Tumor Necrosis Factor Family Research Articles

Nonsurviving Patients with Severe Traumatic Brain Injury Have Maintained High Serum sCD40L Levels

Soluble cluster of differentiation 40 ligand (sCD40L) is a member of the tumor necrosis factor family with proinflamatory and procoagulant effects. A previous study found higher serum sCD40L levels at day 1 of traumatic brain injury (TBI) in nonsurviving than surviving patients. Thus the objective of this study was to compare serum sCD40L levels during the first week of a severe TBI between surviving and nonsurviving patients and to determine whether it could be used as a mortality predictor biomarker. In this multicenter study severe TBI patients (with Glasgow Coma Scale score <9) with an Injury Severity Score in noncranial item <9 were included. Serum sCD40L concentrations at days 1, 4, and 8 of TBI were determined. We performed receiver operating characteristic analyses to determine the capacity of 30-day TBI mortality prediction by serum sCD40L levels at days 1, 4, and 8 of TBI. We found that nonsurviving (n= 34) patients in comparison with surviving (n= 90) patients had higher sCD40L levels on days 1 (P < 0.001), 4 (P= 0.004), and 8 (P<0.001) of TBI. We also found that the areas under curve of serum sCD40L concentrations at days 1, 4, and 8 of TBI to 30-day mortality prediction were 82% (P < 0.001), 72% (P=0.01) and 83% (P < 0.001), respectively. The existence of higher serum sCD40L levels in nonsurviving than surviving patients during the first week of TBI and fact that serum sCD40L levels during the first week of TBI can be used as a mortality predictor biomarker are the new findings of our study.

BAFF promotes T cell activation through the BAFF-BAFF-R-PI3K-Akt signaling pathway

B-cell activating factor from the tumor necrosis factor family (BAFF) has revealed its critical role in B cell proliferation and survival, as well as the pathogenesis of T-cell mediated autoimmune disease. However, the effect and molecular mechanisms of BAFF on T cell physiological function have not been fully elucidated. In this study it was seen that BAFF can promote the vitality of purified T cells, increase the proportion of CD3+CD4+, CD4+CD25+, CD4+CD154+, and CD4+CD69+ subgroups and reduce the proportion of CD4+CD62L+ subgroups. Negating BAFF activity with Atacicept (TACI-Fc) reverses vitality and activation of T cells. Furthermore, immunofluorescence detection revealed that BAFF promotes the expression of BAFF receptor (BAFF-R) and transmembrane activator and CAML interactor (TACI) in T cells. Flow cytometry displayed that BAFF/BAFF-R activates the PI3K-Akt signaling pathway while the application of PI3K inhibitor (wortmannin) illuminated that BAFF induces T cell vitality and activation through the PI3K-Akt signaling pathway. We conclude that BAFF is involved in not only the physiology of B cells, but also that of T cells. BAFF affects physiological T-cell activation through BAFF-R-mediated activation of the PI3K-Akt signaling pathway which mirrors one of the pathological mechanisms of T cell-mediated autoimmune diseases.

Antitumor Effect of Periplocin in TRAIL‐Resistant gastric cancer cells via upregulation of death receptor through activating ERK1/2‐EGR1 pathway

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a member of the tumor necrosis factor family, induces apoptosis in a variety of cancer cells. However, gastric cancer (GC) cells are insensitive to TRAIL usually. In the previous study, we showed that Periplocin could induce apoptosis in GC cells via the activation of ERK1/2-EGR1 pathway. In the present study, we have shown that the combination of Periplocin and TRAIL had a greater inhibitory effect on gastric cancer cell viability in vitro and in vivo than Periplocin or TRAIL alone. Through upregulating the expression of DR4 and DR5 at transcriptional and protein levels, Periplocin enhanced the sensitivity of gastric cancer cells to TRAIL. Furthermore, enhanced activity of ERK1/2-EGR1 pathway was responsible for upregulating of DR4 and DR5 uponPeriplocin treatment, subsequently reducing the expression of Mcl-1 and Bcl2 and activating Bid and caspase-3/8. Collectively, these data implied that Periplocin might act as a sensitizer of TRAIL and could be a potential strategy for the treatment of GC.

