<b>OBJECTIVEb>To investigate the significance of detecting TCR gene clonal rearrangement in the diagnosis of mycosis fungoides (MF) and to optimize the primers used for detecting the TCR gene clonal rearrangement with PCR in paraffin embedded tissues of MF.

<b>METHODSb>Nineteen cases of MF were enrolled into the study. A panel of 10 antibodies were used for immunophenotypic analysis and polymerase chain reaction for TCR-&gamma; and TCR-&beta; gene rearrangement detection in this study.

<b>RESULTSb>TCR gene clonal rearrangements were detected in all 19 cases, in which 84.2% cases (16/19) had TCR-&gamma; gene clonal rearrangements. The positive rates of the primers T(VG)/T(JX), V(2-5)/V(8-12)/JGT(1) and BIOMED-2-TCR-&gamma; were 47.4%, 78.9% and 31.6%, respectively. The positive rate of V(2-5)/V(8-12)/JGT(1) was statistically significantly higher than that of T(VG)/T(JX) and BIOMED-2-TCR-&gamma; (P < 0.05). No TCR gene clonal rearrangement was detected using the primers V(&gamma;11)/V(&gamma;101)/J&gamma;12 and V(&gamma;11)/V(&gamma;101)/J(p12). TCR-&beta; gene clonal rearrangement was detected in 31.6% (6/19) cases.

<b>CONCLUSIONSb>TCR gene clonal rearrangement analysis is a useful tool in the diagnosis of MF and TCR-&gamma; gene is a good target gene for the detection. The primers T(VG)/T(JX), V(2-5)/V(8-12)/JGT(1) and BIOMED-2-TCR-&gamma; can be used in clinicopathologic detection for TCR gene clonal rearrangement and V(2-5)/V(8-12)/JGT(1) may be the first choice.

Adolescent ; Adult ; Aged ; Antigens, CD7 ; metabolism ; Base Sequence ; CD2 Antigens ; metabolism ; CD3 Complex ; metabolism ; CD4 Antigens ; metabolism ; Child ; Child, Preschool ; Female ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Humans ; Leukocyte Common Antigens ; metabolism ; Male ; Middle Aged ; Molecular Sequence Data ; Mycosis Fungoides ; diagnosis ; genetics ; metabolism ; pathology ; Paraffin Embedding ; Receptors, Antigen, T-Cell, alpha-beta ; genetics ; Receptors, Antigen, T-Cell, gamma-delta ; genetics ; Skin Neoplasms ; diagnosis ; genetics ; metabolism ; pathology ; Young Adult

The characterization of the human T-cell receptor (TCR) repertoire has made remarkable progress, with most of the work focusing on the TCRβ chains. Here, we analyzed the diversity and complexity of both the TCRα and TCRβ repertoires of three healthy donors. We found that the diversity of the TCRα repertoire is higher than that of the TCRβ repertoire, whereas the usages of the V and J genes tended to be preferential with similar TRAV and TRAJ patterns in all three donors. The V-J pairings, like the V and J gene usages, were slightly preferential. We also found that the TRDV1 gene rearranges with the majority of TRAJ genes, suggesting that TRDV1 is a shared TRAV/DV gene (TRAV42/DV1). Moreover, we uncovered the presence of tandem TRBD (TRB D gene) usage in ~2% of the productive human TCRβ CDR3 sequences.
Complementarity Determining Regions ; genetics ; DNA Primers ; chemistry ; genetics ; Female ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; genetics ; Gene Rearrangement, delta-Chain T-Cell Antigen Receptor ; genetics ; Genes, T-Cell Receptor beta ; genetics ; Genetic Variation ; High-Throughput Nucleotide Sequencing ; Humans ; Immunoglobulin Joining Region ; genetics ; Immunoglobulin Variable Region ; genetics ; Male ; Receptors, Antigen, T-Cell, alpha-beta ; genetics

<b>OBJECTIVEb>To establish a sensitive and effective method for detection of immunoglobulin and T-cell receptor (Ig/TCR) gene rearrangement,and to explore its role in diagnosis and differential diagnosis of lymphoproliferative disorders.

