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Heteroduplex analysis of T-cell receptor gamma gene rearrangements for
diagnosis and monitoring of cutaneous T-cell lymphomas
M Bottaro, E Berti, A Biondi, N Migone and L Crosti
Centro CNR Immunogenetica e Istocompatibilita, Universita di Torino, Italy.
The possibility to detect markers of T-cell clonality at the T-cell
receptor (TCR) beta and gamma loci in skin biopsy samples has proven to be
helpful for the often difficult clinical and immunohistochemical diagnosis
of cutaneous T-cell lymphoma (CTCL). However, particularly at the early
stage of the neoplastic infiltration, an emerging clonal pattern at
Southern may be obscured by the germline TCR configuration of the
predominant dermal and epidermal cell component. Additionally, multiple TCR
gamma rearranged bands of variable intensity are often observed, either in
the presence or in the absence of a major clone. To overcome these
difficulties, we have investigated the T-lymphocyte clonality in selected
patients with variable signs of CTCL by means of heteroduplex analysis of
the amplified TCR gamma VJ junctions, separated in nondenaturing
polyacrylamide gel. This technique has several advantages over standard
Southern blot because it is simple, rapid, not radioactive, and likely more
sensitive than other polymerase chain reaction-based procedures. In
particular, the cases with uncertain or contradictory TCR beta and gamma
patterns were solved by the heteroduplex analysis, showing homoduplex or
heteroduplex bands of clonal nature. The direct sequence of the VJ
junctions, easily obtained from the homoduplex or heteroduplex bands,
allowed us to confirm the same clonal marker in two apparently different
skin lesions and in different biopsy samples obtained from the same
patients, either at the same or different time points, thus emphasizing the
utility of this method in monitoring CTCL clinical progression.
Volume 83,
Issue 11,
pp. 3271-3278,
06/01/1994
Copyright © 1994 by The American Society of Hematology
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