Constitutively active MyD88/CD40 costimulation enhances expansion and efficacy of chimeric antigen receptor T cells targeting hematological malignancies

Successful adoptive chimeric antigen receptor (CAR) T-cell therapies against hematological malignancies require CAR-T expansion and durable persistence following infusion. Balancing increased CAR-T potency with safety, including severe cytokine-release syndrome (sCRS) and neurotoxicity, warrants inclusion of safety mechanisms to control in vivo CAR-T activity. Here, we describe a novel CAR-T cell platform that utilizes expression of the toll-like receptor (TLR) adaptor molecule, MyD88, and tumor-necrosis factor family member, CD40 (MC), tethered to the CAR molecule through an intentionally inefficient 2A linker system, providing a constitutive signal that drives CAR-T survival, proliferation, and antitumor activity against CD19+ and CD123+ hematological cancers. Robust activity of MC-enhanced CAR-T cells was associated with cachexia in animal models that corresponded with high levels of human cytokine production. However, toxicity could be successfully resolved by using the inducible caspase-9 (iC9) safety switch to reduce serum cytokines, by administration of a neutralizing antibody against TNF-α, or by selecting “low” cytokine-producing CD8+ T cells, without loss of antitumor activity. Interestingly, high basal activity was essential for in vivo CAR-T expansion. This study shows that co-opting novel signaling elements (i.e., MyD88 and CD40) and development of a unique CAR-T architecture can drive T-cell proliferation in vivo to enhance CAR-T therapies.

BLOOD PLATELETS AS ACTIVATORS AND REGULATORS OF INFLAMMATORY AND IMMUNE REACTIONS. PART 2. THROMBOCYTES AS PARTICIPANTS OF IMMUNE REACTIONS

Thrombocytes keep a leading role in conjugating thrombosis, inflammation and congenital immune responses. The platelets provide stable adhesion and interaction with immune cells. Activated platelets express CD40L (CD154), a membrane glycoprotein of tumor necrosis factor (TNF) family. Hence, the platelets are the main source of sCD40L in blood plasma. Platelet CD154 may interact with CD40 receptor on endothelial cells, causing an inflammatory response, and enhancing production of immunoglobulins by B-lymphocytes. Membrane and soluble CD154 of platelets combined with other signals can induce maturation and activation of dendritic cells (DC). The platelets possess functional receptors, e.g., TLR2, TLR4, TLR7 and TLR9 they also bear Fc-receptors, including FcγRIIA, FcεRI and FcαRIA. FcγRIIA on platelets mediate protection against bacteria. Cross-linking of FcαRI on platelets results in production of prothrombotic and pro-inflammatory mediators such as tissue factor and IL-1β. Activation of platelets via FcεR1 causes release of chemokine RANTES and serotonin, which contribute to the pro-inflammatory response of other immune cells. Platelets possess receptors for activated complement components and its fragments (CR2, CR3, CR4, C1q, C1 inhibitor and factors D and H). Activated platelets trigger the complement system through the release of protein kinases and ATP, and also by phosphorylation of C3 and C3b. α-granules of platelets contain chemokines which represent the most numerous group of antimicrobial proteins of platelets (kinocidins), and there is an antimicrobial protein of the defensin family – hBD-1 in the cytoplasm of platelets. Ligand and receptor of the TNF superfamily (TRAIL and LIGHT), the SDF-1 chemokine (CXCL12), the IL-1βinterleukins, IL-8 and the soluble IL-6 receptor (sRIL-6) are recognized as platelet products belonging to the family of cytokines and their receptors. The HMGB-1 protein classified as an inflammatory cytokine, is expressed by activated platelets and causes formation of the extracellular traps by neutrophils. Platelets produce numerous growth factors, including EGF-α and EGF-β1, EGF-β2, TGF-α and TGF-β1, TGF-β2, PDGF, HGF, FGF-β, IGF, pro- and antiangiogenic factors, e.g., VEGF-F and angiopoietins Ang-1 and Ang-2. Fulfillment of immune functions by the platelets is carried out by their interaction with leukocytes, which are attracted to the site of infection and inflammation and retained during the development of an “immune thrombus” under conditions of high shear stress. Platelets can not only maintain and guide the immune response, but also initiate these events. They are able to present the antigen in the context of MHC class I molecules, and activate naїve CD8+T lymphocytes. Potential consequences of platelet interaction with neutrophils, monocytes, dendritic cells and lymphocytes are discussed in the review article.

