Rost Bettina, Hanf Gerald, Ohnemus Ulrich, Otto-Knapp Ralf, Groneberg David A, Kunkel Gert, Noga Oliver
Allergy and Asthma Clinic, Department of medicine Infectious and Respiratory diseases, Charité, Humboldt University, Augustenburger Platz 1, Berlin 13353, Germany.
Regul Pept. 2005 Jan 15;124(1-3):19-25. doi: 10.1016/j.regpep.2004.06.024.
Recent studies have shown that neurotrophins (NTs) are involved in inflammatory processes. Elevated plasma levels of NTs were found allergic diseases with the highest levels in allergic asthma. However, the exact cellular sources involved in the regulation and release of neurotrophins in allergic inflammation are still not well defined.
The aim of this study was to assess whether monocytes of allergic and non-allergic subjects produce, store and release the neurotrophins NGF, BDNF and NT-3.
Monocytes of allergic and non-allergic donors were purified by immunomagnetic selection. APAAP-staining for the presence of NTs and their receptors was performed. RT-PCR and Western blot evaluated the production and storage of NTs. Monocytes were incubated and supernatants were collected for measurement of neurotrophic factors after stimulation with lipopolysaccharide (LPS) as inflammatory stimulus. The neurotrophin content in lysates and cell culture supernatants was determined by ELISA.
Human monocytes express the neurotrophins NGF, BDNF and NT-3 but also their specific receptors TrkA, TrkB and TrkC. RT-PCR amplification of isolated mRNA demonstrated expression of the examined neurotrophins. Proteins were detectable by Western blot. NTs were found in the monocyte lysates and supernatants at different levels in allergic and non-allergic donors. Cell stimulation with LPS leads to release of NGF and NT3.
Monocytes, produce, store and release NGF, BDNF and NT-3. They are a possible source of elevated neurotrophin levels found in allergy and asthma.
最近的研究表明,神经营养因子(NTs)参与炎症过程。在过敏性疾病中发现血浆NTs水平升高,其中过敏性哮喘患者的水平最高。然而,过敏性炎症中神经营养因子调节和释放的确切细胞来源仍未明确。
本研究旨在评估过敏性和非过敏性受试者的单核细胞是否产生、储存和释放神经营养因子NGF、BDNF和NT-3。
通过免疫磁选纯化过敏性和非过敏性供体的单核细胞。进行抗碱性磷酸酶桥联酶标法(APAAP)染色以检测NTs及其受体的存在。逆转录聚合酶链反应(RT-PCR)和蛋白质免疫印迹法(Western blot)评估NTs的产生和储存情况。将单核细胞孵育,在用脂多糖(LPS)作为炎症刺激物刺激后收集上清液以测量神经营养因子。通过酶联免疫吸附测定法(ELISA)测定裂解物和细胞培养上清液中的神经营养因子含量。
人单核细胞表达神经营养因子NGF、BDNF和NT-3,以及它们的特异性受体TrkA、TrkB和TrkC。对分离的信使核糖核酸(mRNA)进行RT-PCR扩增显示所检测的神经营养因子有表达。通过Western blot可检测到蛋白质。在过敏性和非过敏性供体的单核细胞裂解物和上清液中发现不同水平的NTs。用LPS刺激细胞导致NGF和NT-3的释放。
单核细胞产生、储存和释放NGF、BDNF和NT-3。它们可能是在过敏和哮喘中发现的神经营养因子水平升高的一个来源。