bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inflammatory Response to Chronic Opioid Exposure in Humans
Authors: Mathi Kavita , Dobrota Sylvie D , Lee Timothy J , Sayyid Zahra N , Rouholiman Dara , Gamble Jamison G , Encisco Ellen M , Rico Tom , Maecker Holden T , Chu Larry F 
Published: January 2023

Abstract

Objective
In this study, we investigated if chronic opioid exposure leads to changes in immune signaling pathways.

Method
We used phospho-flow cytometry to study intracellular signaling in immune cells of patients with chronic opioid exposure. PBMCs were stimulated with cytokines (IFN-α, IFN-γ, IL-2, IL-6, IL-7, IL-10, IL-21), then stained to analyze 6 lymphocyte subsets and monocytes for phosphorylated STATs (signal transducers and activators of transcription), in particular p-STATs 1, 3 and 5.

Results
After one month of opioid exposure as compared to baseline, IL-2 stimulation of B cells resulted in significantly elevated pSTAT3 levels whereas IFN-γ stimulation resulted in significantly reduced pSTAT5 levels. CD4+ T cell subsets showed reduced pSTAT3 induction in response to multiple cytokine stimuli, and reduced pSTAT1 and pSTAT5 induction with IFN-γ. Similar reductions in pSTAT3 and pSTAT5 induction were seen in CD8+CD45RA+ T cells with several cytokine stimuli. In monocytes, IL-7 and IL-21 stimulation resulted in significantly decreased pSTAT1 levels. No significant changes in pSTAT signaling was found in non-B non-T cells after stimulation with various cytokines. In addition, acute opioid withdrawal via intravenous naloxone resulted in significantly increased pSTAT1 and pSTAT5 levels after IFN-γ stimulation in CD8+ T cells.

Conclusion
One month of opioid exposure significantly reduced cytokine-induced levels of phospho-STAT signaling. These findings suggest that chronic opioid exposure suppresses immune signaling pathways.
Anti-CD3 Mono-Fab co-potentiates TCR antigen recognition in human T cells
Authors: Becher Laura R , Nevala Wendy , Sutor Shari , Abergel Megan , Hoffmann Michele , Parks Christopher , Pease Larry R , Schrum Adam G , Markovic Svetomir N , Pages Diana Gil 
Published: January 2023

Abstract
TCR mediated antigen recognition results in a signaling cascade leading to T cell activation. Antigen binding to its cognate TCR drives receptor aggregation and conformational changes that trigger signaling from the receptor. Conformational change in the CD3 complex (CD3Δc) is required for robust signaling and productive T cell responses. In contrast, weakly recognized antigens do not induce CD3Δc and the resulting T cell response is poor. Thus, T cell responses to weak antigens may be improved by induction of CD3Δc. We previously described an anti-mouse CD3ɛ monovalent Fab (Mono-7D6-Fab) that induced CD3Δc and synergized with weak antigens to enhance the T cell response, a process we termed co-potentiation. However, Mono-7D6-Fab did not induce classical T cell signaling in the absence of antigenic ligation. Mono-7D6-Fab effectively co-potentiated T cells against murine B16-F10 melanoma cells and reduced tumor burden in vivo in a T cell dependent manner. We exploited a model of human cytomegalovirus (CMV) to generate proof of concept that co-potentiation may enhance T cell antigen recognition in humans. Anti-human CD3ɛ monovalent Fab (Mono-OKT3-Fab) improved the CD8 T cell response to the CMV pp65495–503 peptide as measured by increased T cell proliferation, cytokine production, and the ability to kill target cells. Co-potentiation was dependent on antigen specificity, as 1) the effects were dampened when antigen recognition was impaired and 2) sequencing of the TCRs revealed a restricted number of expanded TCR clones. Our results suggest that Mono-OKT3-Fab may have therapeutic clinical applications to achieve a more robust antigen-specific T cell response in the context of diverse human disease, from cancer to chronic infection.
The Role of Circadian Clock Period 2 (per 2) gene and the expression of per2 related gene in atopic dermatitis animal model
Authors: Kang Tae Jin , Prakash Annamneedi Venkata Venkata , Seong Juwon 
Published: January 2023

