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1.
Chinese Journal of Pathology ; (12): 234-238, 2013.
Article in Chinese | WPRIM | ID: wpr-256208

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinicopathologic features and differential diagnosis of splenic B-cell marginal zone lymphoma (SMZL) involving bone marrow.</p><p><b>METHODS</b>The clinical and pathologic features of 22 patients with SMZL were retrospectively studied. Immunophenotypic analysis was carried out by flow cytometry and immunohistochemistry. Immunoglobulin heavy chain rearrangement study was performed using polymerase chain reaction-based method.</p><p><b>RESULTS</b>Villous lymphocytes were found in peripheral blood smears of 11/18 of the patients. In bone marrow aspirates, lymphocytosis (> 20%) was demonstrated in 15 cases (15/18) and villous lymphocytes in 6 cases (6/18). Flow cytometry showed CD19(+) CD20(+) FMC7(+) CD22(+) CD10(-) CD2(-) CD3(-) CD7(-) in 18 cases. Bone marrow biopsies of all the 22 patients revealed various degrees and patterns of neoplastic infiltration, as follows: mild (4 cases, 18.2%), moderate (11 cases, 50.0%) or severe (7 cases, 31.8%); intrasinusoidal (16 cases, 72.7%), interstitial (14 cases, 63.6%), nodular (11 cases, 50.0%) or diffuse (1 case, 4.5%). Reactive germinal center formation (CD23(+) bcl-2(-)) was found in 2 cases (91.0%). Immunohistochemical study showed the following results: CD20(+) PAX5(+) CD3(-) CD5(-) CD10(-) cyclin D1(-) CD23(-) CD43(-) Annexin A1(-) CD11C(-) CD25(-) in all the 22 cases, CD38(+) in 2 cases (9.1%) and CD138(+) in 2 cases (9.1%).</p><p><b>CONCLUSIONS</b>Different and overlapping patterns of bone marrow involvement are observed in SMZL. As the histologic and immunophenotypic features are not specific to SMZL, distinction from other types of mature B-cell lymphomas is necessary.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Antigens, CD20 , Metabolism , Bone Marrow , Pathology , Diagnosis, Differential , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Leukemia, Lymphocytic, Chronic, B-Cell , Metabolism , Pathology , Lymphoma, B-Cell, Marginal Zone , Genetics , Metabolism , Pathology , Lymphoma, Follicular , Metabolism , Pathology , Lymphoma, Mantle-Cell , Metabolism , Pathology , Neoplasm Invasiveness , Retrospective Studies , Splenic Neoplasms , Genetics , Metabolism , Pathology , Waldenstrom Macroglobulinemia , Metabolism , Pathology
2.
Chinese Journal of Pathology ; (12): 234-238, 2012.
Article in Chinese | WPRIM | ID: wpr-241945

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinicopathologic features of primary thymic extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue (MALT).</p><p><b>METHODS</b>The clinical and pathologic findings were evaluated in 3 cases of biopsy confirmed thymic MALT lymphoma. The clincopathologic features, treatment and prognosis were discussed and literatures reviewed.</p><p><b>RESULTS</b>One male and two female patients presented with asymptomatic mediastinal masses with a history of Sjögren syndrome. They were aged 36, 35 and 41 years respectively, and only one patient had B symptoms. Grossly, all three tumors were encapsulated and had multiple variable-sized cysts on cut-surface. Histopathologically, the normal thymic lobular architecture was effaced by abnormal dense lymphoid infiltration. Prominent lymphoepithelial lesions were formed by centrocyte-like cells infiltrating and expanding Hassall's corpuscles and epithelial cyst lining. All cases showed apparent plasmacytic differentiation. Immunohistochemically, the tumor cells were positive for CD20, CD79a, bcl-2 and negative for CD3, CD5, cyclin D1, CD43, CD10, bcl-6, and CD23. The plasma cells showed kappa light chain restriction. Immunoglobulin heavy chain rearrangement in three cases was confirmed by PCR. All patients were at early stage and received routine chemotherapy with or without radiotherapy after surgical removal. All patients achieved complete remission with 24, 18 and 3 months follow-up, respectively.</p><p><b>CONCLUSIONS</b>Primary thymic MALT lymphoma may be a rare distinctive lymphoma. It can be diagnosed by HE and immunohistochemical study and should be differentiated from reactive lymphoid proliferation, other types of lymphoma and mediastinal thymoma.</p>


Subject(s)
Adult , Female , Humans , Male , Antibodies, Monoclonal, Murine-Derived , Therapeutic Uses , Antigens, CD20 , Metabolism , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Cyclophosphamide , Therapeutic Uses , Diagnosis, Differential , Doxorubicin , Therapeutic Uses , Follow-Up Studies , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Immunoglobulin Heavy Chains , Genetics , Keratin-19 , Metabolism , Lymphoma, B-Cell, Marginal Zone , Drug Therapy , Genetics , Metabolism , Pathology , General Surgery , Prednisone , Therapeutic Uses , Pseudolymphoma , Pathology , Thymus Hyperplasia , Pathology , Thymus Neoplasms , Drug Therapy , Genetics , Metabolism , Pathology , General Surgery , Vincristine , Therapeutic Uses
3.
Chinese Journal of Pathology ; (12): 460-464, 2011.
Article in Chinese | WPRIM | ID: wpr-261753

ABSTRACT

<p><b>OBJECTIVE</b>To study the immunophenotype and gene rearrangement pattern of pulmonary lymphomatoid granulomatosis.</p><p><b>METHODS</b>Nine cases of pulmonary lymphomatoid granulomatosis, included 5 cases of open lung biopsy, 3 cases of lobectomy specimen and 1 case of autopsy, were retrospectively analyzed by immunohistochemistry, in-situ hybridization for Epstein-Barr virus-encoded RNA, immunoglobulin and T-cell receptor gene rearrangement studies.</p><p><b>RESULTS</b>The age of patients ranged from 3 to 59 years. The male-to-female ratio was 3: 6. Histologically, all cases showed lymphocytic infiltration surrounding the blood vessels and in the perivascular areas. Most of these lymphoid cells expressed T-cell marker CD3. There were also variable numbers of CD20-positive B cells. The staining for CD56 was negative. According to the WHO classification, there were 4 cases of grade I , 1 case of grade II and 4 cases of grade III lesions. Six cases had gene rearrangement studies performed and 3 of them demonstrated clonal immunoglobulin gene rearrangement (including 1 of the grade II and 2 of the grade III lesions). No T-cell receptor gene rearrangement was detected.</p><p><b>CONCLUSIONS</b>Pulmonary lymphomatoid granulomatosis may represent a heterogeneous group of lymphoproliferative disorders. Some of the cases show B-cell immunophenotype and clonal immunoglobulin gene rearrangement, especially the grade II and grade lesions. They are likely of lymphomatous nature.</p>


