bluuebunny/crossref_metadata_2025 huggingface dataset:
Temporal analysis of the plasma membrane proteome after vaccinia virus infection sheds light on virus strategies to evade the immune response
Authors: Depierreux Delphine M , Altenburg Arwen F , Soday Lior , Fletcher-Etherington Alice , Ferguson Brian J , Weekes Michael P , Smith Geoffrey L
Published: January 2023

Vaccinia virus (VACV) is a poxvirus and the vaccine used to protect against variola virus and related orthopoxviruses. VACV encodes multiple proteins whose function is to evade the host immune response, contributing to an immune suppressive environment at the site of infection. There is, however, an incomplete understanding of how this immune suppression by VACV is consistent with a strong induction of immunological memory. There is especially little known about the relationship between NK cells and VACV, such as which ligands trigger NK activation and whether VACV uses strategies to interfere with the NK cell response. The purposes of this study is to clarify these open questions.

Here, we performed a quantitative proteomic analysis of membrane-associated proteins from VACV-infected cells at various time points. Using tandem mass-tag spectrometry we detected about 70 viral proteins and 900 membrane-associated host proteins. Analysis revealed that VACV infection rapidly altered the expression of proteins with immune functions, downregulated the surface expression of plexins, ephrins, cadherins and selectively downregulated MHC-I proteins. Moreover, VACV prevented the upregulation of stress-induced ligands and death receptors. Finally, the kinetics of expression of VACV proteins at the surface of the infected cell were analysed.

Collectively, these data constitute a valuable resource for the study of VACV and its interaction with the host immune response, provide insights into the interaction of NK cells with VACV, suggest novel VACV immune evasion mechanisms and shed light on which host factors are required to raise a protective immunological memory.
Latent gammaherpesvirus infection licenses age-associated B cells for pathogenicity during EAE and MS
Authors: Mouat Isobel , Allanach Jessica , Shanina Iryna , Vorobeychik Galina , Horwitz Marc S
Published: January 2023

Epstein-Barr virus (EBV) has long been implicated in multiple sclerosis (MS), though its mechanism of contribution remains unknown. Age-associated B cells (ABCs) are known to expand and persist following viral infection and are increased in MS patients. We hypothesize that EBV infection expands the number of ABCs and skews the population towards a Th1 inflammatory phenotype, leading them to function pathogenically in MS. To explore how the EBV-primed ABC population contributes to MS we have utilized the in vivo models of EBV, gammaherpesvirus 68 (gHV68), and MS, and MOG35–55 experimental autoimmune encephalomyelitis (EAE). We observe that ABCs display distinct phenotypes during gHV68 infection and EAE: ABCs secrete anti-viral IFNg during gHV68 infection and IL10 during EAE. Intriguingly, latent gHV68 infection prior to EAE results in a heterogeneous ABC phenotype, with IL10 and IFNg-secreting subsets. As IFNg is pathogenic during EAE and MS, and IL10 is protective, the gHV68-EAE ABCs display a more pathogenic phenotype compared to EAE alone, corresponding to the enhanced clinical course during gHV68-EAE. Knocking out ABCs results in an amelioration of disease in gHV68-EAE, but not EAE alone, further substantiating that gHV68 infection drives ABCs towards pathogenicity. In MS patients we observe an increased number and altered inflammatory phenotype of circulating ABCs compared to age and sex-matched healthy controls. These findings indicate that gHV68 and EBV prime ABCs to contribute pathogenically during EAE and MS and suggest that ABCs may be a therapeutic target.
CX3CR1/CX3CL1 axis in breast cancer pathology and its therapeutic use
Authors: Singh Rajesh , Singh Santosh Kumar , Mishra Manoj K , Lillard James W
Published: January 2023

