Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 117997 Moscow, Russia.
Ecole Polytechnique Federale de Lausanne, 1015 Lausanne, Switzerland.
Cells. 2024 Mar 17;13(6):530. doi: 10.3390/cells13060530.
NK cells play a decisive role in controlling hCMV infection by combining innate and adaptive-like immune reactions. The hCMV-derived VMAPRTLFL (LFL) peptide is a potent activator of NKG2C NK cells. Proposed here is an autologous system of LFL stimulation without T lymphocytes and exogenous cytokines that allows us to evaluate NK-cell hCMV-specific responses in more native settings. In this model, we evaluated LFL-induced IFNγ production, focusing on signaling pathways and the degranulation and proliferation of NK cells orchestrated by microenvironment cytokine production and analyzed the transcriptome of expanded NK cells. NK cells of individuals having high anti-hCMV-IgG levels, in contrast to NK cells of hCMV-seronegative and low-positive donors, displayed increased IFNγ production and degranulation and activation levels and enhanced proliferation upon LFL stimulation. Cytokine profiles of these LFL-stimulated cultures demonstrated a proinflammatory shift. LFL-induced NK-cell IFNγ production was dependent on the PI3K and Ras/Raf/Mek signaling pathways, independently of cytokines. In hCMV-seropositive individuals, this model allowed obtaining NK-cell antigen-specific populations proliferating in response to LFL. The transcriptomic profile of these expanded NK cells showed increased adaptive gene expression and metabolic activation. The results complement the existing knowledge about hCMV-specific NK-cell response. This model may be further exploited for the identification and characterization of antigen-specific NK cells.
自然杀伤 (NK) 细胞通过结合先天和适应性免疫反应，在控制人类巨细胞病毒 (hCMV) 感染方面发挥决定性作用。hCMV 衍生的 VMAPRTLFL（LFL）肽是 NKG2C NK 细胞的有效激活剂。本文提出了一种无需 T 淋巴细胞和外源性细胞因子的自体 LFL 刺激系统，使我们能够在更自然的环境中评估 NK 细胞对 hCMV 的特异性反应。在该模型中，我们评估了 LFL 诱导的 IFNγ 产生，重点关注信号通路以及由微环境细胞因子产生协调的 NK 细胞脱颗粒和增殖，并分析了扩增的 NK 细胞的转录组。与 hCMV 血清阴性和低阳性供体的 NK 细胞相比，具有高抗 hCMV-IgG 水平的个体的 NK 细胞在 LFL 刺激下显示出增加的 IFNγ 产生、脱颗粒和激活水平以及增强的增殖。这些 LFL 刺激培养物的细胞因子谱显示出促炎偏移。LFL 诱导的 NK 细胞 IFNγ 产生依赖于 PI3K 和 Ras/Raf/Mek 信号通路，而不依赖于细胞因子。在 hCMV 血清阳性个体中，该模型允许获得对 LFL 有反应性增殖的 NK 细胞抗原特异性群体。这些扩增的 NK 细胞的转录组谱显示出适应性基因表达和代谢激活的增加。这些结果补充了关于 hCMV 特异性 NK 细胞反应的现有知识。该模型可进一步用于鉴定和表征抗原特异性 NK 细胞。
