Detection of adult T-cell leukemia-associated antigens by direct immunoperoxidase microscopy with Fab'-peroxidase conjugates prepared with a maleimide compound.

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Abstract
Direct immunoperoxidase staining was applied to the detection of adult T-cell leukemia (ATL)-associated antigens (ATLA) in human T-cell leukemia virus (HTLV-I)-producing MT-2 cells. We successfully conjugated Fab'' to peroxidase using a maleimide compound to perform highly sensitive immunohistochemistry with a low background and specific binding. Antisera from three healthy carriers containing antibodies reacting with ATLA were analyzed by Western blotting. ATLA was observed at the light and electron microscopic levels. Positive staining of ATLA was demonstrated in the plasma membrane, endoplasmic reticulum, nuclear membrane, and viral particles, but not in the nuclues nor the mitochondria in frozen-sectioned specimens of MT-2 cells fixed with periodate-lysine-paraformaldehyde (PLP) fixative. Control cells (Molt-4) were not stained. The staining in MT-2 cells and HTLV-I represents the distribution of the viral antigens or their precursors.

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