Department of Pharmacology and Biomedical Sciences, Faculty of Pharmacy and Medical Sciences, University of Petra, Amman 11196, Jordan.
Cell Therapy Center, The University of Jordan, Amman 11942, Jordan.
J Immunol Res. 2022 Feb 10;2022:6031776. doi: 10.1155/2022/6031776. eCollection 2022.
This study is aimed at investigating the immunological response after treating THP-1 cells with gold nanorods conjugated with a phosphatidylinositol 3-kinase (PI3K) inhibitor. Gold nanorods were synthesized and functionalized with cholesterol-PEG-SH moiety, and the treatment groups were as follows: nanocomplex (a drug-conjugated gold nanorods), free drug (phosphatidylinositol 3-kinase (PI3K) inhibitor), and GNR (the nanocarrier; cholesterol-coated gold nanorods). THP-1 cells were differentiated into macrophages and characterized by measuring the expression of macrophage surface markers by flow cytometry. Then, differentiated cells were activated by lipopolysaccharide (LPS). Afterwards, activated macrophages were treated with the different treatments: nanocomplex, free drug, and GNR, for 24 hrs. After treatment, the production of the inflammatory cytokines measured at gene and protein levels by using qPCR and CBA array beads by flow cytometry.
Our results show that THP-1 cells were successfully differentiated into macrophages. For inflammatory cytokine expression response, nanocomplex and free drug showed the same expression level of cytokines at gene level, as the expression of IL-1, IL-6, and TNF- was significantly downregulated ( < 0.0005, < 0.0005, < 0.00005), respectively, while IL-8, IL-10, and TGF- were all upregulated in a significant manner for nanocomplex ( < 0.00005, < 0.00005, < 0.00005) and free drug treatment group ( < 0.00005, < 0.05, < 0.05) compared to the control untreated group. While in the GNR group, IL-6 and TNF- were downregulated ( < 0.005, < 0.00005), and IL-12p40 ( < 0.00005) was upregulated all in a statistically significant manner. While at protein level, cells were treated with our nanocomplex: IL-1, IL-6, TNF-, and IL-12p70 and were significantly decreased ( < 0.00005, < 0.005, < 0.05, < 0.00005), and IL-10 was found to be significantly increased in culture compared to the untreated control group ( < 0.005). For free drug; IL-1 and IL-12p70 were significantly decreased ( < 0.00005, < 0.00005), while a significant increase in the secretion levels of IL-10 only was noticed compared to the untreated group ( < 0.005). For GNR treatment groups, IL-1, TNF-, and IL-12p70 were significantly decreased ( < 0.00005, < 0.05, < 0.00005).
We can conclude that our nanocomplex is a potent effector that prevents tumoral progression by activating three main immunological strategies: switching the surface expression profile of the activated macrophages into a proinflammatory M1-like phenotype, downregulating the expression of proinflammatory cytokines, and upregulating the expression level of anti-inflammatory cytokines.
本研究旨在探讨用磷脂酰肌醇 3-激酶（PI3K）抑制剂偶联的金纳米棒处理 THP-1 细胞后的免疫反应。合成并功能化了金纳米棒与胆固醇-PEG-SH 部分，处理组如下：纳米复合物（药物偶联的金纳米棒）、游离药物（PI3K 抑制剂）和 GNR（纳米载体；胆固醇包被的金纳米棒）。THP-1 细胞分化为巨噬细胞，并通过流式细胞术测量巨噬细胞表面标志物的表达来进行表征。然后，用脂多糖（LPS）激活分化细胞。之后，用不同的处理方法处理激活的巨噬细胞：纳米复合物、游离药物和 GNR，持续 24 小时。处理后，通过 qPCR 和流式细胞术 CBA 阵列珠测量基因和蛋白质水平的炎症细胞因子的产生。
我们的结果表明，THP-1 细胞成功分化为巨噬细胞。对于炎症细胞因子表达反应，纳米复合物和游离药物在基因水平上表现出相同的细胞因子表达水平，因为 IL-1、IL-6 和 TNF-的表达显著下调（<0.0005，<0.0005，<0.00005），而 IL-8、IL-10 和 TGF-在纳米复合物（<0.00005，<0.00005，<0.00005）和游离药物处理组中均呈显著上调（<0.00005，<0.05，<0.05）。与未处理的对照组相比。而在 GNR 组中，IL-6 和 TNF-下调（<0.005，<0.00005），IL-12p40 上调（<0.00005）均有统计学意义。在蛋白质水平上，用我们的纳米复合物处理细胞：IL-1、IL-6、TNF-和 IL-12p70 明显减少（<0.00005，<0.05，<0.05，<0.00005），IL-10 明显增加与未处理的对照组相比（<0.005）。对于游离药物；IL-1 和 IL-12p70 明显减少（<0.00005，<0.00005），而仅观察到 IL-10 的分泌水平显著增加与未处理组相比（<0.005）。对于 GNR 处理组，IL-1、TNF-和 IL-12p70 明显减少（<0.00005，<0.05，<0.00005）。
我们可以得出结论，我们的纳米复合物是一种有效的效应物，通过激活三种主要的免疫策略来防止肿瘤进展：将激活的巨噬细胞的表面表达谱转变为促炎性 M1 样表型，下调促炎性细胞因子的表达，并上调抗炎性细胞因子的表达水平。
