Identification of a mannose‐binding lectin‐like protein recognized by the anti‐CD25 antibody in haemocytes fromCherax quadricarinatus

Abstract

The crustacean haemolymph contains three main cell populations; however, it is not clear which mechanisms participate in the regulation of cells related to innate immunity. This work aimed to identify potential interleukin-like receptors that could regulate cellular responses in Cherax quadricarinatus. By histochemical analysis with murine anti-CD25 staining (targeting the α-chain of the IL-2 receptor), we identified that this antibody recognizes cytoplasmic granules in semigranular and granular haemocytes. In haemocytes stimulated with phorbol myristate acetate (PMA), increased fluorescence was observed in these cytoplasmic granules, whereas staining with a human IL-2 antibody after stimulation with 1–10 ng/ml PMA revealed no overexpression of the receptor or oxidative burst in haemocytes. Two-dimensional Western blot analysis of haemocyte lysates showed that anti-CD25 identified a 27.4-kDa protein with an isoelectric point (pI) of 7.7 and a 46-kDa protein with a pI of 6.9. De novo sequencing of these proteins identified that they had 32% homology with a mannose-binding lectin (MBL) from Pacifastacus leniusculus. Our results indicate that a mannose-binding lectin-like protein could exert a protective effect that prevents damage from other activated immune responses.