bluuebunny/crossref_metadata_2025_split huggingface dataset:
In vivo imaging of caspase-1 activation enables efficient and rapid detection of various inflammatory disorders
Authors: Lee Jaewon , Ko Young ji , Yang Eunjeong , Jang Nayoon , Kim Hye sun , Kwon Ick chan , Cho Nam-hyuk 
Published: January 2023

Abstract
Inflammasome plays a critical role in diverse inflammatory diseases, including cancers and Alzheimer’s disease. It can be activated by a various stimuli and induces secretion of IL-1β after cleavage by active caspase-1, an executing enzymatic component of the inflammasome complex. Therefore, direct imaging of active caspase-1 in vivo may provide enormous advantages for diagnosis, drug discovery, and therapeutic monitoring in various inflammatory disorders. Here, we developed a new imaging probe for in vivo visualization of active caspase-1. For this, we synthesized a caspase-1-specific cleavable peptide bridging a near-infrared (NIR) fluorescence (Cy5.5) dye and a hydrophobic quencher. This novel caspase-1 probe can be efficiently delivered into cells and specifically emit fluorescence upon caspase-1 activation as measured by in vitro and in vivo inflammatory models. We demonstrated efficient in vivo imaging of caspase-1 activation in early stage of various inflammatory conditions in mice models, including endotoxin stimulation, inflammatory bowel disorder, transplanted cancer, and Alzheimer’s disease. Efficient and rapid detection of caspase-1 activity in vivo can revolutionize the current paradigm for diagnosis and therapeutics in various inflammatory disorders.
The anti-inflammatory effects of the Chromogranin A hormone cleavage product catestatin on human monocytes and macrophages
Authors: Muntjewerff Elke M. , Christoffersson Gustaf , Nicolasen Mara J.T. , Mahata Sushil K. , van den Bogaart Geert 
Published: January 2023

Abstract

Objective
High levels of the prohormone chromogranin A (CgA) have been detected in the plasma of patients with various cancers and inflammatory diseases such as rheumatic arthritis, inflammatory bowel disease as well as type 1 and 2 diabetes mellitus (T2DM). In contrast, T2DM and hypertensive patients show reduced levels of the CgA-derived peptide catestatin (CST). Here we show the anti-inflammatory effects of CST on human macrophages and phagocyte migration.

Method
Mouse phagocyte (monocytes and granulocytes) migration was imaged in the cremaster muscle of mice superperfused with buffer containing CST or the chemokine MIP-2. Human blood monocytes were used for in vitro migration to CCL2 and/or CST and macrophage differentiation assays. Macrophages were treated with CST and polarized towards pro-inflammatory or anti-inflammatory phenotypes. Afterwards, cytokine production was measured by ELISA (Il-10, TNF-a, IL-6) and cells were phenotyped by EM and RNA sequencing.

Results
In vivo and in vitro imaging shows that CST works as a chemo-attractant for monocytes. Interestingly, the combination of CST with the well-known chemoattractant CCL2 results in an anti-inflammatory effect. Moreover, CST treatment of inflammatory macrophages inhibited TNF-α and IL-6 production. Additionally, several proinflammatory gene levels in CST treated macrophages were decreased.

Conclusions
CST influences monocyte migration and promotes M1-M2 polarization by decreasing expression of pro-inflammatory cytokines and increasing anti-inflammatory cytokine expression. These factors are important for tissue homeostasis, which makes CST an interesting therapeutic target for treatment and/or diagnosis of various metabolic and immune diseases.
Trauma Induces Emergency Hematopoiesis through IL-1/MyD88-Dependent Production of G-CSF
Authors: Fuchs Anja G , Monlish Darlene , Ghosh Sarbani , Chang Shin-Wen , Bochicchio Grant V , Schuettpelz Laura G , Turnbull Isaiah R 
Published: January 2023

