bluuebunny/crossref_metadata_2025_split huggingface dataset:
Two phase 3 study results of children and adults with mild-to-moderate atopic dermatitis treated with Crisaborole Topical Ointment, 2%, a novel, nonsteroidal, topical, anti-inflammatory, phosphodiesterase 4 inhibitor
Authors: Zane Lee Thomas , Paller Amy S. , Tom Wynnis L. , Lebwohl Mark G. , Blumenthal Robin L. , Boguniewicz Mark , Call Robert S. , Eichenfield Lawrence F. , Forsha Douglass W. , Rees William C. , Simpson Eric L. , Stein Gold Linda F. , Zaenglein Andrea L. , Hebert Adelaide A. 
Published: January 2023

Abstract
Up to 90% of children and adults with atopic dermatitis (AD), a chronic inflammatory skin disease, present with mild-to-moderate disease. Crisaborole Topical Ointment, 2%, is a novel, nonsteroidal, topical, anti-inflammatory, phosphodiesterase 4 inhibitor being studied for the treatment of AD. The efficacy and safety of crisaborole was assessed in 2 identically designed, multicenter, vehicle-controlled, double-blind Phase 3 studies (301 and 302) that enrolled patients ≥2 years old with mild-to-moderate AD affecting ≥5% of body surface area (BSA). Patients were randomized 2:1 to receive crisaborole or vehicle twice daily and evaluated on Days 8, 15, 22, and 29. The primary endpoint defined success in the Investigator’s Static Global Assessment (ISGA) as “almost clear/1” or “clear/0” with ≥2-grade improvement from baseline at Day 29. Secondary endpoints analyzed the time to success and the percentage of patients achieving “almost clear/1” or “clear/0” on ISGA. At Day 29, more crisaborole-treated patients achieved ISGA success than vehicle (301: 32.8% vs 25.4%, P = 0.038; 302: 31.4% vs 18.0%, P &lt; 0.001), with a greater percentage of “almost clear/1” or “clear/0” ISGA scores (301: 51.7% vs 40.6%, P = 0.005; 302: 48.5% vs 29.7%, P &lt; 0.001). Success in ISGA scores was achieved earlier with crisaborole than vehicle (P &lt; 0.001). Treatment-related adverse events (AEs) were usually mild and included upper respiratory tract infection (pooled data, crisaborole vs vehicle: 3.0% vs 3.0%) and application site pain (4.4% vs 1.2%). AE-related discontinuation rates were low for both groups (1.2%). 2 large Phase 3 studies demonstrated crisaborole may represent a novel, safe, and efficacious treatment for patients with mild-to-moderate AD.
Characterization of Allergen Experienced Group 2 Innate Lymphoid Cells: where do they come from, where do they go?
Authors: Mathae Laura , Martinez-Gonzalez Itziar , Takei Fumio 
Published: January 2023

Abstract
We identified group 2 innate lymphoid cells (ILC2s) in mouse lungs. Upon exposure to the protease allergen papain, lung ILC2s produced large amounts of IL-5 and IL-13 and induced eosinophilia and mucus hyperproduction. Intranasal injections of recombinant IL-33 also stimulated lung ILC2s. ILC2s that were previously stimulated by an allergen persisted long after the initial inflammation was resolved and responded to allergen challenge more vigorously than naïve ILC2s. Microarray analyses of the gene expression profiles of ILC2s isolated from naïve and allergen-treated mice showed that allergen-experienced ILC2s have a similar gene profile to that of memory T cells. To further study the process of “memory ILC2” generation, we analyzed the expression of cell surface molecules on ILC2s from untreated mice and those after intranasal IL-33 injections at various time points, which showed differential expression of CD27, CD103, CD41 and the IL-25 receptor (IL-25R). Memory ILC2s expressed higher level of IL-25R than naïve ILC2s, and intranasal injection of IL-25 stimulated memory, but not naïve, lung ILC2s. Intranasal IL-33 injections also increased ILC2 numbers in the lung draining mediastinal lymph node (mLN), peripheral blood, spleen and liver but not in the intestine or skin. Phenotype analysis of those ILC2s suggested that activated lung ILC2s migrated into mLN and the blood, and some settled in the liver. The results indicated that IL-25-responsive memory ILC2s are generated and persist in the lung as well as in mLN while the role of the latter in allergic lung inflammation induced by intranasal allergen challenge in sensitized mice is yet to be determined.
CD4+ T-cell cytokine production within airway mucosal DC-T-cell clusters drives the local recruitment of myeloid effector cells in response to house dust mite allergen.
Authors: Veres Tibor Z , Kopcsányi Tamás , van Panhuys Nicholas , Gerner Michael Y , Liu Zhiduo , Rantakari Pia , Dunkel Johannes , Miyasaka Masayuki , Jalkanen Sirpa , Germain Ronald N 
Published: January 2023

