OBJECTIVETo investigate apoptosis in XG-7, a human myeloma cell line, induced by IL-6 deprivation and the function of three anti-apoptotic Bcl-2 proteins (Bcl-2, Bcl-kappa(L), Mcl-1) in the apoptotic process.

METHODSApoptosis in XG-7 myeloma cells induced by IL-6 withdrawal was determined by flow cytometry with propidium iodide (PI) nuclear staining. Expressions of three Bcl-2 proteins in XG-7 cells were monitored by immunoblotting assay.

RESULTSIn the absence of IL-6 for a certain time, a significant percentage of apoptiotic XG-7 cells can be observed, as well as down-regulated expression of one of the three anti-apoptotic proteins (Mcl-1) in XG-7 cells. IL-6 re-stimulation in XG-7 cells following cytokine removal up-regulated the expression of Mcl-1 and inhibited cell apoptosis.

CONCLUSIONMcl-1,instead of Bcl-2 and Bcl-kappa(L), plays an important role in IL-6 deprivation induced apoptosis in XG-7 human myeloma cells.

Apoptosis ; Humans ; Interleukin-6 ; physiology ; Multiple Myeloma ; pathology ; Myeloid Cell Leukemia Sequence 1 Protein ; Neoplasm Proteins ; analysis ; physiology ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Tumor Cells, Cultured ; bcl-X Protein

PURPOSE: Deterioration of local immunity in the adenoids may make them vulnerable to infection by microorganisms, resulting in otitis media with effusion. To determine the factors associated with this condition, we evaluated adenoid size, mucosal barrier, squamous changes of ciliated epithelium, IgA secretion, and BCL-6 expression in adenoids. MATERIALS AND METHODS: Seventeen children diagnosed with otitis media with effusion (OME group) and 20 children without any history of OME (control group) were enrolled. Their adenoids were sized by lateral view X-ray and stained with hematoxylin and eosin to detect squamous metaplasia. The adenoids were also stained with cytokeratin to evaluate mucosal barriers, and with anti- IgA antibody and anti- BCL-6 antibody to determine expression of IgA and BCL-6. RESULTS: The OME group showed greater incidence of squamous metaplasia, fewer ciliated cells, and lower expression of BCL-6 (p < 0.05 each). Deterioration of the mucosal barrier was detected in the OME group (p > 0.05). IgA secretion and adenoid size were the same for the OME and the control groups. CONCLUSION: These results suggest that increased squamous metaplasia and lower BCL-6 expression in adenoids may be associated with increased susceptibility to OME.
Adenoids/chemistry/*pathology ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin A/analysis ; Immunohistochemistry ; Keratins/analysis ; Male ; Metaplasia ; Mucous Membrane/chemistry/pathology ; Otitis Media with Effusion/metabolism/*pathology ; Proto-Oncogene Proteins c-bcl-6/*analysis

OBJECTIVETo study the clinicopathologic and immunohistochemical features of diffuse large B-cell lymphoma (DLBCL) and the significance of immunohistochemistry in diagnosis and differential diagnosis of DLBCL.

METHODS60 cases of DLBCL were studied and immunohistochemical staining for LCA, L26, BLA-36, CD30, bcl-6 were carried out with the EnVision 2 step method.

RESULTSThe age range of 76.7% (46/60) patients was 40 - 70 years. The location of the lesion includes nodal and extranodal sites. 90.0% (54/60) were in clinical stages of II (24/54), III (21/54), IV (9/54). Histopathologic morphology presented as centroblastic (88.3%, 53/60), immunoblastic (3.3%, 2/60), anaplastic large B cell type (3.3%, 2/60) and T cell rich B cell type (5.0%, 3/60). Immunostaining showed 100% (60/60) DLBCL were positive for LCA, L26, BLA36, 3.3% (2/60) DLBCL positive for CD30, 95% (57/60) expressed bcl-6 protein.

CONCLUSIONSDLBCL is an aggressive lymphoma which shows cytologic variability from case to case. The evaluation of pathologic features and immunohistochemistry in DLBCL are useful and practical for diagnostic purposes, but cannot delineate distinctive morphologic subtypes.

