<b>OBJECTIVEb>To detect potential mutation of the WAS gene in a Chinese family affected with Wiskott-Aldrich syndrome.

<b>METHODSb>Peripheral blood samples were collected from the proband and his family members. All exons and flanking regions of the WAS gene were subjected to PCR amplification - Sanger sequencing as well as restriction endonuclease analysis. Plasma level of B-cell activating factor (BAFF) was also determined for all family members.

<b>RESULTSb>A hemizygous mutation (c.257G>A) of the WAS gene was identified in all patients from the family, for which the patient's mother was heterozygous. The same mutation was not found among healthy members of the family. Compared with unaffected members, all patients had a higher level of BAFF.

<b>CONCLUSIONb>The c.257G>A mutation of the WAS gene probably underlies the Wiskott-Aldrich syndrome in this family.

B-Cell Activating Factor ; blood ; Child, Preschool ; Heterozygote ; Humans ; Male ; Mutation ; Wiskott-Aldrich Syndrome ; genetics ; Wiskott-Aldrich Syndrome Protein ; genetics

Obesity is recognized as a chronic low-grade inflammatory state due to adipose tissue expansion being accompanied by an increase in the production of proinflammatory adipokines. Our group is the first to report that B-cell-activating factor (BAFF) is produced from adipocytes and functions as a proinflammatory adipokine. Here, we investigated how loss of BAFF influenced diet-induced obesity in mice by challenging BAFF-/- mice with a high-fat diet for 10 weeks. The results demonstrated that weight gain in BAFF-/- mice was >30% than in control mice, with a specific increase in the fat mass of the subcutaneous region rather than the abdominal region. Expression of lipogenic genes was examined by quantitative real-time PCR, and increased lipogenesis was observed in the subcutaneous adipose tissue (SAT), whereas lipogenesis in the epididymal adipose tissue (EAT) was reduced. A significant decrease in EAT mass resulted in the downregulation of inflammatory gene expression in EAT, and more importantly, overall levels of inflammatory cytokines in the circulation were reduced in obese BAFF-/- mice. We also observed that the macrophages recruited in the enlarged SAT were predominantly M2 macrophages. 3T3-L1 adipocytes were cultured with adipose tissue conditioned media (ATCM), demonstrating that EAT ATCM from BAFF-/- mice contains antilipogenic and anti-inflammatory properties. Taken together, BAFF-/- improved systemic inflammation by redistributing adipose tissue into subcutaneous regions. Understanding the mechanisms by which BAFF regulates obesity in a tissue-specific manner would provide therapeutic opportunities to target obesity-related chronic diseases.
3T3-L1 Cells ; Adipocytes/drug effects/metabolism ; Adiposity/*genetics ; Animals ; B-Cell Activating Factor/*genetics ; Cells, Cultured ; Culture Media, Conditioned/pharmacology ; Diet, High-Fat ; Disease Models, Animal ; Gene Knockout Techniques ; Inflammation/*genetics ; Lipogenesis/genetics ; Macrophages/metabolism ; Male ; Mice ; Mice, Knockout ; Obesity/*etiology

