Serum BAFF Concentrations Research Articles

B-lymphocyte-activating factor is a potential biomarker associated with susceptibility to Graves’ disease in Iraqi women

BackgroundB-lymphocyte-activating factor (BAFF) is a cytokine involved in regulating the development and maturation of B lymphocyte and has been shown to be up-regulated in patients with Graves’ disease (GD). However, the association of TNFSF13B variants (the gene that encodes BAFF) with the risk of GD has not been well explored. In this case–control study, the aim was to evaluate the role of BAFF, in terms of serum level and polymorphism, in the etio-pathogenesis of GD. Therefore, serum BAFF concentrations were analyzed in Iraqi women with GD and age-matched control women (n = 90 and 93, respectively) using an ELISA kit. In addition, two promoter variants of the TNFSF13B gene, rs9514827 (T > C) and rs9514828 (C > T), were genotyped using a PCR–RFLP-based assay.ResultsMedian BAFF concentrations (interquartile range) were significantly elevated in GD patients compared to controls (1525 [1327–1840] vs. 689 [585–807] pg/mL; probability [p] < 0.001). Elevated BAFF concentrations were a reliable predictor of GD as indicated by the area under the curve of 0.971. BAFF was positively correlated with triiodothyronine (correlation coefficient [rs] = 0.216; p = 0.041) and thyroxine (rs = 0.269; p = 0.01) in GD patients. Mutant alleles, rs9514827 C (odds ratio [OR] = 2.00; p = 0.008; corrected p [pc] = 0.048) and rs9514828 T (OR = 2.15; p = 0.002; pc = 0.012), as well as genotypes, rs9514827 CC (OR = 4.29; p = 0.032; pc = 0.192) and rs9514828 TT (OR = 4.57; p = 0.003; pc = 0.018), were associated with a greater risk of GD. Besides, the C-T haplotype (rs9514827-rs9514828) was also linked to an elevated risk of GD among Iraqi women (OR = 2.71; p = 0.006; pc = 0.024).ConclusionsBAFF showed up-regulated levels in the serum of women with GD. In light of this, BAFF has been proposed as a reliable prognostic biomarker for GD. Regarding its relationship to thyroid hormones, BAFF showed a positive correlation with triiodothyronine and thyroxine. Both variants (rs9514827 and rs9514828) of the TNFSF13B gene showed an association with susceptibility to GD, and rs9514828 may have up-regulatory effects on BAFF levels.

Physiological Effects of Calprotectin and B Cell Activating Factor in COVID-19 Patients

Objectives: This study set out to determine how Calprotectin and B cell activating factor contributes to early COVID-19 patient severity prediction.&#x0D; Methods: The study included 25 healthy controls and 52 patients with SARS-COV2 infection who were clinically diagnosed with COVID-19 illness and were between the ages of 23 and 35. The serum levels of CALP and BAFF were measured using the ELISA method. To gauge CRP levels, an immunoturbidometric assay was performed.&#x0D; Results: Variations in serum levels of CALP and BAFF were found to be statistically insignificant in the study (P=0.7109 &amp; P=0.7575, respectively). When compared to the control group (103.95±36.67 ng/mL; 403.03±1.03), COVID-19 patients had non-significantly raised levels of CALP and BAFF (106.5±4.67 ng/mL; 436.9±12.77 pg/mL, respectively). According to ROC curve analysis, the area under the receiver operating characteristics curve (AUC) for CALP and BAFF was (0.5170) and (0.5259), respectively. (r=0.6923; P=0.0001). There was a significant positive correlation between serum CALP and BAFF levels. The connection between serum CRP levels and CALP (r=0.3010; P=0.1271) and BAFF levels (r=0.2912; P=0.1406) was insignificantly positive.&#x0D; Conclusion: The current study's findings suggested that serum CALP and BAFF concentrations were increased in COVID-19 patients, suggesting that these inflammatory markers may be helpful indicators of the severity of COVID-19.

GSK-3 inhibitor elraglusib (9-ING-41) to enhance tumor-infiltrating immune cell activation in tumor biopsies and synergize with anti-PD-L1 in a murine model of colorectal cancer.

e15138 Background: Glycogen synthase kinase 3 (GSK-3) is a serine/threonine kinase with key roles in myriad biological processes such as tumor progression, and inhibition of GSK-3 using a novel small-molecule elraglusib has shown promising preclinical antitumor activity in multiple tumor types. Methods: We characterize the effects of elraglusib in vitro on tumor and immune cells including cell killing and cytokine profiling , in vivo in combination with checkpoint inhibitors in a syngeneic murine colon carcinoma BALB/c model using MSS cell line CT-26, and in human tumor biopsies and plasma samples from patients with refractory solid tumors of multiple tissue origins enrolled in a Phase 1 clinical trial investigating elraglusib ( NCT03678883 ).We used human plasma samples from patients treated with elraglusib, paired pre- and post-treatment tumor biopsies and performed digital spatial proteomics. Results: Elraglusib promoted immune cell-mediated tumor cell killing, enhanced tumor cell pyroptosis, decreased tumor cell NF-κB-regulated survival protein expression, and increased immune cell effector molecule secretion. Synergy was observed between elraglusib and anti-PD-L1 in an immunocompetent murine model of colorectal cancer. Murine responders had more tumor-infiltrating T-cells, fewer tumor-infiltrating Tregs, lower tumorigenic circulating cytokine concentrations, and higher immunostimulatory circulating cytokine concentrations. Murine responders had lower serum concentrations of BAFF, CCL7, CCL12, VEGF, VEGFR2, and CCL21, and higher serum concentrations of CCL4, TWEAK, GM-CSF, CCL22, and IL-12p70 as compared to non-responders. We utilized human plasma samples from patients treated with elraglusib and correlated cytokine profiles with survival. Elevated baseline plasma levels of proteins such as IL-1 β and reduced levels of proteins such as VEGF correlated with improved PFS and OS. PFS was also found to be positively correlated with elevated plasma levels of immunostimulatory analytes such as Granzyme B, IFN-γ, and IL-2 at 24 hours post-treatment with elraglusib. Several of these secreted proteins correlated with results from the in vivo study where expression of proteins such as IL-1 β, CCL22, CCL4, and TWEAK was positively correlated with improved response to therapy while expression of proteins such as BAFF and VEGF negatively correlated with response to therapy. Using paired tumor biopsies, we found that CD45+ tumor-infiltrating immune cells had lower expression of inhibitory immune checkpoints and higher expression of T-cell activation markers in post-elraglusib patient biopsies. Conclusions: These results introduce several immunomodulatory mechanisms of GSK-3 inhibition using elraglusib, providing a rationale for the clinical evaluation of elraglusib in combination with immunotherapy.