Analysis of serum B cell-activating factor from the tumor necrosis factor family (BAFF) and its soluble receptors in systemic lupus erythematosus.

ObjectivesTo determine the presence and clinical associations of the soluble receptors of B cell‐activating factor from the tumor necrosis factor family (BAFF) in serum of patients with systemic lupus erythematosus (SLE).MethodsSerum BAFF and soluble BAFF receptor (sBAFF‐R) were quantified using ELISA, and soluble B cell maturation antigen (sBCMA) and transmembrane activator and cyclophilin ligand interactor (sTACI) by Luminex, in 87 SLE patients and 17 healthy controls (HC). Disease activity and organ damage were assessed using SLE Disease Activity Index 2000 (SLEDAI‐2K) and Systemic Lupus International Collaborating Clinics (SLICC) SLE Damage Index (SDI), respectively.Results BAFF and all receptors were detectable in all serum samples. Serum sBCMA and sTACI, but not sBAFF‐R, were significantly higher in SLE than in HC. Serum BAFF was also increased in SLE, but this association was attenuated after adjusting for age and ethnicity. Increased serum BAFF was associated with flare and organ damage. Increased serum sBCMA was associated with the presence of anti‐dsDNA, but not with overall or organ‐specific disease activity, flare or organ damage. Neither sTACI nor sBAFF‐R was associated with any SLE clinical parameters in multivariable analysis. While serum BAFF correlated negatively with sBAFF‐R in HC, no statistically significant correlations were observed between BAFF and its receptors in SLE patients.ConclusionSerum BAFF was associated with flare and organ damage independent of the presence of its soluble receptors. While sBCMA was associated with anti‐dsDNA positivity, other soluble BAFF receptors were not associated with SLE clinical indicators.

The TNF Family of Ligands and Receptors: Communication Modules in the Immune System and Beyond.

The tumor necrosis factor (TNF) and TNF receptor (TNFR) superfamilies (TNFSF/TNFRSF) include 19 ligands and 29 receptors that play important roles in the modulation of cellular functions. The communication pathways mediated by TNFSF/TNFRSF are essential for numerous developmental, homeostatic, and stimulus-responsive processes in vivo. TNFSF/TNFRSF members regulate cellular differentiation, survival, and programmed death, but their most critical functions pertain to the immune system. Both innate and adaptive immune cells are controlled by TNFSF/TNFRSF members in a manner that is crucial for the coordination of various mechanisms driving either co-stimulation or co-inhibition of the immune response. Dysregulation of these same signaling pathways has been implicated in inflammatory and autoimmune diseases, highlighting the importance of their tight regulation. Investigation of the control of TNFSF/TNFRSF activities has led to the development of therapeutics with the potential to reduce chronic inflammation or promote anti-tumor immunity. The study of TNFSF/TNFRSF proteins has exploded over the last 30 yr, but there remains a need to better understand the fundamental mechanisms underlying the molecular pathways they mediate to design more effective anti-inflammatory and anti-cancer therapies.

Elevated level of B-cell-activating factor in tissues and serum of patients with vitiligo: a proof of autoimmune theory

Background Several studies strongly supported the involvement of autoimmunity in vitiligo pathogenesis, which proposed the alteration of both humoral and cellular immunity leading to the destruction of melanocytes. B-lymphocyte-activating factor (BAFF) is a member of tumor necrosis factor family that has an important role in peripheral B-cell survival and homeostasis. The increased production of BAFF may lead to autoimmunity. Objective To assess BAFF levels in both serum and tissue in patients with vitiligo to detect its possible role in the disease. Patients and methods The present case–control study included 30 patients with vitiligo and 30 age-matched and sex-matched healthy controls. Overall, 5 ml of blood sample was retrieved, and lesional skin biopsy specimen was obtained from the margin of lesional skin from each patient and site-matched control for estimation of BAFF levels by ELISA. Results A higher level of BAFF was detected in tissues and sera of patients with vitiligo (8.3±5.0 and 1.9±1.0, respectively) compared with healthy controls (1.0±0.3 and 1.1±0.5 with P<0.001 and <0.005, respectively). Significant correlation was found between BAFF levels in either tissue or serum with duration of the disease (r=0.9, P<0.001 and r=0.7, P<0.001, respectively). Moreover, significant correlation was found between BAFF in tissue and BAFF in serum (r=0.7 and P<0.001). Conclusion The present study provides further evidence on the role of BAFF in the pathogenesis of vitiligo.

