Souza Mariana C, Penido Carmen, Costa Maria F S, Henriques Maria Graças
Laboratório de Farmacologia Aplicada, Departamento de Farmacologia Aplicada, Farmanguinhos, Fundação Oswaldo Cruz, Rua Sizenando Nabuco 100, Manguinhos, Rio de Janeiro 21041-250, Brazil.
Infect Immun. 2008 Dec;76(12):5686-93. doi: 10.1128/IAI.00133-08. Epub 2008 Sep 22.
Tuberculous pleurisy is a frequent extrapulmonary manifestation characterized by accumulation of fluid and inflammatory cells in the pleural space. Here, we investigated the mechanisms of T-lymphocyte accumulation in the pleural space by using a murine model of pleurisy induced by Mycobacterium bovis BCG. Intrathoracic (i.t.) injection of BCG (4.5 x 10(5) bacteria/cavity) induced accumulation of T lymphocytes in the pleural cavities of C57BL/6 mice. We observed the presence of CFU in pleural washes conducted 1, 2, 3, 7, and 15 days after pleurisy induction. Pretreatment with fucoidan inhibited T-lymphocyte accumulation at 1 day, but not at 15 days, after BCG-induced pleurisy. Accordingly, adoptive transfer of fluorescein isothiocyanate-labeled blood mononuclear cells to infected mice showed that T lymphocytes migrated into the pleural cavity 1 day (but not 15 days) after BCG injection. Cell-free pleural wash fluids recovered from mice 1 day after BCG i.t. stimulation (day 1 BCG-PW), but not day 7 or day 15 BCG-PW, induced in vitro T-cell transmigration, which was dependent on L-, P-, and E-selectins. In contrast, day 7 BCG-PW (but not day 1 BCG-PW) induced in vitro T-lymphocyte proliferation via interleukin-2 (IL-2) and gamma interferon (IFN-gamma). Accordingly, in vivo IL-2 or IFN-gamma neutralization abolished T-lymphocyte accumulation 7 days after pleurisy induction. Our results demonstrate that pleural infection induced by BCG leads to T-lymphocyte accumulation in two waves. The acute phase depends on selectin-mediated migration, while the second wave of T-lymphocyte accumulation seems to depend on a local proliferation induced by cytokines produced in situ.
结核性胸膜炎是一种常见的肺外表现，其特征为胸腔内积液和炎症细胞积聚。在此，我们利用牛分枝杆菌卡介苗诱导的小鼠胸膜炎模型，研究了T淋巴细胞在胸腔内积聚的机制。向胸腔内（i.t.）注射卡介苗（4.5×10⁵个细菌/腔）可诱导C57BL/6小鼠胸腔内T淋巴细胞积聚。我们在诱导胸膜炎后的第1、2、3、7和15天进行胸腔灌洗，观察到灌洗液中存在集落形成单位（CFU）。在用岩藻依聚糖预处理后，卡介苗诱导的胸膜炎后第1天T淋巴细胞积聚受到抑制，但第15天未受抑制。因此，将异硫氰酸荧光素标记的血液单核细胞过继转移至感染小鼠，结果显示卡介苗注射后第1天（而非第15天）T淋巴细胞迁移至胸腔。卡介苗i.t.刺激后第1天从小鼠回收的无细胞胸腔灌洗液（第1天卡介苗 - PW），而非第7天或第15天的卡介苗 - PW，可诱导体外T细胞迁移，这依赖于L - 、P - 和E - 选择素。相反，第7天的卡介苗 - PW（而非第1天的卡介苗 - PW）通过白细胞介素 - 2（IL - 2）和γ干扰素（IFN - γ）诱导体外T淋巴细胞增殖。因此，体内中和IL - 2或IFN - γ可消除胸膜炎诱导后第7天的T淋巴细胞积聚。我们的结果表明，卡介苗诱导的胸膜感染导致T淋巴细胞分两波积聚。急性期依赖于选择素介导的迁移，而T淋巴细胞积聚的第二波似乎依赖于原位产生的细胞因子诱导的局部增殖。
