bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-1β and caspase-1 drive autoinflammatory disease independently of IL-1α or caspase-8 in a mouse model of Familial Mediterranean Fever
Authors: Sharma Deepika , Sharma Bhesh Raj , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
Mutations in the gene encoding pyrin (MEFV) are associated with autoinflammatory disorder Familial Mediterranean Fever (FMF). A “FMF-knock-in” (FMF-KI) mouse strain that expresses chimeric pyrin protein with a V726A mutation (MefvV726A/V726A) was generated to model human FMF. This mouse strain exhibits an autoinflammatory disorder that is prevented by genetic deletion of IL-1 receptor (IL-1R) or apoptosis-associated speck-like protein containing a CARD (ASC). ASC-mediated cell death leads to the release of IL-1α and IL-1β, both of which signal through IL-1R. Further, caspase-1 and caspase-8 can interact with ASC to mediate secretion of IL-1 cytokines. The specific IL-1 cytokine instigating development of FMF and the enzymatic caspase involved in its secretion are currently unknown. In this study, we demonstrate that the autoinflammation observed in MefvV726A/V726A mice is specifically mediated by IL-1β and not IL-1α. Furthermore, the disorder is dependent on the caspase-1-ASC axis, while caspase-8 is dispensable. Concurrently, aberrant IL-1β release by MefvV726A/V726A monocytes in response to stimulation with lipopolysaccharide is also dependent on the caspase-1-ASC axis. In conclusion, our studies have uncovered a specific role for caspase-1-mediated IL-1β release in the manifestation of FMF.
IL-4 Signal Regulates IL-9-producing Mucosal Mast Cell Differentiation And Function During The Development of Experimental Food Allergy
Authors: Tomar Sunil , Shik Dana , Ganesan Varsha , Smith Andrew , Wang Yui-Hsi 
Published: January 2023

Abstract
The prevalence of food allergy is increasing significantly in developed nations. Accumulating evidences suggest a pivotal role of intestinal mast cells (MCs), TH2 cytokines, and IgE in the pathophysiology of food allergy. We have recently identified the novel IL-9 producing mucosal mast cells (MMC9s), which can produce high levels of IL-9, IL-13, and mast cells mediators. Mice deficient of IL-4 signals fail to develop MMC9s and become resistant to IgE-mediated food allergy. However, it remains unclear whether IL-4 can directly regulate the development and function of MMC9s. In a skin sensitization induced food allergy model, Il4RaF709 mice which have gain-of-function mutations in IL-4Ra were found to produce much higher levels of ovalbumin (OVA) specific IgE and mast cell protease-1 in blood and became more susceptible to experimental food allergy. While the occurrence of MMC9s and type-2 innate lymphoid cells (ILC2s) were comparable between wild type (WT) and Il4RaF709 mice, repeated OVA ingestions induced significant increase of CD4+TH2 cell occurrence and MC numbers in the small intestine of Il4RaF709 mice than those in WT mice. Furthermore, IL-4 treatments induced bone marrow derived mast cells (BMMCs) from Il4RaF709 mice to produce much more IL-13 and TNF-α than those by WT BMMCs. These results suggest that IL-4 signal provided by CD4+TH2 cells may directly potentiate TH2 effector function of MMC9s and promote the differentiation of MMC9s into intestinal mature MCs, resulting in the increased intestinal hypersensitivity to ingested food allergens. Detailed mechanistic studies of IL-4 signaling in MMC9 development and function may provide therapeutic insights into the preventions and treatments of food allergy.
Carbonic anhydrase enzymes regulate mast cell-mediated inflammation
Authors: Henry Everett Kincie , Sy Chandler B , Inclan-Rico Juan M , Siracusa Mark C 
Published: January 2023

Abstract
Type 2 cytokine responses are necessary for the development of protective immunity to helminth parasites but also cause the inflammation associated with allergies and asthma. Recent studies have found that peripheral hematopoietic progenitor cells contribute to type 2 cytokine-mediated inflammation through their enhanced ability to develop into mast cells. Here we show that carbonic anhydrase (Car) enzymes are upregulated in type 2-associated progenitor cells and demonstrate that Car enzyme inhibition is sufficient to prevent murine mast cell responses and inflammation following Trichinella spiralis infection or the induction of food allergy-like disease. Further, we employed CRISPR/Cas9 technology and illustrate that genetically editing Car1 is sufficient to selectively reduce mast cell development. Finally, we demonstrate that Car enzymes can be targeted to prevent human mast cell development. Collectively these studies identify a previously unrecognized role for Car enzymes in regulating mast cell lineage commitment and suggest that Car enzyme inhibitors may possess therapeutic potential that can be employed to treat mast cell-mediated inflammation.
IL-33R signaling in CD4+ T cells regulates intratumoral myeloid cell recruitment.
Authors: Perez-Chanona Ernesto , Smith Carolyne K , Trinchieri Giorgio 
Published: January 2023

