Influence of In Vitro IL-2 or IL-15 Alone or in Combination with Hsp 70 Derived 14-Mer Peptide (TKD) on the Expression of NK Cell Activatory and Inhibitory Receptors on Peripheral Blood T Cells, B Cells and NKT Cells.
Influence of In Vitro IL-2 or IL-15 Alone or in Combination with Hsp 70 Derived 14-Mer Peptide (TKD) on the Expression of NK Cell Activatory and Inhibitory Receptors on Peripheral Blood T Cells, B Cells and NKT Cells.
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DOI：
10.1371/journal.pone.0151535
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Dickinson AM
中科院分区：
文献类型：
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作者：
Hromadnikova I;Li S;Kotlabova K;Dickinson AM
Multhoff等先前的研究报道，质膜Hsp70是活化NK细胞的肿瘤特异性识别结构，用Hsp70和/或Hsp70 n端序列衍生的14聚肽（TKDNNLLGRFELSG， TKD, aa 450-463）培养NK细胞，再加上低剂量的IL-2，可触发NK细胞增殖和迁移，并具有杀死表达Hsp70膜的癌细胞的能力。在此，我们利用流式细胞术测定了IL-2或IL-15单独或联合TKD肽体外刺激健康个体外周血单核细胞对CD94、NK细胞激活受体（CD16、NK2D、NKG2C、NKp30、NKp44、NKp46、NKp80、KIR2DL4、DNAM-1和LAMP1）和NK细胞抑制受体（NKG2A、KIR2DL2/L3、LIR1/ILT-2和NKR-P1A）细胞表面表达的影响，CD3+CD56+ (NKT)、CD3+CD4+、CD3+CD8+和CD19+群体。在NKT、CD8+ T细胞和B细胞中，IL-2或IL-15单独或联合TKD刺激后，NKG2D、DNAM-1、LAMP1和NKR-P1A表达上调。CD94在NKT和CD8+ T细胞中表达上调。同时，观察到表达LIR1/ILT-2的CD8+ T细胞和NKR-P1A阳性的CD4+ T细胞的数量增加。与单独IL-2处理相比，IL-2/TKD处理后表达NKG2D的CD8+ T细胞比例更高。与单独使用IL-15相比，IL-15/TKD处理增加了CD94、LAMP1和NKRP-1A阳性的NKT细胞比例。在B细胞中，与单独IL-15相比，IL-15/TKD对细胞表面NKG2D、LIR1/ILT-2和NKRP-1A表达的影响更大。然而，这种增加没有统计学意义。与单独使用IL-2相比，IL-2/TKD诱导CD8+ T细胞中LAMP1的显著上调。除了NK细胞外，外周血单个核细胞中存在的其他免疫活性细胞也受到低剂量白介素本身或与hsp70衍生（TKD）肽联合治疗的影响。
Previous studies from Multhoff and colleagues reported that plasma membrane Hsp70 acts as a tumour-specific recognition structure for activated NK cells, and that the incubation of NK cells with Hsp70 and/or a 14-mer peptide derived from the N-terminal sequence of Hsp70 (TKDNNLLGRFELSG, TKD, aa 450–463) plus a low dose of IL-2 triggers NK cell proliferation and migration, and their capacity to kill cancer cells expressing membrane Hsp70. Herein, we have used flow cytometry to determine the influence of in vitro stimulation of peripheral blood mononuclear cells from healthy individuals with IL-2 or IL-15, either alone or in combination with TKD peptide on the cell surface expression of CD94, NK cell activatory receptors (CD16, NK2D, NKG2C, NKp30, NKp44, NKp46, NKp80, KIR2DL4, DNAM-1 and LAMP1) and NK cell inhibitory receptors (NKG2A, KIR2DL2/L3, LIR1/ILT-2 and NKR-P1A) by CD3+CD56+ (NKT), CD3+CD4+, CD3+CD8+ and CD19+ populations. NKG2D, DNAM-1, LAMP1 and NKR-P1A expression was upregulated after the stimulation with IL-2 or IL-15 alone or in combination with TKD in NKT, CD8+ T cells and B cells. CD94 was upregulated in NKT and CD8+ T cells. Concurrently, an increase in a number of CD8+ T cells expressing LIR1/ILT-2 and CD4+ T cells positive for NKR-P1A was observed. The proportion of CD8+ T cells that expressed NKG2D was higher after IL-2/TKD treatment, when compared with IL-2 treatment alone. In comparison with IL-15 alone, IL-15/TKD treatment increased the proportion of NKT cells that were positive for CD94, LAMP1 and NKRP-1A. The more potent effect of IL-15/TKD on cell surface expression of NKG2D, LIR1/ILT-2 and NKRP-1A was observed in B cells compared with IL-15 alone. However, this increase was not of statistical significance. IL-2/TKD induced significant upregulation of LAMP1 in CD8+ T cells compared with IL-2 alone. Besides NK cells, other immunocompetent cells present within the fraction of peripheral blood mononuclear cells were influenced by the treatment with low-dose interleukins themselves or in combination with hsp70 derived (TKD) peptide.