Abstract
Atopic dermatitis is a chronic, inflammatory skin disorder associated with defective skin barrier and epidermal hyper reactivity. AD is a Th2 mediated allergic disease. GATA3 is a transcription factor that plays an important role in Th2 differentiation. In this study we investigated the per2 gene role and the expression of per2 related genes in atopic dermatitis. We established the atopic dermatitis animal model in per2 knock out and C57BL/6 by 2,4-dinitrochlorobenzene (DNCB). A subset of mice in both per2 knock out and C57BL/6 were treated with saline as control group. The skin lesions severity score was alleviated in per2 knock out mice DNCB group than wild type group. We examined the TNF-α mRNA expression in skin samples that showed that per2 knock out mice protected from DNCB induced inflammation than wild type mice. We next examined BMAL1 mRNA expression in skin samples that showed BMAL1 functioned as anti-inflammatory molecule in per2 knock out mice and levels are high than wild type mice and GATA3 expression levels were not initiated in per2 knock out mice than wild type mice. According to these results we hypothesized per2 knock out mice protected from allergic atopic dermatitis by increasing anti-inflammatory molecule BMAL1 through supressing GATA3.
Biomarker prediction of pediatric atopic dermatitis severity
Authors: Kaplan Mark H , Engle Sarah , Chang Ching-Yun , Satterwhite Allyson , Ulrich Benjamin , Hayes Tristan , Tepper Robert , Sims Jonathan 
Published: January 2023

Abstract
Atopic dermatitis (AD) is a chronic inflammatory skin disease that affects 25% of children and 1–3% of adults worldwide, and has increased 2- to 3-fold over the past several decades. The pathogenesis of AD may result from complex interactions between environmental and genetic factors, barrier defects and immune dysregulation resulting in epidermal hyperplasia and increased penetration of allergens and microbial pathogens. Although most cases of AD are transient, and AD is often thought of as the first step of the atopic march, some AD patients have active disease throughout life. In this study, we analyzed a population of infants that were at high risk for atopic disease and sampled serum, analyzed peripheral blood mononuclear cells, and clinical parameters upon entry (mean age 10.3 months) and five years later. We tested 161 serum analytes using a combination of single-plex, multiplex, Olink, and Quanterix assays. Broadly, the concentration of many analytes fell over time, while SDF and sCD40L concentration increased at the later time point. Several analytes correlated with AD severity as assessed by SCORAD, and most significantly, IL-13 and MCP-4 correlated with SCORAD at both time points. Subsets of cytokines were significantly correlated with each other, consistent with early disease skewing toward type 2 immune response, and two subsets were correlated with percentages of NKT cells or Th2 cells in the peripheral blood. Importantly, forward selection modeling identified 33 infant serum analytes that could be used to predict AD severity five years later (r2=0.85, p=0.042). This dataset will likely have predictive value for AD persistence past infancy and will be useful in further defining the pathogenesis of atopic disease.
A critical period of innate-immune development: Fetal origins of allergic asthma susceptibility
Authors: Lopez Diego A , Beaudin Anna E. 
Published: January 2023

Abstract
Our lab previously identified a developmentally-restricted hematopoietic stem cell (drHSC) that only exists during fetal development and specifically gives rise to innate-like lymphocytes that persist across the lifespan. The identification of a developmentally-limited cell of origin for innate-like immune cells defines a “critical window” for immune development, in which the phenotype of the developing immune system can be shaped via extrinsic inputs. To test how perturbation during this critical window drives immune dysfunction, we examined underlying changes to innate lymphoid cells (ILCs) in the lung. ILCs are a recently identified family of fetal-derived innate-like lymphocytes that mimic the adaptive T-helper arm of our immune system. In the lung, type-2 innate lymphoid cells (ILC2s) produce IL5 and IL13, cytokines important for eosinophil recruitment, activation and goblet cell hyperplasia during allergic airway inflammation. Surprisingly, a single low-dose injection of poly (i:c) at mid-gestation robustly increased proliferative capacity and cellularity of lung ILC2s in offspring at postnatal day (P)9 and P14, respectively, concomitant with drHSC and common-helper innate lymphoid progenitor (ChILP) cell expansion during fetal development. Additionally, in poly (i:c) perturbed offspring, lung ILC2s exhibited heightened IL5 and IL13 production upon in-vitro stimulation with PMA and ionomycin. Ongoing experiments will examine how perturbation of transient progenitors during fetal development may impact the ChILP cell immune trajectory, altering the establishment and function of neonatal lung ILC2s, and ultimately contributing to allergic asthma susceptibility into adulthood.
Mast cell activation enhances the TLR4-mediated antigen-specific CD4 T cell response
Authors: D’Souza Shaina J , Phong Binh , Immethun Vicki E , Bauer David L , McLachlan James B 
Published: January 2023