Subject(s)
Adult , Child , Child, Preschool , Female , Humans , Male , Middle Aged , Young Adult , Antigens, CD20 , Metabolism , CD3 Complex , Metabolism , Follow-Up Studies , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Immunohistochemistry , Lung Neoplasms , Genetics , Metabolism , Pathology , General Surgery , Lymphomatoid Granulomatosis , Genetics , Metabolism , Pathology , General Surgery , Neoplasm Grading , Pneumonectomy , Methods , Retrospective Studies
4.
Journal of Experimental Hematology ; (6): 379-384, 2011.
Article in Chinese | WPRIM | ID: wpr-244918

ABSTRACT

This study was purposed to investigate the BCL-2/IgH gene rearrangement in major break point region (MBR) and IgH gene rearrangements of patients with non-Hodgkin's lymphoma (NHL), and explore their significance for improving early diagnosis and accurately evaluating chemotherapy effect. DNA for BCL-2/IgH and IgH gene assays was extracted from bone marrow mononuclear cells in 70 cases of lymphoma (60 cases of B-NHL and 10 cases of T-NHL), 7 cases of lymph node inflammatory and 20 healthy controls. The BCL-2/IgH, IgH gene rearrangements were assayed by polymerase chain reaction (PCR), the assayed results were compared with results of pathological biopsy; the factors related with occurrence of these 2 kinds of gene rearrangement were analyzed and the dynamic changes of BCL-2/IgH and IgH gene rearrangements after chemotherapy were compared, the chemotherapy effect was evaluated. The results indicated that (1) BCL-2/IgH gene rearrangement in bone marrow mononuclear cells was observed in 10 cases out of 30 DLBCL cases (33.3%), and was more frequent than that in 30 other B-NHL cases (6.7%), 10 T-NHL cases (0%), 7 lymph nodes inflammatory cases (0%) and 20 healthy controls (5%) (p < 0.05). (2) the quantity of rearranged BCL-2/IgH gene of 8 DLBCL cases reduced from 0.59 to 0.16 (p < 0.05) after 2 courses of R-CHOP chemotherapy and completely disappeared after 6 courses of R-CHOP chemotherapy. (3) 81.8% patients with BCL-2/IgH gene rearrangement showed high serum LDH level, while it was observed in 28.6% patients without this gene rearrangement (p < 0.05). Lymphoma staging, systemic symptoms, β(2)-MG level, bone marrow involvement, infiltration of liver and spleen were not significantly correlated with BCL-2/IgH gene rearrangement. (4) IgH gene rearrangement was found in 9 cases out of 20 DLBCL patients (all newly diagnosed patients) (45%), IgH rearrangement was observed in 14 cases out of 30 other B-NHL (all newly diagnosed or relapsed patients, except patients with DLBCL) (46.7%) and there was no statistical difference between these 2 groups, however IgH rearrangement all were not observed in 20 healthy persons, 10 T-NHL cases and 7 lymph nodes inflammatory cases. (5) the quantity of rearranged IgH gene in 7 DLBCL cases was reduced from 0.42 to 0.13 after one course of R-CHOP chemotherapy (p < 0.05) and completely disappeared after 2 courses of R-CHOP chemotherapy. (6) 90% patients with IgH gene rearrangement had high serum LDH level, while it was found in 30% patients without this gene rearrangement (p < 0.05). Lymphoma staging, systemic symptoms, β(2)-MG levels, bone marrow involvement, infiltrations liver and spleen all were not significantly correlated with IgH gene rearrangement. It is concluded that the BCL-2/IgH and IgH gene rearrangements may be used as specific indicators in early diagnosis and accurate evaluation of therapy efficacy in B-NHL, these 2 kind of rearrangement correlate with LDH level. The BCL-2/IgH gene rearrangement is more specific for in DLBCL.


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , Young Adult , Bone Marrow Cells , Case-Control Studies , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Genetics , Lymphoma, Non-Hodgkin , Blood , Genetics
5.
Chinese Journal of Hematology ; (12): 529-532, 2011.
Article in Chinese | WPRIM | ID: wpr-251495

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the overrepresentation of specific gene segments of immunoglobulin heavy chain variable region (IgVH) among unmutated and mutated chronic lymphocytic leukemia (CLL) patients and its prognostic implication.</p><p><b>METHODS</b>Multiplex PCR was used to identify the expression of IgVH segment and its mutation status in CLL.</p><p><b>RESULTS</b>Analyses were successfully performed in 80 of 85 samples. Marked skewed IgVH families were disclosed. The most commonly used VH was VH3 (40.0%), followed by VH4 (30.0%), VHI (13.8%), VH2 (10.0%) and VH5, VH7 (2.5%). Fifty-six patients (70.0%) had mutated VH, 24 (30.0%) unmutated VH. Nine cases (11.3%) were with 100% germline sequence. Fifteen cases (15/24, 62.5%) in VH4, 29 (29/32, 90.7%) in VH3, and 4 (4/11, 36.3%) in VH1 had mutated VH. The most frequently used IgVH gene was VH4-39 (13.8%), and VH4-34 (8.8%). J4 (36/66, 54.5%) and D3 (25/66, 37.8%) were the most frequently used in J and D genes. The progression-free survival (PFS) was 82 and 17 months (P = 0.000), and the overall survival (OS) was 90 and 41 months (P = 0.009), respectively, for mutated and unmutated cases. Recurrent CDR3 sequences were found in our patients and 2 patients with VH1-69 had CDR3 sequences highly similar to those reported in literature.</p><p><b>CONCLUSION</b>There is difference in IgVH gene segment usage and mutational status in different area CLL patients. Recurrent CDR3 sequences were found in specific IgVH gene segments, which highlights the importance of immunoglobulin mediated stimulation in the development of CLL.</p>


Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Middle Aged , DNA Mutational Analysis , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Genes, Immunoglobulin , Immunoglobulin Heavy Chains , Genetics , Immunoglobulin Variable Region , Genetics , Leukemia, Lymphocytic, Chronic, B-Cell , Genetics , Allergy and Immunology , Pathology , Mutation
6.
Chinese Journal of Pathology ; (12): 296-301, 2010.
Article in Chinese | WPRIM | ID: wpr-333279

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the ancillary diagnostic value of IgH gene rearrangements in those B-cell lymphoproliferative disorder cases whom are difficult in making a final diagnosis.</p><p><b>METHODS</b>IgH gene clonal rearrangements were retrospectively analyzed in a total of 77 diagnostically difficult B-cell lympho-proliferative patients. Standardized BIOMED-2 system IgH gene clonality assay kit targeting FR1, FR2, FR3 was used, followed by heteroduplex-polyacrylamide gel electrophoresis (PAGE) and silver nitrate staining.</p><p><b>RESULTS</b>The final diagnoses of the 77 cases were: 12 cases of reactive lymphoid hyperplasia, 20 cases of atypical lymphoid hyperplasia or suspicious lymphoma, and 45 cases of B-cell lymphoma. Detection rates of at least one positive reaction were 2/12, 11/20 (55%), 36/45 (80%) in the three groups, respectively. In B-cell lymphomas, the clonality detection rate of FR1, FR2 and FR3 was 60% (27/45), 60% (27/45) and 56% (25/45), respectively. The type distribution were: 20 marginal zone lymphomas, including 18 extranodal marginal zone B-cell lymphoma of mucosa-associated lymphoid tissue, 7 diffuse large B-cell lymphomas, 7 follicular lymphomas, 1 mantle-cell lymphoma, 1 Burkitt's lymphoma, 4 plasma cell neoplasms and 5 unclassified B-cell lymphomas. Rearrangements of FR1, FR2 or FR3 were not detected in 9 (20%) of the B cell lymphoma cases, nevertheless, one of them had developed liver lesion later, and was confirmed finally to be B cell lymphoma. Fourteen patients of reactive lymphoid hyperplasia with positive IgH gene clonal rearrangements, and atypical lymphoid hyperplasia had follow-up history available. Four of them were diagnosed as lymphoid malignancies upon further biopsy, and in three of them, clonal IgH gene rearrangements were detected.</p><p><b>CONCLUSIONS</b>B-cell lymphoproliferative disorder requiring a detection of clonal IgH gene rearrangement for making a final diagnosis. Combined detections of three IgH FR1, FR2 and FR3 rearrangements provide important ancillary diagnostic value in confirming suspected B-cell lympho-proliferative disorders. It is important to take an additional biopsy or to follow-up those patients who that have a detectable IgH gene clonal rearrangement but without apparent morphological evidence of lymphoma. For cases with a negative IgH gene rearrangements, it might be necessary to perform clonality analysis for other forms of gene rearrangements including IgH or IgK and IgL in order to further improve the detection sensitivity.</p>


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Young Adult , Follow-Up Studies , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Lymphoma, B-Cell , Diagnosis , Genetics , Pathology , Lymphoma, B-Cell, Marginal Zone , Diagnosis , Genetics , Pathology , Lymphoma, Follicular , Diagnosis , Genetics , Pathology , Lymphoma, Large B-Cell, Diffuse , Diagnosis , Genetics , Pathology , Lymphoproliferative Disorders , Diagnosis , Genetics , Pathology , Neoplasms, Plasma Cell , Diagnosis , Genetics , Pathology , Pseudolymphoma , Diagnosis , Genetics , Pathology , Retrospective Studies
7.
Chinese Journal of Pathology ; (12): 819-824, 2010.
Article in Chinese | WPRIM | ID: wpr-295134

ABSTRACT

<p><b>OBJECTIVE</b>To investigate the molecular genetic features and diagnostic aspects of sporadic Burkitt's lymphoma (BL) in children.</p><p><b>METHODS</b>Tissue microarray was constructed to include 64 cases of pediatric BL and 6 cases of pediatric diffuse large B-cell lymphoma (DLBCL). Immunohistochemistry and fluorescence in-situ hybridization for c-myc, bcl-2, bcl-6, IgH, myc/IgH and bcl-2/IgH gene were performed. Cases of pediatric Burkitt's lymphomas were subclassified into three groups based on their cellular orgins: the germinal center (GC) group, the late-germinal center (late-GC) group and the post-germinal center (post-GC) group.</p><p><b>RESULTS</b>Among 64 Burkitt's lymphomas studied, expression of CD20, CD10, bcl-6, bcl-2 and MUM1 by immunohistochemistry were 100% (64 cases), 98.4% (63 cases), 96.9% (62 cases), 0 (0 cases) and 23.4% (15 cases), respectively. Various gene rearrangements were found involving the c-myc 93.1% (54/58 cases) and IgH 82.8% (48/58 cases). Detailed rearrangements are as follows: 46 cases (85.2%) myc/IgH gene translocation along with c-myc and IgH gene rearrangement; 4 cases (7.4%) c-myc gene rearrangement without IgH and myc/IgH abnormality; 4 cases (7.4%) without c-myc, IgH or myc/IgH gene rearrangement. No case showed bcl-2 gene abnormality (100%). Fifty nine cases showed normal bcl-6 gene status. One case had bcl-6 gene rearrangement and amplification with the pathologic and immunophenotypic characteristics of BL, leading to a revised pathological diagnosis of B-cell lymphoma, unclassifiable with features intermediate between diffuse large B-cell lymphoma and Burkitt's lymphoma (DLBCL/BL). Two cases showed c-myc gene rearrangement. Two cases showed bcl-6 gene amplification and 6 DLBCL cases had a normal status of bcl-2/IgH.</p><p><b>CONCLUSIONS</b>A majority of pediatric sporadic BL arise from the germinal center B cells, most of which have c-myc gene rearrangement. It is useful to distinguish BL and DLBCL by multiple genes detection.</p>


