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Journal of Histochemistry and Cytochemistry, Vol. 49, 139-146, February 2001, Copyright © 2001, The Histochemical Society, Inc.

RAPID COMMUNICATION

In Situ Determination of T-cell Receptor Beta Expression Patterns

Gerard J. Nuovoa, Carl Morrisona, Pierluigi Porcub, Michael A. Caligiurib, and Saul Sustera
a Departments of Pathology, Ohio State University Medical Center, Columbus, Ohio
b Oncology, Ohio State University Medical Center, Columbus, Ohio

Correspondence to: Gerard J. Nuovo, Ohio State Medical Center, Dept. Pathology, Div. Anatomic Pathology, E-411 Doan Hall, 410 W. 10th Ave., Columbus, OH 43210-1228.

A definitive diagnosis of T-cell lymphoma may be contingent on the rearrangement profile of the T-cell receptor. This is most accurately done by molecular analysis of the beta-chain of the T-cell receptor (TCRß) by Southern blotting hybridization that requires unfixed tissue. We describe a reverse transcriptase in situ PCR (RT in situ PCR) method that permits the target-specific direct incorporation of the reporter nucleotide into the different transcripts that comprise the TCRß, using paraffin-embedded, formalin- fixed tissue. Each of the 25 possible Vß segment rearrangments was documented in three lymph nodes with nonspecific lymphadenitis, with clonal expansion evident in a case of metastatic melanoma. Monoclonal expression was documented in seven tissues diagnostic of a T-cell lymphoma. We analyzed five additional tissues for which a definitive diagnosis of T-cell vs B-cell lymphoma could not be rendered on the basis of histological, immunohistological, and flow cytometric analysis. RT in situ PCR for TCRß expression with CD3 co-labeling demonstrated which of these lesions was a B-cell-rich T-cell lymphoma. We conclude that the RT in situ PCR methodology will allow the routine determination of monoclonal vs multiclonal expression patterns of the TCRß using archival paraffin-embedded tissues.

(J Histochem Cytochem 49:139–145, 2001)

Key Words: gene rearrangement, T-cell receptor, lymphoma, in situ PCR


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