OBJECTIVETo study the clinicopathologic features and significance of aberrant CD56 expression in diffuse large B-cell lymphoma (DLBCL).

METHODSThe clinical and pathologic profiles of 10 cases of DLBCL with aberrant expression of CD56 were investigated. Immunohistochemical staining, in-situ hybridization for Epstein-Barr virus encoded RNA and gene rearrangement for IgH and Igκ were carried out.

RESULTSThere were 6 male and 4 female patients. The medium age of patients was 46 years. All of them presented with extranodal lymphoma involvement, with gastrointestinal tract being the commonest site (5/10). Histologic examination showed that most of the atypical lymphoid cells were centroblast-like and demonstrated a diffuse growth pattern. Apoptosis and necrosis were identified in some cases. Immunohistochemical study showed that the tumor cells were positive for CD20 or CD79α and aberrantly expressed CD56. Five cases had the GCB phenotype while the remaining cases had the non-GCB phenotype, according to Hans classification. Bcl-6 was positive in most cases (9/10). All cases showed a high proliferation index by Ki-67. The tumor cells were negative for CD3ε, CD138 and granzyme B. In-situ hybridization for Epstein-Barr virus encoded RNA was performed in 7 cases and none of them showed positive signals. IgH gene rearranged bands were detected in 4 cases (4/6) and Igκ was detected in 3 cases (3/6). Follow-up data were available in 8 patients. Two patients died of disease progression within 5 to 13 months after diagnosis and the other 6 patients were alive 8 to 60 months after therapy.

CONCLUSIONSDLBCL with aberrant expression of CD56 is rare. Most of them present with extranodal involvement, show high frequency of bcl-6 expression and high proliferation index. The patients often have good response to chemotherapy.

Antigens, CD20 ; metabolism ; Apoptosis ; CD56 Antigen ; metabolism ; CD79 Antigens ; metabolism ; Disease Progression ; Female ; Gene Rearrangement ; Granzymes ; metabolism ; Herpesvirus 4, Human ; genetics ; Humans ; Immunophenotyping ; In Situ Hybridization ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Necrosis ; Phenotype ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; RNA, Viral ; analysis

BACKGROUND: Immunohistochemical demonstration of CD20 in diffuse large B-cell lymphoma (DLBCL) is prerequisite not only for the diagnosis but also for assigning patients to rituximab-containing chemotherapy. However, little is known about the impact of abundance of CD20 expression assessed by immunohistochemistry on the clinical outcome of DLBCL. We performed a semi-quantitative immunohistochemical analysis of CD20 expression in DLBCL to examine the prognostic implication of the level of CD20 expression. METHODS: Pre-treatment diagnostic tissue samples from 48 DLBCL patients who were treated with rituximab, cyclophosphamide, doxorubicin, vincristine, and prednisone (R-CHOP) regimen were represented in a tissue microarray and immunostained for CD20. The relative abundance of CD20 expression was semi-quantitatively scored using a web-based ImmunoMembrane plug-in. Receiver operating characteristic curve analysis was used to determine a prognostically relevant cut-off score in order to dichotomize the patients into CD20-high versus CD20-low groups. RESULTS: The levels of CD20 expression were heterogeneous among the patients, with a wide and linear distribution of scores. Patients in CD20-low group showed significantly poor clinical outcome. CONCLUSIONS: The levels of CD20 expression in DLBCL are heterogeneous among the patients with DLBCL. A subgroup of the patients with CD20 expression levels below the cut-off score showed poor clinical outcome.
Antigens, CD20 ; B-Lymphocytes* ; Cyclophosphamide ; Diagnosis ; Doxorubicin ; Drug Therapy ; Humans ; Immunohistochemistry ; Lymphoma, B-Cell* ; Prednisone ; ROC Curve ; Tissue Array Analysis ; Vincristine ; Rituximab

OBJECTIVETo explore the clinical and survival significance of CD20 positive adult patients with B-lineage acute lymphoblastic leukemia (B-ALL).