Skeletal fluorosis is a chronically metabolic bone disease with extensive hyperostosis osteosclerosis caused by long time exposure to fluoride. Skeletal fluorosis brings about a series of abnormal changes of the extremity, such as joint pain, joint stiffness, bone deformity, etc. Differentiation and maturation of osteoblasts were regulated by osteoclasts via Sema4D/Plexin-B1 signaling pathway. Furthermore, the differentiation and maturation of osteoclasts are conducted by osteoblasts via RANKL/RANK/OPG pathway. Both of these processes form a feedback circuit which is a key link in skeletal fluorosis. In this study, an osteoblast-osteoclast co-culture model in vitro was developed to illustrate the mechanism of skeletal fluorosis. With the increase of fluoride concentration, the expression level of Sema4D was decreased and TGF-β1 was increased continuously. OPG/RANKL mRNA level, however, increased gradually. On the basis of that, the inhibition of Sema4D/Plexin-B1/RhoA/ROCK signaling pathway caused by fluoride promoted the level of TGF-β1 and activated the proliferation of osteoblasts. In addition, osteroprotegerin (OPG) secreted by osteoblasts was up-regulated by fluoride. The competitive combination of OPG and RANKL was strengthened and the combination of RANKL and RANK was hindered. And then the differentiation and maturation of osteoclasts were inhibited, and bone absorption was weakened, leading to skeletal fluorosis.
Animals ; Antigens, CD ; genetics ; metabolism ; Cell Proliferation ; drug effects ; Feedback, Physiological ; Fetus ; Fluorides ; pharmacology ; GTPase-Activating Proteins ; genetics ; metabolism ; Gene Expression Regulation, Developmental ; Osteoblasts ; drug effects ; metabolism ; pathology ; Osteoclasts ; drug effects ; metabolism ; pathology ; Osteogenesis ; drug effects ; genetics ; Osteoprotegerin ; genetics ; metabolism ; RANK Ligand ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Receptor Activator of Nuclear Factor-kappa B ; genetics ; metabolism ; Receptors, Cell Surface ; genetics ; metabolism ; Semaphorins ; genetics ; metabolism ; Signal Transduction ; Transforming Growth Factor beta1 ; genetics ; metabolism ; rho-Associated Kinases ; genetics ; metabolism ; rhoA GTP-Binding Protein ; genetics ; metabolism

<b>OBJECTIVEb>To explore the regulating effect of miR-202 on B cell-activating factor, and check whether the regulation influences the growth of multiple myeloma cells.

<b>METHODSb>The potential binding sites of BAFF for miR-202 were predicted using bioinformatics software. Luciferase reporter gene analysis was used to evaluate the regulatory effect of miR-202 on BAFF. Human multiple myeloma U266 cells were transfected with has-miR-202-mimics, has-miR-202-inhibitor, siBAFF and their negative controls, respectively. After above treatments, BAFF mRNA and protein levels were detected by real-time PCR and Western blot analysis, and the proliferation and apoptosis in the multiple myeloma (MM) cells were examined by WST-1 and annexin V-FLUOS assay, respectively.

<b>RESULTSb>The BAFF mRNA expression levels in the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 1.040 ± 0.057, 0.573 ± 0.073, 1.205 ± 0.097 and 0.368 ± 0.052, respectively. BAFF mRNA expressions in U266 cells transfected with has-miR-202-3P-mimics and siBAFF were significantly decreased compared with that in the untransfected group (P < 0.05). The BAFF protein expression level of each group was consistent with the mRNA assay result. The absorbance value in 450 nm of the untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 1.063 ± 0.052, 0.714 ± 0.045, 0.936 ± 0.066 and 0.764 ± 0.053, respectively. In comparison with the untransfected group, the absorbance value at 450 nm of has-miR-202-3P-mimics and siBAFF transfected groups was significantly reduced (P < 0.05). The cell apoptosis rates of untransfected group, has-miR-202-3P-mimics transfected group, has-miR-202-3P-inhibitor transfected group and siBAFF transfected group were 26.2%, 49.6%, 21.1% and 30.7%, respectively. Therefore, the cell apoptosis rate of has-miR-202-3P-mimics transfected group was significantly increased than that of the untransfected group (P < 0.05). p-JNK protein expression level was decreased in the has-miR-202-3P-mimics transfected cells.

<b>CONCLUSIONSb>MiR-202 can inhibit the proliferation and induce apoptosis in MM cells via regulating BAFF. JNK/SAPK signaling pathway is involved in the regulation of BAFF by miR-202.

