目前对天然杀伤细胞（NK细胞）中Toll样受体（TLR）的生物学了解有限。这主要是由于旧观念认为NK细胞无法在没有敏感前特异性清除微生物。相反，已经明确证明它们不仅可以通过与微生物分子结合来激活Toll样受体（TLR），而且这种相互作用还会引起NK细胞释放细胞因子，进而激活先天性和适应性免疫中的其他细胞。因此，基于局部输注TLR配体的免疫疗法目前被认为是治疗实体肿瘤的一种新潜在策略。在此，我们提供了一种有效刺激内质网TLR激动剂并确定内质网TLR基因表达水平的方案。该方案可用于在不同条件下对NK细胞中内质网TLR功能进行体外研究。© 2023. 作者通过排他性许可授予Springer Science+Business Media, LLC的一部分Springer Nature。

Poor knowledge is currently available about the biology of Toll-like receptors (TLRs) in natural killer (NK) cells. This is particularly due to the old belief that NK cells are unable to specifically eliminate microbes without presensitization. On the contrary, it has been clearly demonstrated that not only they can be activated through the engagement of Toll-like receptors (TLRs) by microbial molecules, but also that this interaction induces NK cells to release cytokines that, in turn, activate other cells of both innate and adaptive immunity. For this reason, immunotherapy based on local infusion of TLRs ligands is currently considered as a novel potential strategy to treat solid tumors. Here, we provide a protocol to efficiently stimulate NK cells via endosomal TLRs agonists and to determine endosomal TLRs gene expression level. This protocol can be used for in vitro investigation into endosomal TLRs function in NK cells under different conditions.© 2023. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.