Production of monoclonal antibodies specific to the surface antigens of chicken peripheral blood mononuclear cells |
Jun-Gu Choi1, Haan-Woo Sung1, Sun-Joong Kim2 |
1National Veterinary Research and Quarantine Service 2College of Veterinary Medicine, Seoul National University |
닭의 혈액내 단핵세포 표면항원 특이 단클론성 항체 생산 |
최준구1, 성환우1, 김선중2 |
1국립수의과학검역원 2서울대학교 수의과대학 |
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Abstract |
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 1 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 immunohistochemical 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. |
Key Words:
monoclonal antibody, chicken, peripheral blood mononuclear cell, lymphocyte, monocyte, thrombocyte, macrophage |
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