Department of Dermatology and Allergology, University Hospital RWTH Aachen, 52074 Aachen, Germany.
Biomolecules. 2024 Nov 8;14(11):1427. doi: 10.3390/biom14111427.
The NLRP1 (nucleotide-binding domain, leucine-rich-containing family, pyrin domain-containing-1) inflammasome is the most important inflammasome in human keratinocytes. It plays a crucial role in regulating innate immunity in the skin. This study aimed to evaluate NLRP1 inflammasome activation and the corresponding levels of detection in different keratinocyte cell lines to identify a suitable in vitro model for analyzing inflammasome activation in keratinocytes. We compared NLRP1 inflammasome activation, expression, and cell death among primary keratinocytes and immortalized keratinocyte cell lines HaCaT, HaSKpw, and SVTERT upon stimulation with ultraviolet B (UVB) irradiation or talabostat. The effects of both NLRP1 inducers on cell death and the modification of NLRP1 molecules were examined using fluorescence-activated cell sorting analysis, Western blotting, and an enzyme-linked immunosorbent assay. The key inflammasome components had varied expression levels among the keratinocyte cell models, with the highest expression observed in primary keratinocytes. Moreover, our data showed that both UVB and talabostat triggered cell death, and NLRP1 inflammasome activation was readily detected in primary keratinocytes but not in the analyzed immortalized keratinocyte cell lines. Therefore, we do not recommend the use of the immortalized keratinocyte cell lines HaCaT, HaSKpw, and SVTERT for analyzing inflammasome activation in keratinocytes; we strongly recommend the use of primary keratinocytes for these studies.
NLRP1（核苷酸结合域、富含亮氨酸重复家族、pyrin 域包含蛋白-1）炎性小体是人类角质形成细胞中最重要的炎性小体。它在调节皮肤固有免疫中起着至关重要的作用。本研究旨在评估不同角质形成细胞系中 NLRP1 炎性小体的激活及其相应的检测水平，以确定用于分析角质形成细胞中炎性小体激活的合适体外模型。我们比较了原发性角质形成细胞和永生化角质形成细胞系 HaCaT、HaSKpw 和 SVTERT 在紫外线 B（UVB）照射或 talabostat 刺激下 NLRP1 炎性小体的激活、表达和细胞死亡。使用荧光激活细胞分选分析、Western blot 和酶联免疫吸附试验检测了两种 NLRP1 诱导剂对细胞死亡的影响以及 NLRP1 分子的修饰。角质形成细胞模型中的关键炎性小体成分的表达水平不同，其中原发性角质形成细胞表达水平最高。此外，我们的数据表明，UVB 和 talabostat 均可触发细胞死亡，并且在原发性角质形成细胞中可轻易检测到 NLRP1 炎性小体的激活，但在分析的永生化角质形成细胞系中则无法检测到。因此，我们不建议使用永生化角质形成细胞系 HaCaT、HaSKpw 和 SVTERT 来分析角质形成细胞中的炎性小体激活；我们强烈建议在这些研究中使用原发性角质形成细胞。
