Vanderelst Julie, Rousseau Alexandre, Selvais Nicolas, Biston Patrick, Zouaoui Boudjeltia Karim, Piagnerelli Michaël
Intensive Care, CHU-Charleroi Marie-Curie, Université libre de Bruxelles, Charleroi, Belgium.
Experimental Medicine Laboratory, CHU-Charleroi Vésale, ULB 222 Unit, Université libre de Bruxelles, Montigny-le-Tilleul, Belgium.
Front Med (Lausanne). 2022 Jul 28;9:880657. doi: 10.3389/fmed.2022.880657. eCollection 2022.
During sepsis, red blood cell (RBC) deformability is altered. Persistence of these alterations is associated with poor outcome. Activation of the complement system is enhanced during sepsis and RBCs are protected by membrane surface proteins like CD35, CD55 and CD59. In malaria characterized by severe anemia, a study reported links between the modifications of the expression of these RBCs membrane proteins and erythrophagocytosis. We studied the evolution of RBCs deformability and the expression of RBC membrane surface IgG and regulatory proteins in septic patients.
By flow cytometry technics, we measured at ICU admission and at day 3-5, the RBC membrane expression of IgG and complement proteins (CD35, 55, 59) in septic patients compared to RBCs from healthy volunteers. Results were expressed in percentage of RBCs positive for the protein. RBC shape was assessed using Pearson's second coefficient of dissymmetry (PCD) on the histogram obtained with a flow cytometer technique. A null value represents a perfect spherical shape. RBC deformability was determined using ektacytometry by the elongation index in relation to the shear stress (0.3-50 Pa) applied to the RBC membrane. A higher elongation index indicates greater RBC deformability.
RBCs from 11 septic patients were compared to RBCs from 21 volunteers. At ICU admission, RBCs from septic patients were significantly more spherical and RBC deformability was significantly lower in septic patients for all shear stress ≥1.93 Pa. These alterations of shape and deformability persists at day 3-5. We observed a significant decrease at ICU admission only in CD35 expression on RBCs from septic patients. This low expression remained at day 3-5.
We observed in RBCs from septic patients a rapid decrease expression of CD35 membrane protein protecting against complement activation. These modifications associated with altered RBC deformability and shape could facilitate erythrophagocytosis, contributing to anemia observed in sepsis. Other studies with a large number of patients and assessment of erythrophagocytosis were needed to confirm these preliminary data.
在脓毒症期间,红细胞(RBC)的可变形性会发生改变。这些改变的持续存在与不良预后相关。脓毒症期间补体系统的激活增强,红细胞受到如CD35、CD55和CD59等膜表面蛋白的保护。在以严重贫血为特征的疟疾中,一项研究报道了这些红细胞膜蛋白表达的改变与红细胞吞噬作用之间的联系。我们研究了脓毒症患者红细胞可变形性的演变以及红细胞膜表面IgG和调节蛋白的表达。
通过流式细胞术,我们在重症监护病房(ICU)入院时以及第3 - 5天,测量了脓毒症患者红细胞膜上IgG和补体蛋白(CD35、55、59)的表达,并与健康志愿者的红细胞进行比较。结果以蛋白阳性红细胞的百分比表示。使用流式细胞术技术获得的直方图上的皮尔逊第二不对称系数(PCD)评估红细胞形状。零值代表完美的球形。通过测定红细胞变形性,使用激光衍射法测量施加于红细胞膜的剪切应力(0.3 - 50 Pa)下的伸长指数。较高的伸长指数表明红细胞可变形性更大。
将11例脓毒症患者的红细胞与21名志愿者的红细胞进行比较。在ICU入院时,对于所有剪切应力≥1.93 Pa,脓毒症患者的红细胞明显更呈球形,且红细胞可变形性明显更低。这些形状和可变形性的改变在第3 - 5天持续存在。我们仅在ICU入院时观察到脓毒症患者红细胞上CD35表达显著降低。这种低表达在第3 - 5天仍然存在。
我们在脓毒症患者的红细胞中观察到,保护免受补体激活的CD35膜蛋白表达迅速降低。这些与红细胞可变形性和形状改变相关的变化可能促进红细胞吞噬作用,导致脓毒症中观察到的贫血。需要其他大量患者的研究以及对红细胞吞噬作用的评估来证实这些初步数据。