Establishment of apoptosis-inducing monoclonal antibody 2D1 and 2D1-resistant variants of human T cell lines.

A monoclonal antibody (mAb), 2D1(IgM), was identified for its anti-proliferative effect on human T leukemia cell line, SUP-T13. The cells bound with 2D1 showed DNA ladder patterns of oligonucleosomes, demonstrating apoptosis. Peripheral mononuclear cells activated by phytohemagglutinin or OKT3 induced expression of 2D1 antigen and were growth-inhibited by the antibodies. Among the cell lines tested, T cell lines tended to be growth-inhibited by the antibodies. Epstein-Barr virus-transformed B cells were reactive with 2D1, but were not growth-inhibited by the antibodies. We established stable 2D1-resistant variants LAC2D1R and JKT2D1R from the original SUP-T13 and Jurkat T cell lines, respectively. These variant cells demonstrated phenotypes identical to the original cells, including reactivity to 2D1 and expression of cytoplasmic Bc1-2 protein. The 2D1-resistant cells were as sensitive as the original cells to the other apoptosis-inducing stimuli, such as gamma-irradiation or calcium ionophore A23187. However, the 2D1-resistant variants were also insensitive to anti-Fas, another apoptosis-inducing mAb. Binding of 2D1 was blocked by anti-Fas mAb, suggesting that 2D1 reacts with an epitope of human Fas molecules. The present results demonstrate that a 2D1-reactive, but not 2D1-sensitive, population may exist in highly 2D1-sensitive human leukemia T cells and that pairs of 2D1-sensitive and 2D1-resistant cells are useful in the biochemical analysis of Fas-mediated apoptosis in human T cells.