Kot Kateryna, Kot Yurii, Kurbanov Rustam, Andriiash Hanna, Tigunova Olena, Blume Yaroslav, Shulga Sergiy
Biochemistry Department, V. N. Karazin Kharkiv National University of Ministry of Education and Science of Ukraine, Kharkiv, Ukraine.
Department of Genomics and Molecular Biotechnology, Institute of Food Biotechnology and Genomics National Academy of Science of Ukraine, Kyiv, Ukraine.
Front Neurosci. 2024 Feb 9;18:1325287. doi: 10.3389/fnins.2024.1325287. eCollection 2024.
The leading pathological mechanisms of Alzheimer's disease are amyloidosis and inflammation. The presented work was aimed to study the effect of human peripheral blood mononuclear cells (hPBMcs) cells-matrix adhesion on their pro-inflammatory state . Although direct interaction of Аβ42 to PBMC is not a cellular model of Alzheimer's disease, PBMCs may serve as test cells to detect Аβ42-dependent molecular effects in monitoring disease progression. Peripheral blood mononuclear cells (PBMCs) are used to assess changes in cytokines released in response to diseases or Alzheimer's disease-specific cytotoxic molecules such as Aβ42. The effect of recombinant amyloid β-peptide rАβ42 on the concentration of endogenous amyloid β-peptide Aβ40 and pro-inflammatory cytokines TNFα and IL-1β in human peripheral blood mononuclear cells that were cultured in suspension and immobilized in alginate microcarriers for 24 h were investigated. The localization and accumulation of Aβ40 and rAβ42 peptides in cells, as well as quantitative determination of the concentration of Aβ40 peptide, TNFα and IL-1β cytokines, was performed by intravital fluorescence imaging. The results were qualitatively similar for both cell models. It was determined that the content of TNFα and Aβ40 in the absence of rAβ42 in the incubation medium did not change for 24 h after incubation, and the content of IL-1β was lower compared to the cells that were not incubated. Incubation of cells with exogenous rAβ42 led to an increase in the intracellular content of TNFα and Aβ40, and no accumulation of IL-1β in cells was observed. The accumulation of Aβ40 in the cytoplasm was accompanied by the aggregation of rAβ42 on the outer surface of the cell plasma membrane. It was shown that the basic levels of indicators and the intensity of the response of immobilized cells to an exogenous stimulus were significantly greater than those of cells in suspension. To explore whether non-neuronal cells effects in alginate microcarriers were cell-matrix adhesion mediated, we tested the effect of blocking β1 integrins on proamyloidogenic and proinflammation cellular state. Immobilization within alginate hydrogels after incubation with the β1 integrins blocking antibodies showed a remarkable inhibition of TNFα and Aβ40 accumulation in rAβ42-treated cells. It can be concluded that activation of signal transduction and synthesizing activity of a portion of mononuclear cells of human peripheral blood is possible (can significantly increase) in the presence of cell-matrix adhesion.
阿尔茨海默病的主要病理机制是淀粉样变性和炎症。本研究旨在探讨人外周血单个核细胞（hPBMcs）的细胞-基质黏附对其促炎状态的影响。虽然Aβ42与PBMC的直接相互作用并非阿尔茨海默病的细胞模型，但PBMC可作为检测Aβ42依赖性分子效应以监测疾病进展的测试细胞。外周血单个核细胞（PBMCs）用于评估因疾病或阿尔茨海默病特异性细胞毒性分子（如Aβ42）而释放的细胞因子的变化。研究了重组淀粉样β肽rAβ42对悬浮培养并固定在藻酸盐微载体中24小时的人外周血单个核细胞内源性淀粉样β肽Aβ40以及促炎细胞因子TNFα和IL-1β浓度的影响。通过活体荧光成像对细胞内Aβ40和rAβ42肽的定位和积累以及Aβ40肽、TNFα和IL-1β细胞因子浓度的定量测定进行了研究。两种细胞模型的结果在定性上相似。已确定在孵育培养基中不存在rAβ42的情况下，孵育24小时后TNFα和Aβ40的含量未发生变化，且与未孵育的细胞相比，IL-1β的含量较低。用外源性rAβ42孵育细胞导致TNFα和Aβ40的细胞内含量增加，且未观察到细胞内IL-1β的积累。细胞质中Aβ40的积累伴随着rAβ42在细胞质膜外表面的聚集。结果表明，固定化细胞的指标基础水平和对外源性刺激的反应强度显著高于悬浮细胞。为了探究藻酸盐微载体中的非神经元细胞效应是否由细胞-基质黏附介导，我们测试了阻断β1整合素对淀粉样前体蛋白生成和促炎细胞状态的影响。在用β1整合素阻断抗体孵育后固定在藻酸盐水凝胶中，显示出rAβ42处理的细胞中TNFα和Aβ40积累受到显著抑制。可以得出结论，在存在细胞-基质黏附的情况下，人外周血部分单个核细胞的信号转导和合成活性可能被激活（可显著增加）。