4-1BBL has a Possible Role in Mediating Castration-Resistant Conversion of Prostate Cancer via Up-Regulation of Androgen Receptor.

4-1BB ligand (4-1BBL) was a transmembrane glycoprotein belonging to the tumor necrosis factor family. It was expressed on activated T lymphocytes and function as a co-stimulatory molecule via cross-linking with 4-1BB (a.k.a, CD137). In addition to its role in immune regulation, 4-1BBL transmitted signals into the cells on which it was expressed (reverse signaling). 4-1BBL represented a promising target for enhancing antitumor immune responses. Recent studies indicated that 4-1BBL also expressed in non-immune cells and possessed different functions in various types of cells. Here, we reported that 4-1BBL didn't express in normal prostate tissues and benign prostatic hyperplasia tissues, but it expressed in prostate cancer (PCa) tissues at moderate level. Expression of 4-1BBL was up-regulated during the transition from PCa to castration resistant prostate cancer (CRPC). Increasing expression of 4-1BBL not only promoted expression of androgen receptor (AR), but also augmented proliferation and invasion ability of prostate cancer cells in androgen deprivation environment. These results were further verified by xenograft tumor experiments. Meanwhile, inhibiting AR signal pathway by chemical antagonist was able to significantly reduce 4-1BBL mediated proliferation and invasion of PCa cells. These novel findings indicated that 4-1BBL might mediate prostate cancer progression to castration-resistant prostate cancer via enhancing expression and function of AR.

Effect of Natural Compounds on NK Cell Activation

Natural killer (NK) cells are lymphocytes of the innate immune system that survey the body for stressed and abnormal cells. The integration of signals that they receive through various inhibitory and activating cell surface receptors controls their activation and ability to kill target cells and produce cytokines. In this manner, phenotypically and functionally distinct subsets of NK cells help protect against microbial infections and cancer and shape the adaptive immune response. NK cells can use two different mechanisms to kill their targets, either by cytotoxic granule exocytosis or by induction of death receptor-mediated apoptosis. Death ligands belong to the tumor necrosis factor (TNF) family of ligands. Upon release in close proximity to a cell slated for killing, perforin forms pores in the cell membrane of the target cell through which granzymes and associated molecules can enter and induce apoptosis. NK cells are also involved in antibody-dependent cellular toxicity via the CD16 receptor. In addition to target recognition, NK cells can be also activated by treatment with multiple compounds with stimulatory properties. Apart from interleukins, which belong to the best characterized group of NK cell-stimulating compounds, vitamins and constituents extracted from plants also display the ability to activate NK cells. The current review characterizes several groups of NK cell-activating compounds: vitamins belonging to classes A, B, C, D, and E, polysaccharides, lectins, and a number of phytochemicals used in cancer research, exhibiting stimulatory properties when applied to NK cells. Although in most cases the exact mechanism of action is not known, constituents described in this review seem to be promising candidates for NK cell-stimulating drugs.

A tetrameric form of CD40 ligand with potent biological activities in both mouse and human primary B cells

CD40 ligand (CD40 L) expressed by activated T cells interacts with CD40 on B cells and triggers B cell survival, proliferation and differentiation. Deficiency in CD40 L or CD40 in humans causes hyper IgM syndrome due to a defect in T-B interaction that is essential for Ig gene class switch recombination (CSR). CD40 L belongs to the tumor necrosis factor family and normally forms a homotrimer on the cell surface, which is important for its biological activity. To generate a multimeric CD40 L that can be used to stimulate both mouse and human B cells, we fused the extracellular domain of mouse CD40 L, which is known to also bind human CD40, with streptavidin (SA) that forms a stable tetramer under physiological conditions. As expected, 293 T cells transiently transfected with an SA-CD40 L expression vector secreted tetrameric SA-CD40 L in the culture supernatant. The secreted SA-CD40 L exhibited > 25-fold stronger activities in inducing the survival, activation and proliferation of both mouse and human primary B cells than did an agonistic anti-mouse or anti-human CD40 antibody. In the presence of IL-4, SA-CD40 L also induced efficient CSR and plasma cell differentiation in both mouse and human B cells. Moreover, administration of SA-CD40 L in mice induced activation and proliferation of spleen B cells in vivo. These results demonstrate that the SA-CD40 L fusion protein generated in the present study recapitulates the function of membrane-bound trimeric CD40 L and has potent biological activities in both mouse and human primary B cells.