<b>METHODSb>Fifty-eight lymphoid tissue samples from 54 patients with lymphoproliferations were evaluated by the novel BIOMED-2 multiplex polymerase chain reaction (PCR) for antigen receptor genes rearrangement.

<b>RESULTSb>Multiplex PCR demonstrated monoclonal Ig/TCR gene rearrangements in 22 of 25 (88.0%) B-cell malignancies and 8 of 15 (53.3%) T-cell malignancies. Among 17 benign lymphoproliferations confirmed histopathologically, polyclonal rearrangements were detected in 14 cases (82.4%). In total, the clonality analysis and the final clinico-histopathological diagnosis were concordant in 77.2%. Combination detection of Iglambda and TCR delta gene rearrangements did not increase the detection rate of monoclonal rearrangement of Ig/TCR, but might help to the detection of Iglambda+ or TCR delta+ lymphomas.

<b>CONCLUSIONb>The novel BIOMED-2 multiplex PCR strategy is a rapid, reliable and sensitive approach to detecting clonality in suspected lymphoproliferations, especially in atypical cases.

Female ; Gene Rearrangement, B-Lymphocyte, Light Chain ; Gene Rearrangement, T-Lymphocyte ; Humans ; Lymphoproliferative Disorders ; diagnosis ; genetics ; Male ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity

<b>OBJECTIVEb>To evaluate the efficiency of the BIOMED-2 PCR assay and its implication in the diagnosis of mature B-cell non-Hodgkin's lymphomas.

<b>METHODSb>Clinical, morphological and immunohistochemical features of 72 cases of non-Hodgkin's lymphomas were studied, including 25 reactive lymphoid hyperplasia, 37 diffuse large B cell lymphomas (DLBCL) and 35 extranodal marginal zone lymphomas of mucosa associated lymphoid tissues (MALT lymphoma and in addition, 25 cases of reactive lymphoid hyperplasia were used as the controls). DNA was exacted from the paraffin embedded formalin fixed tissue blocks and the quality of DNA was assessed using the BIOMED-2 specimen control reaction. Adequate samples were then analyzed by BIOMED-2 for immunoglobulin heavy and kappa light chain rearrangements.

<b>RESULTSb>Adequate DNA was obtained in 83 of 97 samples, including 60 mature B cell lymphomas and 23 reactive lymphoid hyperplasia. Clonal B-cell gene rearrangements were detected in 57 of 60 (95%) lymphomas. In contrast, clonal Ig gene rearrangements were not detected in any of the 23 cases of reactive lymphoid hyperplasia.

<b>CONCLUSIONb>BIOMED-2 assay is highly sensitive and specific for the detection of clonal B cell gene rearrangement using routine paraffin embedded formalin fixed specimens.

Antigens, CD20 ; metabolism ; CD79 Antigens ; metabolism ; DNA, Neoplasm ; genetics ; Gene Rearrangement, B-Lymphocyte ; genetics ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; genetics ; Gene Rearrangement, B-Lymphocyte, Light Chain ; genetics ; Genes, Immunoglobulin ; Humans ; Immunophenotyping ; Lymphoma, B-Cell ; genetics ; immunology ; pathology ; Lymphoma, B-Cell, Marginal Zone ; genetics ; immunology ; pathology ; Lymphoma, Large B-Cell, Diffuse ; genetics ; immunology ; pathology ; Paraffin Embedding ; Pseudolymphoma ; genetics ; immunology ; pathology ; Sensitivity and Specificity