Abstract
Allergic asthma develops in the mucosal tissue of small bronchi. Upon allergen exposure, a TH2-type inflammatory response leads to airflow obstruction and wheezing. Recent studies provided a detailed understanding on the role of various cell types and mediators in the allergic response on the level of the whole organism or tissue. However, less well understood is the micro-anatomical organization of the key cellular players, dendritic cells (DCs) and CD4+ T cells within the airway mucosa and the mechanisms by which local T-DC interactions contribute to the ‘downstream’ inflammatory response (e.g. eosinophil recruitment). Here we utilized advanced in vivo dynamic and in situ static imaging to visualize CD4+ T-cell activation and effector function in the tracheal mucosa.
Upon mucosal sensitization to house dust mite (HDM) allergen, newly recruited CD4+ T cells formed discrete cellular clusters and showed prolonged interactions with DCs in an antigen-specific manner. While forming direct cell-cell contacts with MHC-II+ DCs, a small fraction of CD4+ T cells, but not innate lymphoid cells (ILCs), produced IL-13 or IL-17. As a result of local TH2 cytokine signaling, revealed by STAT-6 phosphorylation, eosinophils were locally recruited into these cellular clusters. Neutrophils also infiltrated these clusters in a T-cell dependent manner; however, their distribution was more diffuse. Our findings reveal the focal nature of antigen-driven allergic responses in the airways and define multiple steps with potential for interference with the progression of asthmatic pathology.
This work was supported in part by the Intramural Research Program of NIAID, NIH.
Pulmonary NK cells prevent allergic airway sensitization: Regulation by CB2 cannabinoid receptors
Authors: Jaffar Zeina , Ferrini Maria , Roberts Kevan 
Published: January 2023

Abstract
Allergic asthma is a chronic inflammatory disease involving the interaction between the innate and adaptive immune response. Endogenously generated cannabinoids, acting via CB receptors play important roles in both homeostatic and inflammatory processes. CB2-active compounds are known to impact innate immunity including responses by monocytes and NK cells. However, the contribution of pulmonary NK cells and CB2-active endocannabinoids to the innate events preceding sensitization of the airways to the common house dust mite (HDM) allergen is unclear. We investigated the significance of CB2 activation during allergen-triggered pulmonary inflammation and NK cell effector function. Allergic airway inflammation was induced in mice by sensitization via the airways by intranasal HDM instillation, and responses in wild type (WT) and CB2-deficient (CB2−/−) mice were compared. Mice lacking CB2 receptor exhibited elevated numbers of pulmonary NK cells yet were resistant to the induction of allergic inflammation and showed diminished airway hyperreactivity, pulmonary eosinophilia, TH2 cytokine production and mucus secretion after allergen inhalation. Cellular depletion and adoptive transfer studies were undertaken to dissect the mechanisms involved. Depletion of NK cells restored HDM responsiveness. Conversely, transfer of CB2−/− NK cells into WT mice suppressed the allergic airway inflammation and was associated with a reduction in monocyte-derived dendritic cells but elevated CX3CL1 release and recruitment of Ly6C−CX3CR1+ monocytes. These studies established a crucial role for CB2 activation in allergic lung disease via regulation of NK cell function and identified novel therapeutic targets for treatment of asthma.
Polychromatic Flow: MDR1 Expression on Th17/Th17.1 effector cells in Patients with Autoimmune Disease - Potential Role in Drug Resistance
Authors: Bisconte Angelina , Kisselbach Lynn , Neidinger Erik , Phippard Deborah 
Published: January 2023

