Setrakian J C, Yee J, Christou N V
Department of Surgery, McGill University, Montreal, Quebec.
Arch Surg. 1994 Feb;129(2):187-92. doi: 10.1001/archsurg.1994.01420260083011.
To determine the cytokine response to lipopolysaccharide in patients in the intensive care unit.
Patients in a mixed medical/surgical intensive care unit with fever and a de novo clinical dysfunction of at least one organ system.
Whole blood from patients and from laboratory controls was stimulated with 8 ng/mL of lipopolysaccharide (Escherichia coli 0111:B4) at 37 degrees C, and tumor necrosis factor alpha (TNF-alpha) was measured using enzyme linked immunosorbent assay at 4, 8, and 24 hours. The same subjects' purified monocytes were cultured with 8 ng/mL of lipopolysaccharide in the presence of autologous or pooled control plasma or cocultured with purified autologous polymorphonuclear leukocytes at a polymorphonuclear leukocyte-monocyte ratio of 10:1, and TNF-alpha was measured at 24 hours using the enzyme linked immunosorbent assay.
We detected high (n = 5) and low (n = 5) TNF-alpha responders in whole blood producing a mean (+/- SEM) of 27.2 +/- 6.3 pg/mL per 1000 monocytes vs 0.0 +/- 2.4 pg/mL per 1000 monocytes, respectively (controls, 58.0 +/- 13.0 pg/mL per 1000 monocytes). The kinetics of TNF-alpha production in both groups were comparable. Purified monocytes from both groups of patients cultured with lipopolysaccharide alone produced equivalent TNF-alpha values (42.4 +/- 10.5 vs 40.8 +/- 12.5 pg/mL per 1000 monocytes). Assayable TNF-alpha was not different with autologous vs control serum but was markedly diminished by the presence of polymorphonuclear leukocytes in patients as well as in controls; the two groups of patients did not differ in this polymorphonuclear leukocyte effect.
Lipopolysaccharide stimulation of monocytes in the whole blood results in marked variation of TNF-alpha production. This phenomenon may account for the variable septic response to infection in patients in the intensive care unit.
确定重症监护病房患者对脂多糖的细胞因子反应。
入住内科/外科混合重症监护病房、发热且至少有一个器官系统出现新发临床功能障碍的患者。
将患者和实验室对照的全血在37℃下用8 ng/mL脂多糖（大肠杆菌0111:B4）刺激，在4小时, 8小时和24小时使用酶联免疫吸附测定法测量肿瘤坏死因子α（TNF-α）。将相同受试者的纯化单核细胞与8 ng/mL脂多糖在自体或混合对照血浆存在的情况下培养，或以10:1的多形核白细胞 - 单核细胞比例与纯化的自体多形核白细胞共培养，并在24小时使用酶联免疫吸附测定法测量TNF-α。
我们在全血中检测到高反应者（n = 5）和低反应者（n = 5），每1000个单核细胞产生的TNF-α平均值（±SEM）分别为27.2±6.3 pg/mL和0.0±2.4 pg/mL（对照为每1000个单核细胞58.0±13.0 pg/mL）。两组中TNF-α产生的动力学具有可比性。两组患者单独用脂多糖培养的纯化单核细胞产生的TNF-α值相当（每1000个单核细胞42.4±10.5 vs 40.8±12.5 pg/mL）。可检测的TNF-α在自体血清与对照血清之间没有差异，但在患者和对照中，多形核白细胞的存在使其明显减少；两组患者在这种多形核白细胞效应方面没有差异。
全血中单核细胞受到脂多糖刺激会导致TNF-α产生的显著差异。这种现象可能解释了重症监护病房患者对感染的脓毒症反应的变异性。
