Murine models of pulmonary diseases induced by adoptive transfer of T cell clones.

由于T细胞克隆的维护存在一些困难，我们无法完成上述主题的实验。相反，我们在此报告研究的基本实验结果。我们成功地建立了一个抗原驱动的小鼠肺嗜酸性粒细胞增多症模型。用卵白蛋白（OVA）和明矾（3次，间隔2周）诱导小鼠（BALB/c， 6-8周龄）吸入OVA （50mg/ml，持续20min， 6天），最后一次给药后24小时支气管肺泡灌洗液（BALF）中产生9x10^5/ml的Eo。组织学分析显示细支气管周围和血管周围细胞浸润淋巴细胞和嗜酸性粒细胞。近年来，血栓素A2 （TxA2）合成酶抑制剂（Sl）和受体拮抗剂（RA）已被开发用于预防支气管收缩。这些化合物可以改善支气管高反应性，这表明它们的作用点似乎不仅仅是支气管收缩。为了探讨其对嗜酸性粒细胞（Eo）浸润的影响，我们分别用OKY-046 （TxA2 Sl; 100、10、1mg/kg）和S-1452 （TxA2 RA; 25、2.5、0.25 mg/kg）处理该模型小鼠。用任何一种化合物治疗均能以剂量依赖的方式显著降低BALF中的Eo数，最高剂量可达70%。虽然我们既不能在未处理小鼠的BALF中检测到IL-5，也不能在未处理小鼠的血清中检测到IL-5，但在使用任何一种化合物的小鼠的OVA加脾脏细胞的培养上清中，IL-5的产生以剂量依赖性的方式下降，最高剂量时可达75%。干扰素- γ和IL-2的产生也显著降低。小鼠脾细胞CD3、cd4、cd8阳性的百分率与对照组差异无统计学意义。这些结果表明，抑制TxA2不仅可以减少支气管收缩反应，还可以通过抑制细胞因子的产生来减少肺嗜酸性粒细胞的浸润。

We could not complete the experiments of above theme, because of some difficulties concerning maintenance of T cell clones. Instead, we report herein the results of fundamental experiments of the study. We have successfully made a murine model of antigen-driven pulmonary eosinophilia. Primed mice (BALB/c, 6-8wk old) with ovualbumin (OVA) and alum (three times, 2-wk interval) were challenged with OVA by inhalation (50mg/ml for 20min, 6 days) , yielded 9x10^5/ml Eo in bronchoalveolar lavage fluids (BALF) at 24-hours after the last challenge. Histological analysis revieled peribronchiolar and perivascular cell infiltration of lymphocytes and eosinophils. Recently, thromboxane A2 (TxA2) synthetase inhibitor (Sl) and receptor antagonist (RA) have been developed for the prophylaxis of bronchoconstriction. These compounds can improve pronchial hyperreactivity, indicating that the points of their action seem to be more than bronchoconstriction. To explore their influence on eosinophil (Eo) infiltration, we treated this model mice with OKY-046 (TxA2 Sl ; 100,10,1mg/kg) and S-1452 (TxA2 RA ; 25,2.5,0.25 mg/kg). Treatment with either compound significantly reduced Eo number in BALF in a dose-dependent manner up to-70% at the highest dose. Although we could detect IL-5 neither in BALF nor serum from untreated mice, the production of IL-5 decreased in culture supernatants of OVA plus spleen cells from treated mice with either compound in a dose-dependent manner up to-75% at the highest dose. The production of both interferon-gamma and IL-2 also significantly decreased. The cell differentials and percentages of CD3,4 or 8-positive splenocytes from treated mice were not significantly different from untreated mice. These results suggest that inhibition of TxA2 may reduce not only bronchoconstrictive response but also pulmonary infiltration of eosinophils by inhibiting cytokines production in vivo.