Roth Michael, Pasquali Christian, Stolz Daiana, Tamm Michael
Pulmonary Cell Research, DBM University Basel and Pneumology Clinic, University Hospital Basel, Basel, Switzerland.
OM Pharma SA, Geneva, Switzerland.
PLoS One. 2017 Nov 28;12(11):e0188010. doi: 10.1371/journal.pone.0188010. eCollection 2017.
Bronchial epithelial cells (BEC) are primary target for Rhinovirus infection through attaching to cell membrane proteins. OM-85, a bacterial extract, improves recovery of asthma and COPD patients after viral infections, but only part of the mechanism was addressed, by focusing on defined immune cells.
We therefore determined the effect of OM-85 on isolated primary human BEC of controls (n = 8), asthma patients (n = 10) and COPD patients (n = 9).
BEC were treated with OM-85 alone (24 hours) or infected with Rhinovirus. BEC survival was monitored by manual cell counting and Rhinovirus replication by lytic activity. Immuno-blotting and ELISA were used to determine the expression of Rhinovirus interacting proteins: intracellular adhesion molecule (ICAM), major histocompatibility complex class II (MHC-2), complement component C1q receptor (C1q-R), inducible T-Cell co-stimulator (ICOS), its ligand ICOSL, and myeloid differentiation primary response gene 88 (Myd88); as well as for signal transducers Erk1/2, p38, JNK mitogen activated protein kinases MAPK), and cAMP.
OM-85 significantly reduced Rhinovirus-induced BEC death and virus replication. OM-85 significantly increased the expression of virus interacting proteins C1q-R and β-defensin in all 3 probes and groups, which was prevented by either Erk1/2 MAPK or cAMP inhibition. In addition, OM-85 significantly reduced Rhinovirus induced expression of ICAM1 involving p38 MAPK. In BEC OM-85 had no significant effect on the expression of ICOS, ICOSL and MHC-2 membrane proteins nor on the adaptor protein MyD88.
The OM-85-induced increased of C1q-R and β-defensin, both important for antigen presentation and phagocytosis, supports its activity in host cell's defence against Rhinovirus infection.
支气管上皮细胞(BEC)是鼻病毒通过附着于细胞膜蛋白进行感染的主要靶细胞。细菌提取物OM-85可改善病毒感染后哮喘和慢性阻塞性肺疾病(COPD)患者的恢复情况,但仅通过关注特定免疫细胞阐述了部分机制。
因此,我们确定了OM-85对分离的对照(n = 8)、哮喘患者(n = 10)和COPD患者(n = 9)的原代人BEC的影响。
BEC单独用OM-85处理(24小时)或感染鼻病毒。通过手动细胞计数监测BEC存活情况,通过裂解活性监测鼻病毒复制情况。采用免疫印迹法和酶联免疫吸附测定法测定鼻病毒相互作用蛋白的表达:细胞间黏附分子(ICAM)、主要组织相容性复合体II类(MHC-2)、补体成分C1q受体(C1q-R)、诱导性T细胞共刺激分子(ICOS)、其配体ICOSL和髓样分化初级反应基因88(Myd88);以及信号转导分子细胞外调节蛋白激酶1/2(Erk1/2)、p38、应激活化蛋白激酶(JNK)丝裂原活化蛋白激酶(MAPK)和环磷酸腺苷(cAMP)。
OM-85显著降低鼻病毒诱导的BEC死亡和病毒复制。OM-85显著增加了所有3组样本中病毒相互作用蛋白C1q-R和β-防御素的表达,而Erk1/2 MAPK或cAMP抑制可阻止这种增加。此外,OM-85显著降低了鼻病毒诱导的涉及p38 MAPK的ICAM1表达。在BEC中,OM-85对ICOS、ICOSL和MHC-2膜蛋白的表达以及衔接蛋白Myd88均无显著影响。
OM-85诱导的C1q-R和β-防御素增加,这两者对抗原呈递和吞噬作用都很重要,支持了其在宿主细胞抵御鼻病毒感染中的活性。