Physiological effects of Calprotectin and B Cell activating factor in COVID-19 Patients

Objectives: This study set out to determine how Calprotectin and B cell activating factor contributes to early COVID-19 patient severity prediction. Methods: The study included 25 healthy controls and 52 patients with SARS-COV2 infection who were clinically diagnosed with COVID-19 illness and were between the ages of 23 and 35. The serum levels of CALP and BAFF were measured using the ELISA method. To gauge CRP levels, an immunoturbidometric assay was performed. Results: Variations in serum levels of CALP and BAFF were found to be statistically insignificant in the study (P=0.7109 &amp; P=0.7575, respectively). When compared to the control group (103.95±36.67 ng/mL; 403.03±1.03), COVID-19 patients had non-significantly raised levels of CALP and BAFF (106.5±4.67 ng/mL; 436.9±12.77 pg/mL, respectively). According to ROC curve analysis, the area under the receiver operating characteristics curve (AUC) for CALP and BAFF was (0.5170) and (0.5259), respectively. (r=0.6923; P=0.0001). There was a significant positive correlation between serum CALP and BAFF levels. The connection between serum CRP levels and CALP (r=0.3010; P=0.1271) and BAFF levels (r=0.2912; P=0.1406) was insignificantly positive. Conclusion: The current study's findings suggested that serum CALP and BAFF concentrations were increased in COVID-19 patients, suggesting that these inflammatory markers may be helpful indicators of the severity of COVID-19.

Serum Calprotectin and B-cell activating factor are potential biomarkers for &lt;i&gt;Helicobacter pylori&lt;/i&gt; infection

Humans always mount a robust immune response to the bacterial infection caused by Helicobacter pylori, which causes various gastrointestinal tract infections. Calprotectin (CALP) and B-Cell Activating Factor (BAFF) are inflammatory biomarkers having a role in the gastrointestinal neutrophilic response to bacterial infection. The study was designed to assess serum CALP and BAFF as inflammatory biomarkers in H. pylori infection and peptic ulcer patients. The current study comprised 112 people, including 62 H. pylori-infected patients (34 men and 28 women) who were clinically diagnosed with H. pylori infection via testing positive for the H. pylori stool antigen test; they were compared to a control group of 50 healthy people (34 men and 16 women) who were age and gender- matched to H. pylori-infected patients. The serum level of CALP and BAFF were assayed using the ELISA technique. The biochemical parameters were statistically compared between patients and controls by unpaired Man-Whitney U t-test and Receiver Operating Characteristic (ROC) curve analysis. There was a significant elevation of serum CALP in H. pylori-infected patients [116.4(120.7), p=0.0132] in comparison to healthy controls [99.50(115.8)]. Similarly, there was a significant elevation of serum BAFF concentration in H. pylori-infected patients [485.7(367.1), p=0.0014] in comparison to healthy controls [444.5(513.0)]. The ROC curve analysis suggests serum CALP and BAFF as reasonable inflammatory biomarkers for H. pylori infection with statistically significant (p=0.0135, p=0.0015) area under the ROC curve of (0.6361, 0.6748), respectively. CALP and BAFF are potent inflammatory biomarkers involved in the development and etiology of H. pylori infection. Serum CALP and BAFF levels could be used as biomarkers for chronic inflammation induced by H. pylori. CALP and BAFF biomarkers can be combined to diagnose and predict the prognosis of H. pylori infection.

Serum B-cell activating factor and lung ultrasound B-lines in connective tissue disease related interstitial lung disease.