Abstract
The biological functions governed by IL-33 include the activation of TH1, TH2, CD8+ T cell and NK cell responses, however the contribution of IL-33 to anti-tumor responses remains controversial. One study showed that IL-33 drives the CD8+ T cell-mediated attenuation of lung metastasis of B16 melanomas. Another study, however, demonstrated that the depletion of the IL-33 receptor (IL-33R) enhanced anti-tumor responses of NK cells against 4T1 mammary carcinomas. By eliminating the IL-33R in specific immune compartments, including T cells and myeloid cells, we have begun to unravel the complexity of the intratumoral biology of IL-33. For example, we have observed that deletion of Il33r in T cells using Il33rfl/fl mice crossed with CD4-Cre expressing mice significantly impaired tumor growth of subcutaneous MC38 colorectal carcinomas compared with the floxed controls. After three weeks, CD4+ Gata3+ cells and neutrophils were notably more abundant in the tumors of Il33rΔTcells mice, while the CD8+ cell populations remained unaffected. Myeloid cell populations, such as CD11b+ Ly6Chi SSClo MHCII+ monocyte/macrophages, and CD11b+ Ly6C+Ly6G+ granulocytes infiltrated in larger numbers into the tumors growing in Il33rΔTcells mice than those in control mice. The contribution of IL-33 towards the anti-tumor activity or tumor-promoting capacity of these cells is under investigation. In addition, our ongoing studies suggest the possibility that not only IL-33R signaling mediates the extent of tumor clearance, but it also contributes to cancer-induced morbidities such as cachexia and the therapeutic efficacy of chemotherapies. Thus, our results suggest that IL-33R signaling may be a target for cancer immunotherapy.
Macrophage depletion decreases tumorigenesis and alters gut microbiota in the AOM/DSS mouse model of colon cancer
Authors: Bader Jackie E , Velazquez Kandy T , Enos Reilly T , Carson Meredith S , Cranford Taryn L , Nagarkatti Prakash , Nagarkatti Mitzi , Davis J Mark , Murphy E Angela 
Published: January 2023

Abstract
Macrophages are recognized as major players in the connection between inflammation and cancer; they represent up to 50% of the tumor mass and produce a wide array of inflammatory mediators with pro-tumoral functions. Emerging evidence links gut microbiota to the development of colorectal cancer and host-derived immune and inflammatory responses are important driving forces that can shape microbial community composition. We used a macrophage depletion technique to examine the role of macrophages on colon tumorigenesis and gut microbiota. To induce colorectal cancer, male C57BL/6 mice (n=32) received a single injection of AOM followed by three cycles of DSS supplemented water at concentrations of 2%, 1%, 1% at weeks 1, 4, and 7, respectively. Prior to the final DSS cycle (week 7) and twice weekly until sacrifice, mice (n=16/group) received 200ul i.p. of clodronate (CL2MDP) or PBS encapsulated liposomes to deplete macrophages. Mice were sacrificed at 12 weeks following the initial AOM injection and colon tissue was analyzed for tumor burden, macrophage markers, and inflammatory mediators. Stool samples were collected and DNA was isolated and subsequently sequenced for 16S rRNA. Macrophage depletion decreased tumor number by ~35% and specifically large (≥1mm) tumors by ~35% (p&lt;0.05). This was consistent with a decrease in gene expression of EMR1 (overall macrophage marker) as well as expression of markers associated with M2 macrophages (IL-13, IL-10, TGFβ & CCL17) (p&lt;0.05) but not M1 markers (NOS2, IL-12a & IL-23). Macrophage depletion also increased relative abundance of the Firmicutes phylum (p&lt;0.05) in stool. We demonstrate that macrophages are important mediators of tumor growth and changes in gut microbiota in colorectal cancer.
MiR150 over-expression impairs iNKT cell development, maturation, and function
Authors: Wu Xiaojun , Zhou Li , Wang Jie , Li Guihua , Mi Qing-Sheng 
Published: January 2023