Abstract
The acute inflammatory response to infection or injury dramatically increases the hematopoietic demand on the bone marrow to replace effector leukocytes consumed in the inflammatory response. In the setting of infection, pathogen-associated molecular patterns induce emergency hematopoiesis, activating hematopoietic stem and progenitor cells to proliferate, thereby providing progeny for accelerated myelopoiesis. Sterile tissue injury due to trauma also increases leukocyte demand; however, the effect of sterile tissue injury on hematopoiesis is not well described. We used a mouse model of multisystem injury to investigate the effects of these injuries on bone marrow progenitor frequencies and phenotypes. We find that tissue injury alone induces emergency hematopoiesis in mice subjected to polytrauma. This process is driven by IL-1/MyD88-dependent production of G-CSF. G-CSF induces expansion of hematopoietic progenitors including hematopoietic stem cells and multipotent progenitors and increases the frequency of myeloid-skewed progenitors. These data provide the first comprehensive description of injury-induced emergency hematopoiesis and identify an IL-1/MyD88/G-CSF dependent pathway as the key regulator of emergency hematopoiesis after injury.
Reduced Expression of the DNA-Binding Protein ARID3a, a Mediator of Human Hematopoiesis, in Hematopoietic Progenitors Contributes to Age-Related Transcriptional Changes
Authors: Ratliff Michelle , Garton Joshua W , James Judith A , Webb Carol 
Published: January 2023

Abstract
Population studies estimate that 40% of European and US populations will be over the age of 60 by 2050. Aged individuals exhibit impaired immune responses consisting of poor responses to vaccines and increases in myeloid lineage cells. The reasons for decreased immunity in the elderly haven’t been completely elucidated, making studies of the mechanisms leading to these poor responses important. One possibility is that defects in early hematopoiesis may contribute to poor immune responses in the elderly. Previous studies in our lab indicated that inhibition of the DNA-binding protein ARID3a in hematopoietic cultures of human cord blood led to hematopoietic developmental skewing toward myeloid lineage rather than lymphoid lineage progenitors. Furthermore, we previously reported that numbers of ARID3a-expressing hematopoietic stem cells (HSCs) can be highly variable in some patient subsets as well as in healthy individuals. ARID3a expression levels in hematopoietic progenitor cells have not previously been analyzed in relationship to age. We hypothesized that alterations in ARID3a expression levels in early hematopoietic progenitors could contribute to poor immune responses observed in elderly people. Our data suggest that hematopoietic progenitors obtained from peripheral blood of aged healthy persons show decreased frequencies of ARID3a-expressing HSCs in comparison to HSCs from young individuals. Furthermore, lineage committed progenitor numbers were also altered in aged persons compared young people. Ongoing analyses to investigate the gene expression data in young and aged HSCs in relation to ARID3a expression will assess potential genes regulated by ARID3a that contributes to lineage commitment and development.
Glucocorticoids control allergic inflammation via Foxp3+regulatory T cells
Authors: Nguyen Quang Tam , Min Booki 
Published: January 2023

Abstract
Asthma is a chronic inflammatory disease in the lung characterized by airway inflammation and airway hyper-responsiveness (AHR). Synthetic glucocorticoids such as Dexamethasone (Dex) are a mainstay treatment option for the clinical management of asthma. However, the target cells that mediate their therapeutic effects and the underlying mechanisms remain unclear. Utilizing a cockroach antigen-induced allergic inflammation model in mice, we sought to test the roles of Foxp3+ regulatory T cells (Tregs) during Dex-mediated regulation of allergic inflammation. Administration of Dex significantly reduced the development of airway inflammation and AHR. Accumulation of eosinophils and effector CD4 T cells expressing Th2 cytokines in the lung were dramatically reduced following Dex administration. To our surprise, the Dex-induced reduction was completely abolished in the absence of Tregs. Inflammatory cell accumulation, cytokine production, and AHR development remained unchanged in this condition, suggesting that Tregs may be an important target cell of Dex in vivo. Indeed, adoptive transfer of in vitro generated Tregs into Treg-depleted mice fully restored Dex effects. We performed an RNA-sequencing analysis comparing gene expression profiles in lung infiltrating effector CD4 T cells from mice treated with control, Dex, and Dex with Treg depletion. Genes upregulated or downregulated by Dex treatment remained unchanged by Treg depletion. Taken together, our results uncover a novel function of Tregs during Dex-mediated treatment of allergic inflammation.
Metabolic differences between Th2 and Th17 cells in airway inflammation
Authors: Contreras Diana C , Cephus Jacqueline , Newcomb Dawn , Rathmell Jeffrey C 
Published: January 2023