Subject(s)
Child , Child, Preschool , Female , Humans , Male , Antigens, CD20 , Metabolism , Burkitt Lymphoma , Genetics , Metabolism , Pathology , Diagnosis, Differential , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Genes, myc , Genetics , Immunoglobulin Heavy Chains , Genetics , Lymphoma, Large B-Cell, Diffuse , Genetics , Metabolism , Pathology , Neprilysin , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Proto-Oncogene Proteins c-bcl-6 , Metabolism , Translocation, Genetic
8.
Chinese Journal of Pathology ; (12): 31-34, 2010.
Article in Chinese | WPRIM | ID: wpr-273427

ABSTRACT

<p><b>OBJECTIVE</b>To assess the practical value of BIOMED-2 primers in the diagnosis of ocular adnexal lymphoma by PCR.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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).</p><p><b>CONCLUSIONS</b>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.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Actins , Genetics , DNA Primers , Eye Neoplasms , Diagnosis , Genetics , Pathology , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Gene Rearrangement, B-Lymphocyte, Light Chain , Lymphoma, B-Cell , Diagnosis , Genetics , Pathology , Lymphoma, Follicular , Diagnosis , Genetics , Pathology , Paraffin Embedding , Pilot Projects
9.
Chinese Journal of Pathology ; (12): 177-182, 2010.
Article in Chinese | WPRIM | ID: wpr-273407

ABSTRACT

<p><b>OBJECTIVE</b>To investigate clinicopathologic features of solitary plasmacytoma of bone (SPB) and the role of immuno-phenotype and immunoglobulin gene rearrangement detection in the diagnosis and differential diagnosis of SPB.</p><p><b>METHODS</b>A total of 21 cases of SPB were selected during a period from 1990 to 2008. A retrospective clinicopathologic study and immunohistochemistry (EnVision or EliVision methods) of 17 antigens were performed. In addition, universal IgH (FR3A/LJH/VLJH) primers and BIOMED-2 PCR multiplex tubes were used for IgK and IgL rearrangement analysis.</p><p><b>RESULTS</b>The age of patients ranged from 36 to 72 years with a media of 50 years. Axial skeleton was the most common site of involvement, accounting for 66.7% of the cases (14 of 21), followed by the extremities of 33.3% (7 cases). Low serum level of M-components was found in 5 cases, including two of IgG type (21.4 g/L) and three of IgA type. Clinical manifestations were closely related to the anatomic sites involved, such as pain due to bone destruction, symptoms and signs caused by compression of spinal cord or nerve root, and pathological fracture. All cases presented as a solitary osteolytic lesion. According to the histological grading criteria, grade I tumor was seen in 12 of 21 cases (57.1%). The remaining were grade II (5 cases, 23.8%) and grade III (4 cases, 19.0%). Immunohistochemically, the neoplastic cells expressed two or more plasma cell antigens, including CD138, CD38 and PC, but no CD19 and CD20. CD79a expression detected in 23.8%(5/21) of the cases. Expression of CD56, CD27 and CD44v6 were 57.1% (12/21), 15.0% (3/20) and 23.8% (5/21), respectively. Follow-up data were available in 12 of the 21 patients (57.1%). Five patients were alive and 7 died. Three patients developed multiple myeloma (MM) and died of the tumor.</p><p><b>CONCLUSIONS</b>SPB is a rare tumor with bone pain as the most common presenting symptom due to bone destruction. The diagnosis of EMP can only be established after exclusion of an extramedullay invasion by MM. Immunophenotype and IgH gene rearrangement analysis play important roles in the diagnosis of SPB.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , ADP-ribosyl Cyclase 1 , Metabolism , Bone Neoplasms , Genetics , Metabolism , Pathology , General Surgery , Diagnosis, Differential , Follow-Up Studies , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Immunophenotyping , Lymphoma, Large B-Cell, Diffuse , Metabolism , Pathology , Lymphoma, Large-Cell, Anaplastic , Metabolism , Pathology , Melanoma , Metabolism , Pathology , Multiple Myeloma , Pathology , Plasmacytoma , Genetics , Metabolism , Pathology , General Surgery , Retrospective Studies , Survival Rate , Syndecan-1 , Metabolism
10.
Chinese Journal of Pathology ; (12): 739-744, 2009.
Article in Chinese | WPRIM | ID: wpr-249051

ABSTRACT

<p><b>OBJECTIVE</b>To evaluate the efficiency of the BIOMED-2 PCR assay and its implication in the diagnosis of mature B-cell non-Hodgkin's lymphomas.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>BIOMED-2 assay is highly sensitive and specific for the detection of clonal B cell gene rearrangement using routine paraffin embedded formalin fixed specimens.</p>


Subject(s)
Humans , 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 , Immunophenotyping , Lymphoma, B-Cell , Genetics , Allergy and Immunology , Pathology , Lymphoma, B-Cell, Marginal Zone , Genetics , Allergy and Immunology , Pathology , Lymphoma, Large B-Cell, Diffuse , Genetics , Allergy and Immunology , Pathology , Paraffin Embedding , Pseudolymphoma , Genetics , Allergy and Immunology , Pathology , Sensitivity and Specificity
11.
Chinese Journal of Oncology ; (12): 183-188, 2009.
Article in Chinese | WPRIM | ID: wpr-255534

