Altered intracellular expression of the chemokines MIP-1α, MIP-1β and IL-8 by peripheral blood CD4+ and CD8+ T cells in mild allergic asthma
Altered intracellular expression of the chemokines MIP-1α, MIP-1β and IL-8 by peripheral blood CD4+ and CD8+ T cells in mild allergic asthma
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DOI：
10.1034/j.1398-9995.2003.00035.x
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发表时间：
2003-03-01
期刊：
影响因子：
12.4
通讯作者：
Bisset, LR
中科院分区：
文献类型：
--
作者：
Grob, M;Schmid-Grendelmeier, P;Bisset, LR
背景：趋化因子调节Th1和Th2反应的能力表明在过敏性哮喘等特应性疾病的发病机制中起作用，其中Th2反应已被观察到优势。虽然过敏性哮喘对肺局部趋化因子产生的影响一直是研究的主题，但对疾病进展对外周趋化因子产生的影响知之甚少。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血t细胞趋化因子表达的影响。方法：研究对象包括轻度过敏性哮喘患者（n=7）和非哮喘对照组（n=7）。体外用12-肉豆酸酯phorbol （PMA）和离子霉素刺激外周静脉血后，用流式细胞术估计CD4（+）和CD8(+) T细胞产生多种趋化因子的百分比，这些趋化因子包括巨噬细胞炎症蛋白mip -1 α和mip -1 β， RANTES（受激活调节，T细胞表达和分泌），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。定量ELISA法检测血清mip -1 α、mip -1 β、RANTES、MCP-1、IL-8、ifn - γ、IL-4水平。结果：变应性哮喘患者细胞内CD4（+）和CD8(+) T细胞mip -1 β的表达明显低于非哮喘患者(中位数=2.29% (1.75 ~ 3.50)vs 4.57% (3.38 ~ 6.64)， P=0.05；14.20%(13.18 - -17.88)和44.10% (30.38 - -48.70),P = 0.01)。同样，变应性哮喘患者CD8(+) T细胞mip -1 α的细胞内表达也显著降低(3.67% （1.17-5.42)vs 17.10% (4.97-20.43)， P=0.05）。相反，变应性哮喘患者CD4（+）和CD8(+) T细胞IL-8表达均显著增强(9.93% (7.77 ~ 11.28)vs 4.14% (3.61 ~ 7.11)， P=0.05；8.40%(6.97 - -10.04)和4.98% (3.37 - -6.08),P = 0.05)。细胞内ifn - γ和IL-4的检测显示，变应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，变应性哮喘患者CD8(+) t细胞中ifn - γ的表达明显降低(24.60% （21.08-32.50)vs 48.40% (41.50-55.28)， P=0.01）。结论：轻度过敏性哮喘患者外周血t细胞趋化因子表达提示Th1反应减弱，与提示Th2反应偏倚的细胞因子谱的边际变化相吻合，证实了趋化因子参与过敏性哮喘病因学的重要性。利用全血培养来估计T细胞亚群中趋化因子表达的能力可能最终为评估疾病状态和监测针对趋化因子的早期干预治疗提供实用手段。
Background: 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.Methods: 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.Results: 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).Conclusions: 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.