Vascular Endothelial Growth Inhibitor, a Cytokine of the Tumor Necrosis Factor Family, is Associated With Epithelial-Mesenchymal Transition in Renal Cell Carcinoma.

Previous studies have revealed that the activation of the epithelial-mesenchymal transition (EMT) endows metastatic properties upon cancer cells to promote invasion and migration. In this study, immunohistochemical analysis was performed in 50 cases of clear cell renal cell carcinoma (RCC) and paired normal kidney tissues. We detected the expression of vascular endothelial growth inhibitor (VEGI) and EMT markers (E-cadherin, fibronectin, and Slug) and recorded the clinical, pathologic, and follow-up (median follow-up: 79.0 mo) information. The expression of VEGI and E-cadherin was significantly lower in RCC tissues compared with normal kidney tissues (P<0.001). However, the expression of fibronectin and Slug was higher in RCC tissues (P<0.05). VEGI and EMT marker expression marginally differed in tumor size and stage. Significant differences were found in the pathologic grade (P<0.05). The Spearman correlation analysis suggested a positive correlation between VEGI and E-cadherin (r=0.451, P<0.01). A negative correlation was shown between VEGI and fibronectin (r=-0.465, P<0.01). There was also a negative correlation between VEGI and Slug (r=-0.758, P<0.01). During the 79.0 months (range, 7 to 119 mo) of follow-up, 6 patients died due to RCC, and the tumor-free survival rate was 88% (44/50). We did not find a significant correlation between VEGI/EMT markers (E-cadherin, fibronectin, and Slug) and overall survival (P>0.05). Our findings indicate that VEGI plays an important role in EMT in RCC. It suggests that VEGI may be investigated as a disease biomarker and therapeutic target in RCC.

CSF TACI and BAFF levels in patients with primary CNS lymphoma as novel diagnostic biomarkers

We used an enzyme‐linked immunosorbent assay to measure pretreatment B cell‐activating factor belonging to the tumour necrosis factor family (BAFF) and transmembrane activator and CAML‐interactor (TACI) levels in CSF and serum collected from patients with primary central nervous system lymphoma (PCNSL) and control groups. The decision tree analysis of CSF TACI and BAFF levels for patients with a PCNSL diagnosis showed 100% sensitivity and 100% specificity when we attempted to differentiate PCNSL from glioblastoma and CNS inflammatory diseases. The combination of CSF TACI and BAFF levels may thus be a novel and useful diagnostic biomarker of PCNSL.

Prognostic Significance of Serum BAFF, APRIL, TACI and BCMA Levels in Chronic Lymphocytic Leukemia.

As chronic lymphocytic leukemia (CLL) has a variable disease course, novel prognostic markers and risk assessment models are being developed in order to identify high-risk patients who may need early treatment. The two tumor necrosis factor family proteins BAFF and APRIL and their receptors BAFF-R, TACI and BCMA are considered to play a critical role in the survival of normal B cells. In order to highlight the pathophysiological role of this complicated biological network, we aimed to analyze the potential prognostic effects of BAFF, APRIL, TACI and BCMA in CLL patients. We investigated the prognostic impact of serum BCMA, TACI, BAFF and APRIL levels in 129 newly diagnosed CLL patients [median age: 64 (39-88) years; male/female: 85/44]. Serum BAFF, TACI and BCMA levels were significantly lower in the patient group compared to the control group (p < 0.001), while serum APRIL level did not differ significantly between two groups (p > 0.05). Serum BCMA [(p = 0.029; r = 0.208)] and TACI levels [(p = 0.011; r = 0.241)] were positively correlated with serum free light chain ratio. Serum BAFF [(p = 0.008; r = - 0.236)] and BCMA [(p = 0.042; r = - 0.183)] levels were negatively correlated with Rai stage. Overall survival (OS) was relatively better in patients with low serum BAFF levels [60 (1-187) months vs 39.5 (0-256) months; p = 0.063]. Probability of OS was higher in patients with low BAFF levels when compared to patients with normal levels, without statistical significance (53.6% vs 23.6%; p > 0.05). Large prospective studies are needed to validate the prognostic role of this essential biological pathway in CLL.