Adult ; Aged ; Antigens, CD20 ; analysis ; DNA-Binding Proteins ; analysis ; Female ; Humans ; Immunohistochemistry ; Ki-1 Antigen ; analysis ; Leukocyte Common Antigens ; analysis ; Lymphoma, B-Cell ; metabolism ; pathology ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Middle Aged ; Proto-Oncogene Proteins ; analysis ; Proto-Oncogene Proteins c-bcl-6 ; Transcription Factors ; analysis

OBJECTIVEGene expression profiling of diffuse large B-cell lymphoma (DLBCL) of different immunophenotypes.

METHODSThe study included 156 cases of DLBCL, which were subclassified by immunohistochemistry including CD10, bcl-6 and MUM1. Affymetrix U133 plus2.0 oligonucleotide microarrays were used to obtain differential gene expression profiling of 9 DLBCL (3 representative cases from each immunophenotypical group) and 3 tonsils. Clinical stages of all 9 lymphomas were Ann Arbor stage IV.

RESULTSThe immunohistochemistry subclassified 156 cases of DLBCL into 3 groups: CD10(+) and/or bcl-6(+), MUM1(-) (group 1); CD10(+) and/or bcl-6(+), MUM1(+) (group 2); CD10(-) and bcl-6(-), MUM1(+) (group 3). By gene expression array, 9 lymphomas and 3 tonsils were clustered in an unsupervised fashion into 4 groups (A, B, C and D), which were in accordance with the immunophenotypical groups (group 1, 2, 3 and normal). A total of 81 genes were markedly decreased and 86 genes were over-expressed in all DLBCL groups. Although Group B lymphomas showed mixed immunophenotypical features of both germinal center B-cell-like DLBCL (Group A) and activated B-cell-like lymphomas (Group C), gene profile clustering showed that Group B was dissimilar to Group A or Group C, with 45 over-expressed and 27 uniquely expressed genes.

CONCLUSIONSGene expression profiling indicates that DLBCL can be subgrouped at the molecular level and can be identified by immunophenotyping. The gene expression profile of Group B lymphomas suggests that factors other than the cell-of-origin may contribute to the pathogenesis of DLBCL.

Aged ; Cluster Analysis ; Gene Expression Profiling ; Humans ; Immunohistochemistry ; Immunophenotyping ; methods ; Interferon Regulatory Factors ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; classification ; genetics ; immunology ; pathology ; Middle Aged ; Neprilysin ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; metabolism ; Young Adult

OBJECTIVETo explore the influence of BCL6, MYC, P53 genes abnormalities can on the prognosis of diffuse large B-cell lymphoma (DLBCL), and to identify independent prognostic factors for DLBCL in order to facilitate clinical prognosis and selection of stratification treatment for the patients.

METHODSSixty five newly diagnosed DLBCL pathological specimens were collected from 2009 to 2012. Interphase fluorescence in situ hybridization technique (I-FISH) was used to detect the status of BCL6, MYC and P53 genes. Clinical factors were combined with immunohistochemical results for multiple-factor survival analysis.

RESULTSThe rates of BCL6 gene rearrangement, P53 gene deletion and MYC rearrangement were 21.5% (14/65), 35.4% (23/65) and 7.7% (5/65), respectively. BCL6 rearrangement group has obviously poorer overall survival (OS)(P< 0.05). COX proportional hazards model analysis showed that gender, BCL6 protein, BCL6 rearrangement, Ki67 index were prognosis factors independent of international prognostic index (IPI).

CONCLUSIONBCL6 can influence the prognosis of patients with DLBCL at gene and protein levels and both are independent prognostic factors for DLBCL.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; DNA-Binding Proteins ; genetics ; Female ; Gene Deletion ; Gene Rearrangement ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; diagnosis ; genetics ; mortality ; Male ; Middle Aged ; Mutation ; Prognosis ; Proto-Oncogene Proteins c-bcl-6 ; Proto-Oncogene Proteins c-myc ; genetics ; Survival Analysis ; Survival Rate ; Tumor Suppressor Protein p53 ; genetics ; Young Adult

OBJECTIVETo study the alterations of follicular T helper cells (CD4(+)CXCR5(+)Tfh cells, Tfh) on circulating T lymphocytes in children with asthma, and to study the expression of transcription regulatory factors BCL-6 and BLIMP-1 mRNA.

