Detection of T Cell Clonality at Diagnosis, in Small Samples and Monitoring of Minimal Residual Disaease (MRD) Using TCR-V Beta Repertoire Analysis.
Prashant R Tembhare, Constance M Yuan, Liqiang Xi, John Janik, John Morris, Mark Raffeld, Maryalice Stetler-Stevenson. Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD; Center for Cancer Research, National Cancer Institute, Mark O. Hatfield Clinical Research Center, National Institutes of Health, Bethesda, MD
Background: Flow cytometric (FC) TCR-Vβ repertoire analysis (TCR-Vβ-R) is a sensitive method for detection of T-cell clonality. However, it is rarely being used for evaluation of body fluids or soft tissue invlovment and MRD estimation in T cell lymphoma. We evaluated TCR-Vβ-R in diagnosis of T-cell neoplasia and MRD monitoring. In addition TCR-Vβ-R analysis was applied to samples with small cell numbers, such as fine needle aspirates and cerebrospinal fluid.
Design: Comprehensive FC panel with complete TCR-Vβ-R was used for initial evaluation and abbreviated panel with single clone-specific V-beta cocktail was used in low cellularity specimens and MRD. Diagnostic testing for mature T-cell neoplasia was performed on 46 patients (27 HTLV-I-associated adult T-cell leukemia/lymphoma, 7 cutaneous T-cell lymphoma, 7 large granular lymphocytic leukemia, 4 peripheral T-cell lymphoma-not otherwise specified, 1 T-cell prolymphocytic leukemia). TCR-Vβ-R was applied to 12 FNA and 8 CSF containing low cell number. MRD was monitored in 72 sequential samples (59 PB, 7 FNA, 4 CSF and 2 BM) from 16 patients.
Results: TCR-Vβ-R demonstrated T-cell neoplasia in 46 initial specimens (42 peripheral blood (PB), 3 Bone marrow (BM), 1 fine needle aspirate (FNA)). T cell clonality was confirmed by PCR. MRD was successfully detected in follow up samples from all 16 patients evaluated, including 12 FNA and 8 CSF specimens. Furthermore, minimal residual disease post therapy was accurately quantitated in the 59 PB specimens.This method was highly sensitive in documenting treatment response, allowing detection of MRD as low as 0.8 clonal T cells/ul.
Conclusions: TCR-Vβ-R analysis, a sensitive method for detection of T-cell clonality, is useful for diagnosis and MRD detection in multiple specimen types, including those with low cellularity such as FNA and CSF.
Monday, February 28, 2011 9:30 AM
Poster Session I Stowell-Orbison/Surgical Pathology/Autopsy Awards Poster Session # 192, Monday Morning