Production of monoclonal antibodies specific to the surface antigens of chicken peripheral blood mononuclear cells

Choi, Jun-Gu; Sung, Haan-Woo; Kim, Sun-Joong

Korean Journal of Veterinary Research 2002;42(2):209-217.

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

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