Laboratorio de Fisiología de los Procesos Inflamatorios, Instituto de Medicina Experimental (IMEX)-Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET)/Academia Nacional de Medicina de Buenos Aires, Buenos Aires, Argentina.
J Innate Immun. 2024;16(1):489-500. doi: 10.1159/000541414. Epub 2024 Sep 18.
Epithelial and endothelial cells modulate innate immune responses in the lung, including the arrival of neutrophils (PMN), which are crucial cells for the antibacterial host defense. Cells are exposed to prokaryotic RNA (pRNA) during bacterial infections and different pRNA may promote or attenuate the inflammatory response on different immune cells. Pseudomonas aeruginosa (PAE) can cause severe pneumonia and has several immune-evading mechanisms. The aim of this study was to determine the effects of the RNA from PAE (RNAPAE) on lung epithelial, endothelial cells, and PMN, and its impact on bacterial elimination.
Purified total RNAPAE was used as a stimulus on a human lung epithelial cell line (Calu-6), human microvascular endothelial cell line HMEC-1 and isolated healthy human PMN. Activation and cytokine secretion were evaluated. In addition, PMN elimination of live ECO or PAE was determined in the presence of RNAPAE.
We found that RNAPAE either induced a pro-inflammatory response on Calu-6 and HMEC-1 or PMN. Pre-stimulation of PMN with RNAPAE diminished activation and chemotaxis induced by live bacteria. Moreover, we found that RNAPAE reduced phagocytosis of live ECO. Finally, we also found that non-degraded fragments of small RNA (<200 bp) were responsible for the PMN microbicidal attenuation during PAE elimination.
Our results indicated that short fragments of RNAPAE diminished the immune response of PMN favoring bacterial survival.
上皮细胞和内皮细胞调节肺部的固有免疫反应,包括中性粒细胞(PMN)的到达,PMN 是抗菌宿主防御的关键细胞。细胞在细菌感染期间暴露于原核 RNA(pRNA),不同的 pRNA 可能促进或减弱不同免疫细胞的炎症反应。铜绿假单胞菌(PAE)可引起严重肺炎,并具有几种免疫逃避机制。本研究旨在确定来自 PAE(RNAPAE)的 RNA 对肺上皮细胞、内皮细胞和 PMN 的影响及其对细菌清除的影响。
将纯化的总 RNAPAE 用作人类肺上皮细胞系(Calu-6)、人微血管内皮细胞系 HMEC-1 和分离的健康人 PMN 的刺激物。评估激活和细胞因子分泌。此外,还在存在 RNAPAE 的情况下测定 PMN 对活 ECO 或 PAE 的消除。
我们发现,RNAPAE 要么在上皮细胞和内皮细胞或 PMN 上诱导促炎反应。用 RNAPAE 预先刺激 PMN 会减弱活细菌诱导的激活和趋化作用。此外,我们发现 RNAPAE 减少了活 ECO 的吞噬作用。最后,我们还发现,小 RNA(<200 bp)的非降解片段负责在 PAE 消除过程中减弱 PMN 的杀菌作用。
我们的结果表明,RNAPAE 的短片段减弱了 PMN 的免疫反应,有利于细菌的存活。