To investigate the role of serum B-cell activating factor (BAFF) and lung ultrasound (LUS) B-lines in connective tissue disease related interstitial lung disease (CTD-ILD), and their association with different ILD patterns on high resolution computed tomography (HRCT) of chest. We measured the levels of BAFF and KL-6 by ELISA in the sera of 63 CTD-ILD patients [26 with fibrotic ILD (F-ILD), 37 with non-fibrotic ILD (NF-ILD)], 30 CTD patients without ILD, and 26 healthy controls. All patients underwent chest HRCT and LUS examination. Serum BAFF levels were significantly higher in CTD patients compared to healthy subjects (617.6 ± 288.1 pg/ml vs. 269.0 ± 60.4 pg/ml, p < 0.01). BAFF concentrations were significantly different between ILD group and non-ILD group (698.3 ± 627.4 pg/ml vs. 448.3 ± 188.6 pg/ml, p < 0.01). In patients with ILD, BAFF concentrations were significantly correlated with B-lines number (r = 0.37, 95% CI 0.13-0.56, p < 0.01), KL-6 level (r = 0.26, 95% CI 0.01-0.48, p < 0.05), and Warrick score (r = 0.33, 95% CI 0.09-0.53, p < 0.01), although all correlations were only low to moderate. B-lines number correlated with Warrick score (r = 0.65, 95% CI 0.48-0.78, p < 0.01), and KL-6 levels (r = 0.43, 95% CI 0.21-0.61, p < 0.01). Patients with F-ILD had higher serum BAFF concentrations (957.5 ± 811.0 pg/ml vs. 516.1 ± 357.5 pg/ml, p < 0.05), KL-6 levels (750.7 ± 759.0 U/ml vs. 432.5 ± 277.5 U/ml, p < 0.05), B-lines numbers (174.1 ± 82 vs. 52.3 ± 57.5, p < 0.01), and Warrick score (19.9 ± 4.6 vs. 13.6 ± 3.4, p < 0.01) vs. NF-ILD patients. The best cut-off values to separate F-ILD from NF-ILD using ROC curves were 408 pg/ml for BAFF (AUC = 0.73, p < 0.01), 367 U/ml for KL-6 (AUC = 0.72, p < 0.05), 122 for B-lines number (AUC = 0.89, p < 0.01), and 14 for Warrick score (AUC = 0.87, p < 0.01) respectively. Serum BAFF levels and LUS B-lines number could be useful supportive biomarkers for detecting and evaluating the severity and/or subsets of CTD-ILD. If corroborated, combining imaging, serological, and sonographic biomarkers might be beneficial and comprehensive in management of CTD-ILD.

Low Serum BAFF Concentration Is Associated with Response to TNF Inhibitors in Seropositive Patients with Rheumatoid Arthritis.

We investigated B-cell-activating factor (BAFF) in relation to response to treatment with TNF inhibitors (TNFis) in rheumatoid arthritis (RA). This was a longitudinal study including 158 patients with RA treated with TNFis and followed up for 6 months. Clinical response at 6 months of treatment was defined according to the EULAR criteria for good responders (GRs). BAFF concentration was measured in serum samples, collected at baseline and at 6 months. Associations with EULAR response were evaluated using univariable and multivariable logistic regression models. ROC analysis was performed to determine the optimal threshold of serum BAFF concentration associated with good EULAR response to treatment. After 6 months of TNFi treatment, 24% of patients were GRs. They had a lower BMI, lower baseline DAS28 and lower baseline serum BAFF concentration than non-responders. After 6 months of TNFi treatment, autoantibody-positive patients who attained GR had significantly lower serum BAFF concentrations compared with patients who did not. Serum BAFF < 968 pg/mL at 6 months represented the concentration likely to best discriminate between GR and non-GR at 6 months of TNFi treatment. Autoantibody-seropositive patients who had serum BAFF < 968 pg/mL at 6 months demonstrated a more than four-fold increased probability to be GRs compared with patients with higher BAFF concentrations. In conclusion, serum BAFF concentrations were associated with response to TNFis in seropositive RA patients, corroborating the importance of the B-cell compartment in RA.

AB0030 THE BIOLOGICAL FUNCTION AND MECHANISM EXPLORATION OF TACI IN SJOGREN’S SYNDROME

Backgroundprimary Sjogren’s syndrome (pSS) is a chronic autoimmune disease that affects the lacrimal, salivary and other exocrine glands. More and more studies have shown that B cells play a central role in the pathogenesis of SS.ObjectivesWe intended to explore the expression of the transmembrane activator, calcium modulator and cyclophilin ligand interactor (TACI) on the B cells, its role in pSS, and possible signal transduction pathways.MethodsWe included 34 naive pSS patients who visited the rheumatology department of Peking University First Hospital, and 37 gender- and age-distribution matched healthy controls (HCs). (1) To compare the B cell subsets, expression of TACI and relevant receptors in pSS patients, peripheral blood mononuclear cells were separated to analyze the ratio of B subsets, TACI, and BAFF-R by flow cytometry; ELISA was applied to detect the serum BAFF and soluble TACI (sTACI) concentration. (2) As for functional research of TACI, CD19+ B cells separated by magnetic sorting were treated under in vitro culture circumstances with raw TACI, TACI knocked down by siRNA, and sTACI analog (telitacicept) intervention with varing doses. The apoptosis, proliferation, differentiation and regulatory capacity on T cells were analyzed by flow cytometry, inflammatory cytokines and immunoglobulin levels in the culture supernatants were detected by CBA. (3) As for TACI-associated signaling pathway exploration, based on our previous miRNA data and relevant report of high quality, miRNA associated with TACI with significantly biased expression was confirmed by RT-qPCR. Screening the target gene of the candidate miRNA, and miRNA overexpression and inhibition experiments were conducted to validate the targeted relationship in B cells. And TACI-associated signaling pathway was explored via overexpressing and inhibiting the target gene.Results(1) Compared to HC, peripheral blood B subsets of pSS patients exhibited a significant bias, manifesting as increased proportion of CD19+CD24hiCD38hi Breg and decreased ratio of CD19+CD24+CD38- memory B cells. The expression of TACI in all B subsets was down-regulated, while that of BAFF-R was up-regulated. Both the serum concentration of BAFF and sTACI in pSS patients increased significantly. (2) As for functional research of TACI, in the setting of raw TACI, B cells in the pSS group showed higher apoptosis rate than HC under culture in vitro, where the ligand of TACI (APRIL or BAFF) addition turned the rate comparable; more active proliferation, and impaired capacity of inducing Treg cells to secrete IL-10. When TACI was knocked down by 50%, B cells performed less late apoptosis, significantly increased proliferation, impaired differentiation, significant dysfunction of Breg itself and impaired induction of Treg cells to secrete IL-10. While telitacicept intervention increased early apoptosis rate of B cells, significantly inhibited proliferation in 500ng/mL group and impaired ability of Breg and Treg cells to secrete IL-10. Besides, increased TACI on B cells treated with telitacicept, decreased IgG and increased IgA in the culture supernatants were observed. (3) As for TACI-associated signaling pathway exploration, hsa-miR-30b-5p showed satisfactory correlation between both transmembrane and sTACI. Besides, the expression of hsa-miR-30b-5p was significantly down-regulated, and inhibition its expression in vitro could lead to differentiation retard, impaired secretion of IL-10 by Breg cells. SMAD1 was screened based on database and validated as its target gene by overexpressing and inhibiting hsa-miR-30b-5p in B cell. After targeted up- or down-regulating the transcription of SMAD1 further, the transcription of ID2 downstream the TGF-β/Hippo signaling pathway changed accordingly.ConclusionThe expression of TACI on peripheral blood B cells was deficient in pSS patients. TACI deficiency was closely associated with the downregulation of hsa-miR-30b-5p, activating TGF-β/Hippo pathway mediated by its target gene SMAD1 and taking part in the pathogenesis of pSS.Disclosure of InterestsNone declared