Recent reports from the breast cancer (BrCa) suggests that >90% of patients diagnosed at an early stage are successfully treated with surgery, chemotherapy, and radiation or combination. However, chemoresistance, recurrence and metastasis have been noticed in advanced BrCa patients and still remains a challenge. Chemokine receptor-ligand interaction has been noticed to play a significant role in homing to cancer cells to distant sites. Fractalkine (CX3CL1), a structurally distinct chemokine, and its receptor (CX3CR1), are expressed in majority of the cancer including BrCa. Thus, therapeutic strategies that inhibit tumor immune microenvironment of BrCa cells can improve the quality life of patients. In this study, we analyzed the expression and role of the CX3CR1/CX3CL1 axis interface in the progression and invasiveness of BrCa. In vitro investigation of BrCa cell lines MCF-7 and MDA-MB-231 showed higher expression of CX3CR1 analyzed through flow cytometry, RT-PCR, western blot, and immunofluorescence techniques. This was corroborated by our clinical tissue microarray results that showed significantly (p < 0.001) higher expression of CX3CR1 and CX3CL1 in cancerous tissues compared to normal tissues. Furthermore, treatment of BrCa cells with small molecule antagonist JMS-17-2 abrogated the proliferation and metastasis of cancer cells delineating the role CX3CR1-CX3CL1 chemotaxis mediated metastasis of BrCa. This data demonstrates, both biological and clinical significance of CX3CR1 and CX3CL1 expression that may contribute to BrCa metastasis. Inhibition of CX3CR1/CX3CL1 interaction thus could be a promising avenue for researchers to target or treat BrCa.
Validation of a murine multiplex panel to quantitate cytokine release syndrome biomarkers
Authors: Lehmann Jason S. , Mays-Aboagye Kelli , Liao Wen-Cheih , Zhao Amy , Ji Shaoquan , Ni Jessie
Published: January 2023

Cytokine release syndrome (CRS) is a systemic inflammatory response syndrome caused by adverse effects of monoclonal or CAR T cell immunotherapy. CRS can also arise as a complication during infection in response to sepsis. Regardless of its initial etiology, CRS is characterized by an auto-amplifying cascade of leukocyte activation which drives the production and release of several inflammatory biomarkers.

We have developed a murine-specific bead-based multiplex assay panel that simultaneously quantifies up to 13 CRS biomarkers from one sample volume aliquot using commonly available flow cytometers. This panel is manufactured with highly specific antibodies in order to ensure robust assay sensitivities, analytical accuracy, and reproducibility.

The panel was validated with biological samples generated from both in-vitro and in-vivo models of inflammatory cytokine release. Trends in cytokine expression differences between treatment groups were orthogonally confirmed using single target ELISAs.

Taken together, our cytokine release studies and rigorous performance validation establish this multiplex panel as a reliable tool for researchers seeking to mitigate the potentially fatal complications of CRS during infection and immunotherapy treatment.
Biological embedding from adverse childhood experiences in the development of an altered inflammatory state and poor health outcomes in young adults
Authors: Moore Jessy , Wade Terrance J. , Cairney John , O’Leary Deborah D. , MacNeil Adam J.
Published: January 2023

Exposure to emotional, physical, and/or sexual abuse as a child, cumulatively referred to as Adverse Childhood Experiences (ACEs), has been linked with poor health outcomes in early adulthood. Toward a mechanistic explanation for this link, dysregulated inflammation has garnered attention as a potential ‘social-immunological’ axis. Altered immune profiles are associated with ACEs and may contribute to mental health status, cardiovascular disease, and early mortality. Increases in circulating interleukin-6 (IL-6), C-reactive protein (CRP), and tumor necrosis factor α (TNFα) have been detected in individuals exposed to ACEs and are proposed to signal an immunological link to these outcomes. While this sets the stage in characterizing the inflammatory profile, gaps remain in the cast of mediators involved, preventing a more refined mechanistic view. We thus aim to expand the associated immunological cast. A sample of young adults from the Niagara Longitudinal Heart Study self-reported early life trauma (ACEs), and serum levels of a panel of immune biomarkers were quantified. We confirm that levels of IL-6, CRP and TNFα are altered, but also further identify new immune biomarkers that are significantly correlated (p<0.05) with ACEs, including pentraxin, chitinase, and interferons. This further refines the nature of ACE-dependent immunological dysregulation and offers new insights into contributing mechanisms, potential early risk-factor detection, and/or therapeutic targets in those exposed to ACEs. This evidence encourages promotion of public health initiatives and psychosocial preventative interventions aimed at limiting early adversity and its sequalae as an investment in later health status/quality of life.
UHRF1 Controls Thymocyte Fate Decisions Through the Epigenetic Regulation of EGR1 Expression
Authors: Zhang Yiwen , Chen Yingshi , Ma Rong , Wei Lai , Zhang Hui
Published: January 2023