METHODSThe clinical features and survival of 168 adult patients with B-ALL diagnosed and treated in our department from May 2007 to July 2011 were analyzed retrospectively, 58 expressed CD20 and 110 not.

RESULTSThe sex, distribution of age, anemia, thrombocytopenia, infiltration of liver, spleen and lymph nodes, the expression of myeloid lineage marker, incidence of Ph chromosome, complete remission within 4 weeks showed no significant differences in CD20 positive and negative groups (P>0.05); median white blood cell count at diagnosis and the rate of patients with high white blood cell count in CD20 positive group were 19.2×10⁹/L and 37.9% respectively, which were significantly higher than those of 6.93 × 10⁹/L and 20.9% in CD20 negative group (P<0.05); cumulative incidence of relapse between two groups showed significant difference (P<0.05); multivariable analysis for overall survival and progress-free survival identified CD20 positivity as independent predictor.

CONCLUSIONThe expression of CD20 in adult patients with B-ALL appeared to be associated with high white blood cell count and poor prognosis.

Adult ; Antigens, CD20 ; Cell Lineage ; Disease-Free Survival ; Humans ; Leukocyte Count ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma ; Recurrence ; Remission Induction ; Retrospective Studies ; Survival Analysis

OBJECTIVETo study the clinicopathologic characteristics of thymic epithelial tumors and to evaluate the diagnostic reproducibility and clinical relevance of the 2004 WHO histologic classification system.

METHODSThe morphology and immunophenotype of 52 cases of thymic epithelial tumor were reviewed. The tumors were classified according to the new WHO classification system and the clinical data were analyzed.

RESULTSOf the 52 cases studied, 45 were thymomas and 7 were thymic carcinomas. Amongst the 45 cases of thymoma, 6 (13.4%) were type A, 15 (33.3%) were type AB, 4 (8.9%) were type B1, 9 (20.0%) were type B2, 9 (20.0%) were type B3 and 2 (4.4%) were metaplastic thymoma. Amongst the 7 cases of thymic carcinoma, 6 were squamous cell carcinomas and 1 was neuroendocrine carcinoma. The commonest presentations were cough and chest pain. Some cases were incidentally discovered by routine physical examination. Thirteen cases (25.0%) of thymoma were associated with myasthenia gravis. CT scan showed that 49 cases (94.2%) were located in the anterior mediastinum. All cases of type A, AB and B1 thymoma and most cases of B2 thymoma appeared as well-defined homogeneous mass, whereas a few cases of type B2 thymoma and most cases of type B3 thymoma and thymic carcinoma were poorly demarcated and heterogeneous. According to Masaoka staging system, 20 cases (41.7%) belonged to stage I, 15 cases (31.3%) stage II, 11 cases (22.9%) stage III and 2 cases (4.1%) stage IV. The histologic subtypes of thymic epithelial tumors significantly correlated with the clinical stages (chi(2) = 32.5, P < 0.01).

CONCLUSIONSThe 2004 revision of WHO histologic classification system for thymic epithelial tumors shows a high degree of reproducibility. Correlation with the radiologic, clinical and prognostic parameters is helpful in determining the management strategy for individual patients.

Adult ; Aged ; Antibodies, Monoclonal ; analysis ; Antigens, CD20 ; metabolism ; CD5 Antigens ; metabolism ; Carcinoma, Neuroendocrine ; classification ; diagnostic imaging ; metabolism ; pathology ; Carcinoma, Squamous Cell ; classification ; diagnostic imaging ; metabolism ; pathology ; Female ; Follow-Up Studies ; Humans ; Keratins ; immunology ; Male ; Middle Aged ; Myasthenia Gravis ; Neoplasm Recurrence, Local ; Neoplasm Staging ; Prognosis ; Radiotherapy, Adjuvant ; Retrospective Studies ; Thymoma ; classification ; diagnostic imaging ; metabolism ; pathology ; Thymus Neoplasms ; classification ; diagnostic imaging ; metabolism ; pathology ; Tomography, X-Ray Computed