Abstract
CD1d-restricted invariant natural killer T (iNKT) cells play an important role in the regulation of diverse immune responses. MicroRNAs have been shown to be critically involved in iNKT cell development, maturation, and function, but the roles of specific miRNAs in it remain not completely understood. We had previously shown the dynamic upregulation of miR-150 during thymus iNKT cell maturation and identified the interrupted iNKT final stage maturation and elevated IFNγ production in miR-150KO mice. To determine the requirement of dynamic expressional changes of miR150 during iNKT cell development, we generated thymus-specific miR150 knock in mice (miR-150KI). MiR-150 over expression resulted in reduced overall iNKT cell number in both thymus and periphery, which could be related to increased apoptosis. MiR-150KI thymus iNKT cells showed the blockade of iNKT maturation at multiple maturation transition points. Splenic iNKT with miR-150KI displayed interrupted maturation and reduced cytokine production, including IFNγ, IL-4, and TNFα, upon in vitro stimulation. Furthermore, Bone marrow chimera study confirmed the effect of miR-150KI on iNKT cell development, maturation, and function are cell autonomous. Altogether, our results indicated that the tight control of miR150 expression is required in iNKT cell development and function. Detailed molecular mechanisms of miR-150 mediated iNKT cell regulation in different developmental stages and activation status are currently under investigation.
IDO1 activity in selected immune cells controls antibody responses
Authors: Scalisi Giulia , Gargaro Marco , Turco Antonella , Iorio Alfonso , Puccetti Paolo , Matino Davide , Fallarino Francesca 
Published: January 2023

AbstractIndoleamine 2,3-dioxygenase 1 (IDO1) is an enzyme involved in the initial step of tryptophan degradation along the kynurenine pathway. It has immunosuppressive effects, linked to enzymic and non-enzymic regulatory functions. IDO1 is also crucial for sustaining the function of T regulatory cells (Treg), which are involved in the establishment of peripheral tolerance.We recently found that defective IDO1 induction is associated to anti-FVIII antibody production in patients with severe Hemophilia A1. In hemophilic mice, CpG- rich oligodeoxynucleotides (CpG-ODN) administration induced IDO1 expression in dendritic cells (DCs) that control FVIII antibody response.Recently we discovered that CpG-ODN selectively induced IDO1 in a subset of conventional DCs (cDCs). IDO1 was found to be highly expressed in gut CD11C+CD103+ DCs, which are required for establishment of oral tolerance2. Based on these data we analyzed the impact of IDO1 deficiency in controlling antibody responses, in two different experimental models (FVIII and ovalbumin, OVA, immunization). Interestingly, IDO1 deficiency resulted in a significant increase of FVIII-specific antibody production relative to wild type controls. Similarly, IDO KO mice showed a significant immunoglobulin production in a model of OVA-induced oral tolerance.These data suggest the regulatory role of IDO1 in controlling antibody responses to self and non-self antigens.
Pam3CSK4 induces MMP-9 expression in human monocytic THP-1 cells
Authors: Alrashed Fatema , Kochumon Shihab , Sindhu Sardar , Ahmad Rasheed 
Published: January 2023

Abstract

Background
Matrix metalloproteinase (MMP)-9 is known to degrade the extracellular matrix and increased MMP-9 levels are related with the pathogenesis of many inflammatory conditions including obesity. Pam3CSK4 is a synthetic triacylated lipopeptide (LP) which is a potent activator of immune cells and induces cytokine production. However, it is unclear whether Pam3CSK4 is able to induce MMP-9 expression in monocytic cells. We, therefore, determined MMP-9 production by Pam3CSK4-treated THP-1 cells and also investigated the signal transduction pathway(s) involved.

Methods
MMP-9 expression was determined by real-time qPCR and ELISA and MMP-9 activity was assessed by zymography. THP-1 cells, THP1-XBlue™ cells, THP1-XBlue™-defMyD cells, anti-TLR2 mAb and selective pharmacological inhibitors were used to study signaling pathways involved. Phospho and total proteins were detected by western blotting.

Results
Pam3CSK4 induced MMP-9 expression (P&lt;0.05) at both mRNA and protein levels in human monocytic THP-1 cells. Increased NF-κB/AP-1 activity was detected in Pam3CSK4-treated THP-1 cells and MMP-9 production in these cells was significantly suppressed by pre-treatment with anti-TLR2 neutralizing antibody or by inhibition of clathrin-dependent endocytosis. Also, MyD88−/− THP-1 cells did not express MMP-9 following treatment with Pam3CSK4. Inhibition of JNK, MEK/ERK, p38 MAPK and NF-κB significantly suppressed MMP-9 gene expression (P&lt;0.05).