Increased B Cell-Activating Factor Expression Is Associated with Postoperative Recurrence of Chronic Rhinosinusitis with Nasal Polyps.

Background Chronic rhinosinusitis with nasal polyps (CRSwNP) is a common upper airway inflammatory disease with a high postoperative recurrence rate. This study is aimed at exploring the expression of B cell-activating factor (BAFF) in CRSwNP and its association with postoperative recurrence. Methods A total of 80 CRSwNP patients, including 40 primary CRSwNP patients and 40 recurrent CRSwNP patients, 40 chronic rhinosinusitis without nasal polyps (CRSsNP) patients, and 40 healthy controls (HC) were enrolled in this study, and the serum and tissue samples were collected. The circulating and tissue BAFF expressions were detected by enzyme-linked immunosorbent assay reverse transcription-polymerase chain reaction and immunohistochemistry. Their clinical values for predicting postoperative recurrence of CRSwNP were evaluated. Results We determined serum levels of BAFF were remarkably increased in the CRSwNP group than the CRSsNP and HC groups (P < 0.05), and higher concentrations of BAFF were associated with peripheral eosinophil percentage (r = 0.614, P < 0.001). The serum BAFF concentrations were significantly higher in the recurrent CRSwNP group in comparison with the primary group (P < 0.05). Multivariate analysis and receiver operating characteristic (ROC) curve presented that serum BAFF levels were associated with the postoperative recurrence in CRSwNP patients (P < 0.05). Moreover, tissue BAFF levels were significantly increased in the CRSwNP group than the HC group, especially in the recurrent CRSwNP group (P < 0.05), and enhanced BAFF RNA expressions were correlated with serum BAFF levels (r = 0.703, P < 0.001). Conclusion Our results elucidated that the BAFF expression was enhanced in CRSwNP patients and associated with postoperative recurrence. BAFF could be a serologic biomarker for predicting postoperative recurrence in CRSwNP patients.

Circulating BAFF as novel biomarker in distinguishing chronic rhinosinusitis with nasal polyps endotypes and predicting postoperative recurrence

BackgroundB cell-activating factor (BAFF) is a proinflammatory cytokine involved in inflammatory and allergic diseases, but its role in chronic rhinosinusitis with nasal polyps (CRSwNP) remains unclear. This study aims to explore the predictive value of circulating BAFF in CRSwNP endotypes and postoperative recurrence. MethodsWe recruited 120 CRSwNP patients, including 68 non-eosinophilic CRSwNP (neCRSwNP) patients, 52 eosinophilic CRSwNP (CRSwNP) patients, and 60 healthy controls (HCs). Circulating BAFF levels of all participants were measured by enzyme-linked immunosorbent assay (ELISA), and receiver-operating characteristic (ROC) and logistic regression analyses were applied to assess the predictive ability of BAFF levels in distinguishing CRSwNP endotypes. All CRSwNP patients were followed for more than 3 years, and the predictive value of circulating BAFF for postoperative recurrence was evaluated. ResultsSerum BAFF levels were elevated in CRSwNP patients compared with the HCs (P < 0.01) and significantly higher in eCRSwNP patients. The increased serum BAFF concentrations positively correlated with blood eosinophil counts and percentages, tissue eosinophil counts, and serum total IgE (P < 0.05). The ROC curve showed that serum BAFF exhibited strong discriminative ability for eCRSwNP. Finally, 99 CRSwNP patients completed the follow-up schedule, 65 patients were classified into non-recurrence group and the other 34 patients were categorized into recurrence group. Serum BAFF levels were significantly higher in recurrence group than non-recurrence group (P < 0.001), and the ROC curve suggested a high predictive value of serum BAFF in predicting postoperative recurrence. Moreover, logistic regression and Kaplan-Meier curves showed that serum BAFF was an independent risk factor for postoperative recurrence (P < 0.05). ConclusionOur data suggested that serum BAFF levels were upregulated in CRSwNP patients and correlated with mucosal eosinophil infiltration severity. Serum BAFF seemed to be a novel biomarker for preoperatively distinguishing CRSwNP endotypes and predicting postoperative recurrence.

