Do Tomas, Guran Roman, Jarosova Rea, Ondrackova Petra, Sladek Zbysek, Faldyna Martin, Adam Vojtech, Zitka Ondrej
Department of Chemistry and Biochemistry, Faculty of AgriSciences, Mendel University in Brno, 613 00 Brno, Czech Republic.
Central European Institute of Technology, Brno University of Technology, 612 00 Brno, Czech Republic.
Molecules. 2020 Dec 3;25(23):5723. doi: 10.3390/molecules25235723.
Respiratory infections are a real threat for humans, and therefore the pig model is of interest for studies. As one of a case for studies, (APP) caused infections and still worries many pig breeders around the world. To better understand the influence of pathogenic effect of APP on a respiratory system-lungs and tracheobronchial lymph nodes (TBLN), we aimed to employ matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-TOF MSI). In this study, six pigs were intranasally infected by APP and two were used as non-infected control, and 48 cryosections have been obtained. MALDI-TOF MSI and immunohistochemistry (IHC) were used to study spatial distribution of infectious markers, especially interleukins, in cryosections of porcine tissues of lungs (necrotic area, marginal zone) and tracheobronchial lymph nodes (TBLN) from pigs infected by APP. CD163, interleukin 1β (IL‑1β) and a protegrin-4 precursor were successfully detected based on their tryptic fragments. CD163 and IL‑1β were confirmed also by IHC. The protegrin-4 precursor was identified by MALDI-TOF/TOF directly on the tissue cryosections. CD163, IL‑1β and protegrin‑4 precursor were all significantly ( < 0.001) more expressed in necrotic areas of lungs infected by APP than in marginal zone, TBLN and in control lungs.
呼吸道感染对人类构成了真正的威胁，因此猪模型在研究中具有重要意义。作为研究案例之一，副猪嗜血杆菌（APP）引起的感染仍然困扰着世界各地的许多养猪户。为了更好地了解APP致病作用对呼吸系统——肺和气管支气管淋巴结（TBLN）的影响，我们旨在采用基质辅助激光解吸/电离飞行时间质谱成像（MALDI-TOF MSI）技术。在本研究中，6头猪经鼻感染APP，2头作为未感染对照，共获得48个冷冻切片。利用MALDI-TOF MSI和免疫组织化学（IHC）技术研究感染标志物，尤其是白细胞介素，在感染APP的猪肺组织（坏死区、边缘区）和气管支气管淋巴结（TBLN）冷冻切片中的空间分布。基于其胰蛋白酶片段成功检测到CD163、白细胞介素1β（IL-1β）和一种防御素-4前体。CD163和IL-1β也通过IHC得到证实。防御素-4前体在组织冷冻切片上直接通过MALDI-TOF/TOF鉴定。APP感染的肺坏死区中CD163、IL-1β和防御素-4前体的表达均显著（<0.001）高于边缘区、TBLN和对照肺。