Conclusion
Pam3CSK4 induces MMP-9 production in THP-1 cells through the TLR-2/MyD88-dependent mechanism involving ERK, JNK, p38 MAPK and NF-κB/AP-1 activation.
Toll-Like Receptor (TLR)-2 Mediates Recruitment of Neutrophils During Influenza Infection in a Neonatal Murine Model
Authors: Kumova Ogan K , Nguyen Linda T , Fike Adam J , Carey Alison J 
Published: January 2023

Abstract
Infants less than six months of age are particularly susceptible to influenza infection due to their impaired cytotoxic T cell response. The neonate relies on innate immunity to contain infection without excessive inflammatory response, by using pattern recognition molecules, like collectins and TLRs. TLR2 has been identified as a bacterial recognition receptor. However, little is known about its role in viral recognition and associated pathogenesis, particularly in the neonate. To address this, 3-day old TLR-2−/− and control C57Bl/6 mice were infected with influenza type A virus strain PR/8/34 (H1N1) and tracked for survival. Bronchoalveolar lavage fluid (BALF) was collected and lungs were harvested at post infection day (PID) 1, 3, and 6. BALF was analyzed by flow cytometry for cellular infiltration in the airways. RNA was isolated and analyzed via real-time PCR for viral loads and various cytokines. There was a striking difference in survival between the TLR-2−/− (61%) and control neonates (25%) (p&lt; 0.01). Surprisingly, TLR-2−/− mice had higher viral loads than controls (p &lt; 0.05) on PID 1, which was coupled with lower neutrophil numbers in the BALF (p &lt; 0.02). No significant differences were noted on PID 3 or 6 for neutrophil counts or viral loads. On PID 3, TLR-2−/− mice had more than a 2-fold lower transcription of CXCL-5, a key chemoattractant for neutrophils. Therefore, the resistance of neonatal TLR-2−/− mice to influenza infection was not due to heightened influenza clearance. Rather, there was impaired neutrophil recruitment, which potentially reduced pulmonary inflammation. Future studies using our neonatal murine model could elucidate the mechanism of TLR-2 and CXCL-5 mediated inflammation during neonatal viral infection.
Parenteral-Mucosal prime-boost with a TB subunit vaccine enhances early lung T cells against aerosol M. tuberculosis infection with transient improved protection.
Authors: Woodworth Joshua S , Christensen Dennis , Andersen Peter 
Published: January 2023

Abstract

Background
T cell mediated immunity is critical to control M.tuberculosis (Mtb) infection and tuberculosis (TB) disease. However, Mtb actively delays the adaptive T cell response after initial infection. Thus, a cornerstone of TB vaccine development is to overcome this delay and recruit T cells for direct contact with infected lung phagocytes to control early bacterial growth.
Given parenterally, the H56:CAF01 subunit vaccine results in an early CD4 T cell response in Mtb-infected lungs and significant reduction in CFU. Here, we investigate mucosal prime-boost delivery of H56:CAF01 to elicit a lung-localized/homing memory T cell population for further accelerated lung T cell recruitment and Mtb control.

Methods
CB6F1 mice vaccinated with H56 (Ag85B-ESAT6-Rv2660) adjuvanted with CAF01 (Th1/Th17-skewing liposomes) either 3× s.c., or 2× s.c. + i.n. boost(s) were rested to memory phase and then challenged with aerosol Mtb. Tissue-localized Mtb-specific T cells were enumerated by flow cytometry, and bacterial CFU determined. In vivo intravascular staining identified lung-localized from vasculature-localized T cells.

Results
Prime-boost with H56 resulted in a 10-fold increase in recruited/expanded Mtb-specific T cells in the lung parenchyma early after Mtb challenge over s.c.-only vaccination and a reduction in lung CFU at peak infection. However, no CFU difference was observed the chronic stage. Inhibition of S1P1 did not enhance the early protective effect, suggesting against local Trm as driving protection. Overall, the limited increased protection despite significant lung T cell enhancement suggests a constrained ability of vaccine T cells to control early Mtb replication.
Upregulation of innate signaling upon ligation of FcγRIIIa by immune complexes in Human CD4+ T-cells
Authors: Chauhan Anil K 
Published: January 2023

