Messerschmidt K, Hempel S, Holzlöhner P, Ulrich R G, Wagner D, Heilmann K
Eur J Microbiol Immunol (Bp). 2012 Sep;2(3):231-8. doi: 10.1556/EuJMI.2.2012.3.9. Epub 2012 Sep 10.
Viral proteins are highly antigenic and known as potent stimulators of adaptive immune responses. This mechanism is often used for biotechnological applications in monoclonal antibody production resulting in high-affinity IgG antibodies in most cases. The aim of this study was to increase antigen-specific IgA antibody levels in mice in order to generate monoclonal IgA antibodies by hybridoma technology. For this purpose, hamster polyomavirus (HaPyV) major capsid protein VP1 was used to immunize mice by different routes in order to induce VP1-specific IgA titers. Recombinant HaPyV-VP1 was generated in Escherichia coli and administered intraperitoneally, orally, and intrarectally. VP1-specific antibodies were determined by ELISA in sera and organ culture supernatants. We found a significant increase of HaPyV-VP1-specific IgAs in spleen organ cultures after rectal immunization of mice but not in cultures of mesenteric lymph nodes, colon, or Peyer's patches. In contrast, oral and intraperitoneal immunization did not provide an appropriate specific IgA induction at all. These results show that specific IgA antibodies can be induced by intrarectal immunization in the spleen. The generation of monoclonal IgA antibodies with well-defined properties is a useful tool for the investigation of mucosal immune responses or autoimmune diseases and extends the spectrum of antibodies with specific effector functions.
病毒蛋白具有高度抗原性,是适应性免疫反应的有效刺激物。这种机制常用于单克隆抗体生产的生物技术应用中,在大多数情况下可产生高亲和力的IgG抗体。本研究的目的是提高小鼠体内抗原特异性IgA抗体水平,以便通过杂交瘤技术产生单克隆IgA抗体。为此,使用仓鼠多瘤病毒(HaPyV)主要衣壳蛋白VP1通过不同途径免疫小鼠,以诱导VP1特异性IgA滴度。重组HaPyV-VP1在大肠杆菌中产生,并通过腹腔内、口服和直肠内给药。通过ELISA测定血清和器官培养上清液中的VP1特异性抗体。我们发现,小鼠直肠免疫后,脾脏器官培养物中HaPyV-VP1特异性IgA显著增加,但肠系膜淋巴结、结肠或派伊尔结培养物中未增加。相比之下,口服和腹腔内免疫根本不能诱导出合适的特异性IgA。这些结果表明,直肠内免疫可在脾脏中诱导特异性IgA抗体。产生具有明确特性的单克隆IgA抗体是研究黏膜免疫反应或自身免疫性疾病的有用工具,并扩展了具有特定效应功能的抗体谱。