Mycosis fungoides (MF), the most common type of cutaneous T-cell lymphoma, has various unspecific clinical and histological characteristics. Its early diagnosis is challenging. The application of T-cell receptor (TCR) gene clonal rearrangement to the diagnosis of MF has been widely studied. In this study, we used polymerase chain reaction (PCR) to investigate the diagnostic significance of detecting TCR-&gamma; and -&beta; gene clonal rearrangement in the early diagnosis of mycosis fungoides. PCR for TCR-&gamma; and TCR-&beta; gene rearrangement was performed on 19 patients with suspected early MF, 6 with typical MF, and 6 with chronic dermatitis. Of the 19 patients with suspected early MF, 13 had TCR-&gamma; gene clonal rearrangement, whereas none had TCR-&beta; gene clonal rearrangement. All patients with typical MF had TCR gene clonal rearrangement, in which 4 showed TCR-&gamma; clonal rearrangement, 1 showed TCR-&beta; gene clonal rearrangements, and 1 showed both. No patients with chronic dermatitis had TCR gene clonal rearrangement. These results indicate that TCR gene clonal rearrangement analysis is a useful tool in diagnosing early MF. TCR-&gamma; gene is recommended to the routine analysis, whereas TCR-&beta; gene has potential in combination toward intractable cases.
Adolescent ; Adult ; Aged ; Base Sequence ; genetics ; DNA, Neoplasm ; genetics ; Early Detection of Cancer ; methods ; Female ; Gene Rearrangement, beta-Chain T-Cell Antigen Receptor ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Humans ; Male ; Middle Aged ; Mycosis Fungoides ; diagnosis ; genetics ; Polymerase Chain Reaction ; Skin Neoplasms ; diagnosis ; genetics ; Young Adult

<b>OBJECTIVEb>To evaluate the practical values of PCR detectable T-cell receptor (TCR) gene rearrangement in paraffin embedded tissue samples in the diagnosis of T-cell malignancies using BIOMED-2 PCR multiplex tubes TCRgamma(A+B).

<b>METHODSb>Traditional phenol-chloroform method was used to extract DNA from 55 cases of archival paraffin embedded tissues samples of T-cell malignancies and the DNA quality was evaluated by PCR-based amplification of housekeeping gene beta-globin. The selected BIOMED-2 PCR multiplex tubes TCRgamma(A+B) were used to detect TCR gene rearrangement and comparison with the results of universal TCR primers (T(VG)/T(JX)) was performed.

<b>RESULTSb>Positive detection rates by the BIOMED-2 multiplex tubes TCRgamma(A+B) and the universal primers (T(VG)/T(JX)) were 76.4% and 60.0%, respectively. There were not statistical difference between the methods (P > 0.05).

<b>CONCLUSIONb>BIOMED-2 multiplex tubes TCRgamma(A+B) is suitable for detection of clonal rearrangements of TCR genes in current archival paraffin embedded tissue samples of T-cell malignancies.

DNA Primers ; DNA, Neoplasm ; analysis ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Humans ; Lymphoma, T-Cell ; genetics ; metabolism ; pathology ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; beta-Globins ; metabolism

<b>OBJECTIVEb>To assess the practical value of BIOMED-2 primers in the diagnosis of ocular adnexal lymphoma by PCR.

<b>METHODSb>DNA was extracted from 63 formalin-fixed paraffin-embedded (FFPE) ocular adnexal lymphoma specimens. The DNA quality was evaluated by PCR-based amplification of housekeeping gene beta-actin. IgH(B) and IgK(B) primers of BIOMED-2 standardized clonality analysis system were used to evaluate the immunoglobin gene rearrangements. PCR products were analyzed using capillary electrophoresis and GeneScan software.

<b>RESULTSb>76.2% (48/63) of FFPE samples produced amplifiable DNA for detection of Ig gene rearrangements.Positive detection rates by BIOMED-2 IgH(B) and IgK(B) primers were 79.2% (38/48) and 68.8% (33/48), respectively, with a combined positive detection rate of 91.7% (44/48).