Abstract
IL-17A secreting T cells represent a distinct lineage of CD4+ T effector cells (Th17) that have been suggested to play a role in the pathogenesis of inflammatory/autoimmune diseases such as Crohn’s disease. Precision for Medicine cryopreserves primary Th17 cells (AccuCell®) to provide an easy alternative to isolating fresh cells, for use in screening assays and functional immunological studies.
The human MDR1 gene encodes a membrane protein, P glycoprotein (Pgp), whose function is to export toxic substances or metabolites from the cell. MDR1 expresssion has been demonstrated in tissues of Crohn’s disease patients and on their circulating Th17 cells. Using AccuCell® Th17 cells from healthy and diseased patients we investigate how MDR1 expression differs in the disease state and highlight the challenges of screening drugs using healthy cells as opposed to the targeted disease state.
First we evaluated our cryopreserved Th17 cells to fresh isolations for surface marker, chemokines, intracellular cytokines (IL-17A, IFNγ, IL-22), and RORγt. Full retention of the tested markers were observed in the recovered cryopreserved cells. On our disease state PBMCs phenotypic and functional evaluation of MDR1+ Th17/17.1 cells included activation markers CD5, CD69, CD2, human leukocyte antigen-DR (HLA-DR) and co-stimulatory receptor CD28, compared to normal healthy donors. T-cell receptor dependent proliferation in MDR1+ Th17 cells was compared to healthy donor demographic matched Th17 cells. Utilizing AccuCell® cryopreserved Th17 cells which are fully functional empowered these studies to elucidate mechanisms as discussed above; thus eliminating fresh isolations so researchers can focus on studying the pathogenesis of these cells.
Plasma cell mitochondrial pyruvate import controls the duration of humoral immunity.
Authors: Lam Wing Yu , Becker Amy M , Kennerly Krista , Wong Rachel , Payne Elizabeth , Curtis Jonathan , McCommis Kyle , Fahrmann Johannes , Nunley Ryan , Wolfgang Michael , Patti Gary , Fink Brian , Pearce Erika , Bhattacharya Deepta 
Published: January 2023

Abstract
Durable antibody production after vaccination or infection is mediated by long-lived plasma cells. Pathways that specifically allow long-lived plasma cells to persist remain unknown. Through bioenergetic profiling, we demonstrate that human and mouse long-lived plasma cells possess much greater mitochondrial spare respiratory capacity than do short-lived plasma cells. Inhibition of mitochondrial pyruvate import in long-lived plasma cells attenuates spare, but not basal respiration. Deletion of Mpc2, which encodes an essential component of the mitochondrial pyruvate carrier, leads to a progressive loss of long-lived plasma cells and vaccine-specific antibodies in vivo. Long-lived plasma cells uptake more glucose than do their short-lived counterparts, and subsequent catabolism to pyruvate is essential for spare respiratory capacity. IL-4 or IFNg treatment of activated B lymphocyte precursors to plasma cells promotes glucose uptake. Thus, signals provided to precursors, glucose uptake, and mitochondrial pyruvate transport allow long-lived plasma cells to provide enduring antibody-mediated immunity.
Checkpoint Regulator VISTA plays a role in Suppression of B-Cell Responsiveness by Monocytic Myeloid Derived Suppressor Cells from LP-BM5 retrovirus-infected Mice
Authors: Green Kathy A , Noelle Randolph J , Green William R , Wang Li 
Published: January 2023

