Rombout J H, Taverne N, van de Kamp M, Taverne-Thiele A J
Department of Experimental Animal Morphology and Cell Biology, Agricultural University, Wageningen, The Netherlands.
Dev Comp Immunol. 1993 Jul-Aug;17(4):309-17. doi: 10.1016/0145-305x(93)90003-9.
Electrophoretic analysis did not reveal clear differences between skin mucus and serum immunoglobulin (Ig) of carp. The majority of both Igs were tetrameric (+/- 760 kDa) and composed of 25 kDa light (L) chains and 70 kDa heavy (H) chains, but dimeric and monomeric forms were found as well. Monoclonal antibody (mAb) WCI 12 produced from serum Ig appeared to react with the H chain of both molecules. After immunisation of mice with purified mucus Ig, mAbs could be selected that were reactive with mucus Ig only. Two of these mAbs (WCI M1 and WCI M2) were immunoreactive with the H chain of mucus Ig and not or hardly immunoreactive with the H chain of serum Ig, indicating differences in the composition of the H chains of both molecules. Because WCI M2 appeared to recognize a carbohydrate determinant, differences seem to occur in the protein as well as carbohydrate composition of mucus and serum Ig. Flow cytometric results showed that both mAbs were reactive with the same subpopulation of WCI 12-positive B cells. Immunohistochemical reactions on cryosections also showed a limited reaction by these mAbs compared with WCI 12; only epithelium of skin and bile ducts and capillaries in the liver were strongly positive with these mAbs. The presence of mucus Ig at these locations is discussed. Our results indicate structural and functional differences between mucus and serum Ig, which may explain the mucosal immune responses reported for fish. Such a specific mucosal defense system can be very important for fish, living in a pathogen-rich environment.
电泳分析未揭示鲤鱼皮肤黏液和血清免疫球蛋白(Ig)之间的明显差异。两种免疫球蛋白大多为四聚体(±760 kDa),由25 kDa的轻链(L)和70 kDa的重链(H)组成,但也发现了二聚体和单体形式。由血清Ig产生的单克隆抗体(mAb)WCI 12似乎与两种分子的重链发生反应。用纯化的黏液Ig免疫小鼠后,可以筛选出仅与黏液Ig反应的单克隆抗体。其中两种单克隆抗体(WCI M1和WCI M2)与黏液Ig的重链具有免疫反应性,而与血清Ig的重链无反应或几乎无反应,这表明两种分子重链的组成存在差异。由于WCI M2似乎识别一种碳水化合物决定簇,黏液和血清Ig在蛋白质以及碳水化合物组成上似乎都存在差异。流式细胞术结果表明,两种单克隆抗体均与WCI 12阳性B细胞的同一亚群发生反应。与WCI 12相比,对冰冻切片的免疫组织化学反应也显示这些单克隆抗体的反应有限;只有皮肤、胆管上皮以及肝脏中的毛细血管对这些单克隆抗体呈强阳性。讨论了这些部位黏液Ig的存在情况。我们的结果表明黏液和血清Ig之间存在结构和功能差异,这可能解释了鱼类报道的黏膜免疫反应。这样一种特定的黏膜防御系统对生活在富含病原体环境中的鱼类可能非常重要。