This study was aimed to investigate the effects of rituximab (RTX), a chimeric human anti-CD20 monoclonal antibody, on lymphoma cell injury induced by X ray irradiation. The human Burkitt EBV-infected and moderate radioresistance lymphoma cells (Namalwa) were used in the this study. Cytotoxicity of rituximab combined with X ray irradiation on Namalwa cells was measured by sulforhodamine B (SRB)-staining; the apoptosis of Namalwa cells was detected by flow cytometry with FITC-Annexin V/PI double staining; the morphologic changes of cells were observed under transmission electron microscope (TEM) and the change of intracellular free calcium level ([Ca(2+)]i) in response to irradiation and rituximab was determined by means of the fluorescent dye fluo-3 and confocal microscopy. The results showed that the growth inhibition in Namalwa cells exposed to irradiation was enhanced by treatment with rituximab. Compared with irradiation alone, rituximab combined with irradiation significantly induced the cell apoptosis and a sustained rise of intracellular free calcium ([Ca(2+)]i) level in Namalwa cells; the serial apoptotic appearances of cells could be observed under TEM. It is concluded that rituximab can enhance the sensitivity of lymphoma cells on X ray irradiation as to induce cell more apoptosis, in this process the intracellular free calcium ([Ca(2+)]i), as an intracellular signaling molecule probably plays an important role.
Antibodies, Monoclonal, Murine-Derived ; immunology ; pharmacology ; Antigens, CD20 ; immunology ; Apoptosis ; Calcium ; analysis ; Cell Line, Tumor ; Humans ; Lymphoma ; drug therapy ; metabolism ; pathology ; Radiation Tolerance ; drug effects ; Rituximab

The aim of this study was to explore the clinical features and survival of adult patients with CD20 positive B-lineage acute lymphoblastic leukemia (B-ALL). The clinical manifestations, examination results, therapeutic effect and survival rate of 119 adult B-ALL patients diagnosed and treated in our hospital from May 2004 to January 2008 were analyzed retrospectively. The results showed that among 119 cases, CD20 positive B-ALL accounted for 40 cases (33.61%), CD20 negative B-ALL patents accounted for 79 cases (66.39), the percentage of male patients in CD20 positive and negative groups were 72.50% and 50.63%, the leukocyte counts at diagnosis in these two groups were (27.35+/-30.29)x10(9)/L and (0.11+/-81.72)x10(9)/L, respectively, there were significant differences (p<0.05), whereas the distribution of age, infiltration of liver, spleen, lymph nodes and central nervous system, the hemoglobin and platelet levels, the expression of myeloid lineage marker, the incidence of Ph chromosome, the ratio of hyperdiploid and normal karyotype, the complete remission rate within 4 weeks, induction death rate and relapse rate and so on in CD20 positive and negative groups showed no significant differences (p>0.05). The analysis of Kaplan-Meier curve on survival rate demonstrated that the median overall survival time and 3-year overall survival rate of adult B-ALL patients in CD20 positive and negative groups were 11.0 months and 12.0 months, 28% and 20% respectively, there were no statistical differences (p=0.832). It is concluded that the expression of CD20 in adult B-ALL appears to be associated with sex and leukocyte count, but not associated with other clinical features, which indicates no significant influence on the prognosis of patients.
Adolescent ; Adult ; Antigens, CD20 ; metabolism ; Female ; Humans ; Leukemia, B-Cell ; mortality ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; mortality ; Prognosis ; Retrospective Studies ; Survival Analysis ; Survival Rate ; Young Adult

OBJECTIVETo explore the diagnostic implication of immunophenotyping and gene rearrangement in subcutaneous panniculitis-like T-cell lymphoma (SPTL).

METHODSAccording to the selection criteria of 2005 WHO-EORTC classification for cutaneous lymphomas, 20 SPTL patients were enrolled in this study. A 10-antibody panel was used for immunophenotyping and in addition, polymerase chain reaction for TCR gamma and IgH gene rearrangement and in situ hybridization for EBER1/2 were also employed.

