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Emerging evidence indicates B-cell activating factor (BAFF, Tnfsf13b) to be an important cytokine for antitumor immunity. In this study, we generated a BAFF-overexpressing B16.F10 melanoma cell model and found that BAFF-expressing tumors grow more slowly in vivo than control tumors. The tumor microenvironment (TME) of BAFF-overexpressing tumors had decreased myeloid infiltrates with lower PD-L1 expression. Monocyte depletion and anti-PD-L1 antibody treatment confirmed the functional importance of monocytes for the phenotype of BAFF-mediated tumor growth delay. RNA sequencing analysis confirmed that monocytes isolated from BAFF-overexpressing tumors were characterized by a less exhaustive phenotype and were enriched for in genes involved in activating adaptive immune responses and NF-κB signaling. Evaluation of patients with late-stage metastatic melanoma treated with inhibitors of the PD-1/PD-L1 axis demonstrated a stratification of patients with high and low BAFF plasma levels. Patients with high BAFF levels experienced lower responses to anti-PD-1 immunotherapies. In summary, these results show that BAFF, through its effect on tumor-infiltrating monocytes, not only impacts primary tumor growth but can serve as a biomarker to predict response to anti-PD-1 immunotherapy in advanced disease. Significance: The BAFF cytokine regulates monocytes in the melanoma microenvironment to suppress tumor growth, highlighting the importance of BAFF in antitumor immunity.

This interim analysis from the atopic dermatitis registry TREATgermany shows robust long-term efficacy, favourable safety and high persistence of dupilumab under real life conditions.

Real-World data on effectiveness, safety and drug survival of dupilumab: An analysis from the TREATgermany registry. Table 1: Effectiveness outcomes of dupilumab and cyclosporine to month 3, 6 and 12

Real-World data on effectiveness, safety and drug survival of dupilumab: An analysis from the TREATgermany registry. Table 1: Effectiveness outcomes of dupilumab and cyclosporine to month 3, 6 and 12

Real-World data on effectiveness, safety and drug survival of dupilumab: An analysis from the TREATgermany registry. Table 1: Effectiveness outcomes of dupilumab and cyclosporine to month 3, 6 and 12

Real-World data on effectiveness, safety and drug survival of dupilumab: An analysis from the TREATgermany registry. Table 1: Effectiveness outcomes of dupilumab and cyclosporine to month 3, 6 and 12

Real-World data on effectiveness, safety and drug survival of dupilumab: An analysis from the TREATgermany registry. Table 1: Effectiveness outcomes of dupilumab and cyclosporine to month 3, 6 and 12

Additional file 1.

Additional file 1: Table S1. Affymetrix® expression analysis of NHDF treated with HA vs. control showing the 50 most upregulated genes (FC = fold change). Table S2. Affymetrix® expression analysis of NHDF treated with HA vs. control showing the 50 most downregulated genes (FC = fold change). Table S3. Affymetrix® expression analysis of NHDF treated with medium-sized HA vs. control showing the 50 most upregulated genes (FC = fold change). Table S4. Affymetrix® expression analysis of NHDF treated with medium-sized HA vs. control showing the 50 most downregulated genes (FC = fold change). Table S5. Affymetrix® expression analysis of NHDF treated with HYAL vs. control showing the 50 most upregulated genes (FC = fold change). Table S6. Affymetrix® expression analysis of NHDF treated with HYAL vs. control showing the 50 most downregulated genes (FC = fold change). Figure S1. (A, C) HAS1, HAS3 gene expression levels in normal human dermal fibroblasts (NHDF) after stimulation with 1 mg/ml HA, 1.5 U/ml HYAL and HA + HYAL co-stimulation for 2 h, 4 h, 12 h and 24 h, (B, D) HAS1, HAS3 gene expression levels of NHDF after stimulation with 15 U/ml, 1.5 U/ml, 0.15 U/ml and 0.015 U/ml HYAL for 24 h. Asterisks above columns indicate statistical significant differences compared to their respective medium controls. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 (t-test, two-sided). Figure S2. (A, C, E) HAS1, HAS2, HAS3 gene expression levels in primary human keratinocytes after stimulation with 1 mg/ml HA, 1.5 U/ml HYAL and HA + HYAL co-stimulation for 2 h, 4 h, 12 h and 24 h, (B, D, F) HAS1, HAS2, HAS3 gene expression levels in keratinocytes after stimulation with 15 U/ml, 1.5 U/ml, 0.15 U/ml and 0.015 U/ml HYAL for 24 h, (G, H) HA amount (ng/ml) measurement by means of ELISA in supernatants of NHDF treated as described in A–F. Asterisks above columns indicate statistical significant differences compared to their respective medium controls. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001 (t-test, two-sided).