Abstract
Mast cells are important immune mediators located at host environment interfaces and are most well-known as the major mediators of allergy and asthma. When activated, they have the ability to release prestored granules that contain immune modulating compounds, such as histamine and inflammatory cytokines. In skin, mast cell activation happens via crosslinking of the high-affinity FcɛRI receptor which is activated by the antibody IgE on the mast cell surface binding to a pathogen or allergen inducing intracellular signaling. While there is some evidence that mast cell activation alone does not induce expansion of endogenous antigen-specific CD4 helper T cells, it is not clear whether mast cells have the capacity to fine tune the expansion of T cells that are responding to other stimuli. To assess this, we focused on determining the effects of mast cell activation through FcɛRI alone or in combination with bacterial products on the generation of antigen specific CD4 T cells. Using an in vivo skin model of mast cell activation, combined with MHC class II tetramers, we show that crosslinking FcɛRI alone does not induce antigen-specific CD4 T cell immunity; however, cross-linking FcɛRI in the presence of the Toll-like receptor 4 (TLR4) agonist LPS provides a synergistic co-stimulatory signal that significantly increases T cell expansion. This effect is lost when stimulating with CpG, a TLR9 agonist. We also show that this synergy is lost when dendritic cells are ablated prior to activation. Further, we demonstrate that TLR4 expression by the mast cells themselves is essential to maintain this synergy. This study provides insights into the ability of mast cells to alter antigen-specific CD4 T cell responses in vivo in response to inflammatory stimuli.
Establishment of airway tolerance in early life to inhaled house dust mite involves mitochondrial H2O2
Authors: Yuan Huijuan , Chen Jie , Hu Sanmei , Sinha Ayan , Ray Prabir , Ray Anuradha 
Published: January 2023

Abstract

BACKGROUND
Exposure to innocuous inhaled foreign antigens such as dust mite and pollen triggers aberrant immune responses in the respiratory tract of individuals genetically predisposed to asthma. Immune tolerance restricts such overreaction and plays a key role in preventing unwanted inflammatory responses. Yet the mechanisms underlying this protection are still largely unclear. Our studies of immune tolerance in regulation of experimental asthma and those of others in the context of cancer demonstrated PPARγ-facilitated fatty acid oxidation (FAO) resulting in mitochondrial H2O2 production in dendritic cells (DCs) in promotion of immune tolerance. However, neither study addressed whether mitochondrial H2O2 is essential for suppression of inflammation in response to antigens.

OBJECTIVE
In this study, we sought to establish a mouse model of airway tolerance to inhaled house dust mite (HDM) allergen in newborn mice and study the role of mitochondrial H2O2 in tolerance establishment.

METHODS AND RESULTS
Repeated exposure of wild type (WT) mice to a low dose of the common allergen HDM in early life suppressed airway inflammation and airway hyperreactivity when the mice were challenged by a high dose of the allergen in adulthood. This protective effect was abolished in MCAT transgenic mice, which overexpress the catalase gene in their mitochondria. The inflammatory phenotype in the MCAT mice was associated with a high TH2 response facilitated by lung dendritic cells (DCs).

CONCLUSIONS
Our study illustrates a crucial role of mitochondrial H2O2 in the establishment of airway tolerance in early life, which suppresses DC-mediated TH2 response.
Plasmactoid dendritic cells express EphA2 to negatively regulate type I interferon production in response to viral infection
Authors: FANG MINGLI , XING JUNJI , ZHANG ZHIQIANG 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (pDCs) are primary producers of type I interferon (IFN-I) in response to viral infection. But how such responses are negatively regulated remains poorly defined. Here, we report that the EPH receptor 2 (EphA2), a member of the protein-tyrosine kinase subfamily of ephrin receptors, is selectively expressed in pDCs. Knockout of EphA2 enhanced IFN-I production in pDCs from infection with influenza virus or DNA virus of HSV-1. The expression of Epha2 was dramatically down-regulated after pDCs activation. Surprisingly, challenge of EphA2−/− mice with HSV-I resulted in lethal inflammation and tissues including stomach, intestine, lung and liver damage due to massive production of IFN-I, while depletion of pDCs, partially blocking of IFN-I receptor (IFNAR) signaling, or transplantation of wild type bone marrow cells into irradiated EphA2−/− mice protected mice from this lethal infection. Mechanistically, EphA2 phosphorylates tumor necrosis factor receptor (TNFR)-associated factor 6 (TRAF6), a key adaptor protein in pDCs function, and inhibits IRF7 activation to reduce IFN-I production in pDCs. These data demonstrate that pDCs expressed EphA2 limits excessive IFN-I production in response to viral infection to avoid uncontrolled inflammation and multiple organ failure. And this finding may provide attractive opportunities for therapeutic targeting pDCs mediated acute inflammation.
Differences in NK and Memory CD8 T cell responses to antigen-nonspecific stimulation by interleukin-15
Authors: Judge Sean J , Dunai Cordelia , Le Catherine T. , Khuat Lam T. , Vick Logan V. , Stoffel Kevin M. , Monjazeb Arta M. , Canter Robert J. , Murphy William J. 
Published: January 2023

