Long-term proliferation of functional human NK cells, with conversion of CD56dim NK cells to a CD56bright phenotype, induced by carcinoma cells co-expressing 4-1BBL and IL-12
Long-term proliferation of functional human NK cells, with conversion of CD56dim NK cells to a CD56bright phenotype, induced by carcinoma cells co-expressing 4-1BBL and IL-12
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DOI：
10.1007/s00262-011-1122-3
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发表时间：
2012-05-01
影响因子：
5.8
通讯作者：
Searle, Peter F.
中科院分区：
文献类型：
--
作者：
Dowell, Alexander C.;Oldham, Kimberley A.;Searle, Peter F.
4-1BB结扎共刺激T细胞活化，激动抗体已进入临床试验。自然杀伤（NK）细胞在激活后也表达4-1BB，并与4-1BB刺激小鼠的抗肿瘤效果有关；然而，人类NK细胞对4-1BB刺激的反应尚不明确。表达4-1BB配体（4-1BBL）或IL-12的OVCAR-3细胞刺激非贴壁PBMC导致NK细胞群优先扩增，而4-1BBL + IL-12的组合对来自健康供体和肾细胞或卵巢癌患者的功能性NK细胞的激活和增殖更有利，支持长期（21天）NK细胞增殖。扩增的NK细胞主要是CD56（亮），我们发现分离的CD56（暗）CD16(+) NK细胞可以转换为CD56（亮）CD16（-）表型，并在4-1BBL + IL-12的作用下增殖。NK细胞响应4-1BBL的4-1BB上调需要其他PBMC的“帮助”，IL-12可以在分离的NK细胞上诱导4-1BB上调，但仅在靶细胞（OVCAR-3）存在的情况下。用表达4-1BBL + IL-12的OVCAR-3细胞进行初始刺激并静息至第21天，NK细胞主要保持CD56（明亮），并保持对K562靶点的高细胞毒能力，以及相对于PBMC中NK细胞产生IFN γ的能力增强。这些数据支持NK细胞可以促进人类4-1BB激动剂的抗肿瘤活性的概念，并表明将4-1BB刺激与IL-12结合可能有利于NK细胞的体外或体内扩增和激活，以用于癌症免疫治疗。
4-1BB ligation co-stimulates T cell activation, and agonistic antibodies have entered clinical trials. Natural killer (NK) cells also express 4-1BB following activation and are implicated in the anti-tumour efficacy of 4-1BB stimulation in mice; however, the response of human NK cells to 4-1BB stimulation is not clearly defined. Stimulation of non-adherent PBMC with OVCAR-3 cells expressing 4-1BB ligand (4-1BBL) or IL-12 resulted in preferential expansion of the NK cell population, while the combination 4-1BBL + IL-12 was superior for the activation and proliferation of functional NK cells from healthy donors and patients with renal cell or ovarian carcinoma, supporting long-term (21 day) NK cell proliferation. The expanded NK cells are predominantly CD56(bright), and we show that isolated CD56(dim)CD16(+) NK cells can switch to a CD56(bright)CD16(-) phenotype and proliferate in response to 4-1BBL + IL-12. Whereas 4-1BB upregulation on NK cells in response to 4-1BBL required 'help' from other PBMC, it could be induced on isolated NK cells by IL-12, but only in the presence of target (OVCAR-3) cells. Following primary stimulation with OVCAR-3 cells expressing 4-1BBL + IL-12 and subsequent resting until day 21, NK cells remained predominantly CD56(bright) and retained both high cytotoxic capability against K562 targets and enhanced ability to produce IFN gamma relative to NK cells in PBMC. These data support the concept that NK cells could contribute to anti-tumour activity of 4-1BB agonists in humans and suggest that combining 4-1BB-stimulation with IL-12 could be beneficial for ex vivo or in vivo expansion and activation of NK cells for cancer immunotherapy.