Differences in mucus and serum immunoglobulin of carp ( Cyprinus carpio L.)

Abstract

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 pathogenrich environment.