Characterization of Distinct Subpopulations of Japanese Flounder Lymphocytes with Monoclonal Antibody against Serum Immunoglobulin

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  • Tokuda Yuki
    Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University
  • Toyohara Haruhiko
    Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University
  • Kina Tatsuo
    Department of Immunology, Institute for Frontier Medical Sciences. Kyoto University
  • Sakaguchi Morihiko
    Division of Applied Biosciences, Graduate School of Agriculture, Kyoto University

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Monoclonal antibodies (rnAb) FB17 was produced against Japanese flounder Paralichthys olivaceus serum immunoglobulin. Western blotting analysis showed that FB17 rnAb recognized the H chain of Japanese flounder IgM. Furthermore, FB17 mAb was reacted with surface immunoglobulin (slg) expressed on lymphocytes. FB17 mAb was applied to characterize lymphocytes in peripheral blood and thymocytes of Japanese flounder according to the expression of slg. Flow cytometric analysis showed that about 40% of the lymphocyte-enriched fraction were slg-positive and that almost all thymocytes were slg-negative.<br> Peripheral blood leucocytes (PBL) separated according to their adherence to a nylon fiber column were examined as to the expression of slg. As a result, 91% of the adherent cell population expressed slg, while only 3.2% of the non-adherent population did. The slg+ cells found in the non-adherent population were stained more dimly than those in the adherent population. These results suggest that Japanese flounder slg+ cell consists of at least two distinct subpopulations; one which expresses more slg molecules and adheres to nylon fiber and the other which expresses less slg molecules and does not adhere to nylon fiber. These characteristics of Japanese flounder slg+ cells were similar to those of murine B cells.

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