ABSTRACT

<p><b>OBJECTIVE</b>To explore the feasibility of semi-nested PCR technique for detection of immunoglobulin heavy chain (IgH) clonal rearrangement in bone marrow of B-cell lymphoma patient and to further evaluate its clinicopathological value.</p><p><b>METHODS</b>Gene clonal rearrangement of IgH was detected by semi-nested PCR using primers of FR2 & FR3A in 105 bone marrow samples of patients with B-cell lymphoma. The PCR detection results were compared with the cytomorphology of bone marrow aspiration biopsy. The correlation between PCR detection results and clinicopathological factors were evaluated.</p><p><b>RESULTS</b>Among 105 cases of B-cell lymphoma, bone marrow involvement was detected by PCR technique in 48 cases (45.7%), while only 22 cases (21.0%) were detected by bone marrow cytological analysis. There was a significant difference between two methods (P < 0.05), and the concordance rate was 71.4%. The incidence of bone marrow involvement at the time of initial diagnosis detected by PCR technique was 30.8% for diffuse large B cell lymphoma (DLBCL), 25.0% for follicular lymphoma (FL), and 100.0% for small lymphocytic lymphoma (SLL), respectively. Bone marrow involvement detected by PCR detection correlated with Ann Arbor stage. Rate of clonal IgH gene rearrangement by PCR in early B-cell lymphoma was lower than that in advanced stage B-cell lymphoma patients (P = 0.02). There was no statistically significant difference in efficacy between patients with positive and negative results detected by PCR (P > 0.05). But difference in complete response (CR) rate (23.3% and 46.3%) had significant difference (P = 0.019).</p><p><b>CONCLUSION</b>Semi-nested PCR analysis may be an effective method for detection of abnormalities in bone marrow in patients with B-cell lymphoma and is superior to cytomorphology. The positive rate in patients with advanced Ann Arbor stage is higher than that in patients with early Ann Arbor stage, and patients with PCR negative result have more chances to achieved CR after treatment.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Antineoplastic Combined Chemotherapy Protocols , Therapeutic Uses , Biopsy , Methods , Bone Marrow , Pathology , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Immunoglobulin Heavy Chains , Genetics , Leukemia, Lymphocytic, Chronic, B-Cell , Drug Therapy , Genetics , Pathology , Lymphoma, Follicular , Drug Therapy , Genetics , Pathology , Lymphoma, Large B-Cell, Diffuse , Drug Therapy , Genetics , Pathology , Neoplasm Staging , Polymerase Chain Reaction , Methods , Remission Induction
12.
Scientific Journal of Iranian Blood Transfusion Organization Research Center [The]. 2008; 5 (2): 157-166
in Persian | IMEMR | ID: emr-90312

ABSTRACT

Rearrangement of V, D, and J segments of immunoglobulin heavy chain gene with inserted or deleted nucleotides within rearranged segments makes unique hypervariable regions [CDR-3]. These regions can be used for evaluation of B cell clonality for the purpose of molecular diagnosis of Non-Hodgkin Lymphoma [NHL] and for confirmatory diagnosis in suspicious cases. In this study, samples of 42 patients were collected from Taleghani, Baqhiyatalah, and Aliasghar hospitals; out of this number, there were 22 patients with diagnosis of B cell NHL, 10 with reactive hyperplasia, and 10 with malignant lymphoma. After DNA extraction from formalin fixed paraffin embedded tissues, PCR was done using consensus primers for amplification of CDR-3 region. PCR products were analyzed after heteroduplex analysis using polyacrylamide gel electrophoresis and silver stain. Results Clonal patterns in group 1 [B cell NHL], 2 [reactive and follicular hyperplasia], and 3 [morphological diagnosis without immunohistochemistry] were observed in 77.2%, 0%, and 70% of patients, respectively. Our findings are compatible with other international studies with minor differences. The diagnosis of B-cell lymphoid malignancy can frequently be substantiated by detecting clonal immunoglobulin heavy chain [IGH] gene rearrangement


Subject(s)
Humans , Lymphoma, Non-Hodgkin/diagnosis , Immunoglobulin Heavy Chains , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Molecular Diagnostic Techniques , Polymerase Chain Reaction
13.
Journal of Southern Medical University ; (12): 1964-1967, 2008.
Article in Chinese | WPRIM | ID: wpr-321781

ABSTRACT

<p><b>OBJECTIVE</b>To analyze and optimize the gene rearrangement primers of different frame regions (FR) of immunoglobulin heavy chain (IgH) genes by bioinformatic methods and explore the application of these primers in the detection of paraffin-embedded lymphoma tissues.</p><p><b>METHODS</b>Three pairs of primers from IgH FR1, FR2 and FR3 regions (P1c, P2A and P31, respectively) were selected as the B cell gene rearrangement primers after comparison of the gene fragments in 44 IgH variable and 6 joining regions. Using one pair of T cell receptor (TCR) gamma primer as the T cell gene rearrangement primer, 101 histopathologically confirmed lymphoproliferative samples including 80 B cell lymphomas, 14 T cell lymphomas, and 7 reactive proliferative lymph nodes were examined by PCR for gene arrangement. The DNAs from DG75 and Jurkat cell lines were used as the positive controls for B and T cell lymphoma, respectively, with those from reactive proliferative lymph nodes as the negative control.</p><p><b>RESULTS</b>The positivity rates of IgH primers (P1c, P2A and P31) in the 80 B cell lymphomas were 37.5% (30/80), 52.5% (42/80) and 70.0% (56/80), respectively, and only one of the 14 T cell lymphoma cases was positive for the primers, suggesting significant differences in the detection rates of B cell lymphomas by the 3 primers. The detection rate was increased to 83.9% by combining the results by P31 and P2A primers. No positivity was found in the proliferative reaction tissues.</p><p><b>CONCLUSION</b>Primers from IgH FR3 region genes are more sensitive than that from the FR1 and FR2 regions in the detection of gene rearrangement in paraffin-embedded lymphoma tissues. The detection rates can be increased by combining the results with the primers for IgH FR3 with that of FR2.</p>


Subject(s)
Humans , Male , DNA Primers , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Genetics , Immunoglobulin Heavy Chains , Genetics , Lymphoma, B-Cell , Diagnosis , Genetics , Pathology , Lymphoma, Non-Hodgkin , Diagnosis , Genetics , Pathology , Lymphoma, T-Cell , Diagnosis , Genetics , Pathology , Paraffin Embedding
14.
J. bras. pneumol ; 33(6): 625-634, nov.-dez. 2007. ilus, tab
Article in English, Portuguese | LILACS | ID: lil-471283