Urinary B-cell-activating factor of the tumour necrosis factor family (BAFF) in systemic lupus erythematosus.

We examined the clinical relevance of urinary concentrations of B-cell-activating factor of the tumour necrosis factor family (BAFF) and a proliferation-inducing ligand (APRIL) in systemic lupus erythematosus (SLE). We quantified urinary BAFF (uBAFF) by enzyme-linked immunosorbent assay in 85 SLE, 28 primary Sjögren syndrome (pSS), 40 immunoglobulin A nephropathy (IgAN) patients and 36 healthy controls (HCs). Urinary APRIL (uAPRIL) and monocyte chemoattractant protein 1 (uMCP-1) were also quantified. Overall and renal SLE disease activity were assessed using the Systemic Lupus Erythematosus Disease Activity Index 2000. uBAFF was detected in 12% (10/85) of SLE patients, but was undetectable in HCs, IgAN and pSS patients. uBAFF was detectable in 28% (5/18) of SLE patients with active nephritis vs 5/67 (7%) of those without ( p = 0.03), and uBAFF was significantly higher in active renal patients ( p = 0.02) and more likely to be detected in patients with persistently active renal disease. In comparison, uAPRIL and uMCP-1 were detected in 32% (25/77) and 46% (22/48) of SLE patients, respectively. While no difference in proportion of samples with detectable uAPRIL was observed between SLE, HCs and IgAN patients, both uAPRIL and uMCP-1 were significantly detectable in higher proportions of patients with active renal disease. uBAFF was detectable in a small but a significant proportion of SLE patients but not in other groups tested, and was higher in SLE patients with active renal disease.

AB007. OA01.07: CD70 in thymic carcinoma: a promising diagnostic marker

Background: CD70, one of tumor necrosis factor family protein, is reported to be expressed in a large spectrum of solid tumors and hematological malignancies. CD70 is an attractive target for antibody-based therapy and several clinical trials targeting CD70 are being conducted. We previously demonstrated CD70 expression was frequently observed on neoplastic epithelial cells of thymic squamous cell carcinoma (TSCC) via immunohistochemistry (IHC) with antibody for frozen section in the limited number of samples. There have been no reports of CD70 expression focusing on formalin fixed paraffin embedded (FFPE) thymic tumors. In this study, we investigated the utility of a CD70 antibody applied on routine FFPE tissues by assessing a larger number of thymic epithelial tumors, thymic carcinoids and lung squamous cell carcinomas (LSCCs).

The changes and significance of the soluble B cell-activating factor in the peripheral blood of patients with chronic human immunodeficiency virus infection

Objective To elaborate the changes of the soluble B cell-activating factor of the tumor necrosis factor family (BAFF) in the peripheral blood of chronic human immunodeficiency virus (HIV)-infected patients, and to study the correlation between the soluble BAFF in HIV-infected patients and the progressions of acquired immune deficiency syndrome (AIDS). Methods Fifty untreated HIV outpatients and 30 healthy controls were recruited. According to the counts of CD4+ T lymphocytes, HIV-infected patients were divided into three groups, 350 cells/μL group. B cell counts and the BAFF levels were compared among the three groups and the healthy controls. The correlation analysis was conducted for the levels of BAFF, the counts of CD4+ T lymphocytes and B cells, and viral load in HIV-infected patients. The value of BAFF in staging of HIV disease was identified by receiver operating characteristic (ROC) curve. Results The B cell counts were (90.3±43.1) cells/μL in 350 cells/μL group and (307.1±97.0) cells/μL in healthy controls, which was significantly different among the four groups (F=47.92, P<0.05). The concentrations of BAFF in the four groups were (1 737.5±719.7), (962.8±341.1), (859.8±270.4), and (456.9±163.7) ng/L, with significant difference among the groups (F=36.72, P<0.05). The level of BAFF was negatively correlated with both B cell counts and CD4+ T lymphocyte counts (r=-0.722 and -0.568, respectively; both P<0.05), and positively correlated with viral load (r=0.607, P<0.05). The area under the ROC curve was 0.881. If the level of BAFF was 1 281.5 ng/L, the sensitivity and specificity to predict the period of AIDS were 74.1% and 87.0%, respectively. Conclusion The levels of soluble BAFF in HIV-infected patients are significantly increased and related with the reduction of B cell counts and disease progression. Key words: HIV-1; B-lymphocytes; BAFF