Regulation of the functional activity of peripheral B lymphocytes in pregnant women with a history of recurrent miscarriage

BACKGROUND:The frequency of recurrent miscarriage is up to 5 % of all desired pregnancies and is mainly due to immunological disorders. Dysfunction in the regulation of the functional activity of B lymphocytes is the pathogenetic link in multiple obstetric complications, including habitual miscarriage.&#x0D; AIM:The aim of this study was to characterize the regulation of the functional activity of peripheral B lymphocytes in pregnant women with threatened spontaneous miscarriage and a history of habitual miscarriage.&#x0D; MATERIALS AND METHODS:We examined 88 women aged 18-40 years at a gestation period of 5-12 weeks. The main group consisted of 36 patients with threatened spontaneous miscarriage at the time of examination and a history of habitual miscarriage. The control group included 28 women with uncomplicated pregnancy. The comparison group consisted of 24 primary pregnant patients with threatened spontaneous abortion at the time of examination. BAFF and APRIL levels in the blood serum were determined by enzyme immunoassay. The content of CD19+BAFFR+B lymphocytes in the lymphocyte gate was evaluated in the peripheral blood by flow cytometry using monoclonal antibodies. Akt mRNA expression was assessed using real-time reverse-transcription quantitative polymerase chain reaction. CD19+В lymphocytes were isolated by direct magnetic separation.&#x0D; RESULTS:In the main group, there was an increase in expression of BAFF receptors on peripheral CD19+B lymphocytes and a decrease in the serum BAFF concentration compared to the parameters in the other study groups. We also found a pronounced trend towards a decrease in the serum APRIL level in the main and comparison groups of patients compared to healthy pregnant women. Besides, Akt mRNA expression in peripheral CD19+B lymphocytes was increased in the main group.&#x0D; CONCLUSIONS:Threatened habitual abortion is associated with the deficit of the regulatory influence of BAFF and APRIL, which is expressed in the disruption of B cell homeostasis and the weakening of humoral effector mechanisms.

Inhibitory effect of HTLV-1 infection on the production of B-cell activating factors in established follicular dendritic cell-like cells.

IntroductionThe low frequency of ectopic germinal center in labial salivary glands of patients with human T‐cell leukemia virus type 1 (HTLV‐1) antibody‐positive Sjögren's syndrome (SS) suggests that HTLV‐1 has some effects on follicular dendritic cells (FDCs).MethodsWe used flow cytometry, immunofluorescence, and enzyme‐linked immunosorbent assays (ELISAs) to investigate the direct effect of HTLV‐1 on B‐cell activating factors produced by established FDC like cells obtained from excised human tonsils. We then measured the serum B‐cell activating factor (BAFF) and C‐X‐C motif ligand (CXCL) 13 concentrations of the HTLV‐1‐seropositive SS patients and the HTLV‐1‐seronegative SS patients by ELISA.ResultsAmong the 31 isolated specimens, 22 showed morphological characteristics of FDCs. Day 2‐cultured specimens showed expressions of CD14, CD23, CD40, intracellular adhesion molecule‐1 (ICAM‐1), and vascular cell adhesion molecule‐1. After 2 weeks, 12 of these specimens expressed ICAM‐1, FDC, and fibroblast cell marker. Intracellular BAFF and CXCL13 were constitutively expressed regardless of stimulation. After direct coculture with the HTLV‐1‐infected T‐cell line HCT‐5 or MT‐2, the BAFF and CXCL13 expressions on the FDC‐like cells were decreased in accord with the increased number of HCT‐5 and MT‐2 cells with styliform change and without HTLV‐1 Gag protein expression. Interferons upregulated the concentration of BAFF (but not CXCL13) in the culture supernatant, which showed a declining trend under the presence of HCT‐5 or MT‐2. The serum concentrations of BAFF and CXCL13 in the HTLV‐1‐seropositive SS patients were lower than those of the HTLV‐1 seronegative SS patients.ConclusionsHTLV‐1 partially inhibited the BAFF and CXCL13 expressions of established FDC‐like cells.

Altered expression profile of BAFF receptors on peripheral blood B lymphocytes in Graves\u2019 disease

BackgroundB lymphocyte activating factor (BAFF) is a growth factor regulating B lymphocytes survival and maturation. Serum BAFF levels were elevated in patients affected with autoimmune thyroid diseases (AITD), including Graves’ disease (GD) and Hashimoto’s thyroiditis (HT). The aim of this study is to explore the association of expression levels of BAFF and its receptors with AITD.MethodsFifty-two GD patients, 39 Hashimoto’s thyroiditis (HT) patients and 23 healthy controls (HC) were recruited in this study. Serum BAFF levels were measured by ELISA. Expression of BAFF receptors, including BAFF receptor 3 (BR3) and transmembrane activator and calcium-modulating and cyclophilin ligand interactor (TACI), on B lymphocytes were analyzed by flowcytometry. Effects of steroids on serum BAFF levels and expression of BR3 and TACI were also observed in 10 patients with Graves’ orbitopathy (GO) receiving steroids therapy.ResultsSerum BAFF levels were significantly elevated from 0.93 ± 0.24 ng/ml in HC to 1.18 ± 0.33 ng/ml in GD (P = 0.0027) and 1.02 ± 0.24 ng/ml in HT (P = 0.0331). BR3 expression on peripheral B lymphocytes were elevated in GD (mean MFI: 4.52 ± 2.06 in GD vs. 3.00 ± 0.87 in HC, P = 0.0015), while TACI expression on peripheral B lymphocytes were decreased in GD without significance (mean MFI: 7.96 ± 4.06 in GD vs. 9.10 ± 3.37 in HC, P = 0.1285). Expression of BR3 and TACI was not changed significantly in HT patients. Steroids significantly suppressed serum BAFF concentrations (from 1.18 ± 0.27 ng/ml to 0.97 ± 0.10 ng/ml, P = 0.0364) and BR3 expression in GO patients (mean MFI from 6.26 ± 4.91 to 4.05 ± 1.58, P = 0.0083).ConclusionsAltered expression of BAFF and its receptor may mediate the autoimmunity in GD. Restoring the normal expression profile of receptors for BAFF could be a new strategy to treat GD.