ABSTRACT

OBJETIVO: Determinar a utilidade, na prática rotineira, da análise da clonalidade dos linfócitos T e B nos tecidos pulmonares por reação em cadeia da polimerase no diagnóstico das doenças linfoproliferativas pulmonares. MÉTODOS: Avaliaram-se, mediante análise imunohistoquímica e rearranjo molecular dos genes, 8 casos de pneumonia intersticial linfocítica (PIL) e 7 casos de doenças linfoproliferativas pulmonares. RESULTADOS: Todos os 8 casos de PIL expressaram imunocoloração moderada a forte para CD3, em contraste com apenas 2 casos de linfoma e 1 caso de pseudolinfoma. Rearranjo gênico foi detectado em 4 de 8 casos de PIL, o que mudou o diagnóstico de PIL para linfoma, indicando, assim, a importância da detecção de rearranjo gênico em casos de PIL. Nesta situação, rearranjo gênico usando-se os pares de primers VH/JH e Vgama11/Jgama12 foi detectado em 3 e 1 casos de PIL, respectivamente, e não foram detectadas anormalidades gênicas usando-se as pares Dbeta1/Jbeta2 e Vgama101/Jgama12. Uma associação positiva foi detectada entre a intensidade de imunoexpressão CD20 e CD68 e rearranjo gênico usando-se o par de primers VH/JH. Antes do rearranjo gênico, 4 pacientes com PIL morreram rapidamente, enquanto que, após o rearranjo gênico, apenas 1 paciente com PIL morreu. CONCLUSÕES: A detecção de células B e T monoclonais por imunofenotipagem e reação em cadeia da polimerase mostrou impacto no diagnóstico de linfomas pulmonares em pacientes previamente diagnosticados com PIL. Portanto, imunofenotipagem e reação em cadeia da polimerase devem ser incluídas como métodos de 'padrão ouro' na rotina diagnóstica.


OBJECTIVE: To determine the usefulness, in routine practice, of using polymerase chain reaction to analyze B and T lymphocyte clonality in pulmonary tissue as a tool for the diagnosis of pulmonary lymphoproliferative disorders. METHODS: Immunohistochemistry and molecular gene rearrangement analysis were performed in order to assess 8 cases of lymphoid interstitial pneumonia (LIP) and 7 cases of pulmonary lymphoproliferative disorders. RESULTS: All 8 cases of LIP presented moderate to strong immunostaining for CD3, compared with only 2 cases of lymphoma and 1 case of pseudolymphoma (p = 0.02). Gene rearrangement was detected in 4 of the 8 cases, which changed the diagnosis from LIP to lymphoma, showing the importance of gene rearrangement detection in cases of LIP. In this situation, gene rearrangement using the VH/JH and Vgamma11/Jgamma12 primer pairs was detected in 3 cases and 1 case, respectively, and no gene abnormalities were found using the Dbeta1/Jbeta2 and Vgamma101/Jgamma12 primer pairs in any of the cases. A significant positive association was found between the intensity of CD20 and CD68 expression and gene rearrangement using the VH/JH primer pair. Prior to the gene rearrangement, 4 patients with LIP died quickly, whereas only one patient with LIP died after the gene rearrangement. CONCLUSIONS: Detection of monoclonal B and T cells by immunophenotyping and polymerase chain reaction had an impact on the diagnosis of pulmonary lymphomas in patients previously diagnosed with LIP. Therefore, immunophenotyping and polymerase chain reaction should be used as 'gold standard' techniques in routine practice.


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child, Preschool , Female , Humans , Male , Middle Aged , Gene Rearrangement , Immunophenotyping , Lung Diseases, Interstitial/immunology , Lung Neoplasms/immunology , Lymphoma/immunology , Antigens, CD/analysis , Case-Control Studies , Diagnosis, Differential , DNA Primers , Feasibility Studies , Gene Rearrangement, B-Lymphocyte, Heavy Chain/genetics , Gene Rearrangement, B-Lymphocyte, Heavy Chain/immunology , Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor/genetics , Gene Rearrangement, gamma-Chain T-Cell Antigen Receptor/immunology , Lung Diseases, Interstitial/diagnosis , Lung Diseases, Interstitial/genetics , Lung Neoplasms/diagnosis , Lung Neoplasms/genetics , Lymphoid Tissue/pathology , Lymphoma/diagnosis , Lymphoma/genetics , Lymphoproliferative Disorders/diagnosis , Lymphoproliferative Disorders/genetics , Lymphoproliferative Disorders/immunology , Polymerase Chain Reaction , Pseudolymphoma/diagnosis , Pseudolymphoma/genetics , Pseudolymphoma/immunology , Retrospective Studies
15.
Scientific Journal of Iranian Blood. 2007; 4 (2): 123-136
in Persian | IMEMR | ID: emr-99413

ABSTRACT

Diversity of IgH and IgK molecules is generated during B and T Lymphocyte differentiation through the rearrangement of variable, diversity, junction and constant gene segments. Additionally, random insertion and deletions of nucleotides between gene segments make unique sequences which are cell or clone specific. Similar IgH and IgK genes rearranged in normal cells of lymphoid leukemia cases can be used as a marker of clonality and for evaluation of minimal residual disease [MRD]. The purpose of this study is to evaluate the pattern of IgH chain and IgK gene rearrangements using polymerase chain reaction [PCR] in beta-precursor acute lymphoblastic leukemias [ALL] to follow the MRD at day 14, day 28 [end of remission induction], week 10, 3-6 months and 6-12. month after the initiation of treatment. In our prospective study bone marrow aspirates of 183 children at the mean age of 63.6 months with diagnosis of acute leukemia were collected at admission before any chemotherapy. After reviewing cytomorphology and immunophenotyping, only 140 cases with diagnosis of beta-precursor ALLs were selected for study. Mononuclear cells including leukemic blasts were isolated by density gradient. After DNA extraction, IgH and IgK [V[K] I-IV / Kde] were amplified by consensus primers using PCR. PCR products were analyzed after heteroduplex analysis and polyacrylamide gel electrophoresis [silver stain]. The DNA sequences were compared and aligned with the sequences homologous for IgH and IgK published by Gene Bank. The follow up specimens were collected at day 14, day 28 [end of remission induction], day 45-month 3, and 3-6 months and 6-12 months after initiation of treatment. After routine cytomorphologic analysis, similar PCR was done on follow up extracted DNAs in parallel with diagnosis DNA. MRD was considered to be approved positive if bands similar to those at the time of diagnosis were present. Statistical analysis using SPSS software [version 11.5] was performed. 90.5% of patients had clonal IgH gene rearrangements. Monoclonal, biclonal and oligoclonal patterns were observed in 57.8%, 34.9% and 5.5% of patients with IgH [CDR III] rearrangement, respectively. Clonal patterns of IgK-Kde were detected in 59 [67%; n: 88] of BP-ALLs. According to cytomorphology about 92% of patients were in complete remission. MRD positivity decreased from more than 90% to 20% using different gene rearrangements in defined time points. Four patients who relapsed during follow up were MRD positive using 1-3 rearrangements and all except one were in clinical remission. Clonal rearrangement of IgH had a pattern similar to other populations. IgK was slightly more frequent than previously reported and the VKI [25%] was the most common type. These differences can be explained by different techniques, DNAs and clonality markers. According to the results, these clonal markers can be used in diagnosis and follow up of MRD