Thymocyte differentiation is a highly complex process that is accompanied by epigenetic changes. Ubiquitin-like containing PHD ring finger 1 (UHRF1) is a critical epigenetic modifier involved in various cellular processes. Here, we demonstrated that it is highly expressed in T-cell precursors of the thymus. Further, its deficiency results in significantly-reduced thymocyte cellularity and thymus size. Through systematic analysis based on single-cell RNA-sequencing, we found that UHRF1 deficiency thwarts αβ T cell lineage development while biasing γδ lineage differentiation, dampens the progression of immature single-positive cells (ISPs). Further, the analysis of gene-regulatory networks demonstrated that UHRF1 controls the expression of early growth response 1 (EGR1). UHRF1 interacts with DNA methyltransferase 1 (DNMT1) at the CpG promoter region of Egr1 loci and affects the nearby chromatin modifications of H3K9me3 and H3K4me3. Taken together, our results demonstrate that UHRF1 is a key factor that mediates the epigenetic regulation of EGR1 and consequently thymocyte fate decisions.
Tissue plasminogen activator mobilizes neutrophils and T cells that exacerbate hemorrhagic transformation in stroke
Authors: Shi Kaibin , Liu Qiang , Jia Dong-Mei , Yang Xiaoxia , Zou Ming , Shi Samuel , Dong Jing-fei , Sheth Kevin N , Wang Xiaoying , Shi Fu-Dong
Published: January 2023

Hemorrhagic complications represent a major limitation of thrombolytic therapy with tissue plasminogen activator (tPA) in patients with ischemic stroke. Here we report that tPA induces a swift surge of circulating neutrophils and T cells in both ischemic stroke patients and a rat model of focal embolic stroke. tPA exacerbates endothelial injury and increases adhesion and migration of neutrophils and T cells, which are associated with brain hemorrhage in rats subjected to embolic stroke. Genetic ablation of annexin A2 in neutrophils and T cells diminishes the effect of tPA on these cells. Decoupling the interaction between mobilized neutrophils/T cells and the neurovascular unit, achieved by using a sphingosine-1-phosphate receptor 1 modulator RP101075 and a CCL2 synthesis inhibitor bindarit, which block lymphocyte egress and myeloid cell recruitment, respectively, attenuates hemorrhagic transformation and improves neurological function after tPA thrombolysis. Our study suggests that immune invasion of the neurovascular unit represents a previously unrecognized mechanism governing the emergence of tPA-mediated brain hemorrhage, which can be overcome by precise immune modulation during thrombolytic therapy.
Monocytes and monocyte-derived macrophages as mediators of sex differences in allergic asthma
Authors: Heller Nicola M , Diaz Mireya Becerra
Published: January 2023

Allergic asthma is a chronic, immune-mediated lung disease mediated that affects more than 200 million people worldwide. Alveolar macrophages (AM) and monocytes are important cellular contributors to asthma. During allergic lung inflammation, AM polarize to a predominantly M2 phenotype. Recruited monocyte-derived macrophages (MDM), rather than resident AM, have been linked to worse allergic lung inflammation by inducing inflammatory cell recruitment, tissue remodeling and lung fibrosis. There are sex differences in the prevalence and severity of asthma. Women have worse symptoms, longer hospital stays, more asthma-related deaths, and they are more refractory to therapy. Here, we wanted to determine if differences in M2 polarization and chemokine receptors by monocytes and MDM from women compared to men could account for sex differences in asthma pathogenesis. We obtained blood monocytes from healthy and asthmatic female and male donors in parallel. We analyzed the expression of chemokine and IL-4/IL-13 receptors, and in M2 macrophage genes in monocytes and MDM stimulated with IL-4 and IL-13. We found robust differences in CCR2, CCR5, and γC between healthy and asthmatic donors. Moreover, we found sex differences in M2 gene expression between asthmatic women and men. Female cells, in general, induced higher M2 gene expression in response to IL-4. Female monocytes expressed more chemokine receptors that would facilitate recruitment to the asthmatic lung. Our findings provide novel insights into how sex affects monocyte and macrophage polarization in asthmatic patients. These sex differences may represent a new therapeutic avenue for managing pathogenic M2 polarization and preventing monocyte recruitment to the allergic lung.
Mucosal IgE antibody secreting cell (ASC) repertoire analysis suggests an extrafollicular developmental program
Authors: Ramonell Richard Paul , Woodruff Matthew Charles , Saney Celia Lorenza , Duan Meixue , Lee Eun-Hyung
Published: January 2023