METHODSSixty-four children with asthma and 25 healthy controls were enrolled in this study. On the basis of the disease, the children with asthma were classified into acute phase group (n=36) and remission phase group (n=28). The flow cytometry was used to detect the proportion of CD4(+)CXCR5(+)Tfh cells on CD4(+)T lymphocytes. Real-time PCR was performed to detect the levels of BCL-6 mRNA and BLIMP-1 mRNA. The double -antibody Sandwich ELISA was used to detect plasma concentrations of total IgE, IL-2, IL-6 and IL-21.

RESULTSThe proportion of CD4(+)CXCR5(+)Tfh cells was significantly higher in the acute group than in the control group and the remission group (P<0.05). Transcription levels of BCL-6 mRNA were significantly higher, while the inhibitory factors BLIMP-1 mRNA was significantly lower in the acute group than in the remission group and control group (P<0.05). The plasma concentration of IL-6 in the acute group increased significantly compared with the control group (P<0.05). Plasma concentrations of total IgE and IL-21 increased significantly, in contrast, plasma IL-2 concentration decreased significantly in the acute group, compared with the control group and the remission group (P<0.05). Correlation analysis showed that both IL-21 and IL-6 concentrations were positively correlated with the proportion of CD4(+)CXCR5(+)Tfh cells (r=0.76, r=0.46 respectively; P<0.05), while IL-2 level was negatively correlated with the proportion of Tfh cells (r=-0.68, P<0.05).

CONCLUSIONSThe abnormal proportion of CD4(+)CXCR5(+)Tfh cells might be involved in the immunological pathogenesis of acute asthma in children. The increased expression of BCL-6 mRNA and decreased expression of BLIMP-1 mRNA as well as the alterations of plasma total IgE, cytokines IL-2, IL-6 and IL-21 in microenvironment might be account for the increased proportion of CD4(+)CXCR5(+)Tfh cells in children with acute asthma.

Asthma ; immunology ; Child ; Child, Preschool ; DNA-Binding Proteins ; genetics ; Female ; Humans ; Immunoglobulin E ; blood ; Infant ; Interleukins ; blood ; Male ; Positive Regulatory Domain I-Binding Factor 1 ; Proto-Oncogene Proteins c-bcl-6 ; RNA, Messenger ; analysis ; Receptors, CXCR5 ; analysis ; Repressor Proteins ; genetics ; T-Lymphocytes, Helper-Inducer ; immunology

Antigens ; analysis ; Cervical Intraepithelial Neoplasia ; metabolism ; Citric Acid ; Cyclin-Dependent Kinase Inhibitor p21 ; analysis ; Edetic Acid ; Formaldehyde ; Hot Temperature ; Humans ; Hydrogen-Ion Concentration ; Immunohistochemistry ; Intestinal Mucosa ; immunology ; Ki-67 Antigen ; analysis ; Microwaves ; Palatine Tonsil ; metabolism ; Proto-Oncogene Proteins c-bcl-6 ; analysis

OBJECTIVETo investigate the association of 3q27 chromosome rearrangement with bcl-6 gene amplification and the molecular classification, therapeutic efficacies, and clinical stages in diffuse large B cell lymphoma (DLBC).

METHODSThe newly invented cell microarray was used to detect 3q27 chromosome rearrangement and bcl-6 gene amplification in 60 cases of DLBCL by fluorescence in situ hybridization (FISH). The molecular classification of germinal center B-cell-like (GCB) and non-germinal center B-cell-like (non-GCB) was investigated by analyzing the expression of CD20, CD10, bcl-6 and MUM1 simultaneously by immunohistochemical S-P method and tissue microarray. The information of therapeutic efficacies and clinical stages was obtained by analyzing clinical cases. The relationships among the factors were analyzed by statistics.

