Hibino Y, Konishi Y, Koike J, Tabata T, Ohashi Y, Sugano N
Cell Biology Division, Faculty of Pharmaceutical Sciences, Toyama Medical & Pharmaceutical University, Japan.
Immunopharmacology. 1994 Jul-Aug;28(1):77-85. doi: 10.1016/0162-3109(94)90041-8.
A xylose-rich heteroglycan-protein fraction (LAP1) was prepared from a solid culture medium in which Lentinus edodes mycelia were growing actively. Mouse splenic cells (SPs) were incubated with [3H]TdR in the presence of LAP1. The incubated SPs were fractionated into plastic adherent splenic cells (ADs), nylon-column effluent splenic cells (NEs) and sIg-expressed splenic cells (SIs), which are rich in Mac-1+, Thy-1.2+ and Ly-5+ cells, respectively. The incorporation of [3H]TdR in response to LAP1 was enhanced in each of the fractionated cell populations. Northern or dot blot hybridization showed that productions of IFN-gamma and its receptor mRNAs are induced predominantly in NEs. In another experiment, SPs were fractionated into ADs, NEs and SIs. Then, NE-AD, SI-AD and NE-SI mixtures were prepared and incubated in the same manner. A significant incorporation of [3H]TdR was shown only in the NE-AD mixture. The enzyme-linked immunosorbent assay showed that IFN-gamma production in response to LAP1 is induced in SPs or in the NE-AD mixture, but not in NEs alone. The level of the production was about 5 times higher in the mixture than in SPs after a 72 h incubation. Moreover, LAP1 was capable of inducing NO2- production in SPs. Thus, the present studies imply that this heteroglycan-protein fraction stimulates productions of IFN-gamma and nitrite in mouse splenic cells, augmenting antitumor immune response(s).
从香菇菌丝体在其中活跃生长的固体培养基中制备了富含木糖的杂聚糖 - 蛋白质组分（LAP1）。将小鼠脾细胞（SPs）在LAP1存在的情况下与[³H]TdR一起孵育。孵育后的SPs被分离为塑料贴壁脾细胞（ADs）、尼龙柱流出脾细胞（NEs）和表面免疫球蛋白（sIg）表达的脾细胞（SIs），它们分别富含Mac - 1⁺、Thy - 1.2⁺和Ly - 5⁺细胞。在每个分离的细胞群体中，对LAP1反应的[³H]TdR掺入量均增加。Northern或斑点印迹杂交显示，IFN - γ及其受体mRNA的产生主要在NEs中被诱导。在另一项实验中，将SPs分离为ADs、NEs和SIs。然后，制备NE - AD、SI - AD和NE - SI混合物并以相同方式孵育。仅在NE - AD混合物中显示出显著的[³H]TdR掺入。酶联免疫吸附测定表明，对LAP1反应的IFN - γ产生在SPs或NE - AD混合物中被诱导，但单独在NEs中未被诱导。孵育72小时后，混合物中的产生水平比SPs中的高约5倍。此外，LAP1能够诱导SPs中NO₂⁻的产生。因此，本研究表明这种杂聚糖 - 蛋白质组分刺激小鼠脾细胞中IFN - γ和亚硝酸盐的产生，增强抗肿瘤免疫反应。