Subject(s)
Humans , /genetics , Prospective Studies , Polymerase Chain Reaction , Silver Staining , Electrophoresis, Polyacrylamide Gel , Neoplasm, Residual/diagnosis , Neoplasm, Residual/genetics , Gene Rearrangement, B-Lymphocyte, Light Chain , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Child
16.
Journal of Experimental Hematology ; (6): 1236-1239, 2007.
Article in Chinese | WPRIM | ID: wpr-318750

ABSTRACT

The study was purpose to evaluate the value of real time quantitative-PCR for monitoring IgH level in patients with B-cell malignancy after hematopoietic stem cell transplantation (HSCT). Quantification of IgH levels was performed on bone marrow mononuclear cells from 9 patients with B-cell malignancy before and after HSCT by PCR using the consensus JH TaqMan probe in combination with an allele-specific oligonucleotide (ASO) upstream primer. The IgH levels was normalized by control gene GAPDH. The results indicated that the reproducible sensitivity of RQ-PCR was 1 copy, the significant reduction of IgH copies was observed in bone marrow samples of 9 patients at one month post HSCT (6.67x10(3)/10(6) GAPDH vs 29/10(6) GAPDH, p<0.01). 3 out of 9 patients who achieved complete clinical and molecular cytogenetic remission (CCyR) contained persistently measurable low IgH level of 10(2)/10(6) GAPDH within 15 months and no detectable IgH at 18 months post HSCT. Whereas 5 out of 9 patients whose IgH copies were less than 10(2)/10(6) GAPDH within 3 months and less than 10(3)/10(6) GAPDH 3 months post HSCT achieved a sustained complete remission (CR). IgH copies in one patient were 4.5x10(3)/10(6) GAPDH at 3 months post HSCT, who relapsed at 4 months post HSCT. The median levels of tumor contamination in the stem cell harvests from 8 patients measured by RQ PCR were 3.68x10(2) (0-1720)/10(6) GAPDH. RQ PCR showed that PBPC harvests were less contaminated than BM harvests [75 (0-890)/10(6) GAPDH vs 1.1x10(3) (527-1720)/10(6) GAPDH, p<0.05]. 8 patients whose stem cell harvest were avaiable for RQ PCR were still in CR despite of the tumor contamination. The level of tumor contamination in stem cell harvest well correlated with IgH levels at diagnosis and one month after HSCT (r=0.810, r=0.708, p<0.05). It is concluded that RQ PCR can effectively monitor the IgH levels in patients with B-cell malignancy after auto-HSCT. 10(3)/10(6) GAPDH within 3 months post HSCT may be a cut-off level of IgH copies, which may be used to evaluate different prognoses of patients.


Subject(s)
Adolescent , Adult , Female , Humans , Male , Young Adult , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Hematopoietic Stem Cell Transplantation , Immunoglobulin Heavy Chains , Genetics , Neoplasm, Residual , Precursor B-Cell Lymphoblastic Leukemia-Lymphoma , Genetics , Therapeutics , Reverse Transcriptase Polymerase Chain Reaction
17.
Chinese Journal of Pathology ; (12): 736-741, 2007.
Article in Chinese | WPRIM | ID: wpr-350026

ABSTRACT

<p><b>OBJECTIVE</b>To analyze the clonal relationship between transformed and non-transformed components in mucosa-associated lymphoid tissue (MALT) lymphoma.</p><p><b>METHODS</b>Six cases of MALT lymphoma with high-grade transformation were studied. Immunohistochemical study was carried out by EliVision using bcl-10 antibody. Reverse transcription-polymerase chain reaction was used to detect the presence of API2-MALT1 fusion gene transcripts. The target cells were selected by laser microdissection and studied by polymerase chain reaction and sequence analysis for rearrangement of immunoglobulin heavy chain gene.</p><p><b>RESULTS</b>In the 6 cases of MALT lymphoma with high-grade transformation, nuclear and cytoplasmic expression of bcl-10 was demonstrated in 1 case, while API2-MALT1 fusion gene was present in 2 cases. Identical fragments of rearranged immunoglobulin heavy chain gene were detected in both transformed and non-transformed components in each of the 6 cases, except for the differences at 2 nucleotide positions in N or D regions in 2 cases.</p><p><b>CONCLUSION</b>The tumor cells from both transformed and non-transformed components in MALT lymphoma derive from the same clone.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Adaptor Proteins, Signal Transducing , Metabolism , B-Cell CLL-Lymphoma 10 Protein , Base Sequence , Cell Nucleus , Metabolism , Cell Transformation, Neoplastic , Cloning, Molecular , Cytoplasm , Metabolism , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Genetics , Lymphoma, B-Cell, Marginal Zone , Genetics , Metabolism , Pathology , Oncogene Proteins, Fusion , Metabolism , Polymerase Chain Reaction
18.
Chinese Journal of Pathology ; (12): 600-604, 2007.
Article in Chinese | WPRIM | ID: wpr-347720

