Franceschetti Marta, Pievani Alice, Borleri Gianmaria, Vago Luca, Fleischhauer Katharina, Golay Josée, Introna Martino
Laboratory of Cellular Therapy G. Lanzani, USC of Hematology, Ospedali Riuniti Bergamo, Bergamo, Italy.
Exp Hematol. 2009 May;37(5):616-628.e2. doi: 10.1016/j.exphem.2009.01.010.
Cytokine-induced killer cells (CIK) are CD3(+)CD56(+) T cells with natural killer (NK)-like cytotoxic activity used for the immunotherapy of tumors. We aimed to fully characterize CIK cells and define their ontogeny.
CIK were generated in vitro by stimulation of peripheral blood mononuclear cells or T-cell subsets with interferon-gamma, anti-CD3 and interleukin-2. They were fully characterized in terms of phenotype, cytotoxic activity, and gene expression with respect to circulating CD3(+)CD56(+) cells, NK cells, and CD56(-) T cells present in CIK cultures.
We demonstrate that CIK are terminally differentiated CD8 T cells that derive from proliferating CD3(+)CD56(-)CD8(+) T cells. They express polyclonal T-cell receptor Vbeta chains and have acquired CD56, NKG2D, and large granular lymphocyte morphology, but lack expression of most NK-specific activating (NKp30, NKp44, NKp46) and inhibitory (KIR2DL1, KIR2DL2, KIR3DL1, NKG2A, CD94) receptors, and can kill K562 targets. Circulating CD3(+)CD56(+) cells are also CD8(+)CD16(-), but are oligoclonal, poorly cytotoxic for K562, and express lower levels of CD56 and NKG2D. Gene profiling of CIK, CD56(-) T and NK cells present at the end of culture shows that differences are much more limited between CIK and CD56(-) T compared to CIK and NK cells. Most of the genes upregulated in CIK cells compared to CD56(-) T cells are part of the tumor necrosis factor gene network.
The CIK phenotype, that is CD45RA(+), CCR7(-), CD62L-weakly positive, CD11a(+), CD27(+), CD28(-), macrophage inflammatory protein 1alpha(+), perforin(+), Fas ligand(+) coincides almost exactly with that described for the T RA(+) effector memory CD27 single positive subset of terminally differentiated human memory T cells.
细胞因子诱导的杀伤细胞（CIK）是具有自然杀伤（NK）样细胞毒性活性的CD3(+)CD56(+) T细胞，用于肿瘤免疫治疗。我们旨在全面表征CIK细胞并确定其起源。
通过用γ干扰素、抗CD3和白细胞介素-2刺激外周血单个核细胞或T细胞亚群在体外生成CIK。根据其表型、细胞毒性活性以及与CIK培养物中存在的循环CD3(+)CD56(+)细胞、NK细胞和CD56(-) T细胞相关的基因表达对它们进行全面表征。
我们证明CIK是终末分化的CD8 T细胞，来源于增殖的CD3(+)CD56(-)CD8(+) T细胞。它们表达多克隆T细胞受体Vβ链，获得了CD56、NKG2D和大颗粒淋巴细胞形态，但缺乏大多数NK特异性激活（NKp30、NKp44、NKp46）和抑制（KIR2DL1、KIR2DL2、KIR3DL1、NKG2A、CD94）受体的表达，并且可以杀伤K562靶细胞。循环CD3(+)CD56(+)细胞也是CD8(+)CD16(-)，但为寡克隆性，对K562的细胞毒性较差，并且表达较低水平的CD56和NKG2D。培养结束时存在的CIK、CD56(-) T细胞和NK细胞的基因谱分析表明，与CIK和NK细胞相比，CIK和CD56(-) T细胞之间的差异要小得多。与CD56(-) T细胞相比，CIK细胞中上调的大多数基因是肿瘤坏死因子基因网络的一部分。
CIK的表型，即CD45RA(+)、CCR7(-)、CD62L弱阳性（CD62L-weakly positive）、CD11a(+)、CD27(+)、CD28(-)、巨噬细胞炎性蛋白1α(+)、穿孔素(+)、Fas配体(+)几乎与终末分化的人类记忆T细胞的T RA(+)效应记忆CD27单阳性亚群所描述的表型完全一致。