Abstract
Natural killer (NK) cells are innate lymphoid cells that exert immediate functions which can be further augmented and sustained with immunostimulatory cytokines. Memory CD8 T cells, due to expression of CD132 and CD122, can be activated by similar cytokines in the absence of TCR engagement (termed “bystander” activation). This results in activation and proliferation but necessitates high amounts of cytokine as high-affinity IL2R complexes (CD25) are not induced. Interestingly, both cell types can then elicit similar effector functions via NKG2D-mediated target cell recognition. As these cell types can fill a similar immunologic niche, we set out to compare NK and memory CD8 T cell responses following IL-15 exposure in vitro from healthy human donors. Cell analysis was performed by flow cytometry and qRT-PCR. At baseline, CD25 expression is negligible at &lt;5% on both human NK and memory CD8 T cells. Surprisingly, culture with rhIL-15 (10 ng/mL) for 4–6 days resulted in marked CD25 upregulation on CD56+CD3− NK cells but not bystander-activated CD45RA-CD95+ CD8 T cells (72±9.2% vs 11±3.8%, P=0.003) despite comparable expansion. Additionally, cytokine-activated NK cells expressed higher levels of inhibitory receptor TIGIT (85±4% vs 57±2%, P=0.01) and activation marker CD69 (99±1% vs 27±9%, P=0.008). Functionally, NK cells had increased expression of granzyme B compared to bystander-activated CD8 T cells. Thus, although NK and bystander CD8 T cells can fill a similar immunologic niche regarding target cell killing, there are significant differences in expression of critical markers following activation. These differences may have consequences in the regulation of these cell types and impact anti-viral and anti-tumor responses.
NATURAL KILLER CELL KILLING OF EXTRACELLULAR PSEUDOMONAS AERUGINOSA
Authors: Feehan David D , Jamil Khusraw , Mody Christopher H 
Published: January 2023

Abstract
Pseudomonas aeruginosa is an opportunistic pathogen that commonly infects individuals with the genetic illness, Cystic Fibrosis and contributes to airway blockage and loss of lung function. NK cells are cytotoxic, granular lymphocytes that are part of the innate immune system. NK cell secretory granules contain the cytolytic proteins granulysin, perforin and granzymes. NK cells, in addition to their cytotoxic effects on cancer and virally infected cells have been shown to play a role in an innate defense against microbes.

Hypothesis
NK cells will kill P. aeruginosa using cytolytic effector protein(perforin, granulysin or granzymes) alone or working synergistically.

Results
Live-cell imaging of a co-culture of YT cells, a human NK cell line, incubated with GFP P. aeruginosa in the presence of the viability dye, propidium iodide, demonstrated that YT cell killing of P. aeruginosa is contact-mediated. CRISPR knockout of granulysin or perforin in YT cells had no significant affect on NK cell killing of P. aeruginosa, as determined by CFU counts. Pre-treatment of YT and NK cells with the serine protease inhibitor 3,4-Dichloroisocoumarin (DCI) to inhibit granzymes, resulted in an inhibition of killing. CRISPR knockout of granzyme B in YT cells, in addition to treatment with the granzyme A inhibitor Futhan, significantly inhibited killing of P. aeruginosa, as determined by CFU counts. These results suggest that NK cells induce membrane damage in P. aeruginosa through a contact dependent process, and that killing requires granzymes.
The PB1-F2 virulence protein regulates host ILC2 responses during influenza
Authors: Barman Tarani Kanta , Califano Danielle , Huber Victor C. , Metzger Dennis W. 
Published: January 2023

