Plasma component inhibits platelet aggregation caused by the extracellulal products of Streptococus mitis

血浆成分抑制轻链球菌细胞外产物引起的血小板聚集

• 批准号: 08670322
• 负责人: OHKUNI Hisashi
• 金额: $ 1.41万
• 依托单位: Nippon Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08670322/
• 关键词: Streptococcus mitis; pletelet aggregation factor; Inhibition factor

We demonstrated that culture supernatant concentrate of S.mitis, strain Nm-65, powerfully aggregated blood platelet obtained from one volunteer (YY). A human platelet aggregation factor was purified from the extracellular products (ECP) of S.mitis, strain Nm-65, by sequential chromatography on DEAE-Sepgarose CL-6B, hydroxyapatite and Superdex 75 column. An aggregation factor was tentatively designated S.mitis-derived human platelet aggregation factor (Sm-hPAF). A preliminary study showed that there are considerable individual differences among the platelet-rich plasma (PRP) obtained from healthy adult volunteers with respect to susceptibility to Sm-hPAF.Twenty (77%) PRP specimens derived from 26 healthy volunteers were aggregated by Sm-hPAF, but the remaining 6 (23%) were not reactive. Continuous study, was clearly demonstrated the presence of an inhibitory factor (IF) against Sm-hPAF in the plasma from non -reactive donor, and partial characterization of the IF was studied. The results obtained were as follows :1)IF was a sugar protein with a molecular weight of 55 kDa on SDS-PAGE,and the electrophoretic mobility of IF was associated with alpha-globulin.2)The monoclonal antibody (mAb) to IF, obtained by immunization of BALB/c mouse with an electrophoretically purified IF preparation, inhibited the platelet aggregation by Sm-hPAF, and the mAb directly reacted with Sm-hPAF on nitrocellulose membrane.3)The IF was found in the culture supernatant of human hepatocyte cell line (He cells) on Western blot analysis using the mAb to IF.4)IF in plasms was mesured by Sandwich ELISA using the mAb and polyclonal Ab to IF.When the IF levels in plasma from non-reactive donor against Sm-hPAF were compared with that in plasma from reactive donor, the IF IN plasma of non-reactive donor had significantly higher levels than that of reactive donor.

我们证明了米氏链球菌Nm-65菌株的培养上清浓缩物对一名志愿者(YY)的血小板有很强的聚集作用。用DEAE-Sepgarose CL-6B、羟基磷灰石和Superdex 75柱层析，从米氏链霉菌Nm-65胞外产物中分离纯化出一种人血小板聚集因子。一种聚集因子暂定名为S.mitis来源的人血小板聚集因子(Sm-hPAF)。初步研究表明，健康成年志愿者富含血小板血浆(PRP)对Sm-hPAF的易感性存在较大的个体差异。26名健康志愿者的20份(77%)PRP标本可被Sm-hPAF聚集，其余6份(23%)无反应。连续研究表明，非反应性供者血浆中存在Sm-hPAF抑制因子(IF)，并对IF的部分特性进行了研究。结果表明：1IF是一种分子量为55 kDa的糖蛋白，其凝胶迁移率与α-球蛋白有关。2)用纯化的IF制剂免疫BALB/c小鼠获得的抗IF的单抗能抑制Sm-hPAF诱导的血小板聚集。3)抗Sm-hPAF的单抗与硝酸纤维素膜上的Sm-hPAF直接作用。4)用抗Sm-hPAF的单抗和多克隆抗体进行夹心ELISA法测定血浆中的IF。

项目成果

Hisashi Ohkuni et al.: "Purification and partial characterization of a novel human platelet aggregation factor in the extracellular products of Streptococcus mitis,strain Nm-65" FEMS Immunol.Med.Microbiol.17・2. 121-129 (1997)

Hisashi Ohkuni 等人：“轻链球菌菌株 Nm-65 细胞外产物中新型人血小板聚集因子的纯化和部分表征”FEMS Immunol.Med.Microbiol.17・2（1997）。

Hisashi Ohkuni, er al.: "Purification and partial characterization of a novel human platelet aggergation factor in the extyracellular products of Streptococcus mitis, strain Nm-65" FEMS Immunol.Med.Microbiol.17(2). 121-129 (1997)

Hisashi Ohkuni 等人：“轻症链球菌 Nm-65 株细胞外产物中新型人血小板聚集因子的纯化和部分表征”FEMS Nutritionol.Med.Microbiol.17(2)。