Immunocytochemical detection and cytomorphology of lymphocyte subpopulations in a teleost fish Dicentrarchus labrax

Abstract.

The monoclonal antibodies DLT15 and DLIg3 directed against thymocytes and serum immunoglobulins of the sea bass (Dicentrarchus labrax, L.) were used to study cells from the thymus, head kidney, spleen, gut-associated lymphoid tissue and peripheral blood leukocytes of this fish by immunofluorescence and pre-embedding immunoelectron microscopy. Immunofluorescence and flow cytometry of leukocyte fractions revealed a large number of DLT15-positive cells in the thymus (∼80%) and intestine (∼55%) and fewer cells in the spleen (∼7%), head kidney (∼6%) and peripheral blood (∼3%). DLT15-positive cells had two main morphologies, both detectable among thymocytes: a large round heterochromatic nucleus with light sparse cytoplasm (type a) and an irregular and heterocromatic nucleus with cytoplasm rich in polysomes and mitochondria (type b). Type b was most represented in spleen, head kidney, intestine and blood. We suggest that the type b morphology represents more differentiated lymphocytes. Flow cytometry revealed numerous DLIg3-positive cells in the head kidney (∼33%), spleen (∼30%) and peripheral blood leukocytes (∼21%) and fewer positive cells in the intestine (∼3%) and thymus (∼2%). DLIg3-positive cells had the morphology of lymphocytes (with a large round nucleus) or macrophages in all tissues. Plasma cells lacked membrane immunoreactivity. This is the first ultrastructural characterisation of putative T- and B-lymphocyte subpopulations in a fish species; these subpopulations are differentially distributed in teleost lymphoid organs.