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IgA antibody production by intrarectal immunization of mice usingrecombinant major capsid protein of hamster polyomavirus

机译:通过使用小鼠的直肠内免疫产生IgA抗体仓鼠多瘤病毒的重组主要衣壳蛋白

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摘要

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 usefultool for the investigation of mucosal immune responses or autoimmune diseasesand extends the spectrum of antibodies with specific effector functions.
机译:病毒蛋白具有高度抗原性,被称为适应性免疫反应的有效刺激物。在大多数情况下,这种机制通常用于产生单克隆抗体的生物技术应用中,从而产生高亲和力的IgG抗体。这项研究的目的是增加小鼠中抗原特异性IgA抗体的水平,以便通过杂交瘤技术产生单克隆IgA抗体。为此,仓鼠多瘤病毒(HaPyV)主要衣壳蛋白VP1用于通过不同途径免疫小鼠,以诱导VP1特异性IgA效价。重组HaPyV-VP1在大肠杆菌中产生,并经腹膜内,口服和直肠内给药。通过ELISA在血清和器官培养上清液中确定VP1特异性抗体。我们发现,在对小鼠进行直肠免疫后,脾脏器官培养物中的HaPyV-VP1特异性IgA显着增加,而肠系膜淋巴结,结肠或淋巴集结的培养物中,则没有。相反,口服和腹膜内免疫根本没有提供适当的特异性IgA诱导。这些结果表明,可以通过脾脏的直肠内免疫来诱导特异性的IgA抗体。具有明确特性的单克隆IgA抗体的产生是有用的用于研究粘膜免疫反应或自身免疫疾病的工具并扩展具有特定效应子功能的抗体的范围。

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