This Article Order Full text via Infotrieve Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Greiner, T. C. Articles by Jaffe, E. S. Search for Related Content PubMed PubMed Citation Articles by Greiner, T. C. Articles by Jaffe, E. S.

American Journal of Pathology, Vol 146, 46-55, Copyright © 1995 by American Society for Investigative Pathology

REGULAR ARTICLES

Analysis of T cell receptor-gamma gene rearrangements by denaturing gradient gel electrophoresis of GC-clamped polymerase chain reaction products. Correlation with tumor-specific sequences

TC Greiner, M Raffeld, C Lutz, F Dick and ES Jaffe
Hematopathology Section, National Cancer Institute, Bethesda, Maryland.

We describe a modified denaturing gradient gel electrophoresis (DGGE) procedure with a 40-nucleotide GC clamp in the polymerase chain reaction to improve resolution in amplifying T cell receptor-gamma (TCR- gamma) rearrangements. DNA from 46 cases of lymphoblastic leukemia/lymphoma, 5T cell lines, 2 B cell lines, 7 normal lymphocytes, and 3 cases of Hodgkin's disease was amplified by polymerase chain reaction. In addition, 20 cases of paraffin-embedded T cell lymphomas and 5 cases of reactive hyperplasia were also studied. Clonal TCR-gamma rearrangements were identified on DGGE by the presence of a predominant band. Results obtained from 5 T cell lines and 12 lymphoblastic leukemia/lymphomas containing known TCR-gamma gene rearrangements revealed 100% concordance in detecting clonal rearrangements between DGGE and traditional Southern blot analysis. Of the remaining 34 lymphoblastic leukemia/lymphoma cases studied by DGGE alone, 30 were positive. DGGE analysis of 10 lymphoblastic leukemia/lymphoma cases with known group IV gamma to J gamma 1 or J gamma 2 rearrangement sequences confirmed that the electrophoretic migration was dependent on the tumor-specific rearranged TCR-gamma sequence. In addition, 17 of 20 cases of paraffin-embedded T cell lymphomas were positive by DGGE, 6 of which had the clonal population also identified in fresh tissue DNA. DGGE analysis of GC-clamped polymerase chain reaction products can provide a way to more accurately detect TCR-gamma clonality of lymphoid tumors and can be applied to archival tissues.

This article has been cited by other articles:

				O. A. Gra, J. V. Sidorova, E. A. Nikitin, A. Y. Turygin, S. A. Surzhikov, A. L. Melikyan, A. B. Sudarikov, A. S. Zasedatelev, and T. V. Nasedkina Analysis of T-Cell Receptor-{gamma} Gene Rearrangements Using Oligonucleotide Microchip: A Novel Approach for the Determination of T-Cell Clonality J. Mol. Diagn., April 1, 2007; 9(2): 249 - 257. [Abstract] [Full Text] [PDF]

				F. C. Kuo, D. Hall, and J. A. Longtine A Novel Method for Interpretation of T-Cell Receptor {gamma} Gene Rearrangement Assay by Capillary Gel Electrophoresis Based on Normal Distribution J. Mol. Diagn., February 1, 2007; 9(1): 12 - 19. [Abstract] [Full Text] [PDF]

				M Christensen, A D Funder, K Bendix, and F B Soerensen Comparative investigations of T cell receptor {gamma} gene rearrangements in frozen and formalin-fixed paraffin wax-embedded tissues by capillary electrophoresis J. Clin. Pathol., June 1, 2006; 59(6): 645 - 654. [Abstract] [Full Text] [PDF]

				A. Molina, J. Zain, D. A. Arber, M. Angelopolou, M. O'Donnell, J. Murata-Collins, S. J. Forman, and A. Nademanee Durable Clinical, Cytogenetic, and Molecular Remissions After Allogeneic Hematopoietic Cell Transplantation for Refractory Sezary Syndrome and Mycosis Fungoides J. Clin. Oncol., September 1, 2005; 23(25): 6163 - 6171. [Abstract] [Full Text] [PDF]

				X. Y. Yang, D. Xu, J. Du, H. Kamino, J. Rakeman, and H. Ratech Rapid Detection of Clonal T-Cell Receptor-{beta} Gene Rearrangements in T-Cell Lymphomas Using the LightCycler-Polymerase Chain Reaction with DNA Melting Curve Analysis J. Mol. Diagn., February 1, 2005; 7(1): 81 - 88. [Abstract] [Full Text] [PDF]