Abstract
Asthma is an airway inflammatory disease that is mediated by T effector (Teff) cells, specifically Th2 and Th17 cells. As disease increases in severity, there is a shift towards a higher Th17 response. Treatment of asthmatic patients include the use of glucocorticoid (GC) steroids. These drugs are effective at controlling inflammation in mild cases but as disease severity increases towards neutrophilic disease there is resistance by Th17 cells to effects of the drugs. A key therapeutic objective is to identify either alternative treatments for asthma or targets that make Th17 cells more susceptible to GCs. Studies have shown cells which have increased glycolysis and oxidative phosphorylation are more resistant to the effects of GCs. In this study we show, that in the murine model of airway inflammation there are differences in the expression of metabolic proteins between Th2 and Th17 cells that are present in the lung. This has been seen in cells that have higher amounts glucose uptake. These results imply that the higher glucose utilization of Th17 cells and their metabolic flexibility may contribute to their resistance to the effects of GCs. We also show by CyTOF of PBMCs that individuals with asthma have increased expression of metabolic associated proteins compared to control individuals in their CD4 cells. These data support the metabolic shift observed in our in vitro studies.
The function of the circulating innate lymphoid cells is dysregulated in patients with mild asthma
Authors: Drake Li Y , Bachman Kay , Kita Hirohito 
Published: January 2023

Abstract
Group 2 innate lymphoid cells (ILC2s) play crucial roles in type 2 immunity in the lungs and in the development of asthma pathology. While ILC2s are residents in mucosal tissues, they also circulate in peripheral blood. Evidence suggests that blood ILC2s can be used as an indirect but useful biomarker to monitor the activity of ILC2s in mucosal tissues, such as the lungs. To investigate both the innate immune response and the corresponding ILC2 cell levels in the peripheral blood of asthma patients, we collected peripheral blood mononuclear cells (PBMCs) from 11 adult patients with mild asthma (MA) and 12 adult healthy control (HC) subjects. The innate type 2 immune responses of PBMCs and the number of blood ILC2s were examined by in vitro culture and flow cytometry. We found that in response to IL-25 or IL-33 stimulation, PBMCs from MA subjects produced significant higher levels of IL-5 than PBMCs from HC subjects. However, there were no significant differences in ILC2 numbers or proportions between MA and HC groups. These results suggest that the function, but not the number, of blood ILC2s may serve as a general biomarker for human asthma.
Prolonged activation in CD4+ T cells results in extensive mitochondrial remodeling despite the metabolic dominance of aerobic glycolysis
Authors: Akkaya Billur , Roesler Alexander , Theall Brandon P , Al Souz Jafar A , Miozzo Pietro , Traba Javier , Smelkinson Margery , Kabat Juraj , Dorward Dave , Pierce Susan K , Akkaya Munir 
Published: January 2023

Abstract
Activation of CD4+ T cells to proliferate drives cells toward aerobic glycolysis for energy production while using mitochondria primarily for macromolecular synthesis. In addition, the mitochondria of activated T cells increase production of reactive oxygen species, providing an important second messenger for intracellular signaling pathways. To better understand the critical changes in mitochondria that accompany prolonged T cell activation, we carried out an extensive analysis of mitochondrial remodeling using a combination of conventional strategies and a novel high-resolution imaging method. We show that for four days following activation, mouse CD4+ T cells sustained their commitment to glycolysis facilitated by increased glucose uptake through increased expression of GLUT transporters. Despite their limited contribution to energy production, mitochondria were active and showed increased reactive oxygen species production. Despite elevated ROS production, it was successfully counterbalanced by antioxidant enzymes, hence the mitochondria did not display signs of oxidative stress as evidenced by the healthy cristae structure and overall morphology as well as the maintained mitochondrial reserve. Moreover, prolonged activation of CD4+ T cells led to increases in mitochondrial content and volume, in the number of mitochondria per cell and in mitochondrial biogenesis. Thus, during prolonged activation, CD4+ T cells continue to obtain energy predominantly from glycolysis but also undergo extensive mitochondrial remodeling, resulting in increased mitochondrial activity.
A cognate interaction between T cell-expressed BTLA and B cell-expressed HVEM regulates mucosal immunity in the gut
Authors: Stienne Caroline , Moshayedi Aref , Chappell Elizabeth , Ware Carl F. , Sedy John R. 
Published: January 2023

