Grob M, Schmid-Grendelmeier P, Joller-Jemelka H I, Ludwig E, Dubs R W, Grob P J, Wüthrich B, Bisset L R
Clinical Immunology, Department of Internal Medicine, University Hospital, Zürich, Switzerland.
Allergy. 2003 Mar;58(3):239-45. doi: 10.1034/j.1398-9995.2003.00035.x.
The ability of chemokines to regulate Th1 and Th2 responses suggests a role in the pathogenesis of atopic disorders such as allergic asthma where Th2 response dominance has been observed. Although the impact of allergic asthma on local chemokine production in the lung has been the subject of investigation, little is know about the influence of disease progression on peripheral chemokine production. We now report use of whole blood culture and flow cytometry to assess the influence of mild allergic asthma on peripheral T-cell chemokine expression.
Study participants included patients with mild allergic asthma (n = 7) and nonasthmatic controls (n = 7). Following in vitro stimulation of peripheral venous blood with phorbol 12-myristate acetate (PMA) and ionomycin, flow cytometry was used to estimate the percentage of CD4+ and CD8+ T cells producing a number of chemokines, including macrophage inflammatory proteins MIP-1alpha and MIP-1beta, RANTES (regulated on activation, T-cell expressed and secreted), monocytic chemotactic protein-1 (MCP)-1, and interleukin (IL)-8, or the cytokines interferon (IFN)-gamma and IL-4. Serum levels of MIP-1alpha, MIP-1beta, RANTES, MCP-1, IL-8, IFN-gamma and IL-4 were also assessed by quantitative ELISA.
Intracellular expression of MIP-1beta by CD4+ and CD8+ T cells from allergic asthmatics was significantly reduced in comparison to that observed for nonasthmatics (median = 2.29% (1.75-3.50) vs 4.57% (3.38-6.64), P = 0.05; 14.20% (13.18-17.88) vs 44.10% (30.38-48.70), P = 0.01). Similarly, intracellular expression of MIP-1alpha by CD8+ T cells from allergic asthmatics was also significantly lower (3.67% (1.17-5.42) vs 17.10% (4.97-20.43), P = 0.05). Conversely, IL-8 expression by both CD4+ and CD8+ T cells from allergic asthmatics demonstrated significant enhancement (9.93% (7.77-11.28) vs 4.14% (3.61-7.11), P = 0.05; 8.40% (6.97-10.04) vs 4.98% (3.37-6.08), P = 0.05). Examination of intracellular IFN-gamma and IL-4 revealed no significant difference in the expression of either cytokine by CD4+ T-cells from allergic asthmatics and nonasthmatics. In contrast, expression of IFN-gamma was significantly reduced in CD8+ T-cells from allergic asthmatics (24.60% (21.08-32.50) vs 48.40% (41.50-55.28), P = 0.01).
The occurrence in mild allergic asthma of peripheral T-cell chemokine expression suggestive of a diminished Th1 response, coinciding with marginal change in cytokine profiles indicative of a Th2 response bias, confirms the importance of chemokine involvement in the etiology of allergic asthma. The ability to use whole blood culture to estimate chemokine expression in T cell subsets may ultimately provide a practical means to evaluate disease status and to monitor early intervention therapies which target chemokines.
趋化因子调节Th1和Th2反应的能力表明其在特应性疾病如过敏性哮喘的发病机制中发挥作用，在过敏性哮喘中已观察到Th2反应占主导。尽管过敏性哮喘对肺内局部趋化因子产生的影响已成为研究对象，但关于疾病进展对外周趋化因子产生的影响却知之甚少。我们现在报告使用全血培养和流式细胞术来评估轻度过敏性哮喘对外周T细胞趋化因子表达的影响。
研究参与者包括轻度过敏性哮喘患者（n = 7）和非哮喘对照者（n = 7）。用佛波酯12 - 肉豆蔻酸酯乙酸盐（PMA）和离子霉素体外刺激外周静脉血后，使用流式细胞术估计产生多种趋化因子的CD4 +和CD8 + T细胞的百分比，这些趋化因子包括巨噬细胞炎性蛋白MIP - 1α和MIP - 1β、调节激活正常T细胞表达和分泌因子（RANTES）、单核细胞趋化蛋白 - 1（MCP） - 1和白细胞介素（IL） - 8，或细胞因子干扰素（IFN） - γ和IL - 4。还通过定量ELISA评估MIP - 1α、MIP - 1β、RANTES、MCP - 1、IL - 8、IFN - γ和IL - 4的血清水平。
与非哮喘患者相比，过敏性哮喘患者CD4 +和CD8 + T细胞中MIP - 1β的细胞内表达显著降低（中位数 = 2.29%（1.75 - 3.50）对4.57%（3.38 - 6.64），P = 0.05；14.20%（13.18 - 17.88）对44.10%（30.38 - 48.70），P = 0.01）。同样，过敏性哮喘患者CD8 + T细胞中MIP - 1α的细胞内表达也显著降低（3.67%（1.17 - 5.42）对17.10%（4.97 - 20.43），P = 0.05）。相反，过敏性哮喘患者CD4 +和CD8 + T细胞中IL - 8的表达均显著增强（9.93%（7.77 - 11.28）对4.14%（3.61 - 7.11），P = 0.05；8.40%（6.97 - 10.04）对4.98%（3.37 - 6.08），P = 0.05）。对细胞内IFN - γ和IL - 4的检测显示，过敏性哮喘患者和非哮喘患者CD4 + T细胞中这两种细胞因子的表达均无显著差异。相比之下，过敏性哮喘患者CD8 + T细胞中IFN - γ的表达显著降低（24.60%（21.08 - 32.50）对48.40%（41.50 - 55.28），P = 0.01）。
轻度过敏性哮喘外周T细胞趋化因子表达提示Th1反应减弱，同时细胞因子谱有轻微变化提示Th2反应偏向，这证实了趋化因子参与过敏性哮喘病因的重要性。使用全血培养估计T细胞亚群中趋化因子表达的能力最终可能提供一种评估疾病状态和监测以趋化因子为靶点的早期干预治疗的实用方法。
