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KMID : 0578319960060020161
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Molecules and Cells
1996 Volume.6 No. 2 p.161 ~ p.168
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Characterization of Immunological and Moelcular properties of an Anti-Human 4-1BB Monoclonal Antibody
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Lee, Soo-Hyun
Lee, Na-Gyong/suh, So young/Kim, Joong Gon/Choi, Eung-Chil/Kang, Chang yuil
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Abstract
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4-1BB was first identified to be expressed on activated murine T cells as a 55 kDa homodimer and to belong to the tumor necrosis factor receptor family of integral membrane proteins. The ligand for 4-1BB (4-1BBL) was found on activated macrophages and mature B cells. 4-1BB and 4-1BBL interaction may be important in costimulation of T lymphocyte activation. The gene encoding human 4-1BB (h4-1BB) was recently isolated from a cDNA library of activated human peripheral T cells. In this study, we generated a monoclonal antibody (MAb) against h4-lBB and characterized its immunological and molecular properties. MAbs generated against recombinant h4-lBB protein were assayed by ELISA for affinity to 4-1BB, and one of the clones with the highest affinity was designated 4B4-1-1. Using flow cytometry, we found that 4B4-1-1 had a high affinity to 4-1BB on phytohemagglutinin-activated human CD4^(+) and CD8^(+) T cells. The 4B4-1-1 heavy chain variable region (V_(H)) and light chain variable region (V_(L)) genes were cloned using PCR. The cloned V_(H) gene coded for 118 amino acid residues and the deduced amino acid sequence shared the highest homology with that of rheumatoid factor-binding antibody (A5´CL) (86. 2% identity, 90.5% similarity). The V_(H) segment belonged to the subgroup II (B) and the D and J_(H) segments originated from DFL16.2 and the J_(H3) gene, respectively. The V_(L) gene coded for 107 amino acids and the deduced amino acid sequence was very similar to VK23.32´CL (93.8% identity, 94.8% similarity). The V_(L) segment and J_(K) segment of the 4B4-1-1 V_(L) gene belonged to the subgroup V and MUSJK1 gene, respectively. Since MAb 4B4-1-1 has been shown to have immunosuppressive properties, information and materials obtained in this study will be valuable in development of chimeric and humanized antibodies for the treatment of autoimmune diseases.
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KEYWORD
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