Abstract
Mucosal immunity in the gut is achieved in part through cellular connections between lymphoid cells such as regulatory-T cells (Treg), T follicular helper cells (Tfh), T follicular regulatory cells (Tfr) and Germinal Center B cells (GC B). Btla (B and T Lymphocyte Attenuator) is a receptor mainly expressed on B and T cells that is activated by its ligand Hvem (Herpesvirus entry mediator; Tnfrsf14) to inhibit cellular activation. However, the role of these specific immune checkpoints in the regulation of mucosal immunity in the gut is still unknown.
In mice containing genetic ablation of Btla in T cells, we observed an increased frequency of Tfh and GC B cells in Peyer’s Patches (PP), and a decreased frequency of Treg and Tfr cells in these tissues. Interestingly, in mice with a B-cell-specific deletion of the Btla ligand Tnfrsf14 (Hvem), we observe similar increases in GC B cells and Tfh cells and decreased in Treg. Moreover, these mice are characterized by an increase of mucosal IgA bound to bacteria in fecal pellets, indicating a functional outcome of Btla regulation of the GC. Finally, treating wild-type mice with an antibody agonist for Btla increases the frequency of regulatory T cells in PP demonstrating the potential therapeutic effect of activating Btla inhibitory signaling in mucosal immunity.
Together, we show that T cell-expressed Btla interacts with B cell-expressed Hvem to regulate GC responses in Peyer’s Patches. A disruption of the balance effector T cell/regulatory T cell is responsible for many inflammatory autoimmune diseases. It will be crucial to understand the role of Btla and its ligand in these populations in mucosal tissues to understand how immunity is regulated, and to develop potential novel therapeutics to treat disease.
The skin: a site of extralymphoid IgM production
Authors: McGettigan Shannon E , Wilson R. Paul , Dang Van Duc , Kumar Anil , Nikbakht Neda , Stohl William , Debes Gudrun F 
Published: January 2023

Abstract
Antibodies are critical to cutaneous host defense and inflammation, but the mechanisms by which skin antibodies are maintained are poorly described. In this study, we found extralymphoid antibody production in unperturbed human and mouse skin. IgM was the predominant isotype produced by plasma cells in healthy murine skin and was also secreted in healthy human skin. In mice, cutaneous IgM plasma cells were independent of T cells and microbiota. Importantly, chronic inflammation caused the massive accumulation of IgM secreting cells that was dependent on APRIL or BAFF in skin but not in other sites. In addition, skin immunization caused cutaneous accumulation of both T cell-dependent and -independent antigen-specific IgM secreting cells. Thus, we reveal the skin as a novel niche for plasma cells with important implications for the regulation of cutaneous antibodies. We propose that skin plasma cells supply natural and adaptive IgM to the cutaneous environment, thereby supporting homeostatic skin barrier functions in addition to providing defense against pathogen intrusion.
Pressure enabled drug delivery of anti-CEA CAR-T cells increases intra-hepatic CAR-T tumor penetration and therapeutic index in a murine model of liver metastasis
Authors: Hardaway John C , Chai Louis F. , Lopes Mikayla , Rabinowitz Benjamin , Guha Prajna , Jaroch David , Cox Bryan , Knight Robert , Zeldis Jerome , Espat N. Joseph , Katz Steven C 
Published: January 2023

Abstract

Introduction
We have demonstrated safety and clinical activity with anti-CEA CAR-T (Sorrento/TNK Therapeutics, Inc.) regional infusions for treatment of liver metastases (LM) using a pressure-enabled drug delivery device (PEDD™, TriSalus™ Life Sciences, Inc.) to increase CAR-T penetration. To further explore the potential benefit of PEDD, we developed a pre-clinical murine LM model that enables correlation of infusion pressure with response.

Methodology
Mice with CEA+ LM were infused with anti-CEA CAR-T into the portal vein with specified rate and pressure levels. Flow cytometry (FC), bioluminescence imaging (BLI), and serum liver function tests (LFTs) were conducted at multiple time points.

Results
Flow-pressure relationship was established with ΔP of 7.23 mmHg vs 2.33 mmHg (p=0.01) in high-pressure (10 mL/min, HP) and low-pressure (1 mL/min, LP) cohorts. FC assessment of hepatic non-parenchymal cells 1d after infusion demonstrated 15.9% CD3+CAR+ in HP, compared to 5.1% in LP (p=0.0004). Tumor BLI at day 3 (geometric mean relative to baseline) showed significantly enhanced therapeutic effect in HP, −18% relative signal intensity (RSI), compared to +148% RSI in LP (p=0.05) or +178% RSI in controls (p=0.04). LFTs were similar in LP and HP groups with AST 302 vs 437 U/L (p=0.60) and total bilirubin 0.25 vs 0.7 mg/dL (p=0.33), respectively.

