4处理后,这些细胞系均未显示增殖有任何变化。这些研究将白细胞介素-4受体确立为髓母细胞瘤相关靶点,有望用于肿瘤导向性癌症治疗。有必要进一步研究原发性髓母细胞瘤肿瘤中白细胞介素-4受体的表达以及体外和体内对cpIL-4PE的敏感性。