Abstract
MDSC inhibition of tumor directed T cell responses is well described. We have reported an increase of monocytic MDSC (M-MDSC) during infection of B6 mice by LP-BM5 immunodeficiency causing retrovirus. These M-MDSCs suppressed T, and B cell responsiveness ex vivo. M-MDSC inhibition of stimulated T-cell proliferation and IFN-gamma production was ~100% iNOS/NO dependent; whereas suppression of B-cells was only ~50% dependent on iNOS/NO. An additional mechanism(s) for M-MDSC inhibition of B cell responsiveness involved V-domain Ig Suppressor of T cell Activation (VISTA), negative checkpoint regulator. Using anti-VISTA blocking mAb, ~50% of total MDSC suppression of B-cell responses was dependent on MDSC-expressed VISTA. The combination of iNOS/NO inhibitors and VISTA lead to additive, if not synergistic, blockade of M-MDSC suppression of B cell responsiveness. Regarding the LP-BM5-infection dependency of M-MDSCs, spleens from uninfected mice yielded ~3-fold fewer enriched M-MDSCs; and on a per-cell basis, the M-MDSCs from uninfected mice were also substantially less suppressive (4.3 fold); than those from infected mice. Consistent with a direct role of VISTA, in the absence of M-MDSCs, a VISTA-Ig fusion protein also partially inhibited B-cell responsiveness. Initial experiments show differential VISTA expression for the LP-BM5 M-MDSC CD11b+hiLy6C+hi subpopulations we have recently defined (O’Connor et al, 2015, Virology). Thus, LP-BM5 infected, when compared to analogous subsets from uninfected mice, contain substantially more VISTA+ cells. These results suggest an unique role for M-MDSCs in LP-BM5 induced immunodeficiency, and highlight multiple suppressive pathways in the area of MDSC suppression of B cell responses.
Protein tyrosine kinase Fyn regulates TLR4-elicited responses on mast cells controlling the function of a PP2A-PKC a/b signaling node leading to TNF secretion
Authors: Gonzalez-Espinosa Claudia , Ibarra-Sánchez Alfredo , Ávila Alejandro Martín , Medina-Tamayo Jaciel , Vázquez-Victorio Genaro , Castillo-Arellano Jorge Iván , Hernández-Mondragón Alma Cristal , Rivera Juan , Madera-Salcedo Iris Karina , Blank Ulrich , Macías-Silva Marina 
Published: January 2023

Abstract
Signaling pathways leading to cytokine production after TLR-4 triggering in mast cells (MC) are not well described. In this work, the participation of the Src family kinase Fyn on the production of TNF after stimulation with LPS was evaluated MCs from WT and Fyn-deficient mice. Fyn−/− cells showed higher LPS-induced secretion of preformed and de novo synthesized TNF and the processes underlying that phenotype were investigated. In both cell types, TNF co-localized with VAMP3-positive compartments. Although LPS addition provoked coalescence of VAMP3 and the interaction of VAMP3 with SNAP23 in WT and Fyn−/− cells, the apparent size of VAMP3-containing aggregates was larger in Fyn−/− cells and increased formation of VAMP3-SNAP23 complexes was observed compared to WT. LPS-addition to Fyn−/− cells lead to higher activation of TAK-1, ERK1/2 and IKK, which was reflected on increased nuclear translocation of transcription factors such as NFkB and CEBPd. Increased LPS responsiveness in Fyn−/− cells was associated to a lower activity of Protein Phosphatase 2A(PP2A) and augmented phosphorylation of key signaling kinases, such as PKCa/b, which controlled TLR4-induced VAMP3 coalescence. Reconstitution of MC-deficient Wsh mice with Fyn−/− MC produced an increased LPS-dependent production of TNF in peritoneal cavity. Our data show that Fyn kinase is activated after TLR-4 triggering and exerts an important negative control on LPS-dependent TNF production in MC controlling the inactivation of PP2Ac and PKC a/b phosphorylation necessary for the secretion of TNF by VAMP3-positive carriers.
Supported by Grant ANR-Conacyt 188565 to UB, CGE and MMS.
Programming of RIG-I signaling through co-factor interactions
Authors: Dewey Elyse C , Tan Min Jie Alvin , Hibberd Martin , Gale Michael J 
Published: January 2023

