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This study was performed to produce monoclonal antibodies (mAb) specifically reacting with chicken leukocyte surface antigens. Popliteal lymph node cells of BALB/c mice previously immunized through foot-pad with peripheral blood mononuclear cells (PBMC) of chickens separated by Ficoll-Histopaque method. They were fused with P3X63Ag14 mouse myeloma cells. A total of 34 hybridomas secreted antibodies specifically binding to the PBMC. According to the reactivity patterns with PBMC, the mAbs were divided into 4 groups. Group Ⅰ mAbs (ⅡB3, ⅡB10, ⅡE10) specifically reacted with non-adherent lymphocytes but not with adherent cells which were mainly composed of thrombocytes and monocytes in PBMC culture. These mAbs were reactive with 25-59% of thymus cells and 42-64% of spleen cells of chickens. They did not show any significant reactivity with cells in the bursa of Fabricius, T-cell (MDCC-MSB1) and B-cell (LSCC-1104B1) lines. These results indicate that Group Ⅰ mAbs specifically reacted with T-lymphocyte subpopulation. Monoclonal antibodies in Group Ⅱ (ⅠC6, ⅠG2-2 and ⅡD9) showed specific reactivity with monocytes but not with thrombocytes or non-adherent cells in PBMC culture. These mAbs, though not reacted with the chicken macrophage cell line, HD11, also bound to macrophages of the spleen and lung in immnohistochemical staining. Five mAbs in Group Ⅲ showed characteristics of binding to lymphocytes and monocytes, but not to thrombocytes. Twenty-three mAbs in Group Ⅳ showed specific reactivity to lymphocytes, monocytes, and thrombocytes. Two mAbs (ⅠC3 and ⅠE9) in Group Ⅳ reacted with most of PBMC.
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