利用单核细胞-巨噬细胞检测法预测红细胞抗体的重要性

IF 1.2 4区 综合性期刊 Q3 MULTIDISCIPLINARY SCIENCES
Lindsay K Hillis, Selena Cen, Clemence Salou, Yeniley Ruiz Noa, Donald R Branch
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引用次数: 0

摘要

巨噬细胞来源于骨髓中的单核细胞,是一种巨大的先天免疫细胞,在清除死细胞、碎片、肿瘤细胞和外来病原体中起主要作用。单核细胞与巨噬细胞的吞噬能力是一个尚不清楚的概念。在这里,我们的目的是研究单核细胞与巨噬细胞,特别是M1/M2巨噬细胞,对各种调理红细胞的吞噬作用的差异,使用已建立的单核细胞单层试验(MMA)的改进和更新版本。外周血单个核细胞(PBMCs)从供体黄皮毛中分离。利用纯化的单核细胞,通过体外培养和极化产生炎性M1和抗炎M2巨噬细胞。收集M1/M2细胞并用于mma样测定,我们称之为M-MA,以破译临床显着的各种红细胞抗体的吞噬作用。吞噬指数(PI) bb0.5被认为是临床显著的单核细胞吞噬。使用M1/M2巨噬细胞时,吞噬指数(PI)为bbbb12被认为具有临床显著的吞噬作用。与单核细胞和m1细胞相比,M2巨噬细胞吞噬活化红细胞的能力增强。同样的弱抗体(抗s)只对M2巨噬细胞(PI=43)产生显著的吞噬作用,而对m1 (PI=2)或单核细胞(PI=0)则没有作用,这一点在各种抗体中反复得到证实。使用M2巨噬细胞代替单核细胞可以获得更准确的结果,因为这些细胞更具有吞噬性,为该试验提供了进一步的临床相关性。对不同红细胞抗体的进一步研究,包括对具有已知临床意义的抗体的单核细胞-巨噬细胞测定(M-MA)的验证,可能表明M-MA更有助于预测具有临床意义的红细胞同种异体抗体和输血反应。该方法将促进输血医学和免疫学领域的发展。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Prediction of Red Blood Cell Antibody Significance Using the Monocyte-Macrophage Assay.

Derived from monocytes in the bone marrow, macrophages are large, innate immune cells that play a major role in clearing dead cells, debris, tumor cells, and foreign pathogens. The phagocytic capacity of monocytes versus macrophages is a concept that is not well understood. Here, we aim to examine a difference in the phagocytosis of monocytes versus macrophages, specifically M1/M2 macrophages, against various opsonized red cells using a modified and updated version of the established monocyte monolayer assay (MMA). Peripheral blood mononuclear cells (PBMCs) were isolated from donor buffy coats. Using purified monocytes, inflammatory M1 and anti-inflammatory M2 macrophages were produced by in vitro culture and polarization. M1/M2 cells were harvested and used in an MMA-like assay, which we refer to as the M-MA, to decipher clinically significant phagocytosis of various red cell antibodies. A phagocytic index (PI) > 5 was deemed clinically significant phagocytosis with the use of monocytes. A phagocytic index (PI) > 12 was deemed clinically significant phagocytosis with the use of M1/M2 macrophages. M2 macrophages demonstrate an increased ability to phagocytose opsonized RBCs compared to monocytes and M1s. The same weak antibody (anti-S) yields significant phagocytosis with only M2 macrophages (PI=43) but not M1s (PI=2) or monocytes (PI=0), and this was demonstrated repeatedly using various antibodies. The use of M2 macrophages instead of monocytes may allow for more accurate results as these cells are more phagocytic, offering further clinical relevance to the assay. Further studies with different antibodies to red blood cells, including validation of the monocyte-macrophage assay (M-MA) with antibodies having known clinical significance, may show the M-MA more useful to help predict clinically significant red cell alloantibodies and transfusion reactions. This method will advance the field of transfusion medicine and immunology.

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来源期刊
Jove-Journal of Visualized Experiments
Jove-Journal of Visualized Experiments MULTIDISCIPLINARY SCIENCES-
CiteScore
2.10
自引率
0.00%
发文量
992
期刊介绍: JoVE, the Journal of Visualized Experiments, is the world''s first peer reviewed scientific video journal. Established in 2006, JoVE is devoted to publishing scientific research in a visual format to help researchers overcome two of the biggest challenges facing the scientific research community today; poor reproducibility and the time and labor intensive nature of learning new experimental techniques.
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