Immunocytochemical detection of Ig-positive cells in blood, lymphoid organs and the gut associated lymphoid tissue of the turbot (Scophthalmus maximus)

Abstract

The present study was designed to search for the sites of the B-cell lineage in the different lymphoid organs of turbot (Scophthalmus maximus) by immunoperoxidase staining with a rabbit polyclonal antiserum against deglycosylated turbot IgM (TUDG-6). A turbot immunoglobulin (Ig) fraction, isolated by protein A, was checked for purity by gel filtration and SDS-PAGE under reducing conditions. The turbot IgM was deglycosylated and used to raise an antiserum. The antiserum titre was evaluated in ELISA. It was then used to analyse turbot peripheral blood leucocytes for membrane and cytoplasmic Ig and for immunohistochemistry with turbot lymphoid tissues. Very low numbers of Ig+cells were found in thymus sections. In sections of spleen, Ig+cells were observed in white pulp, around ellipsoids but were mostly concentrated and associated with melanomacrophage centers (MMCs). The lymphoid Ig+cells in the kidney tended to be dispersed among haematopoietic and granulopoietic cell populations and were in intimate association with the MMCs and blood vessels. This association between MMCs and Ig+cells in the spleen and the kidney, is discussed with respect to the role played by these organs in the immune system of fish. Last, the lymphoid population in the gut associated lymphoid tissue (GALT) of turbot was characterised with respect to staining for Ig. Immunoreactive cells were rarely detected in the epithelial layer although many lymphocytes were present, but they were frequently observed in the lamina propria, presumably as part of the GALT and involved in mucosal immune responses.