RESULTSThere were 9 males and 11 female with a mean age of 29.5 years. Immunophenotypic study showed that all the patients expressed one to three T-cell associated antigens (CD2, CD3 or CD45RO), 18 patients were positive for beta F1, 18 for CD8, 20 for TIA-1 and 16 for granzyme B. None of the patients expressed CD4, CD20 and CD56. TCR gamma gene rearrangement was found in 16 of 20 cases (80.0%) and none for IgH gene rearrangement. The positive rate of EBER1/2 was 25.0% (5/20).

CONCLUSIONSSince the majority of SPTL patients show clonal TCR gene rearrangements, correlations among clinical presentation, histological features, immunophenotype and gene rearrangement data are considered important in confirming a diagnosis of SPTL.

Adolescent ; Adult ; Antigens, CD20 ; immunology ; CD56 Antigen ; analysis ; immunology ; Child ; Female ; Gene Rearrangement ; genetics ; Humans ; Immunophenotyping ; methods ; In Situ Hybridization ; Lymphoma, T-Cell ; classification ; diagnosis ; genetics ; immunology ; Lymphoma, T-Cell, Cutaneous ; classification ; diagnosis ; genetics ; immunology ; Male ; Middle Aged ; Panniculitis ; RNA, Viral ; immunology ; isolation & purification ; Skin Neoplasms ; immunology ; Subcutaneous Tissue ; Young Adult

OBJECTIVETo study the clinicopathologic features of intravascular large B-cell lymphoma (IVLBCL).

METHODSTwo autopsy cases of IVLBCL were retrieved from the archival file. The clinicopathologic features, immunohistochemistry and molecular findings were studied.

RESULTSThe deceased were 70-year-old and 50-year-old males. Both of them had complained of a sudden onset of weakness and numbness of lower extremities. The clinical course deteriorated rapidly, with multi-organ failure. They died 85 days and 44 days after the presentation, respectively. Post-mortem examination did not reveal any mass lesion, except the presence of multiple skin and epicardium nodules, ranging from 0.5 cm to 2.5 cm in diameter, in the first patient. Pericardial effusion, ascites and pleural effusion were also observed. Histologically, neoplastic lymphoid cells filled up the small vessel lumina in many organs, including brain, hypophysis, spinal cord, spinal nerve roots, heart, lungs, kidneys, liver, spleen, digestive tract, pancreas, adrenal, thyroid, testes and lymph nodes. The tumor cells were relatively monotonous and of medium to large in size with round vesicular nuclei and 1 to 3 small basophilic nucleoli. Immunohistochemical study showed that the lymphoma cells expressed B-cell markers CD20 and CD79a, occasionally positive for CD5 and bcl-2 but negative for CD3, bcl-6, CD10, CD30, myeloperoxidase and cytokeratin. In-situ hybridization for Epstein-Barr virus-encoded RNA was negative. The proliferative index, as demonstrated by Ki-67 staining, was about 80%. Molecular study showed the presence of immunoglobulin heavy chain gene rearrangement in both cases, T-cell receptor-gamma gene rearrangement was not found.

CONCLUSIONSIVLBCL may present as neurological disturbance and carries distinctive morphologic characteristics, immunophenotype and molecular findings. The prognosis of this disease is often dismal.

Aged ; Antigens, CD20 ; analysis ; Autopsy ; B-Lymphocytes ; pathology ; virology ; CD79 Antigens ; analysis ; Herpesvirus 4, Human ; isolation & purification ; Humans ; Immunohistochemistry ; Lymphoma, B-Cell ; immunology ; pathology ; virology ; Lymphoma, Large B-Cell, Diffuse ; immunology ; pathology ; virology ; Male

OBJECTIVETo investigate the clinicopathological characteristics of primary Burkitt's lymphoma (BL) in the spermatic cord.

METHODSA case of BL of the spermatic cord was studied by histopathology and immunohistochemical techniques. The clinical data and the related literature were reviewed.

