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

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. [ABSTRACT FROM AUTHOR]