				J. Willers, R. Dummer, W. Kempf, T. Kundig, G. Burg, and M. E. Kadin Proliferation of CD30+ T-Helper 2 Lymphoma Cells Can Be Inhibited by CD30 Receptor Cross-Linking with Recombinant CD30 Ligand Clin. Cancer Res., July 1, 2003; 9(7): 2744 - 2754. [Abstract] [Full Text] [PDF]

				L. C. Lawnicki, R. J. Rubocki, W. C. Chan, D. M. Lytle, and T. C. Greiner The Distribution of Gene Segments in T-Cell Receptor {gamma} Gene Rearrangements Demonstrates the Need for Multiple Primer Sets J. Mol. Diagn., May 1, 2003; 5(2): 82 - 87. [Abstract] [Full Text] [PDF]

				T. C. Greiner and R. J. Rubocki Effectiveness of Capillary Electrophoresis Using Fluorescent-Labeled Primers in Detecting T-Cell Receptor {gamma} Gene Rearrangements J. Mol. Diagn., August 1, 2002; 4(3): 137 - 143. [Abstract] [Full Text] [PDF]

				K. KUROSU, N. YUMOTO, W. N. ROM, Y. TAKIGUCHI, J. JAISHREE, K. NAKATA, K. TATSUMI, A. MIKATA, T. KURIYAMA, and M. D. WEIDEN Oligoclonal T Cell Expansions in Pulmonary Lymphoproliferative Disorders . Demonstration of the Frequent Occurrence of Oligoclonal T Cells in Human Immunodeficiency Virus-Related Lymphoid Interstitial Pneumonia Am. J. Respir. Crit. Care Med., January 15, 2002; 165(2): 254 - 259. [Abstract] [Full Text] [PDF]

				V. S. Meier, A. Rufle, and F. Gudat Simultaneous Evaluation of T- and B-Cell Clonality, t(11;14) and t(14;18), in a Single Reaction by a Four-Color Multiplex Polymerase Chain Reaction Assay and Automated High-Resolution Fragment Analysis : A Method for the Rapid Molecular Diagnosis of Lymphoproliferative Disorders Applicable to Fresh Frozen and Formalin-Fixed, Paraffin-Embedded Tissues, Blood, and Bone Marrow Aspirates Am. J. Pathol., December 1, 2001; 159(6): 2031 - 2043. [Abstract] [Full Text] [PDF]

				D. A. Arber, R. M. Braziel, A. Bagg, and K. E. Bijwaard Evaluation of T Cell Receptor Testing in Lymphoid Neoplasms: Results of a Multicenter Study of 29 Extracted DNA and Paraffin-Embedded Samples J. Mol. Diagn., November 1, 2001; 3(4): 133 - 140. [Abstract] [Full Text] [PDF]

				D. A. Arber Molecular Diagnostic Approach to Non-Hodgkin's Lymphoma J. Mol. Diagn., November 1, 2000; 2(4): 178 - 190. [Full Text]

				C. Assaf, M. Hummel, E. Dippel, S. Goerdt, H.-H. Muller, I. Anagnostopoulos, C. E. Orfanos, and H. Stein High detection rate of T-cell receptor beta chain rearrangements in T-cell lymphoproliferations by family specific polymerase chain reaction in combination with the GeneScan technique and DNA sequencing Blood, July 15, 2000; 96(2): 640 - 646. [Abstract] [Full Text] [PDF]

				J. Z. Gong, S. Zheng, R. Chiarle, C. De Wolf-Peeters, G. Palestro, G. Frizzera, and G. Inghirami Detection of Immunoglobulin {kappa} Light Chain Rearrangements by Polymerase Chain Reaction : An Improved Method for Detecting Clonal B-CellLymphoproliferative Disorders Am. J. Pathol., August 1, 1999; 155(2): 355 - 363. [Abstract] [Full Text] [PDF]

				T. C. Greiner Advances in Molecular Hematopathology : T-Cell Receptor {gamma} and bcl-2 Genes Am. J. Pathol., January 1, 1999; 154(1): 7 - 9. [Full Text] [PDF]

				S. Signoretti, M. Murphy, M. G. Cangi, P. Puddu, M. E. Kadin, and M. Loda Detection of Clonal T-Cell Receptor {gamma} Gene Rearrangements in Paraffin-Embedded Tissue by Polymerase Chain Reaction and Nonradioactive Single-Strand Conformational Polymorphism Analysis Am. J. Pathol., January 1, 1999; 154(1): 67 - 75. [Abstract] [Full Text] [PDF]