Jang Hyeweon, Lee Geun Woo, Kim Yang-Hoon, Min Jiho
Graduate School of Semiconductor and Chemical Engineering, Jeonbuk National University, 567 Baekje-daero, Deokjin-gu, 54896, Jeonju-si, Jeollabuk-do, Republic of Korea.
School of Biological Sciences, Chungbuk National University, Chungdae-Ro, Seowon-Gu, 28644, Cheongju, South Korea.
AMB Express. 2021 Oct 29;11(1):146. doi: 10.1186/s13568-021-01308-3.
Histamine is an immune mediator that is mainly secreted when an immediate, rapid response is needed in the body, and an excessive secretion of histamine or lack of enzymes that degrade histamine can result in various side effects. Histamine binding protein (HBP) is secreted by a mite species to prevent the host's histamine-induced immune responses by binding the histamine molecule in the blood. Cloning was performed to express HBP on the yeast surface (MBTL-GWL-1), and immunofluorescence (IF) and western blot was performed to confirm the expression of the recombinant protein. The histamine inhibitory ability of GWL-1 cells was tested according to the cell concentration. The highest inhibitory ability of 1.30 × 10 CFU/ml of GWL-1 cells was of about 60 %. The GWL-1 cell concentration and the degree of histamine inhibition were confirmed to be dose-dependent, and dead cell debris was shown to have a histamine inhibitory effect, although not as much as that of whole cells. Phagocytosis assays were performed to determine whether histamine affected the RAW 264.7 cell's phagocytosis, and to indirectly confirm the GWL-1 cell's histamine inhibition. By confirming that, we found that GWL-1 captures histamine. Therefore, it can be expected to become a competitive material in the anti-allergy market.
组胺是一种免疫介质,主要在身体需要即时、快速反应时分泌,组胺分泌过多或降解组胺的酶缺乏会导致各种副作用。组胺结合蛋白(HBP)由一种螨虫分泌,通过结合血液中的组胺分子来阻止宿主的组胺诱导的免疫反应。进行克隆以在酵母表面表达HBP(MBTL - GWL - 1),并进行免疫荧光(IF)和蛋白质印迹以确认重组蛋白的表达。根据细胞浓度测试GWL - 1细胞的组胺抑制能力。GWL - 1细胞浓度为1.30×10 CFU/ml时的最高抑制能力约为60%。证实GWL - 1细胞浓度与组胺抑制程度呈剂量依赖性,并且死细胞碎片显示出组胺抑制作用,尽管不如全细胞的抑制作用强。进行吞噬试验以确定组胺是否影响RAW 264.7细胞的吞噬作用,并间接确认GWL - 1细胞的组胺抑制作用。通过确认这一点,我们发现GWL - 1捕获组胺。因此,有望成为抗过敏市场上具有竞争力的材料。