DETECTION OF HTLV-1 BY POLYMERASE-CHAIN-REACTION IN-SITU HYBRIDIZATION IN ADULT T-CELL LEUKEMIA LYMPHOMA/

A method for nonradioactive polymerase chain reaction in situ hybridiz ation mas developed acid used to determine the distribution of human T -lymphotropic virus type I (HTLV-I) proviral DNA in paraffin-embedded surgical specimens of adult T-cell leukemia/lymphoma (ATLL). As contro ls, me used biopsy samples of five cases of mycosis fungoides, cells o f an HTLV-I-infected cell line (MT2), as well as HTLV-1-negative cells (YAS). We successfully detected the amplicon of the HTLV-1 tax sequen ce in the nuclei of the cutaneous infiltrating lymphoid cells in 90% ( 9/10) of ATLL cases. Studies also revealed the existence of HTLV-1 pro virus DNA in nuclei of sweat gland epithelial cells and vascular endot helial cells as well as lymphoid cells in ATLL patients. Mycosis fungo ides and YAS cells were negative for the HTLV-I tax sequence, but MT2 cells mere strongly positive. The results indicated that this techniqu e mas more sensitive in detecting intracellular amplicons than was the previous in situ hybridization method. Through its use, me were able to easily determine the distribution of HTLV-I-positive cells among th e various cells and tissues of paraffin-embedded archival materials.