Abstract
Influenza remains a major public health concern. The H1N1 PR8 and CA04 influenza viruses are both pandemic strains, yet they have distinct origins and pathogenicity. In PR8, the PB1-F2 viral protein promotes apoptosis of immune cells and negatively influences viral clearance. In the less virulent CA04 strain, however, the PB1-F2 protein is truncated and nonfunctional. Our aim was to examine the role of PB1-F2 in regulating group II innate lymphoid cell (ILC2) function and host susceptibility to viral infection. We found that in the absence of IFN-γ, ILC2s stimulated by CA04 challenge expressed a robust IL-5 response whereas following PR8 challenge, ILC2s predominantly produced IL-13. We observed increased resistance to CA04 challenge in IFN-g−/− mice, which was dependent on ILC2 production of IL-5 and improved tissue integrity. Conversely, during PR8 infection, ILC2s produced only limited amounts of IL-5, with no effect on tissue integrity. We next investigated infection with PB1-F2 recombinant CA04 and PR8 viruses that were created by gene reassortment. CA04 expressing recombinant PR8 PB1-F2 and PR8 expressing recombinant CA04 PB1-F2 induced an ILC2 phenotype and tissue pathogenicity that were similar to the PB1-F2 donor strains, indicating a detrimental role of full-length PB1-F2 protein on ILC2 function and host resistance. Our results suggest that activation of ILC2s can improve host survival following CA04 infection and that the PR8 PB1-F2 inhibits this ILC2-mediated protection. Our findings suggest the concept of a strain-specific contribution of PB1-F2 and ILC2s in protection against influenza virus infection. (supported by NIH Grant R01 HL140496).
The expression of NKG2D ligand is induced in the cutaneous ischemia-reperfusion injury and mitigates pressure ulcer via NKG2D-NKG2D ligand interaction
Authors: Otsuka Noriyuki , Makita Keishi , Tomaru Utano , Kasahara Masanori 
Published: January 2023

Abstract
NKG2D is an activating receptor on NK cells, NKT cells, and several types of T-cells. The ligands for NKG2D are expressed on distressed cells, and NKG2D-NKG2D ligand interaction plays a major role in the surveillance against tumors and viral infection. Several studies have reported the physiological role of NKG2D-positive cells, such as NK cells, iNKT cells, and dendritic epidermal T-cells (DETCs) in skin tissue repair and wound healing. It has been proposed that a major etiology of pressure ulcers is reperfusion injury following ischemia, though the role of NKG2D and its ligands in the process remains to be clarified. Here we report the involvement of NKG2D-NKG2D ligand interaction in the cutaneous ischemia-reperfusion (I-R) injury. The wound size of cutaneous I-R injury was expanded in NKG2D-deficient mice, compared with wild-type and TCRδ knock-out mice. Histopathological evaluation revealed that the accumulation of macrophages and neutrophils was prominent at the periphery of the I-R injury site in NKG2D-deficient mice. The transcripts and the protein expression of murine NKG2D ligands, retinoic acid early inducible gene 1 (RAE-1), were detected in fibrocytes at the I-R injury site. The expression of RAE-1 transcripts was accelerated in primary fibroblasts cultures with sodium arsenite treatment. These results suggest that NKG2D ligand expression is induced by the oxidative stress at the I-R injury site. The pressure ulcer formation by the I-R injury is mitigated via NKG2D-NKG2D interaction. This process may be mediated by the influx of NKG2D-positive cells, which regulate the degree of acute inflammatory cells accumulation at the I-R injury site.
Protective role of lung mesenchymal stem cells in pneumonia caused by the Carbapenem-resistant Klebsiella Pneumoniae.
Authors: Rangasamy Tirumalai , Le John , Ghimire Laxman , Jin Liliang , Jeyaseelan Samithamby 
Published: January 2023

Abstract

Rationale
Although the therapeutic benefits of bone marrow-derived mesenchymal stem cells have been extensively studied in host protection against bacterial pneumonia, the role of lung derived mesenchymal stem cells in bacterial pneumonia has not been explored.

Methods
We purified the stem cell antigen 1+ mesenchymal stem cells from the lung (Sca1+ LMSCs) of C57BL/6J mice using fluorescence activated cell sorter. We subsequently determined the colony forming units, differentiation capacity, proliferation potential, ultrastructural features, and phagocytotic capacity of Sca1+ LMSCs. We also investigated the therapeutic potential of LMSCs in pneumonia caused by the carbapenem resistant K. pneumoniae (CRKP) using a pre-clinical (mouse) model.

Results
Flow cytometric analysis revealed the expression of different mesenchymal stem cell markers in Sca1+ LMSCs. The Sca1+ LMSCs formed colonies, differentiated into osteogenic and adipogenic lineages, and exhibited high proliferation potential in vitro. SEM and TEM images show that the Sca1+ LMSCs are morphologically similar to mouse embryonic fibroblasts but ultra structurally they differ. LMSCs are potent in intracellular killing of CRKPs than BMDMs. Pre-treatment with 3-Methyladenine enhanced the replication of CRKP in both Sca1+ LMSCs and BMDMs. Intriguingly, administration of Sca1+ LMSCs attenuated the infiltration of inflammatory cells (predominantly neutrophils) in the airways and decreased bacterial burden in the lung and in extrapulmonary organs of CRKP-infected mice.

Conclusions
These results show for the first time that the phagocytic capacity and intracellular bacterial killing ability, as well as protective role of LMSCs in pneumonia caused by CRKP.