Abstract
Retinoic acid-inducible gene I (RIG-I) functions as a cytosolic pathogen recognition receptor by binding the pathogen associated molecular patterns (PAMPs) in viral RNA. The molecular signature for RIG-I recognition has been identified as 5′-triphosphate-containing short double stranded or poly-uridine RNA (5′pppRNA) structures found in RNA species that amass in the cytoplasm during virus infection. PAMP-mediated activation of RIG-I triggers many signaling cascades that mediates its effector functions in innate immunity and apoptosis. In the innate immune response to virus infection, RIG-I sets off both IRF3/7 and NF-κB signaling to induce type I interferon (IFN) and pro-inflammatory cytokine production respectively. PAMP-induced RIG-I activation can also induce apoptosis in cancer cells. Since PAMP RNA can act as the activator of RIG-I’s pro-apoptotic and innate immune functions, we hypothesized that the induction of these separate pathways is programmed through differential RIG-I-cofactor interactions. We have identified and characterized novel co-factors of RIG-I by using a systems proteomic approach. These co-factors serve to program apoptotic versus innate immune signaling actions through their interaction with RIG-I. These co-factors and their biologic functions will be presented.
Natural TFH arise in the thymus and periphery of young naive mice
Authors: Adkins Elisabeth B , Wang Xulong , Sproule Thomas J , Christianson Gregory G , Park Giljun , Lane-Reiticker Sarah-Kate , Jain Shweta , Carter Gregory W , Morse Herbert C , Roopenian Derry C 
Published: January 2023

Abstract
T follicular helper cells (TFH) localize to B cell follicles and secrete Interleukin 21 (IL21), a cytokine that is vital for driving the proliferation of antigen-stimulated B cells within germinal centers and their subsequent differentiation into memory B cells and antibody-secreting plasma cells. Much of the current understanding of TFH is limited to studies following immunization of adult mice. Therefore, the ontological processes by which TFH naturally develop and express IL21 are poorly understood. Here we take advantage of a novel IL21-venus fluorescent protein reporter mouse to show that a major fraction of the earliest activated CD4 T cells that develop spontaneously in the thymi and the peripheries of young naive mice express IL21 within 2 weeks of birth. This population is rivaled in frequency only by FoxP3 natural T regulatory cells (nTREG) cells. Results from RNAseq profiling, their partial developmental dependence on IL6 and IL21 signaling and durability after adoptive transfers are all consistent with the generation of a novel, natural TFH (nTFH) population. This nTFH population is functionally defined by IL21 and is able to persist in that differentiation state in naive mice. We further show that thymic development of nTFH requires AIRE, while FoxP3 nTREG act in the periphery to restrain nTFH. We conclude that the precocious and robust presentation of nTFH in naive mice is caused by inherent self-reactivity that is developmentally driven by encounter with promiscuous self-antigens through AIRE. TREG are therefore the primary means by which nTFH are prevented from exercising their pathogenic potential in the periphery.
Sensitivity to restimulation-induced cell death is linked to glycolytic metabolism in human T cells
Authors: Larsen Sasha E , Bilenkin Abegail , Snow Andrew L 
Published: January 2023