Allergic fungal rhinosinusitis (AFRS) is a subset of chronic rhinosinusitis with nasal polyposis (CRSwNP). AFRS is primarily associated with a type 2 inflammatory profile including eosinophilic mucosal infiltration and high serum and mucosal titers of IgE. Patients with AFRS have increased frequencies of IgE ASCs within NP structures, although it is unclear if these cells originate from a germinal center (GC) reaction or undergo local mucosal extrafollicular differentiation and class switch recombination (CSR). To better understand the origin of these cells, CD19+ and CD19− ASCs from human NPs (n = 3) were isolated for integrated single cell RNAseq and adaptive immune receptor repertoire (AIRRseq) profiling. Resulting AIRRseq data were piped into the ImMunoGeneTics (IMGT) V-Quest platform for comprehensive B cell receptor analysis. Interestingly, mutational frequency analysis showed that IgE ASCs have a lower V region mutation frequency (7.4 +/− 2.7%) compared with IgG (9.2 +/− 3.5%) and IgA (9.4 +/− 3.4%) ASCs from the same individuals. Additionally, IgE ASCs have an intermediate frequency of mutations in activation-induced deaminase WRC hotspots when compared with other isotypes. Finally, IgH repertoire lineage analysis does not indicate clonal expansion of NP IgE ASCs. These data suggest NP IgE ASCs compared to IgG and IgA ASCs from patients with AFRS may undergo a distinct developmental program that includes extrafollicular differentiation and CSR.
Characterization of milk-specific immunoglobulins of oral immunotherapy participants
Authors: Zhao Wei Wendy , Ke Danbing , Cohen Casey , Lejtenyi Duncan , Ben-Shoshan Moshe , Grunebaum Eyal , Upton Julia , Mazer Bruce
Published: January 2023

Since the standard treatment for food allergies such as cow’s milk allergy is strict avoidance and epinephrine injection upon anaphylaxis, alternative treatments are needed. Allergic reactions including anaphylaxis are presumed to be due to high-affinity immunoglobulin (Ig)E antibodies against cow’s milk proteins (CMP) such as casein. Currently, our laboratory is leading a pan-Canadian randomized controlled trial of milk oral immunotherapy (OIT). Participants in the OIT receive increasing doses of cow’s milk until 200ml of milk is tolerated without allergic reactions, followed by 12 months of follow-up while continuing to ingest milk and dairy at least twice a week. Using enzyme-linked immunosorbent assay (ELISA) to detect antibodies specific for casein, we determined that female participants have higher casein-specific IgE and IgG4 than male participants. We then aimed to assess whether the sex-dependent change in the affinity of immunoglobulins throughout OIT. Using a modified ELISA that assays for relative binding affinity, we found that the affinity of IgE for casein decreases in approximately half of the participants following the 12-months observation period, compared to the baseline. This is associated with an increase in tolerance and significant protection against minor symptoms or adverse reactions following milk ingestion. The results suggest that oral immunotherapy may induce changes in the affinity binding of allergen-specific IgE and contribute to the improvement of symptoms associated with allergy desensitization to milk.
Safe, rapid suppression of IgE-mediated allergy with monovalent (mv) anti-human (hu) FceRIa monoclonal antibody (mAb)
Authors: Khodoun Marat , Peipp Matthias , Morris Suzanne C , Angerman Elizabeth , Potter Crystal , Herr Andrew , Mulloy James , Wunderlich Mark , Finkelman Fred D
Published: January 2023

Background

Mast cells and FceRI, the high affinity IgE receptor (R), are central to allergy pathogenesis. Strong FceRI crosslinking induces severe allergy while low-level crosslinking removes IgE from mast cells and desensitizes them.

Hypothesis

IgE-mediated allergy can be safely suppressed by a mv anti-FceRIa mAb that persistently crosslinks FceRI to a limited extent.

Methods

We tested our hypothesis with: 1) divalent (dv) anti-huFceRIa mAbs that bind FceRI regardless of its association with IgE; 2) mv Fab/Fc derivatives of these mAbs, which do not directly crosslink FceRI; 3) mice that express hu, instead of mouse FceRIa; hu FceRIa mice that also have a mutant, hyperresponsive IL-4 receptor; and immunodeficient, human stem cell factor/IL-3/GM-CSF-secreting mice that develop large numbers of activated human mast cells following hu cord blood cell reconstitution; and 4) IL-4 and propranolol treatment that increases sensitivity to mast cell-released mediators.

Results

1) A single injection of mv anti-huFceRIa mAb protects our mice against IgE-mediated anaphylaxis and food allergy. 2) A rapid desensitization approach increases the safety of administration of this mv mAb. 3) The mv mAb decreases mast cell responsiveness to FceRI crosslinking and causes loss of most IgE. 4) Protection is maintained and increased by repeated injection of mv anti-huFceRIa mAb. 5) FcgRs are required for FceRI crosslinking by mv anti-FceRIa mAb. 6) Rapid desensitization with mv anti-FceRIa mAb provides a better combination of safety and efficacy than rapid desensitization with dv anti-FceRIa mAb or with a relevant allergen.

