Janský L, Reymanová P, Kopecký J
Faculty of biology, University of South Bohemia, Branisovká, Ceské Budejovice, Czech Republic.
Physiol Res. 2003;52(6):593-8.
To specify the role of individual cytokines in the immune response to pyrogens, isolated and cultivated human peripheral blood mononuclear cells (PBMC) were used for the experiments. Different pyrogens (lipopolysaccharide from Escherichia coli - LPS and live Borrelia afzelii) were applied and the time course of changes in concentrations of different cytokines in the medium was followed using the ELISA method. It was found that nonstimulated human PBMC proliferate under in vitro conditions and produce IL-6, TNF-alpha, IL-10 and finally also IL-1 beta. Productions of IL-12 and INF-gamma are not changed. Proliferation of PBMC is potentiated after incubation with LPS or live Borrelia. PBMC stimulated by LPS increase the net production (stimulated minus unstimulated) of IL-1 beta and TNF-alpha significantly, while production of IL-6 was smaller. A delayed increase in the production of IL-10 was also observed. Productions of IL-12 and INF-gamma were not influenced. In contrast to LPS, stimulation of PBMC with live Borrelia, increases also the production of IL-12 and IFN-gamma, besides IL-1 beta, TNF-alpha, IL-6 and IL-10. Productions of IL-1 beta, IL-6 and TNF alpha increased immediately after incubation with both LPS and Borrelia, while productions of IL-12 and INF-gamma begin to increase 8 hours and production of IL-10 12 hours after stimulation. Data indicate that stimulation with different pyrogens may activate the cells of the immune cascade in a different way. Stimulation of BPMC by LPS seems to activate the initial steps of the immune response (macrophages and granulocytes) only, while infection with live Borrelia also stimulates the later phase of the immune response, probably due to effect of initially produced cytokines.
为明确个体细胞因子在对热原免疫反应中的作用，实验采用分离培养的人外周血单个核细胞（PBMC）。应用不同热原（大肠杆菌脂多糖-LPS和活的阿氏疏螺旋体），采用ELISA法追踪培养基中不同细胞因子浓度的变化时间进程。发现未受刺激的人PBMC在体外条件下增殖并产生IL-6、TNF-α、IL-10，最终也产生IL-1β。IL-12和INF-γ的产生无变化。PBMC与LPS或活的疏螺旋体孵育后增殖增强。LPS刺激的PBMC显著增加IL-1β和TNF-α的净产生量（刺激后减去未刺激），而IL-6的产生量较小。还观察到IL-10产生的延迟增加。IL-12和INF-γ的产生未受影响。与LPS相反，用活的疏螺旋体刺激PBMC，除了IL-1β、TNF-α、IL-6和IL-10外，还增加IL-12和IFN-γ的产生。与LPS和疏螺旋体孵育后，IL-1β、IL-6和TNF-α的产生立即增加，而IL-12和INF-γ的产生在刺激后8小时开始增加，IL-10的产生在刺激后12小时开始增加。数据表明，用不同热原刺激可能以不同方式激活免疫级联反应的细胞。LPS刺激BPMC似乎仅激活免疫反应的初始步骤（巨噬细胞和粒细胞），而活的疏螺旋体感染也刺激免疫反应的后期阶段，可能是由于最初产生的细胞因子的作用。