Apoptosis ; B-Cell Activating Factor ; genetics ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; HEK293 Cells ; Humans ; Luciferases ; metabolism ; MAP Kinase Signaling System ; MicroRNAs ; genetics ; metabolism ; Multiple Myeloma ; metabolism ; pathology ; Plasmids ; RNA, Messenger ; metabolism ; Transfection

3T3-L1 adipocytes express the B-cell-activating factor (BAFF) and three different BAFF receptors (BAFF-Rs). Furthermore, BAFF expression is regulated by inflammatory modulators, such as tumor necrosis factor-alpha and rosiglitazone. Here we investigated the function of BAFF in 3T3-L1 adipocytes and RAW 264.7 macrophages. We examined adipokine expression in 3T3-L1 adipocytes treated with 10 ng ml-1 BAFF. We also examined inflammatory molecule expression in RAW 264.7 macrophages treated with 10 or 100 ng ml-1 BAFF. We examined BAFF expression in the coculture of 3T3-L1 adipocytes and RAW 264.7 macrophages, as well as in white adipose tissue (WAT) of diet-induced obese (DIO) mice. We found that BAFF decreases leptin and adiponectin expression, but increases the expression of proinflammatory adipokines monocyte chemotactic protein-1, interleukin-6 (IL-6), cyclooxygenase-2 (COX-2) and haptoglobin. Coculturing the two cell types resulted in increased BAFF mRNA and protein expression, as well as modulation of BAFF-R mRNA expression in both cell types. These data indicate that BAFF might mediate adipocyte and macrophage interaction. When RAW 264.7 macrophages were treated with BAFF, BAFF-R expression was modulated as in coculture, and nitric oxide synthase and IL-6 expression increased. BAFF expression also increased in WAT of DIO mice. We propose that BAFF can regulate adipokine expression and possibly mediate adipocyte and macrophage interaction.
3T3-L1 Cells ; Adipocytes/drug effects/*metabolism ; Adipokines/genetics/*metabolism ; Adiponectin/genetics/metabolism ; Animals ; B-Cell Activating Factor/*metabolism/pharmacology ; Chemokine CCL2/genetics/metabolism ; Coculture Techniques ; Gene Expression Regulation/drug effects ; Haptoglobins/genetics/metabolism ; Inflammation Mediators/metabolism ; Interleukin-6/genetics/metabolism ; Leptin/genetics/metabolism ; Macrophages/drug effects/*metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Obese ; RNA, Messenger/genetics/metabolism

B cells play an important role in the pathogenesis of rheumatoid arthritis (RA). High levels of B cell activating factor (BAFF) are detected in autoimmune diseases. BAFF and BAFF receptor (BAFF-R) are expressed in B and T cells of RA synovium. The study was undertaken to identify the NF-kappaB signal pathway involved in the induction of BAFF-R in human B cells. Immunohistochemical staining of NF-kappaB p65, NF-kappaB p50, BAFF, and BAFF-R was performed on sections of synovium from severe and mild RA and osteoarthritis (OA) patients. Peripheral blood mononuclear cells (PBMCs) were isolated from control and RA patients and B cells were isolated from controls. BAFF-R was analyzed by flow cytometry, realtime PCR and confocal staining after treatment with NF-kappaB inhibitors. NF-kappaB p65, NF-kappaB p50, BAFF, and BAFF-R were highly expressed in severe RA synovium relative to mild RA synovium or OA synovium. BAFF-R expression was reduced by NF-kappaB inhibitors in PBMCs and B cells from normal controls. We also showed reduction in expression of BAFF-R via inhibition of the NF-kappaB pathway in PBMCs of RA patients. BAFF/BAFF-R signaling is an important mechanism of pathogenesis in RA and that BAFF-R reduction by NF-kappaB blocking therapy is another choice for controlling B cells in autoimmune diseases such as RA.
Arthritis, Rheumatoid/genetics/*metabolism/pathology/physiopathology ; B-Cell Activating Factor/genetics/metabolism ; B-Cell Activation Factor Receptor/genetics/*metabolism ; B-Lymphocytes/*drug effects/immunology/metabolism/pathology ; Cell Separation ; Cells, Cultured ; Disease Progression ; Enzyme Inhibitors/pharmacology ; Flow Cytometry ; Gene Expression Regulation/immunology ; Humans ; Immunohistochemistry ; NF-kappa B/*metabolism ; Signal Transduction/immunology ; Synovial Membrane/*pathology ; T-Lymphocytes/drug effects/immunology/metabolism/pathology ; Transcriptional Activation/drug effects