ABSTRACT

<p><b>OBJECTIVE</b>To study the genetic aberrations and their pathologic significance in follicular lymphoma (FL).</p><p><b>METHODS</b>Paraffin-embedded tissue samples of 55 cases of FL, 28 cases of other small B-cell lymphomas and 10 cases of reactive follicular hyperplasia were retrieved. Nested polymerase chain reaction (PCR) was used to detect clonal rearrangement of immunoglobulin heavy chain gene (IgH) in FL and other small B-cell lymphomas. The translocation t (14; 18) was studied by PCR and dual-color fluorescence in-situ hybridization (FISH) in FL. Cases of reactive follicular hyperplasia were used as controls.</p><p><b>RESULTS</b>Amongst the 55 cases studied, 49 cases were nodal and 6 cases were extranodal. There were 33 males and 22 females. The male-to-female ratio was 1.5:1. The median age of the patients was 57 years. Twenty-five cases belonged to histologic grade 1, while 19 cases were grade 2 and 11 cases were grade 3. Beta-actin DNA was detected in 50 cases of FL. Amongst those 50 cases, clonal IgH rearrangement was present in 34 (68%). Twenty-four cases (48%) and 25 cases (50%) were positive for FR3A and FR2 respectively. Fifteen cases (30%) showed dual positivity for both FR3A and FR2. Thirty-four cases (68%) demonstrated clonal IgH rearrangement. As for other small B-cell lymphomas, 25 cases were positive for beta-actin. FR3A and FR2 were detected in 18 and 17 cases respectively. Clonal IgH rearrangement was demonstrated in 24 cases. In contrast, none of the 4 cases of reactive follicular hyperplasia showed the clonal rearrangement pattern. Amongst the 44 cases of nodal FL analyzed, t (14; 18) was detected in 15 cases (with 14 cases in MBR and 1 case in mcr). In general, FISH was superior to PCR in detecting t (14; 18) using paraffin-embedded tissue samples.</p><p><b>CONCLUSIONS</b>The detection rate of clonal IgH rearrangement in FL is lower than that in other small B-cell lymphomas. Demonstration of t (14; 18) in paraffin-embedded tissue samples by FISH helps in diagnosis of FL. FISH is superior to PCR, as the technique is more sensitive and less labor intensive.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Actins , Metabolism , Chromosomes, Human, Pair 14 , Genetics , Chromosomes, Human, Pair 18 , Genetics , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Genetics , In Situ Hybridization, Fluorescence , Methods , Lymphoma, B-Cell , Genetics , Metabolism , Lymphoma, Follicular , Genetics , Metabolism , Paraffin Embedding , Polymerase Chain Reaction , Methods , Translocation, Genetic
19.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery ; (12): 932-935, 2006.
Article in Chinese | WPRIM | ID: wpr-315561

ABSTRACT

<p><b>OBJECTIVE</b>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.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSIONS</b>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.</p>


Subject(s)
Adolescent , Adult , Aged , Child , Female , Humans , Male , Middle Aged , Young Adult , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Gene Rearrangement, T-Lymphocyte , Lymphoma, Non-Hodgkin , Diagnosis , Pathology , Nasal Cavity , Nose Neoplasms , Diagnosis , Pathology
20.
Chinese Journal of Pathology ; (12): 529-534, 2006.
Article in Chinese | WPRIM | ID: wpr-268908

ABSTRACT

<p><b>OBJECTIVE</b>To study the clinicopathologic features of diffuse large B-cell lymphoma (DLBCL) with expression of anaplastic lymphoma kinase (ALK) protein.</p><p><b>METHODS</b>Nine hundred and forty-five (945) cases of DLBCL (including 177 consultation cases) diagnosed according to the 2001 World Health Organization classification of tumors of hematopoietic and lymphoid tissues were enrolled into the study. Immunohistochemical study for anti-ALK-11 was performed using LSAB technique. The ALK-positive cases were further confirmed by immunohistochemical study using EnVision technique. Only ALK-positive cases by EnVision technique were further analyzed by immunostaining for antigens including CD20, CD3, CD30, EMA, granzyme-B, TIA-1 and PC. Immunoglobulin heavy chain gene rearrangement study was also performed and follow-up data collected.</p><p><b>RESULTS</b>There were altogether 5 (4 males and 1 female) cases of DLBCL showing expression of ALK protein. The age of the patients ranged from 34 to 72 years. All were primary nodal DLBCL. One case belonged to clinical stage I, 2 in stage II and 2 in stage III. The duration of follow up ranged from 4 to 32 months. Three patients subsequently died and the longest survival was 32 months. Morphologic subtypes included centroblastic 2, anaplastic 1, immunoblastic with plasmacytoid differentiation 1 and plasmablastic 1. Immunohistochemically, 4 cases were CD20 positive (including 2 centroblastic, 1 anaplastic and 1 immunoblastic cases). The plasmablastic case expressed kappa light chain and was negative for CD20. Rearrangement of immunoglobulin heavy chain gene was demonstrated in all 5 cases studied. As for ALK protein staining, a mixed membranous and cytoplasmic (1 immunoblastic case), granular cytoplasmic (2 centroblastic and 1 anaplastic cases) and mixed nuclear and cytoplasmic (1 plasmablastic case) patterns were observed.</p><p><b>CONCLUSIONS</b>Expression of ALK protein is a rare phenomenon in DLBCL and can be seen in centroblastic, anaplastic, immunoblastic and plasmablastic subtypes. It is often associated with aggressive clinical behavior and worse prognosis. A new pattern of ALK protein expression, mixed membranous and cytoplasmic, is reported.</p>


Subject(s)
Adult , Aged , Female , Humans , Male , Middle Aged , Antigens, CD20 , Metabolism , Follow-Up Studies , Gene Rearrangement, B-Lymphocyte, Heavy Chain , Genetics , Immunoglobulin kappa-Chains , Metabolism , Immunohistochemistry , Lymphoma, Large B-Cell, Diffuse , Genetics , Metabolism , Pathology , Neoplasm Staging , Polymerase Chain Reaction , Prognosis , Protein-Tyrosine Kinases , Metabolism , Receptor Protein-Tyrosine Kinases
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