RESULTSIn 60 cases of DLBCL, 48.3%(29/60) were GCB and 51.7%(31/60) were non-GCB. The 3q27 chromosome rearrangement and bcl-6 gene amplification were present in 15 and 22 cases respectively. In 15 cases with 3q27 rearrangement, bcl-6 protein expression was positive in 3(20.0%), which was significantly different from that in cases without 3q27 rearrangement (P=0.017). In 60 cases of DLBCL, bcl-6 gene amplification was present in 22 cases, in which 5(22.7%) were GCB and 17(77.3%) were non-GCB, which was significantly different from that in cases without bcl-6 gene amplification (P=0.003). In 36 cases undergoing the normal CHOP program treatment, bcl-6 gene amplification was present in 15 cases and the rates of the complete remission, partial remission and no change were 4(26.7%), 4(26.7%) and 7(46.7%) respectively, and again it was significantly different from that in cases without bcl-6 gene amplification (P=0.016). There were no statistical significances among bcl-6 gene, BCL-6 protein expression, and clinical stages. Cases with BCL-6 protein positive and negative expression were not correlated with therapeutic efficacies and clinical stages.

CONCLUSIONThere is lower expression of BCL-6 protein in cases with bcl-6 gene fragmentation. Cases with bcl-6 gene amplification are non-GCB with worse therapeutic results and later clinical stages. There may be other genes near chromosome 3q27 associated with DLBCL prognosis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; B-Lymphocytes ; metabolism ; Chromosome Aberrations ; Chromosomes, Human, Pair 3 ; genetics ; DNA-Binding Proteins ; genetics ; Female ; Gene Amplification ; Gene Expression Regulation, Neoplastic ; Germinal Center ; pathology ; Humans ; In Situ Hybridization, Fluorescence ; Lymphoma, Large B-Cell, Diffuse ; genetics ; metabolism ; pathology ; therapy ; Male ; Middle Aged ; Neoplasm Staging ; Proto-Oncogene Proteins c-bcl-6 ; Tissue Array Analysis ; Treatment Outcome

OBJECTIVETo investigate the mutation of 5' non-coding region of bcl-6 gene in germinal center B-cell (GCB) subtype of diffuse large B-cell lymphoma (DLBCL).

METHODSt(14;18) detection and immunohistochemical staining (EnVision method) were performed in 60 cases of DLBCL, which were divided into GCB and non-GCB subtypes. Polymerase chain reaction (PCR), single-strand conformation polymorphism and direct DNA sequencing were used to identify mutations in the 5' non-coding region of the bcl-6 gene.

RESULTSSeven of 60 cases showed t(14;18) translocation in the major breakpoint region. Using minimally acceptable criteria, 18 of 60 cases were probably to be germinal centre derived. Bcl-6 mutations were detected in 12 of 60 cases (20.0%) of DLBCL, with a significantly higher frequency in the GCB subgroups (7/18) than in the non-GCB subgroups (11.9%, 5/42). Bcl-6 mutations occurred most frequently in +363 and +469 sites. An association of bcl-6 mutation and GCB subgroup was obtained.

CONCLUSIONSThe 5' regulatory region of the bcl-6 gene underwent less frequent somatic hypermutation during lymphomagenesis than the results of previous reports. Bcl-6 mutation occurred mostly in the GCB subtype and detection of t(14;18) seems helpful in the classification of DLBCL.

Adult ; Aged ; Aged, 80 and over ; B-Lymphocytes ; pathology ; Chromosomes, Human, Pair 14 ; Chromosomes, Human, Pair 18 ; DNA Mutational Analysis ; DNA-Binding Proteins ; genetics ; metabolism ; Female ; Germinal Center ; pathology ; Humans ; Lymphoma, Large B-Cell, Diffuse ; genetics ; pathology ; Male ; Middle Aged ; Point Mutation ; Proto-Oncogene Proteins c-bcl-6 ; Translocation, Genetic ; Young Adult

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