This study was aimed to explore the expression of B cell-activating factor of TNF family (BAFF) and B cell lymphoma/leukemia-2 (BCL-2) in bone marrow mononuclear cells (BMMNC) of multiple myeloma (MM) and the significance of BAFF and BCL-2 for occurrence, development and prognosis of MM. The bone marrow of 40 cases of MM and 10 healthy persons was collected, the mononuclear cells (MNC) were isolated, the expression of BAFF and BCL-2 mRNA in BMMNC was detected by real-time PCR; the plasma was simultaneously collected and the β2-MG level was determined; the clinical staging of MM patients was performed according to Durie-Salmon (D-S) staging criterion. The results indicated that the expression level of BAFF and BCL-2 mRNA in MM patients increased, as compared with normal controls, the difference was statistical significant (p < 0.05); the expression level of BAFF and BCL-2 mRNA in plateau stage after treatment obviously decreased. The expression level of BAFF and BCL-2 mRNA in relapsed/refractory MM patients was significantly higher than that in normal controls and patients reached plateau stage (p < 0.05), there was no statistically significant difference between newly diagnosed and relapsed/refractory MM patients (p > 0.05). The expression of BAFF and BCL-2 mRNA related with D-S staging and β2-MG level. It is concluded that the expression levels of BAFF and BCL-2 mRNA increase, moreover the expression levels of BAFF and BCL-2 mRNA in newly diagnosed and relapsed/refractory MM patients are higher than those in patients reached plateau stage, which suggest the BAFF and BCL-2 may be involved in occurrence and development of MM; the relation of expression level of BAFF and BCL-2 mRNA to MM load is positive, which indicates the expression level of BAFF and BCL-2 mRNA may be a new indicator for evaluating the prognosis of MM patients.
Adult ; Aged ; B-Cell Activating Factor ; genetics ; metabolism ; Case-Control Studies ; Female ; Humans ; Lymphoma, B-Cell ; genetics ; Male ; Middle Aged ; Multiple Myeloma ; diagnosis ; genetics ; Prognosis ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; genetics

The study was purposed to detect BAFF/APRIL gene expression changes in bone marrow mononuclear cells (BMMNC) and myeloma cell line U266 after interference with glucocorticoid and bortezomib. After separation of BMMNC from 7 patients with multiple myeloma, BAFF/APRIL mRNA expression in BMMNC and U266 cell line was detected by real-time PCR after treated with dexamethasone 100, 200 µg/ml, methylprednisolone 100, 200 µg/ml, bortezomib 0.1 µg/ml alone and dexamethasone or methylprednisolone combined with bortezomib respectively for 48 hours. The results showed that U266 cells and BMMNC of untreated MM patients highly expressed BAFF/APRIL genes. When dexamethasone, methylprednisolone or bortezomib was added to U266 cells or BMMNC alone, BAFF/APRIL gene expression decreased as compared with the blank control (p < 0.01). The inhibiting effect of bortezomib to BAFF/APRIL expression was obviously strong(p < 0.05). When dexamethasone or methylprednisolone combined with bortezomib, the BAFF/APRIL gene expression further decreased compared with dexamethasone or methylprednisolone alone (p < 0.01). As compared with the group of methylprednisolone combined with bortezomib, BAFF/APRIL gene expression decreased in dexamethasone combined with bortezomib with a statistically significant difference (p < 0.05). It is concluded that the expression of BAFF/APRIL gene is down-regulated after bing treated with glucocorticoids and bortezomib, which suggests that besides the glucocorticoid receptor and proteasomes targets, BAFF/APRIL and their receptor sites may be new targets of glucocorticoids and bortezomib.
B-Cell Activating Factor ; genetics ; metabolism ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; drug effects ; Glucocorticoids ; pharmacology ; Humans ; Multiple Myeloma ; metabolism ; Pyrazines ; pharmacology ; RNA, Messenger ; genetics ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; genetics ; metabolism