Conclusions
HP delivery correlated with enhanced CAR-T delivery and control of tumor growth, without an increase in liver toxicity. Follow up to this proof of concept study will investigate multiple infusions with combinatorial approaches with the goal of durable tumor eradication. An ongoing clinical study will further explore CAR-T PEDD in patients.
Mouse Models for Immunology Research available from The Jackson Laboratory Repository
Authors: Valenzuela Alicia , Fancher Karen , Rockwood Stephen , Lutz Cathleen 
Published: January 2023

Abstract
The Jackson Laboratory Repository (JAX) serves as a centralized facility for the development, distribution and cryopreservation of mouse models of human biology and disease. JAX distributes more than 10,500 strains to the scientific community, many with applications in immunology research, including mouse models for autoimmune disease, infectious disease, immunodeficient platforms for PDX studies, and multi-purpose “tool strains” (such as CRISPR cas9-expressing lines), recombinase expressing strains, as well as conditional and inducible expression lines.
One of our newest strains is a conditional knock-out (KO) of STING/MYPS, a pathogen sensor involved in the activation of IL6 and IFNb. Additions to our extensive set of interleukin alleles include a SMART-17A knock-in (KI) expressing human NGFR on IL17A expressing cells and a KI/KO of Il9-expressing iCre/EYFP. The latest reporter and cre-expressing strains include an inducible cre KO/KI to Lyz2, and a KI/KO strain with dual reporters of chemokine Ccr2RFP and Cx3cr1GFPactivity, and two strains expressing the INDIA apoptosis reporter, one with widespread conditional expression and the other with expression in germinal center B cells. KOMP mice, offered via the MMRRC portion of the JAX Mouse Repository, provide a growing number of targeted immunological mutations.
Donating a strain to the Repository fulfills NIH’s requirements for sharing mice. Researchers wishing to have mice considered for inclusion in the Repository are encouraged to submit strains: www.jax.org/donate-a-mouse.
This work is supported by NIH, HHMI and private foundations.
Novel therapeutic strategies to mitigate erectile dysfunction in a rat pelvic trauma model
Authors: Hertz Alexandria M , George Evalyn , Hibbert Joseph , Reed-Maldonado Amanda , Brand Timothy , Salgar Shashikumar 
Published: January 2023

Abstract

Introduction
Battlefield pelvic injuries can result in erectile dysfunction (ED) in young soldiers. We investigated whether phosphodiesterase-5-inhibitor (PDE5i), low-energy shock wave (LESWT) and/or mesenchymal stem cell (MSC) therapy could improve erectile function (EF) after pelvic injury.

Methods
Lewis rats 10–12 weeks old were divided into nine groups (n=6/group) - Sham, Injury (control), PDE5i, LESWT, MSC, LESWT+PDE5i, LESWT+MSC, MSC+PDE5i, and LESWT+PDE5i+MSC. Bilateral cavernous nerve crush injury and internal pudendal bundle ligation were performed. Bone marrow-derived MSCs (2x106cells) were injected in to the corpora cavernosum. Rats received LESWT (1000 pulses at 0.06 mJ/mm2, 3Hz, 3 times/week) or sildenafil (PDE5i) in drinking water (~10 mg/kg/day) for four weeks. At 6th week intracavernous pressure (ICP; mmHg) was recorded as EF outcome measure.

Results
Pelvic injury (control group) showed decrease (p&lt;0.001) in EF based on maximum ICP (88±16), average ICP (81±11), and change in ICP (81±13) compared to sham (maximum137±25; average 125±13.0; change 55±24). EF was improved (P&lt;0.001) with PDE5i+LESWT therapy, maximum ICP (131±14), average ICP (116±9), and change in ICP (63±15) compared to control. LESWT and MSC alone improved EF (P&lt;0.05). RT-PCR demonstrated lower (21–87%) BCL-2, eNOS, epiregulin and connective tissue growth factor relative gene expression in control group compared to sham. MSC or LESWT+PDE5i treatment showed increased (69–87%) epiregulin (marker for smooth muscle) expression compared to control; eNOS was markedly higher in LESWT+PDE5i group. Histology/Molecular studies are ongoing.

Conclusion
LESWT, MSC and/or PDE5i combination therapy appears to be promising in EF recovery.