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通过嗜酸性粒细胞存活试验和酶联免疫吸附测定法检测支气管哮喘患者痰液中的转化生长因子-β

Detection of transforming growth factor-beta in sputum from patients with bronchial asthma by eosinophil survival assay and enzyme-linked immunosorbent assay.

作者信息

Adachi T, Motojima S, Hirata A, Fukuda T, Kihara N, Makino S

机构信息

Department of Respiratory Medicine, Kihara Hospital, Tokyo, Japan.

出版信息

Clin Exp Allergy. 1996 May;26(5):557-62.

PMID:8735868
Abstract

BACKGROUND

We have shown that interleukin-5 (IL-5) and granulocyte-macrophage colony-stimulating factor (GM-CSF) are present in sputum from patients experiencing acute asthma attacks, by eosinophil survival assay. The viability of guinea-pig eosinophils was significantly increased in the presence of such sputum extracts after 3 days' culture, and it was inhibited by the addition of anti-IL-5 and anti-GM-CSF antibodies. However, the contribution of IL-5 to the increase in eosinophil viability was less than expected from the values of IL-5 measured by enzyme-linked immunosorbent assay (ELISA). Therefore, we speculated that something in sputum inhibited the function of IL-5.

OBJECTIVE

Transforming growth factor-beta (TGF-beta) was the only cytokine we tested that inhibited the prolongation of survival of guinea-pig eosinophils induced by IL-5. The objective of this study is to detect TGF-beta in the same sputum.

METHODS

Guinea-pig eosinophils were cultured with or without anti-TGF-beta antibody in the presence of sputum extracts, and the eosinophil viability was counted after 3 days. Measurement of TGF-beta 1 in sputum was performed by ELISA.

RESULTS

Eosinophil viabilities with and without anti-TGF-beta antibody were 79.7 +/- 2.9% and 69.0 +/- 2.7%, respectively, and the difference between them was statistically significant (P < 0.05, n = 9). The concentration of TGF-beta 1 in the sputum was 21.7 +/- 3.3 ng/mL (n = 9).

CONCLUSION

These observations suggest that TGF-beta is present in sputum from patients with bronchial asthma.

摘要

背景

我们通过嗜酸性粒细胞存活试验表明,白细胞介素-5(IL-5)和粒细胞-巨噬细胞集落刺激因子(GM-CSF)存在于急性哮喘发作患者的痰液中。豚鼠嗜酸性粒细胞在含有此类痰液提取物的情况下培养3天后,其活力显著增加,而添加抗IL-5和抗GM-CSF抗体可抑制这种增加。然而,IL-5对嗜酸性粒细胞活力增加的贡献低于通过酶联免疫吸附测定(ELISA)测得的IL-5值所预期的水平。因此,我们推测痰液中的某种物质抑制了IL-5的功能。

目的

转化生长因子-β(TGF-β)是我们测试的唯一一种能抑制IL-5诱导的豚鼠嗜酸性粒细胞存活延长的细胞因子。本研究的目的是检测同一痰液中的TGF-β。

方法

在有或无抗TGF-β抗体的情况下,将豚鼠嗜酸性粒细胞与痰液提取物一起培养,3天后计算嗜酸性粒细胞活力。通过ELISA测定痰液中TGF-β1的含量。

结果

有和无抗TGF-β抗体时嗜酸性粒细胞活力分别为79.7±2.9%和69.0±2.7%,两者差异有统计学意义(P<0.05,n=9)。痰液中TGF-β1的浓度为21.7±3.3 ng/mL(n=9)。

结论

这些观察结果表明,支气管哮喘患者的痰液中存在TGF-β。

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