Cold atmospheric plasma induces apoptosis of melanoma cells via Sestrin2-mediated nitric oxide synthase signaling.

Cold atmospheric plasma (CAP) represents a promising therapy for selectively cancer killing. However, the mechanism of CAP-induced cancer cell death remains unclear. Here, we identified the tumor necrosis factor-family members, especially Fas, and overloaded intracellular nitric oxide participated in CAP induced apoptosis in A375 and A875 melanoma cell lines, which was known as extrinsic apoptosis pathway. This progress was mediated by antagonistic protein of reactive oxygen species, Sestrin2. The over expression of Sestrin2 induced by plasma treatment resulted in phosphorylation of p38 mitogen-activated protein kinase (MAPK), followed by increased expression of nitric oxide synthase (iNOS), Fas and Fas ligand. Depletion of Sestrin2 reduced iNOS and Fas expression, which was associated with reduction of plasma-induced apoptosis. In contrast, inhibition of iNOS activity and phosphorylation of p38 did not alter Sestrin2 expression in plasma-treated melanoma cells. Taken together, cold atmospheric plasma increases Sestrin2 expression and further activates downstream iNOS, Fas and p38 MAPK signaling to induce apoptosis of melanoma cell lines. These findings suggest a previously unrecognized mechanism in melanoma cells response to cold atmospheric plasma therapy.

Abstract B62: Immune gene expression profiling identifies predictors of relapse in childhood acute myeloid leukemia