<b>CONCLUSIONSb>IgH(B) and IgK(B) primers of BIOMED-2 are suitable for the detection of clonal rearrangements of Ig gene using FFPE specimens of ocular adnexal lymphomas.

Actins ; genetics ; Adult ; Aged ; DNA Primers ; Eye Neoplasms ; diagnosis ; genetics ; pathology ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Gene Rearrangement, B-Lymphocyte, Light Chain ; Humans ; Lymphoma, B-Cell ; diagnosis ; genetics ; pathology ; Lymphoma, Follicular ; diagnosis ; genetics ; pathology ; Male ; Middle Aged ; Paraffin Embedding ; Pilot Projects ; Young Adult

<b>OBJECTIVEb>To evaluate the significance of immunoglobulin heavy chain (IgH) gene rearrangement for B-cell lymphoma and T-cell receptor (TCR) gene rearrangement for T-cell lymphoma and NK/T-cell lymphoma in diagnosing and typing of primary non-Hodgkin's lymphoma (NHL) in nasal cavity.

<b>METHODSb>Semi-nested polymerase chain reaction ( PCR) with two pairs of primers was used to detect monoclonal IgH gene rearrangement in paraffin-embedded tissues from 11 patients with B-cell lymphoma, and one-stepped PCR with two pairs of primers was used to detect T-cell receptor gene rearrangement from 23 patients with NK/T-cell lymphoma and 20 patients with T-cell lymphoma. Ten patients with nasal polyp were detected with all the primers by PCR respectively.

<b>RESULTSb>Among the 54 patients with an evaluable PCR results, 10 of 11 (90. 9% ) B-cell lymphomas were positive for monoclonal IgH gene rearrangement, 17 of 20 (85. 0% ) T-cell lymphomas and 10 of 23 (43. 5% ) NK/T-cell lymphomas were positive for monoclonal TCR gene rearrangement. Ten patients with nasal polyp were negative for all detection.

<b>CONCLUSIONSb>Detecting gene rearrangement was an efficient method in auxiliary diagnosing and typing of primary NHL in nasal cavity; Using semi-nested PCR or one-stepped PCR with two pairs of primers can enhance the positive rate of gene rearrangement detection.

Adolescent ; Adult ; Aged ; Child ; Female ; Gene Rearrangement, B-Lymphocyte, Heavy Chain ; Gene Rearrangement, T-Lymphocyte ; Humans ; Lymphoma, Non-Hodgkin ; diagnosis ; pathology ; Male ; Middle Aged ; Nasal Cavity ; Nose Neoplasms ; diagnosis ; pathology ; Young Adult

Diagnosis, Differential ; Gene Rearrangement, B-Lymphocyte ; Gene Rearrangement, T-Lymphocyte ; Humans ; Lymphoma ; diagnosis ; genetics ; Lymphoma, Follicular ; diagnosis ; genetics ; Lymphoproliferative Disorders ; diagnosis ; genetics ; Polymerase Chain Reaction ; methods

No abstract available.
Antibodies, Monoclonal, Murine-Derived/therapeutic use ; Antineoplastic Combined Chemotherapy Protocols/therapeutic use ; B-Cell-Specific Activator Protein/metabolism ; Bone Marrow/metabolism/*pathology ; Cyclophosphamide/therapeutic use ; Doxorubicin/therapeutic use ; Endoscopy, Digestive System ; Female ; Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor ; Genetic Loci ; Humans ; Liver/metabolism/pathology ; Lymphocytes/cytology/immunology ; Lymphoma, Large B-Cell, Diffuse/complications/*diagnosis/drug therapy ; Lymphoma, T-Cell, Peripheral/complications/*diagnosis/drug therapy ; Middle Aged ; Prednisone/therapeutic use ; Receptors, Antigen, T-Cell, gamma-delta/genetics ; Tomography, X-Ray Computed ; Vincristine/therapeutic use