RESULTSThe patient was a 4-year-old boy, who was accidentally found with a bump in the scrotum. Surgery showed it to be a tumor located in the left spermatic cord and 5 cm x 3 cm x 2 cm in size, gray and fish-like on cross-sectional imaging. Histologically, it was characterized by monotonous infiltration of medium-sized cells with round nuclei, coarse chromatin, 2-5 basophilic nucleoli, and an appreciable rim of basophilic cytoplasm, in a typically starry-sky pattern imparted by interspersed tangible-body macrophages. Immunohistochemically, the tumor cells were diffused, positive for CD20 and CD79, some for CD10 and about 95% with the nuclear expression of Ki-67, but negative for CD3, CD43, bcl-2 and TdT as well as for EBER in situ hybridization.

CONCLUSIONPrimary spermatic cord BL is extremely rare, highly aggressive and with poor prognosis. Diagnosis of the tumor relies on its pathological characteristics and immunohistochemical staining. It is essential to differentiate BL from other types of lymphomas and malignant small-cell tumors of the non-lymphatic system.

Antigens, CD20 ; analysis ; Burkitt Lymphoma ; metabolism ; pathology ; CD79 Antigens ; analysis ; Child, Preschool ; Genital Neoplasms, Male ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; analysis ; Male ; Neprilysin ; analysis ; Spermatic Cord

BACKGROUND: The aberrant, leukemia-associated antigen expression patterns allow us to discriminate leukemic blasts from normal precursor cells. Our major goal was to determine a guideline for the detection of minimal residual disease using CD20+/CD34+ and myeloid Ag+/CD19+ combination in the bone marrow of acute leukemia in complete remission (CR) after chemotherapy. METHODS: Bone marrow samples from 117 patients with acute leukemia in complete remission after chemotherapy and from 22 healthy controls were immunophenotyped by triple staining and measured by flow cytometry. RESULTS: The CD20+/CD34+ cells in the large lymphocyte gate (R1) ranged from 0% to 3.24% (0.8+/-0.82%, P=0.000) in CD20+/CD34+ B-lineage ALL CR (N=31), from 0.03% to 4.2% (0.7+/-0.83%, P=0.000) in CD20-/CD34- B-lineage ALL CR (N=66), from 0.1% to 0.96% (0.45+/-0.32%, P=0.016) in T-ALL CR (N=10), and from 0.02% to 0.48% (0.18+/-0.15%, P=0.776) in AML CR (N=10). The CD13,33+/CD19+ cells in R1 gate ranged from 0% to 2.69% (0.37+/-0.48%, P<0.001) in CD13,33+/CD19+ B-lineage ALL CR (N=31), from 0% to 1.8% (0.31+/-0.28%, P<0.001) in CD13,33-/CD19+B-lineage ALL CR (N=65), from 0.02% to 0.64% (0.29+/-0.22%, P=0.071) in T-ALL CR (N=9), and from 0% to 0.17% (0.07+/-0.09%, P=0.341) in AML CR (N=3). CONCLUSIONS: Using an immunophenotypic method for the detection of early relapse or minimal residual disease of B-lineage ALL bone marrow in CR after chemotherapy, different cutoff values should be applied according to antigen combination and gating. When the proportion of aberrant antigen combination was less than 5% in large lymphocyte gate, the results should be interpreted with caution.
Acute Disease ; Antigens, CD/*metabolism ; Antigens, CD19/metabolism ; Antigens, CD20/metabolism ; Antigens, CD34/metabolism ; Antigens, Differentiation, Myelomonocytic/analysis/metabolism ; Bone Marrow Cells/*classification/metabolism ; Flow Cytometry ; Hematopoietic Stem Cells/classification/metabolism ; Humans ; Immunophenotyping ; Leukemia/*diagnosis/drug therapy ; Leukemia, Myeloid, Acute/diagnosis/drug therapy ; Neoplasm, Residual ; Remission Induction ; Tumor Markers, Biological/immunology