Zedler S, Bone R C, Baue A E, von Donnersmarck G H, Faist E
Department of Surgery, Klinikum Grosshadern, Ludwig-Maximilians-University Munich, Germany.
Crit Care Med. 1999 Jan;27(1):66-72. doi: 10.1097/00003246-199901000-00028.
To obtain further insight into the constitutional, phenotype-dependent changes of T-helper-1 and T-helper-2 signature lymphokine synthesis after trauma.
Prospective, descriptive study.
Intensive care unit of a burn center in a community hospital.
Ten patients 1, 3, 5, and 7 days after major burn injury and 15 healthy individuals.
Peripheral blood mononuclear cells were separated and incubated (5 hrs) for cytokine production induced by the accessory cell-independent stimulus of ionomycin and phorbol 12-myristate 13-acetate. After fixation and permeabilization, cell samples were immunofluorescently stained for cell surface antigens (CD4 and CD8), intracellular interferon (IFN)-gamma, and interleukin (IL)-4 synthesis. Results were correlated with corresponding enzyme-linked immunosorbent assay measurements of the culture supernatants.
The phenotypic analysis of the composition of the helper (CD4) and suppressor/cytotoxic (CD8) T-cell subset demonstrated that patients suffering from major burns and healthy controls express these antigens in similar percentages. The ratio of CD4 positive to CD8 positive/CD16 negative T-cell subsets showed no significant changes after trauma compared with controls. The production of IL-4 was excessively up-regulated while the release of IFN-gamma was only slightly increased. The predominant cell source of IL-4 after burn trauma was the CD8+ cell with nearly five-fold increased production on day 5 (7.2+/-2.6%) vs. 1.5+/-0.4% in controls. While CD8+ cells are also capable of enhancing their IFN-gamma synthesis under stress by about 60% due to the significant participation of the naive CD45RA+ subset, the CD4+ IFN-gamma release remained largely unchanged. With this study, we demonstrated that in nonsurvivors the number of CD8+ IL-4-producing cells was significantly higher compared with controls; also, the number of IFN-gamma-releasing memory/effector CD45RO+ cells was lower compared with survivors.
In previous experiments, we show that a shift to T(H)2 dominated phenotypes increases the risk for postburn infection. The current study confirms that major burns induce a significant shift of cytokine response in the T(H)2 direction and demonstrates that the CD8+, rather than the CD4+ phenotype, is present. Increased IL-4 production is associated with the T(H)2 lymphocyte. These diagnostic tests may help to differentiate patients with compensatory anti-inflammatory response syndrome and immunosuppression from those patients in the proinflammatory state associated with the systemic inflammatory response syndrome. The profile described in this article is associated with immunosuppression and may contraindicate attempts at anti-inflammatory therapy for sepsis.
进一步了解创伤后辅助性T细胞1（Th1）和辅助性T细胞2（Th2）特征性淋巴因子合成的体质性、表型依赖性变化。
前瞻性描述性研究。
社区医院烧伤中心重症监护病房。
10例重度烧伤后1、3、5和7天的患者以及15名健康个体。
分离外周血单个核细胞，并用离子霉素和佛波酯12-肉豆蔻酸酯13-乙酸酯这种不依赖辅助细胞的刺激物进行孵育（5小时）以诱导细胞因子产生。固定和通透处理后，细胞样本用免疫荧光法染色检测细胞表面抗原（CD4和CD8）、细胞内干扰素（IFN）-γ和白细胞介素（IL）-4的合成。结果与培养上清液相应的酶联免疫吸附测定结果相关联。
辅助性（CD4）和抑制性/细胞毒性（CD8）T细胞亚群组成的表型分析表明，重度烧伤患者和健康对照者中这些抗原的表达百分比相似。与对照组相比，创伤后CD4阳性与CD8阳性/CD16阴性T细胞亚群的比例无显著变化。IL-4的产生过度上调，而IFN-γ的释放仅略有增加。烧伤创伤后IL-4的主要细胞来源是CD8+细胞，第5天时其产生量增加近5倍（7.2±2.6%），而对照组为1.5±0.4%。虽然由于初始CD45RA+亚群的显著参与，CD8+细胞在应激状态下也能够将其IFN-γ合成提高约60%，但CD4+细胞IFN-γ的释放基本保持不变。通过本研究，我们证明在非存活者中，产生IL-4的CD8+细胞数量与对照组相比显著更高；此外，与存活者相比，释放IFN-γ的记忆/效应CD4RO+细胞数量更低。
在先前的实验中，我们表明向以Th2为主导的表型转变会增加烧伤后感染的风险。当前研究证实重度烧伤会导致细胞因子反应显著向Th2方向转变，并表明存在的是CD8+而非CD4+表型。IL-4产生增加与Th2淋巴细胞相关。这些诊断测试可能有助于区分具有代偿性抗炎反应综合征和免疫抑制的患者与处于与全身炎症反应综合征相关的促炎状态的患者。本文所描述的特征与免疫抑制相关，可能提示脓毒症抗炎治疗的禁忌。