Abstract
Restimulation-induced cell death (RICD) helps to regulate immune responses by setting an upper threshold for effector T cell expansion and limiting nonspecific damage to the host. RICD is triggered by re-engagement of the T cell receptor (TCR) on a cycling, activated T cell, resulting in apoptosis. However, it remains unclear how RICD sensitivity is calibrated in T cells derived from different individuals and subsets. As effector T cells expand and become responsive to RICD, they have also switched from a quiescent metabolic state into active aerobic glycolysis (Warburg effect). We hypothesized that in addition to facilitating proliferation and effector function, metabolic programming directly influences RICD sensitivity. We found that secretion of L-lactate, the end product of glycolysis, positively correlated with greater RICD sensitivity in CD8+ effector T cells from different human donors. Strikingly, replacing glucose in culture media for galactose, which results in almost exclusive use of oxidative phosphorylation (OXPHOS), rendered T cells significantly less sensitive to RICD. Consistent with this, blocking glycolysis with 2-deoxy-glucose (2-DG) significantly reduced RICD sensitivity, while inhibiting OXPHOS increased RICD sensitivity. Glycolysis promoted RICD sensitivity in part through mTOR signaling. Moreover, we found that active glycolysis facilitated induction of pro-apoptotic effector molecules like Fas ligand upon TCR restimulation, which was blocked with 2-DG treatment. Collectively, these data suggest that RICD susceptibility is dependent on glycolytic metabolism in effector T cells, and that switching back to OXPHOS may help shield cells transitioning into the memory pool from RICD.
ERK2 regulation of mitophagy is important for T cell activation
Authors: Tudyk Julie , Forsthuber Thomas 
Published: January 2023

Abstract
Critical signals necessary for T cell activation are transduced into the cell, in part, by the MAPK pathway, for which extracellular regulated kinase 2 (ERK2) is a key member. Upon activation, changes to mitochondria location and metabolism mediate T cell priming; however the signaling pathways that modulate mitochondria in T cells are largely unknown. A primary mechanism regulating mitochondria is a form of selective autophagy termed mitophagy. Importantly, ERK2 is required for mitophagy in other cell types and has been shown to co-localize at the autophagosome during formation. These data suggest that ERK2 may have a unique role regulating mitophagy in T cells. Notably, dysregulation of mitochondria in T cells can result in impaired immunity and has also been associated with autoimmune disease.
Here, we investigated the role of ERK2 for mitophagy in CD4+ T cells using a mouse model with T cell-specific deletion of ERK2. Our results show that Erk2−/− CD4+ T cells have impaired autophagosome formation and increased cell death upon activation, a phenotype consistent with mitophagy blockade. Further, we are working to identify key mitophagy proteins that are regulated by ERK2 and to characterize mitochondria signaling metabolites during T cell activation. By elucidating the signaling pathways that connect extracellular immune stimuli with mitochondria regulation, we will obtain important information that may lead to new therapeutic targets for T cell mediated diseases and dysfunction.
The microRNA-212/132 cluster regulates B cell development by setting a threshold for Sox4 expression
Authors: Mehta Arnav , Mann Mati , Baltimore David 
Published: January 2023

Abstract
MicroRNAs play unique roles as logic elements in B cell differentiation by allowing for fine-tuning of differentiation checkpoints. We have found that the miR-212/132 cluster is induced in B cells in response to anti-IgM stimulation and that it can regulate B cell differentiation by establishing a threshold of Sox4 expression necessary for progression of progenitor B cells. Over-expression of miR-132 blocks the transition of pre-pro-B cells to pro-B cells, in part by inducing apoptosis in primary bone marrow B cells. The deletion of miR-212/132 results in increased B cell output under conditions of inflammatory stress. We validate Sox4 as a novel target of miR-132, and show that co-expression of Sox4 with miR-132 can rescue the phenotype observed with over-expression of miR-132 alone. We further demonstrate that over-expression of miR-132 can inhibit the development of B cell leukemia in mice that have B cells expressing the c-myc oncogene. We have therefore described miR-212/132 as a novel regulator of B cell development through its regulation of Sox4 expression.