Astiz M, Saha D, Lustbader D, Lin R, Rackow E
St. Vincent's Hospital and Medical Center, New York, NY 10011, USA.
J Lab Clin Med. 1996 Dec;128(6):594-600. doi: 10.1016/s0022-2143(96)90132-8.
Exposure to endotoxin produces a state of macrophage hyporesponsiveness on subsequent stimulation. Monocytes in patients with septic shock demonstrate a similar hyporesponsiveness to endotoxin. The purpose of this study was to examine whether this state of hyporesponsiveness extends to other inflammatory stimuli and the relationship of this state to cell surface receptor expression and the release of anti-inflammatory cytokines. Twelve normal volunteers, 10 patients with severe sepsis, and 9 patients with septic shock were included in the study. Monocytes from each subject were isolated and stimulated with lipopolysaccharide (LPS), staphylococcal enterotoxin B (SEB), and phorbol myristate acetate (PMA). Tumor necrosis factor-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) were measured in the supernatants by enzyme-linked immunosorbent assay (ELISA). Serum levels of transforming growth factor-beta1 (TGF-beta1), prostaglandin E2 (PGE2), and interleukin-10 (IL-10) were also measured by ELISA. The expression of monocyte CD14 and HLA-DR in whole blood were measured by flow cytometry. Patients with septic shock demonstrated significantly decreased TNF-alpha and IL-1beta release as compared with normal subjects in response to LPS. In response to SEB, patients with sepsis and patient with septic shock demonstrated significantly decreased release of TNF-alpha and IL-1beta. Significant decreases in TNF-alpha release were found in the patients with septic shock after PMA stimulation. There were no significant differences in the monocyte response to the different stimuli between patients with gram-positive sepsis and gram-negative sepsis. HLA-DR expression was significantly decreased in patients with septic shock (58 +/- 9 fluorescence units (flU)) as compared with normal subjects (102 +/- 14 flU) (p < 0.05). No differences in CD14 expression were observed. IL-10 levels were significantly increased in patients with sepsis (16 +/- 4 pg/ml) and in patients with septic shock (42 +/- 15 pg/ml) and were detectable in 1 normal subject. TGF-beta1 levels were decreased in patients with septic shock (25 +/- 6 pg/ml) as compared with those in normal subjects (37 +/- 2 pg/ml)(p < 0.05). PGE2 levels were significantly increased in patients with septic shock and patients with sepsis. These data are consistent with a more generalized monocyte hyporesponsiveness to bacterial toxins that may be related to altered cell surface receptor expression and the release of anti-inflammatory cytokines.
接触内毒素会导致巨噬细胞在后续刺激时出现低反应状态。感染性休克患者的单核细胞对内毒素也表现出类似的低反应性。本研究的目的是检验这种低反应状态是否会扩展到其他炎症刺激，以及这种状态与细胞表面受体表达和抗炎细胞因子释放之间的关系。该研究纳入了12名正常志愿者、10名严重脓毒症患者和9名感染性休克患者。分离出每个受试者的单核细胞，并用脂多糖（LPS）、葡萄球菌肠毒素B（SEB）和佛波酯肉豆蔻酸酯（PMA）进行刺激。通过酶联免疫吸附测定（ELISA）检测上清液中的肿瘤坏死因子-α（TNF-α）和白细胞介素-1β（IL-1β）。还通过ELISA检测血清中转化生长因子-β1（TGF-β1）、前列腺素E2（PGE2）和白细胞介素-10（IL-10）的水平。通过流式细胞术检测全血中单核细胞CD14和HLA-DR的表达。与正常受试者相比，感染性休克患者在LPS刺激下TNF-α和IL-1β的释放显著减少。在SEB刺激下，脓毒症患者和感染性休克患者的TNF-α和IL-1β释放显著减少。PMA刺激后，感染性休克患者的TNF-α释放显著降低。革兰氏阳性脓毒症患者和革兰氏阴性脓毒症患者的单核细胞对不同刺激的反应没有显著差异。与正常受试者（102±14荧光单位（flU））相比，感染性休克患者的HLA-DR表达显著降低（58±9 flU）（p<0.05）。未观察到CD14表达的差异。脓毒症患者（16±4 pg/ml）和感染性休克患者（42±15 pg/ml）的IL-10水平显著升高，1名正常受试者中也可检测到。与正常受试者（37±2 pg/ml）相比，感染性休克患者的TGF-β1水平降低（25±6 pg/ml）（p<0.05）。脓毒症患者和感染性休克患者的PGE2水平显著升高。这些数据与单核细胞对细菌毒素更普遍的低反应性一致，这可能与细胞表面受体表达改变和抗炎细胞因子释放有关。