Abstract Tumor phenotypes are dictated not only by the neoplastic cell component, but also by the tumor microenvironment (TME), which includes immune and inflammatory cells. Acute myeloid leukemia (AML) is the second most common leukemia of childhood. Although intensive multi-agent chemotherapy in conjunction with improved supportive care has increased survival rates to 70%, 30-40% of children with AML relapse and only one-third of them will survive to adulthood. Investigation of new therapeutic strategies for high-risk AML, including immunotherapy, remains a priority and is being actively pursued. Importantly, AML-induced immune suppression poses a fundamental barrier to successful immune-based interventions. We previously showed that in vitro treatment of primary AML cells with interferon (IFN)-γ induces functional indoleamine 2,3-dioxygenase-1 (IDO1), a molecule with potent immunosuppressive properties, in 50% of unselected cases (IFN-γ responders). With a median follow-up of 8 years, IFN-γ responders experienced a significantly shorter event-free survival and overall survival (OS). Herein, we aimed to get insights into the immune landscape of childhood AML using multi-scale immune profiling strategies with the aim to identify molecular circuits that can be targeted to revert leukemia-induced immune dysfunction and improve clinical outcome. We employed a novel high-throughput digital platform, the nCounter system (nanoString Technologies, Seattle, USA), and Optimized Multi-color Immuno-phenotyping Panels (OMIPs) to comprehensively characterize the bone marrow immune infiltrate in 34 children diagnosed with non-promyelocytic AML (17 males, 17 females; median age at diagnosis=10 years; 28 de novo AML, 4 infant leukemia, 1 secondary AML, 1 congenital leukemia; 7 patients with favorable-risk cytogenetics, 21 patients with intermediate-risk cytogenetics and 6 patients with unfavorable-risk cytogenetics). The RNA Pan-Cancer Immune Profiling Panel, which includes 109 cell surface markers for 24 immune cell types, 30 cancer testis antigens, and &amp;gt;500 immune response genes, allowed us to measure immune gene expression levels on the clinic-ready nCounter® FLEX platform (nanoString Technologies). Transcriptomic data were normalized and analyzed using the nSolver software package. Relapse-free survival (RFS) and OS were selected as covariates. For immune phenotyping studies, viable bone marrow mononuclear cells were labeled with fluorochrome-conjugated antibodies and then analyzed on a 10-color Gallios flow cytometer (Beckman Coulter Life Sciences, Buckinghamshire, UK) with standard settings. Hierarchical clustering of immune gene expression data highlighted patient subgroups with heightened expression of T-cell and natural killer (NK)-cell function genes, as well as Toll-like receptor (TLR) genes, interleukin genes and tumor necrosis factor (TNF) family genes. Patients with high levels of CD8A mRNA also expressed IFNG, CXCL9, CXCL10, FOXP3 and negative checkpoints, including LAG3, CTLA4, IDO1 and CD279 (PD-L1), consistent with T-cell inflamed phenotypes that might underpin the establishment of adaptive immune resistance mechanisms of immune escape. In-depth phenotyping studies showed higher percentages of bone marrow-resident naïve B cells, plasmablasts, intermediate and non-conventional monocytes, plasmacytoid dendritic cells (DCs) and CXCR3-expressing Th1 cells at diagnosis compared with complete remission (CR). By contrast, both type 1 and type 2 myeloid DCs were more represented in the TME of patients with AML in CR. The frequencies of innate lymphoid cells (ILCs) and Vγ9+Vδ2+ T cells were not significantly different when comparing diagnosis and CR samples. Relapses occurred in 71.4% of children with “inflamed” AML compared with 28.6% of patients with “non-inflamed” AML (p=0.012; two-sided Fisher’s exact test). Importantly, RFS was remarkably shorter in patients with inflamed compared with non-inflamed AML (median RFS=394 days vs. undefined; hazard ratio = 0.266; 95% CI=0.11-0.64; p=0.0039, log-rank test). A non-significant association with OS was also observed. In conclusion, inflamed or “hot” AMLs are associated with shorter RFS and might be amenable to immunotherapy approaches tailored to the bone marrow TME, including PD-1/PD-L1 blockade and/or small-molecule IDO1 inhibitors. Grant support: Roger Counter Foundation, Dorset, UK and Qatar National Research Fund (grant #NPRP8-2297-3-494). Citation Format: Jayakumar Vadakekolathu, Gemma A. Foulds, Tasleema Patel, Stephen Reeder, Alan Graham Pockley, Sarah K. Tasian, Sergio Rutella. Immune gene expression profiling identifies predictors of relapse in childhood acute myeloid leukemia [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2017 Oct 1-4; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2018;6(9 Suppl):Abstract nr B62.

Association of systemic lupus erythematosus activity with serum levelsof sTWEAK and CD160: A cross-sectional study

Introduction: Systemic lupus erythematosus (SLE) is a relatively common disease among the patients referred to the rheumatology clinics. Tumor necrosis factor (TNF)-like weak inducer of apoptosis (TWEAK), as a cytokine, is a member of TNF family, and CD160 is an essential natural killer (NK) cell activator, both of which have been argued to be associated with SLE activity. Here, we aimed to evaluate the serum levels of sTWEAK and CD160 and their association with SLE activity. Methods: In a descriptive cross-sectional study, 48 patients with SLE, as the case group, and 40 healthy subjects, as controls, were enrolled. SLE activity was assessed using SLE Disease Activity Index (SLEDAI) in the case group. Moreover, the serum levels of sTWEAK and CD160 were determined using enzyme linked immunosorbent assays (ELISA) method in both groups. Results: Mean serum level of sTWEAK was 19.09% lower in the control group compared to the case group (730.15 ± 170.21 pg/ml vs. 895.39 ± 451.25 pg/ml, respectively). Further, mean serum level of CD160 was 47.31% lower in the healthy subjects than that of SLE patients (206.16 ± 88.97 pg/ml vs 391.30 ± 283.46 pg/ml, respectively). The differences in both occasions were found to be significant (P = 0.013 and P =0.001, respectively). Mean SLEDAI in the patients was 8.68 ± 4.00. There was no significant correlation between serum levels of sTWEAK and CD160 with SLE activity. Conclusion: The serum levels of sTWEAK and CD160 markers in patients with SLE are significantly higher than those of healthy subjects. However, we found no correlation of these markers with the disease activity.