Mao Hui, Wang Zeng-li, Liu Chun-tao
Department of Respiratory Medicine, The West China Hospital, Sichuan University, Chengdu 610041, China.
Zhonghua Jie He He Hu Xi Za Zhi. 2003 Mar;26(3):152-6.
To study the possible role of bone marrow-derived hematopoietic cells expressing CD(34)(+) and IL-5 receptor messenger RNA (IL-5R mRNA(+)) in asthmatic airway inflammation.
Balb/c mice were sensitized and challenged by ovalbumin (OVA) to establish the asthmatic model. The control mice were sensitized and exposed to sterile saline. The mice were killed at different time points after challenged by OVA and sterile saline, and bronchoalveolar lavage (BALF), peripheral blood (PB) and bone marrow (BM) were prepared. Eosinophils (EOS) in PB and BALF, and nuclear cells in PB and BM were counted. The percentage of CD(34)(+) cells to nuclear cells (CD(34)(+)%) in PB and BM, and the relative number of CD(34)(+) cells (CD(34)(+)) in PB and BM were calculated by flow cytometry. Immunocytochemistry and in situ hybridization were used to observe the hematopoietic cells with co-localized expression of CD34 and IL-5R mRNA (CD(34)(+)/IL-5R mRNA(+)) in BM. The percentage of BM CD(34)(+)/IL-5R mRNA(+) to BM CD(34)(+) was calculated.
(1) At 6 h after OVA challenge, the number of BALF EOS [(2.67 +/- 1.00) x 105/L] was significantly increased as compared to the number in controls [(0.46 +/- 0.06) x 105/L] (P < 0.01). At 12 h after OVA-challenge, the numbers of BALF EOS [(7.74 +/- 1.98) x 105/L] and PB EOS [(2.91 +/- 0.64) x 108/L] were significantly higher than those in the controls (P < 0.01). At 24 h after OVA-challenge, the numbers of BALF EOS[(19.43 +/- 3.69) x 105/L], PB EOS[(3.93 +/- 0.51) x 108/L] and BM CD(34)(+)/IL-5R mRNA(+) [(300.50 +/- 90.02) per thousand] were increased to the highest levels. The differences were significant as compared to the corresponding parameters in the controls (P < 0.01). At 48 h after OVA-challenge, the numbers of BALF EOS [(12.05 +/- 5.31) x 105/L] and BM CD(34)(+)/IL-5R mRNA(+) [(220.80 +/- 53.41) per thousand] were decreased, but were still significantly different compared to the numbers in the controls (P < 0.01), while other markers returned to the normal levels. (2) The number of BM CD(34)(+)/IL-5R mRNA(+) in the 60 mice was closely correlated with BALF EOS, PB EOS, BM CD(34)(+) and BM CD(34)(+) (P < 0.05).
CD(34)(+) cells expressing IL-5R mRNA, which may favor eosinophilopoiesis and eosinophilic airway inflammation, were increased in the BM of this mouse asthmatic model. A feedback mechanism between the lungs and the bone marrow likely exists, which may be involved in the development and persistence of asthmatic airway inflammation.
研究表达CD(34)(+)和白细胞介素-5受体信使核糖核酸(IL-5R mRNA(+))的骨髓源性造血细胞在哮喘气道炎症中的可能作用。
用卵清蛋白(OVA)致敏并攻击Balb/c小鼠以建立哮喘模型。对照小鼠用无菌生理盐水致敏并暴露。在OVA和无菌生理盐水攻击后不同时间点处死小鼠,制备支气管肺泡灌洗液(BALF)、外周血(PB)和骨髓(BM)。计数PB和BALF中的嗜酸性粒细胞(EOS)以及PB和BM中的核细胞。通过流式细胞术计算PB和BM中CD(34)(+)细胞占核细胞的百分比(CD(34)(+)%)以及PB和BM中CD(34)(+)细胞的相对数量(CD(34)(+))。采用免疫细胞化学和原位杂交观察BM中CD34和IL-5R mRNA共定位表达的造血细胞(CD(34)(+)/IL-5R mRNA(+))。计算BM CD(34)(+)/IL-5R mRNA(+)占BM CD(34)(+)的百分比。
(1)OVA攻击后6小时,BALF中EOS数量[(2.67±1.00)×105/L]与对照组[(0.46±0.06)×105/L]相比显著增加(P<0.01)。OVA攻击后12小时,BALF中EOS数量[(7.74±1.98)×105/L]和PB中EOS数量[(2.91±0.64)×108/L]显著高于对照组(P<0.01)。OVA攻击后24小时,BALF中EOS数量[(19.43±3.69)×105/L]、PB中EOS数量[(3.93±0.51)×108/L]和BM CD(34)(+)/IL-5R mRNA(+) [(300.50±90.02)‰]增加到最高水平。与对照组相应参数相比差异显著(P<0.01)。OVA攻击后48小时,BALF中EOS数量[(12.05±5.31)×105/L]和BM CD(34)(+)/IL-5R mRNA(+) [(220.80±53.41)‰]减少,但与对照组数量相比仍有显著差异(P<0.01),而其他指标恢复到正常水平。(2)60只小鼠中BM CD(34)(+)/IL-5R mRNA(+)数量与BALF中EOS、PB中EOS、BM CD(34)(+)和BM CD(34)(+)密切相关(P<0.05)。
在该小鼠哮喘模型的BM中,表达IL-5R mRNA的CD(34)(+)细胞增加,这可能有利于嗜酸性粒细胞生成和嗜酸性气道炎症。肺和骨髓之间可能存在反馈机制,这可能参与哮喘气道炎症的发生和持续。