行政院國家科學委員會專題研究計畫 成果報告
Osteopontin 基因在急性呼吸窘迫症候群, 結核性肋膜炎及
肺外結核病人之表現
計畫類別： 個別型計畫 計畫編號： NSC93-2314-B-002-221- 執行期間： 93 年 08 月 01 日至 94 年 07 月 31 日 執行單位： 國立臺灣大學醫學院檢驗醫學科 計畫主持人： 李麗娜 報告類型： 精簡報告 處理方式： 本計畫可公開查詢中 華 民 國 94 年 11 月 7 日
行政院國家科學委員會專題研究計畫成果報告
Osteopontin 基因在結核性肋膜炎及粟粒性結核病
人之表現
Expression of osteopontin gene in patients with
tuberculous pleurisy and miliary tuberculosis
計畫編號：NSC 93-2314-B-002-221
執行期限：93 年 8 月 1 日至 94 年 7 月 31 日
主持人：李麗娜
國立台灣大學醫學院檢驗醫學科
一、中文摘要 osteopontin 是由巨噬細胞與 T 淋巴球產生的介 質 (matrix) 蛋白. 過去已有報告顯示, 在結核病 造成的肉芽腫 (granuloma), osteopontin 之表 現很強. 亦有報告顯示, 肺結核病人血液中的 osteopontin 濃度比正常人血液中的值高出很多. 本計畫旨在研究結核性肋膜炎及粟粒性結核病人 血液中 osteopontin 的濃度, 並與正常人血液中的 值比較, osteopontin 的濃度測定是使用 ELISA 法. 研究結果顯示, 結核性肋膜炎及粟粒性結核病人血 漿中的 osteopontin 比正常人血漿中的值, 有顯 著之上升. 關鍵詞：osteopontin, 結核菌, 肋膜炎, 粟粒性, 肉芽腫 AbstractOsteopontin, a noncollagenous matrix protein produced by macrophages and T lymphocytes, is expressed in granulomatous lesions caused by Mycobacterium tuberculosis infection. Circulating osteopontin levels have been found to be elevated in patients with pulmonary tuberculosis, and correlate with severity of pulmonary tuberculosis. In the present study we investigated plasma osteopontin levels in patients with tuberculous pleurisy, those with miliary tuberculosis, as there was no reports of such
study, and compared them with those of normal control subjects. We found that plasma osteopontin levels, as measured by an ELISA assay, were significantly higher in patients with tuberculous pleurisy, those with miliary tuberculosis, than in healthy control subjects.
Keywords: osteopontin, Mycobacterium
tuberculosis, pleurisy, miliary,
granuloma
INTRODUCTION
Osteopontin is a phophorylated acidic glycoprotein, which contains an
arginine-glycine-aspartate-binding motif to the integrin family of adhesion molecules (1-3). It is abundantly produced by T cells and macrophages during the formation of granulomatous lesions in tuberculosis (4). Ashkar and coworkers (5) found a new role for OPN, acting as a proinflammatory cytokine for inducing type 1 T helper (Th1) cell-mediated immune responses. Using osteopontin-knockout mice, they showed that osteopontin was essential for the
†八十六年度及以前的一般 國科會專題計畫(不含產學 合作研究計畫)亦可選擇適 用，惟較特殊的計畫如國科 會規劃案等，請先洽得國科 會各學術處同意。
development of Th1 responses and
granuloma formation through induction of interleukin (IL)-12 production by
macrophages.
Host defense against M. tuberculosis consists largely of Th1 cell-mediated mechanism (6). In the present study we investigated the pathogenic role of osteopontin in tuberculosis by measuring plasma osteopontin levels in patients with tuberculous pleurisy and those with miliary tuberculosis.
MATERIALS AND METHODS
We studied 7 male and 3 female patients with tuberculous pleurisy, and 3 male
patients with miliary tuberculosis. Their ages ranged from 40 to 72 years (61.012.4). M. tuberculosis was isolated from sputum of all patients. Plasma samples were collected before anti-tuberculous chemotherapy was started, and were frozen at -80oC until use.
We also studied 15(10 males) healthy volunteers who had no history of pulmonary tuberculosis or other infection, and their chest radiographs showed no evidence of respiratory diseases. Their ages ranged from 31 to 65 years (54.210.3).
Measurement of osteopontin
The concentration of osteopontin was measured by an antigen-capture ELISA (7).
Results
Plasma osteopontin concentrations in patients with tuberculous pleurisy and those with miliary tuberculosis
To investigate the pathogenic role of osteopontin in tuberculosis, we compared the plasma concentrations of this cytokine in
patients with tuberculous pleural effusion, and those with miliary tuberculosis with healthy control subjects. As shown in Table 1, plasma osteopontin concentrations in patients with tuberculous pleurisy (n=10) were
significantly higher than in control subjects (n=15). Plasma osteopontin concentrations in patients with miliary tuberculosis (n=3) was also significantly higher than in control subjects.
Table 1 Plasma osteopontin levels in patients with tuberculous pleurisy, patients with miliary tuberculosis, and healthy controls.
Diagnosis Plasma osteopontin p value (MSD)(ng/ml) TB pleurisy 739.7321.7 < 0.001 (n=10) Miliary TB 867.3442.9 < 0.001 (n=3) Healthy 227.593.4 (n=15) DISCUSSION
In the present study, we measured the levels of circulating osteopontin in patients with tuberculous pleurisy, miliary
tuberculosis and healthy control subjects. We found that plasma osteopontin levels were significantly higher in patients with tuberculous pleurisy, those with miliary tuberculosis, than in healthy controls.
infection of human alveolar macrophages and a macrophage cell line with M. tuberculosis caused a substantial increase in osteopontin gene expression. The expression of
osteopontin was elevated in pathologic sections from patients with various
granulomatous diseases including pulmonary tuberculosis (4,8). The observations
suggested the possible involvement of osteopontin in the pathogenesis, especially the generation of granulomas, of
tuberculosis.
Kiguchi and colleagues (10) also reported that plasma osteopontin levels were significantly higher in patients with
tuberculosis than in control subjects. They also observed that osteopontin levels were higher in smear-positive than in
smear-negative patients. The severity of tuberculosis, as evaluated by the size of pulmonary lesions on chest radiographs, correlated with plasma osteopontin concentrations. Osteopontin levels decreased in parallel with clinical improvement after treatment with
antituberculous agents. These observations strongly support the role of osteopontin in the pathogenesis associated with pulmonary tuberculosis.
There was, however, no reports of osteopontin concentrations in patients with tuberculous pleurisy or miliary tuberculosis in the past. The results of our preliminary
study were compatible with those involving patients with pulmonary tuberculosis. We need to measure circulating osteopontin levels in patients with other pulmonary diseases, such as malignant pleural effusion, parapneumonic effusions, autoimmune disease associated pleural effusions, uremic effusions, effusions associated pulmonary infaction, and miliary carcinomatosis, before deciding if circulating osteopontin levels can be of help in diagnosing tuberculous pleurisy or miliary tuberculosis.
CONCLUSION
Plasma osteopontin levels were significantly higher in patients with tuberculous pleurisy, those with miliary tuberculosis, than in healthy control subjects.
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