Exploring the role of BAFF as biomarker in the detection of uveal melanoma metastases.

While B-cell activating factor (BAFF) was identified to promote the invasion in other malignancies, its role in the progression of uveal melanoma (UM) still remains unexplored. Here, we analysed the serum level of BAFF in UM patients with regard to its significance as biomarker for the metastases. In this retrospective study, serum BAFF levels in 173 UM patients (36 with metastases and 137 without), and 23 healthy controls were measured with a multiplexed sandwich ELISA system and then correlated with clinicopathological characteristics such as primary tumor size, tumor location, histological cell type, sex, cancer stage, cytogenetic alterations of chromosome 3, and the metastatic burden. Immunohistochemical staining of 50 UM tissue specimens was also performed to evaluate the expression of BAFF and its receptors BAFF-R and TACI. The metastatic patients were identified to have significantly higher serum BAFF levels (mean ± SD, 1520.8 ± 1182.1pg/ml) than those without metastases (950.4 ± 494.6pg/ml) and controls (810.3 ± 140.5pg/ml). While no distinctions were detected with regard to tumor location, histological cell type, gender, and monosomy 3, the patients in cancer stages II, III, and IV displayed higher serum BAFF levels than those in stage I. The serum BAFF level was significantly correlated with the metastatic burden. The serum BAFF level of 1120pg/ml was identified to have the best performance for distinguishing the metastatic patients from non-metastatic patients. In the kinetic study, we noticed that 20.8% of the analysed patients already demonstrated elevated serum BAFF concentrations before the clinical diagnosis of metastases. Positive BAFF staining was detected in the cytoplasm of single tumor cells (in 13 specimens), macrophages (in 12 specimens), and tumor-infiltrating lymphocytes (TILs) (in 13 specimens). The expressions of BAFF-R and TACI were also observed in 17 and 36 of the 50 tested UM specimens, respectively. Our study first suggests that BAFF might be a promising serum marker for the detection of UM metastases.

Change of serum B-cell activating factor level in patients with positive antiphospholipid antibodies and previous adverse pregnancy outcomes and its significance.

Background:B-cell activating factor (BAFF) is vital for B cell survival. Serum BAFF levels are elevated in thrombotic antiphospholipid syndrome, but little is known about levels in patients with positive antiphospholipid antibodies (aPLs) and previous adverse pregnancy outcomes (APOs). We aimed to analyze serum BAFF concentrations of these patients in early pregnancy along with different pregnancy outcomes.Methods:Thirty-six pregnant patients positive for aPLs and previous APOs (patient group), 25 healthy pregnant females (HP group) and 35 healthy non-pregnant females (HNP group) from the Peking University Third Hospital, between October 2018 and March 2019, were enrolled in this study. Serum of HNP and serum of patients as well as HP in the first gestational trimester were collected. Enzyme-linked immunosorbent assay kits were used to measure serum BAFF and interferon-alpha (IFN-α) concentrations. Cytometric bead array analysis was used to measure serum concentrations of cytokines. The patient group was further divided into APOs and non-APOs (NAPOs) group, fetal loss and live birth group according to pregnancy outcomes. The Mann-Whitney U-test was used to assess significance between and within groups. Spearman rank-order was used to evaluate correlation coefficients between BAFF and related cytokines.Results:The serum BAFF level in HP group was significantly lower than HNP group (245.24 [218.80, 265.90] vs. 326.94 [267.31, 414.80] pg/mL, Z = −3.966, P < 0.001). The BAFF level was obviously elevated in patient group compared to that in HP group (307.77 [219.86, 415.65] vs. 245.24 [218.80, 265.90] pg/mL, Z = −2.464, P = 0.013). BAFF levels in APOs group tended to be higher than that in NAPOs group (416.52 [307.07, 511.12] vs. 259.37 [203.59, 375.81] pg/mL, Z = −2.718, P = 0.006). Compared to HP group, concentrations of IFN-α, interleukin (IL-6) and tumor necrosis factor were higher in patient group (33.37 [18.85, 48.12] vs. 13.10 [6.85, 25.47] pg/mL, Z = −2.023, P = 0.043; 39.16 [4.41, 195.87] vs. 3.37 [2.92, 3.90] pg/mL, Z = −3.650, P < 0.001; 8.23 [2.27, 64.46] vs. 1.53 [1.25, 2.31] pg/mL, Z = −3.604, P < 0.001, respectively). Serum BAFF levels had a positive correlation with the concentrations of both IL-6 and IL-10 (IL-6: r = 0.525, P = 0.002; IL-10: r = 0.438, P = 0.012).Conclusions:Serum BAFF levels are increased in patients with positive aPLs and previous APOs as compared to healthy pregnant females and tend to be higher in individuals with current APOs. The BAFF levels have a positive correlation with serum IL-6 and IL-10.