<b>OBJECTIVEb>To detect the B cell activating factor (BAFF) and explore its significance in patients with warm autoimmune hemolytic anemia (WAIHA).

<b>METHODSb>The levels of serum soluble BAFF (sBAFF) and BAFF mRNA in peripheral blood mononuclear cells (PBMCs) in 30 healthy volunteers (control group) and 43 patients with WAIHA were measured by ELISA and real-time quantitative polymerase chain reaction (RT-qPCR) respectively.

<b>RESULTSb>The levels of serum sBAFF and BAFF mRNA in PBMCs in pretreatment group \[2311 (825 approximately 6523) ng/L and 884 (463 approximately 2346) ng/L\] was significanly higher than those in posttreatment group\[1205(358 approximately 5014) ng/L and 446(138 approximately 2699) ng/L\] and control group\[1128 (590 approximately 3201) ng/L and 341 (102 approximately 965) ng/L\] (both P < 0.01), the difference between the posttreatment group and control group was not statistically significant. There was no significant difference between therapy responsive and nonresponsive groups before treatment. There was a significant difference between the pre- and post-treatment resuets in responsive group (P < 0.01), but not in nonresponsive group (P > 0.05). The serum levels of sBAFF was positively correlated with the levels of the BAFF mRNA in PBMCs both in pre- and post therapy group (both P < 0.05).

<b>CONCLUSIONb>The levels of serum sBAFF and BAFF mRNA in PBMCs are increased in patients with WAIHA, their dynamic alterations may contribute to the development of WAIHA.

Anemia, Hemolytic, Autoimmune ; B-Cell Activating Factor ; genetics ; Humans ; Interleukin-4 ; Leukocytes, Mononuclear ; RNA, Messenger ; genetics

<b>OBJECTIVEb>To analyze the polymorphisms of B cell activating factor (BAFF) gene and the plasma levels of BAFF in patients with idiopathic thrombocytopenic purpura (ITP), and to investigate their roles in the pathogenesis of ITP.

<b>METHODSb>Alleles specific polymerase chain reaction (AS-PCR) and agarose gel electrophoresis were used to identify polymorphisms -871C/T of BAFF promotor in 133 ITP patients and 117 healthy controls. The plasma levels of BAFF were assayed by ELISA.

<b>RESULTSb>In ITP group, the frequency of C/C, C/T and T/T was 33.1%, 42.1% and 24.8%, respectively, the corresponding frequency in control group was 55.6%, 33.3% and 11.1%, respectively. The allele frequency of T in ITP and control groups was 45.9% and 27.4%, respectively. There was a significant difference in the BAFF -871C/T genotypic frequency between the ITP and control groups (P < 0.05). BAFF antigen in untreated ITP, treated patients and controls was 875.86 pg/ml, 502.59 pg/ml and 736.88 pg/ml, respectively, being also a significant difference among the three groups (P < 0.05). BAFF antigen in homozygous T/T was higher than that in homozygous C/C and heterozygous C/T, but the difference was not statistically significant (P > 0.05).

<b>CONCLUSIONSb>Over expression of BAFF may be a risk factor for ITP patients. There is a correlation of the BAFF promotor polymorphism -871C/T with ITP, but the polymorphism does not affect the expression of BAFF.

B-Cell Activating Factor ; genetics ; Gene Frequency ; Humans ; Interleukin-4 ; Polymorphism, Genetic ; Purpura, Thrombocytopenic, Idiopathic ; immunology