Additional file 1: Figure S1. B16F10 cells were treated (10 μM-78 nM) for 72 h and IC50 compound values determined. Compounds for which IC50 values were not determinable or were above 10 μM were placed on the 10 μM line. Figure S2. (A) Treatment at the indicated time points with TM (3 μM for B16F10 and A375, 5 μM for RPMI and SH4) did not induce changes in CAMP levels (n = 3-4). (B) Treatment with TM for 18 h did not induce changes in serotonin responsive genes. Expression was normalized to GAPDH (n = 3–4). Figure S3. TM did not affect the MAPK pathway. Melanoma cells were treated as indicated with increasing concentrations of TM (representative immunoblots of n = 3-5 are shown). Blots are quantified in (B). Figure S4. (A) Changes in phospho-S6 (p-S6) at Ser235/6 in MeWo and MEL-JUSO melanoma cells (n = 5–6) following TM treatment are shown. (B) Changes in p-S6 (Ser240/244) (n = 2–3) and p-PDK1 (Ser241) (n = 3–4) following treatment with TM are shown. Figure S5. Dose response curves following treatment with the PI3K inhibitor ZSTK474, pan-Akt inhibitor MK-2206 and mTORC1/mTORC2 inhibitor KU-0063794 are shown (n = 3–4). Figure S6. (A) TM-treated mice as described in Fig. 4a. experienced a small decrease in body weight (n = 6–8). (B) Treatment with TM did not alter liver damage parameters (n = 3). (C) Quantification of the active Caspase-3 staining for Fig. 4d is shown (n = 6 mice). (D) Immunohistochemical staining of tumor tissue for cleaved Caspase-8 is shown and quantified (n = 6 mice). (E) Tumor lysates probed for cleaved Caspase-8 are shown (n = 6–9 mice, with 3 mice shown). Error bars in all experiments indicate SEM. *P < 0.05 as determined by a Student’s t-test (unpaired, 2 tailed) or a one-way ANOVA with a Dunnett’s post-hoc test.

BackgroundWound healing represents a dynamic process involving directional migration of different cell types. Chemokines, a family of chemoattractive proteins, have been suggested to be key players in cell-to-cell communication and essential for directed migration of structural cells. Today, the role of the chemokine network in cutaneous wound healing is not fully understood. Unraveling the chemokine-driven communication pathways in this complex process could possibly lead to new therapeutic strategies in wound healing disorders.MethodsWe performed a systematic, comprehensive time-course analysis of the expression and function of a broad variety of cytokines, growth factors, adhesion molecules, matrixmetalloproteinases and chemokines in a murine cutaneous wound healing model.ResultsStrikingly, chemokines were found to be among the most highly regulated genes and their expression was found to coincide with the expression of their matching receptors. Accordingly, we could show that resting and activated human primary keratinocytes (CCR3, CCR4, CCR6, CXCR1, CXCR3), dermal fibroblasts (CCR3, CCR4, CCR10) and dermal microvascular endothelial cells (CCR3, CCR4, CCR6, CCR8, CCR9, CCR10, CXCR1, CXCR2, CXCR3) express a distinct and functionally active repertoire of chemokine receptors. Furthermore, chemokine ligand-receptor interactions markedly improved the wound repair of structural skin cells in vitro.ConclusionTaken together, we here present the most comprehensive analysis of mediators critically involved in acute cutaneous wound healing. Our findings suggest therapeutic approaches for the management of wound closure by targeting the chemokine network.

Interleukin 17-producing helper T cells (T(H)17 cells) have a major role in protection against infections and in mediating autoimmune diseases, yet the mechanisms involved are incompletely understood. We found that interleukin 26 (IL-26), a human T(H)17 cell-derived cytokine, is a cationic amphipathic protein that kills extracellular bacteria via membrane-pore formation. Furthermore, T(H)17 cell-derived IL-26 formed complexes with bacterial DNA and self-DNA released by dying bacteria and host cells. The resulting IL-26-DNA complexes triggered the production of type I interferon by plasmacytoid dendritic cells via activation of Toll-like receptor 9, but independently of the IL-26 receptor. These findings provide insights into the potent antimicrobial and proinflammatory function of T(H)17 cells by showing that IL-26 is a natural human antimicrobial that promotes immune sensing of bacterial and host cell death.

BackgroundAlthough the cytokine IL-31 has been implicated in inflammatory and lymphoma-associated itch, the cellular basis for its pruritic action is yet unclear.ObjectiveWe sought to determine whether immune cell-derived IL-31 directly stimulates sensory neurons and to identify the molecular basis of IL-31-induced itch.MethodsWe used immunohistochemistry and quantitative real-time PCR to determine IL-31 expression levels in mice and human subjects. Immunohistochemistry, immunofluorescence, quantitative real-time PCR, in vivo pharmacology, Western blotting, single-cell calcium imaging, and electrophysiology were used to examine the distribution, functionality, and cellular basis of the neuronal IL-31 receptor α in mice and human subjects.ResultsAmong all immune and resident skin cells examined, IL-31 was predominantly produced by TH2 and, to a significantly lesser extent, mature dendritic cells. Cutaneous and intrathecal injections of IL-31 evoked intense itch, and its concentrations increased significantly in murine atopy-like dermatitis skin. Both human and mouse dorsal root ganglia neurons express IL-31RA, largely in neurons that coexpress transient receptor potential cation channel vanilloid subtype 1 (TRPV1). IL-31-induced itch was significantly reduced in TRPV1-deficient and transient receptor channel potential cation channel ankyrin subtype 1 (TRPA1)-deficient mice but not in c-kit or proteinase-activated receptor 2 mice. In cultured primary sensory neurons IL-31 triggered Ca(2+) release and extracellular signal-regulated kinase 1/2 phosphorylation, inhibition of which blocked IL-31 signaling in vitro and reduced IL-31-induced scratching in vivo.ConclusionIL-31RA is a functional receptor expressed by a small subpopulation of IL-31RA(+)/TRPV1(+)/TRPA1(+) neurons and is a critical neuroimmune link between TH2 cells and sensory nerves for the generation of T cell-mediated itch. Thus targeting neuronal IL-31RA might be effective in the management of TH2-mediated itch, including atopic dermatitis and cutaneous T-cell lymphoma.