Usefulness of rheumatoid factor as an immunological and prognostic marker in PSS patients

IntroductionThe rheumatoid factor (RF) is present in numerous autoimmune disorders, although its role in many of them remains a subject of research. The study assesses the role of RF as an immunological and prognostic factor in the primary Sjögren’s syndrome (pSS).MethodsSeventy-five pSS patients (mean age 50.03 ± 15.1), 65 (87%) females, and 10 (13%) males. WBC, CRP, RF, ESR, gammaglobulins, C4, C3 component of complement, cryoglobulins, ANA, anti-SS-A, and anti-SS-B antibodies were determined. The disease activity assessed with ESSDAI. Minor salivary gland biopsy (focus score and immunochemistry) was conducted. Results were analyzed with U Mann-Whitney (continuous variables) tests, correlations between quantitative variables assessed with the Spearman correlation coefficient with statistical significance set at p < 0.05. The approval of the Bioethics Committee was obtained.ResultsTwo subgroups I-RF(+) (61%) and II-RF(−) (39%) were established, with lower WBC (p = 0.012) and higher ESR (p = 0.016), gammaglobulin concentration (p = 0.007) in group I. Conjunctivitis sicca was more severe in group I. There was positive correlation between RF and lnANA (rho = 0.496), anti-SS-A, anti-SS-B antibodies (rho = 0.448; rho = 0.397 respectively). There was higher disease activity ESSDAI in group I than in group II (Me, 3.0 vs 2.0; p < 0.003). RF correlated negatively with WBC (rho = − 0.374). RF did not correlate with serum concentrations of BAFF, APRIL, CRP, and C3, C4 and with CD19+, CD3+, CD4+, CD 21+, and CD35+.ConclusionsRF should be considered as a prognostic, but not diagnostic, factor in patients with pSS, as it is associated with more severe disease course (sicca eye symptoms, ESSDAI) and parameters (production of gammaglobulins, ANA, anti SS-A, anti-SS-B autoantibodies) indicating increased B cell activity.

Depletion of B cell-activating factor attenuates hepatic fat accumulation in a murine model of nonalcoholic fatty liver disease

Obesity-induced adipose-tissue dysfunction is a critical contributor to the pathogenesis of nonalcoholic fatty liver disease (NAFLD). B cell-activating factor (BAFF) is an adipokine related to impaired insulin sensitivity, and the serum BAFF concentration is associated with NAFLD severity. In this study, we aimed to determine the direct in vivo role of BAFF in the development of insulin resistance, adipocyte dysfunction, and hepatic steatosis using BAFF−/− mice fed a high-fat diet (HFD). HFD-fed BAFF−/− mice exhibited significantly improved insulin sensitivity despite their increased weight gain and adiposity relative to HFD-fed wild-type mice. Moreover, inflammation, especially the accumulation of CD11c+ adipose-tissue macrophages, and fibrosis of epididymal adipose tissue were reduced, contributing to healthy adipose-tissue expansion in obese BAFF−/− mice. In line with metabolically healthy obesity, hepatic steatosis also decreased, and we observed attenuated de novo lipogenesis in both the livers and hepatocytes of BAFF−/− mice. Our data revealed that BAFF serves as a potential stimulator of unhealthy adipose-tissue expansion by triggering inflammation and fibrosis and ultimately leading to enhanced insulin resistance and NAFLD. Therefore, these results suggest that BAFF is a promising target for diabetes and NAFLD treatment.

B cells in Graves' Orbitopathy: more than just a source of antibodies?

B cells have multiple actions on different phases of an immune reaction, mainly resulting in B and T cell-interaction (help), production of cytokines, regulation of dendritic cells and downregulation of regulatory B cells. The effectiveness of B cell depletion therapy is probably due to blockade of the antigen-presenting function of B cells, occurring very early in the setting of autoimmune reactions. B cells undergo a maturation process from stem cells during which the CD 20 antigen, which is the target of rituximab (RTX), is expressed from the stage of pre-B cells to mature and memory B cells, but not on plasma cells. During the maturation process, the cytokine B cell stimulating factor (BAFF) induces maturation of B cells and expansion of clones to produce plasma cells and eventually antibodies. The effect of RTX in GO is rather rapid, with significant improvement of the disease already 4-6 weeks after the first RTX infusion. Based on the evidence of significant lymphocytic infiltration in the orbits of patients with active GO, it is reasonable to postulate that RTX may cause depletion of B cells and block their antigen-presenting cell mechanism. Since it has been reported that serum BAFF concentrations are elevated in hyperthyroid GD patients and that BAFF is expressed on the thyrocytes of patients with either autoimmune disease or nodular goiter, the hypothesis that belimumab, an anti-BAFF monoclonal antibody, may be effective in patients with active GO his currently being tested in a randomized controlled trial.

