Development of the polymeric antibody specific to Salmonella as antibiotic substitution

肠炎沙门氏菌(Salmonella Enteritidis，SE)是一种通过人的肠道黏膜感染鸡蛋和肉类，引起食物中毒的病原菌。我们重视利用肠道中对消化酶具有耐受性的多聚体抗体(IgA、IgM)作为预防沙门氏菌感染而不依赖抗生素的方法之一。本研究的目的是通过细胞融合的方法，从小鼠脾细胞与骨髓瘤细胞的细胞融合杂交瘤细胞中连续克隆出5种抗沙门氏菌抗原的杂交瘤细胞，获得5种针对沙门氏菌抗原的抗体。抗SE免疫球蛋白单抗2Ga80k是由SE野毒株鼻腔免疫和腹膜免疫，NRA1、NRA2和NRA3三种抗SEIg A单抗和…联合免疫制备的经口服感染临床分离的SE株，可获得更多的ti-SE-IgM单抗ORM1。2Ga80k为IgG2a型抗体，具有κ链，识别约80 kDa的抗原和脂多糖的表面糖链，并显示出阻止约50-86%的黏附于Caco-2细胞的作用。ORM1为IgM型抗体，可识别42 kDa的糖链抗原，也可被沙门氏菌鞭毛抗体识别。后者提示ORM1可以识别沙门氏菌鞭毛。NRA1、NRA2和NRA3是结构上具有αH链的DiGa抗体，可识别50-80 kDa的糖链抗原和脂多糖。观察到荧光素标记的ORM1、NRA1、NRA2和NRA3异硫氰酸酯连接在SE的细胞表面，并阻止沙门氏菌与Caco-2细胞的黏附。结果表明，聚合型抗SE-IgA和抗SE-IgM抗体对SE具有较好的黏附功能。腹水中注射杂交瘤细胞后产生大量的单抗，产生相应的单抗。然而，这种抗体的量还不足以实际使用。这一点将作为未来的一个大课题。较少

Salmonella enteritidis (SE) is a pathogenic germ, which infect via man's intestinal tract mucosa with chicken egg and meat, and causes food poisoning. We paid our attention to using the polymeric antibody (IgA, IgM) which shows tolerance to digestive enzyme in intestine as one of the methods of defending Salmonella infection without being dependent on antibiotics. The purpose of this research is selecting polymeric antibody specific to SE by cell fusion method, and clarifying the characteristics.Five kinds of monoclonal antibodies specific to Salmonella antigen were produced by successive cloning from hybridoma made through cell fusion between spleen cells of the mouse, which were immunized by three different methods, with a myeloma cell. Anti-SE IgG monoclonal antibody, 2Ga80k, was produced by combined use of nasal and peritoneal immunization of wild strain of SE, three kinds of anti-SE IgA monoclonal antibodies, NRA1, NRA2, and NRA3, by nasal immunization of wild strain of SE, and an … More ti-SE-IgM monoclonal antibody, ORM1, by the oral infecting method of clinical strain of SE. 2Ga80k was IgG2a type of antibody, which has κ chain, recognized the surface sugar chain with about 80 kDa of antigen and LPS, and showed the effect, which obstructed about 50-86% adhesion into a Caco-2 cell. ORM1 was IgM type of antibody, recognized sugar chain antigen of 42 kDa, which was also recognized by antibody to flagellum of Salmonella. The latter suggest that ORM1 can recognize flagellum of salmonella. NRA1, NRA2, and NRA3 showed to be the dIgA antibody having α type H chain structurally, and recognized sugar chain antigen of 50-80 kDa and LPS. It was observed that ORM1, NRA1, NRA2 and NRA3, which were labeled with fluorescein, isothiocyanate was connected in cell surface of SE, and that they obstructed adhesion of a salmonella to the Caco-2 cell. It was shown the polymeric anti-SE-IgA and anti-SE-IgM antibodies have superior adhesion function for SE. Large quantities of monoclonal antibodies were comparatively produced from ascites after peritoneal injection of the hybridoma, which produced the respective kind of monoclonal antibody. However, the amount of this antibody was not enough to use practically. This point will be left as a big subject of the future. Less