Abstract
Over the past two decades, role of Fc-receptor (FcR) signaling in CD4+ T-cell responses has been overlooked. Whether FcRs signaling in CD4+ T-cells contribute to tolerance breakdown and nucleic acid sensing remain an open question. We earlier reported that the FcγRIIIa cosignaling differentiates human naïve CD4+ T-cells in the absence of CD28 cosigning (JBC, 5128: 2016). We now show that the FcγRIIIa signaling upregulate the expression of nucleic acid sensing toll-like receptors (NA-TLRs) in naïve human CD4+ T-cells. TLR9 accumulate as a full length 120-kDa protein on the cell surface (revision in JI). TLR9 bound to CpG ODN 2006 and localized with FcγRIIIa on the cell surface. Subcellular location of the TLR9 discriminate between modified self-nucleic acid from encapsulated viral nucleic acids. FcγRIIIa signal relocated MyD88 and HMGB1 proteins underneath cell surface with TLR9 forming “myddosomes”. RNA-seq data showed upregulation of several key pathways, which are normally observed during innate immune responses. FcγRIIIa signal upregulated RNA transcripts encoding for IL-1α, IL-1β, IL-2, IL-17A, IL-22, IL-23, IL-31RA, and IL-34 cytokines. Transcripts from IFN subsets, IL-27, IFNι-2, IFNι-3, IFNω, IFNβ, IFNα and IFN inducible genes were upregulated. TLR9 signaling drives the production of IFN-γ, IL-17A, IL-21, ICOS and Bcl6 proteins. Statistically significant increase in the genes from cellular response to DNA damage; RNA-binding; and ERK-signaling was observed. lncRNAs populate exosomes and also regulate TLRs, which were unregulated by FcγRIIIa signal. Our data show that the FcγRIIIa drives innate signaling pathways often observed during inflammation, which may contribute to the naive CD4+ T-cell differentiation.
Functional plasticity of human Tc17 cells is dependent on cytokine milieu
Authors: WU MEI-YAO , YEN HUNG-RONG 
Published: January 2023

Abstract
Human Tc17 cells (IL-17-producing CD8+ T cells) have been identified, but little is known about their functional characteristics and mechanism of their plasticity. We aimed to optimize the differentiation of Tc17 cells and then investigate whether the reprograming of Tc17 to Tc1 cells can promote the anti-tumor cytotoxicity. In this study, we isolated CD8+ T cells from the peripheral blood of healthy donors to induce Tc17 differentiation. Human CD8+ T cells were stimulated with anti-CD3/anti-CD28 antibodies, TGF-β, IL-1β, IL-6, IL-23, IL-2, anti-IFN-γ, and anti-IL-4 antibodies. After 10 days in culture, Tc17 cells were about 10% IL-17+. Although Tc17 cells had upregulated anti-apoptotic proteins, the expression of cytotoxic molecules and lytic function of Tc17 cells were weaker than Tc1 cells. Interestingly, Tc17 cells could be converted to Tc1 cells under the stimulation of specific cytokine milieu, and their lytic function improved after conversion. Given these findings of significant enhancement of cytotoxicity, we suggest that reprogramming of human Tc17 cells can be used as an immunotherapeutic strategy against cancer.
Assessing metabolic demands in B cells
Authors: Mielenz Dirk , Urbanczyk Sophia , Stein Merle , Reimer Dorothea , Winkler Thomas , Jäck Hans-Martin 
Published: January 2023

Abstract
Assessing metabolism during early B cell development we show here that large pre B cells are superior in their metabolic activity but small pre B cells down-regulate glycolysis significantly, both ex vivo and in IL-7 cultures. The mitochondrial Ca2+ binding protein Swiprosin-2/EFhd1 (EFhd1) has been proposed to balance mitochondrial Ca2+ and the mitochondrial membrane potential. We show here that EFhd1 is expressed in pro B cells but becomes downregulated by surface expression of the pre BCR. Conversely, EFhd1 becomes up-regulated again in plasma cells. A transgenic mouse model, where EFhd1 cannot be downregulated by the pre BCR anymore (EFhd1tg), revealed that the mitochondrial metabolic regulator PGC-1a is upregulated by EFhd1. This lead to mitochondrial dysfunction in EFhd1tg pre B cells, to a disadvantage for EFhd1tg pro and pre B cells in mixed bone marrow chimeras and to an increased mitochondrial proton leak in EFhd1tg LPS blasts. In accordance, EFhd1 transfected into 293 cells is enriched in the mitochondrial fraction and co-localizes with mitochondria. On the other hand, CRISPR-Cas9 mediated knock-out of EFhd1 in the pro B cell line 38B9 as a surrogate for the pre BCR signal increased glycolysis, glycolytic capacity and reserve, providing a mechanism for increased metabolic activity in large pre B cells. Thus, proper regulation of EFhd1 is important to equilibrate mitochondrial and glycolytic metabolism during B cell differentiation. To gain more insight into metabolic demands and mechanisms during B cell development, activation and differentiation we generated EFhd1KO mice. First experiments with EFhd1KO mice revealed an important function for EFhd1 during B cell development, differentiation and function.