BAFF-R and TACI expression on CD3+ T cells: Interplay among BAFF, APRIL and T helper cytokines profile in systemic lupus erythematosus

BackgroundSystemic lupus erythematosus (SLE) is the prototype of systemic autoimmune disease, characterized by loss of immune tolerance against self-antigens where autoantibody production is the hallmark of disease. B-cell-activating factor (BAFF) and A proliferation-inducing ligand (APRIL) are cytokines that promote autoreactive cell survival, immunoglobulin-class switching and autoantibody responses in human and mouse SLE models. BAFF and APRIL exert their functions through interactions with their receptors BAFF-R and TACI that are differentially expressed in B lymphocyte subsets, monocytes, dendritic cells and T lymphocytes. BAFF stimulation favors T lymphocyte activation and cytokine production through BAFF-R, which could contribute to the Th1, Th17 and/or Th2 response dysregulation observed in SLE patients. ObjectiveTo evaluate the expression of the cytokines BAFF and APRIL and their association with the receptors BAFF-R and TACI on CD3+ T cells and to evaluate Th1/Th2/Th17 cytokine profile in patients with SLE. MethodsFifteen healthy controls (HC) and 36 SLE patients were included, and their demographic and clinical data were assessed. The disease activity index (Mex-SLEDAI) and damage index (SLICC) were applied to the SLE patients. BAFF-R and TACI expression on CD3+ T cells were evaluated by flow cytometry. Serum BAFF and APRIL concentrations were measured by enzyme-linked immunosorbent assays (ELISA). Cytokine levels of Th1 (IL-12, IL-2, IFN-γ, TNF-α), Th2 (IL-4, IL-6, IL-10, IL-13) and Th17 (IL-1β e IL-17) were quantified with a multiplex assay (MAGPIX). Statistical analysis was performed using PASW Statistics v.20 and GraphPad Prism v.6 software. ResultsNo differences in BAFF-R or TACI expression on the CD3+ T cells of SLE and HC were observed. BAFF-R expression correlates inversely with disease activity (r = −0.538, p < 0.01), while TACI correlates with disease activity (r = 0.530, p < 0.05). Serum BAFF and APRIL levels were high in SLE patients and correlated with the disease activity index Mex-SLEDAI (r = 0.621, p < 0.01 and r = 0.416, p < 0.05). SLE patients were found to have significantly higher levels of IL-12, IFN-γ, TNF-α, IL-6, IL-10, IL-13, IL-1β and IL-17 compared to HC (p < 0.05). Cytokines IL-17 (r = 0.526) and TNF-α (r = 0.410) correlate with disease activity (p < 0.05), while APRIL (r = 0.477), IL-10 (r = 0.426) and IFN-γ (r = 0.440) levels were associated with organ damage (p < 0.01). Serum BAFF expression levels correlate with IL-4 (r = 0.424; p < 0.05), IL-6 (r = 0.420; p < 0.05) and IL-10 (r = 0.459; p < 0.01), whereas APRIL levels correlate with IL-2 (r = 0.666; p < 0.01), IL-12 (r = 0.611; p < 0.01) and TNF-α (r = 0.471; p < 0.05) cytokines. A subgroup of SLE patients with high serum BAFF levels (>2 ng/mL) also showed increased APRIL, IL-2, IL-6 and IL-10 levels (p < 0.05). Finally, BAFF, IL-4 and TNF-α serum levels were associated with high titers of antinuclear antibodies. ConclusionsThe study demonstrates an imbalance in the Th1/Th2 cytokine profile, with increased proinflammatory cytokines, as well as BAFF and APRIL serum levels. Associations of BAFF with Th2 profile cytokines and disease activity, as well as APRIL with Th1 profile cytokines and organ damage, suggest that BAFF and APRIL generated in the autoimmunity context could through still unknown mechanisms, modulate the microenvironment, and perpetuate the inflammatory response, autoantibody production and organ damage observed in SLE patients.

B-Cell Activating Factor (BAFF) and Its Receptors in Systemic Lupus Erythematosus, Rheumatoid Arthritis

Objective: The aim of this study was to determine and compare the serum level of the BAFF and the level of its three receptors in patients with RA and SLE and also to correlate its serum level with disease activity indices and severity. Patients and methods: This study included 60 {RA (n=30), SLE (n=30)} Egyptian patients with collagen diseases from outpatient clinics of the Internal Medicine and Rheumatology Departments of Banha University Hospitals between December 2014 and December 2015 as well as 25 apparently healthy individuals who were either on the medical staff or were patient’s relatives that served as the healthy controls. All participants underwent history taking, clinical examination, laboratory and radiological investigations, and disease activity score estimation. The serum BAFF concentration, the serum TACI concentration, serum BAFFR concentration, serum BCMA concentration were determined with a quantitative sandwich enzyme immunoassay (ELISA) kits. Results: BAFF, BAFF-R, & TACI levels showed statistically significant increase in SLE & RA Groups as compared to control group (P<0.05) and BCMA level was statistically significant decrease in SLE & RA Groups as compared to control group (P<0.05). Further, BAFF, BAFF-R levels were statistically significantly increased in RA Group when compared to SLE Group. A significant positive correlation was found between the BAFF level and BAFF-R levels in both groups and SLE (r=0.854, p<0.001), RA (r=0.614, p<0.001), also between BAFF, BAFF-R and SLEDAI in SLE group (r=0.413, 0.419, p<0.023, 0.021) respectively, and between BAFF, BAFF-R and DAS28 in RA group (r=0.642, 0.430, p<0.001,0.018) respectively. The current study raveled that the best biomarker for diagnosis of RA &SLE were BAFF-R & BAFF as they had the Roc curve with more sensitivity and specificity. Conclusion: The BAFF serum levels and its three receptors levels are increased in patients with SLE and RA versus the controls. They also have a positive correlation with disease severity in SLE and RA, which suggests that BAFF may play a role in the pathogenesis and activity of these diseases. These results may pose the possibility that a human monoclonal antibody drug which selectively inhibits BAFF biological activity may be useful in the treatment of active resistant cases.