Conclusion

Desensitization with mv anti-huFceRIa mAb is a promising therapy for the suppression of hu IgE-mediated allergy.
A Randomized, Double-Blind, Placebo-Controlled Study of Daily Cannabidiol for the Treatment of Canine Osteoarthritis
Authors: Halpert Matthew , Verrico Chris , Wesson Shonda , Konduri Vanaja , Hofferek Colby , Vazquez-Perez Jonathan , Blair Emek , Dunner Kenneth , Salimpour Pedram , Decker William K
Published: January 2023

Hemp-derived cannabidiol (CBD) is the major non-psychoactive component of cannabis and has been promoted as a potential treatment for a wide variety of disparate inflammatory conditions. Here we evaluated CBD for its ability to modulate the production of pro-inflammatory cytokines in vitro and in murine models of induced inflammation and further validated the ability of a liposomal formulation to increase bioavailability in mice and in humans. Subsequently, the therapeutic potential of both naked and liposomally-encapsulated CBD was explored in a 4-week, randomized placebo-controlled, double-blinded study in a spontaneous canine model of osteoarthritis. In vitro and in mouse models, CBD significantly attenuated the production of pro-inflammatory cytokines IL-6 and TNF-a while elevating levels of anti-inflammatory IL-10. In the veterinary study, CBD significantly decreased pain and increased mobility in a dose-dependent fashion among animals with an affirmative diagnosis of osteoarthritis. Liposomal CBD (20 mg/day) was as effective as the highest dose of non-liposomal CBD (50 mg/day) in improving clinical outcomes. Hematocrit, comprehensive metabolic profile, and clinical chemistry indicated no significant detrimental impact of CBD administration over the four week analysis period. This study supports the safety and therapeutic potential of hemp-derived CBD for relieving arthritic pain and suggests follow-up investigations in humans is warranted.
Lipopolysaccharide (LPS) protects from a normally lethal dose of respiratory viral infection and affects post-airway disease in a dose dependent manner
Authors: Resiliac Jenny , Rohlfing Michelle , Santoro Jennifer , Hussain Syed-Rehan A , Grayson Mitchell
Published: January 2023

Rationale

Respiratory diseases are major public health concerns and leading causes of death and disability in the world. Understanding anti-viral immune responses is crucial to alleviate morbidity and mortality of respiratory infections. Previous data using our Sendai virus (SeV, murine parainfluenza virus type 1) mouse model suggested pretreatment of C57BL6 wild-type (WT) mice with 3μg lipopolysaccharide (LPS) reduced mortality from a normally lethal dose of SeV (2×106pfu) by half. This study was undertaken to identify the minimal effective dose for survival from the normally lethal SeV and determine its effects on subsequent airway disease.

Methods

WT mice were intranasally (i.n.) inoculated with 30 μL of 0.01–3 μg LPS or PBS 24 hours before lethal dose of SeV i.n. and survival determined. In surviving mice, invasive measurement of airway hyper-reactivity (AHR) was determined on d21 post-viral inoculation.

Results

LPS mediated survival and AHR were dose dependent (p<0.0001 comparing all groups, Mantel-Cox for survival and ANOVA for AHR). Mice receiving lower doses of LPS (0.03 and 0.1 μg) had increased survival (92±8% and 67±10% survival, respectively) and decreased AHR compared to those with the highest doses (n≥5 mice per dose with up to 4 experimental repeats).

Conclusion

LPS mediated survival from a lethal paramyxoviral infection is greatest with lower doses of LPS (0.1–0.03 μg). These doses have been associated with development of atopic disease (Eisenbarth SC, J Exp Med. 2002). We are currently undertaking studies to determine the mechanism(s) through which low dose LPS inhibits mortality from normally lethal SeV dose and how this reduces post-SeV atopy.
Microbial exposure increases the magnitude and diversity of peritoneal inflammatory cell infiltrates and cytokine storm during sepsis
Authors: Martin Matthew D , Akal Peter Kassis , Schelzel George , Swanson Whitney , Sjaastad Frances V , Kucaba Tamara A , Badovinac Vladimir P , Griffith Thomas S
Published: January 2023

Sepsis – defined as a life-threatening organ dysfunction caused by a dysregulated host response to infection - remains a major healthcare burden and cause of death in the United States and worldwide. While there has been a vast expansion in knowledge regarding the impact of sepsis on the immune system, virtually all preclinical sepsis research to date has relied on using mice housed under specific pathogen-free (SPF) conditions. Environmental pathogen exposure is one important difference between basic human and laboratory mouse biology that must be considered when using mice to evaluate immune system fitness. Interestingly, B6 mice that have been cohoused with pet store mice (to permit microbial transfer and immune system maturation; “CoH” mice) exhibit exaggerated acute hyperinflammation, morbidity, and mortality during cecal ligation and puncture (CLP)-induced polymicrobial sepsis compared to SPF B6 mice. To understand the mechanism of this increased acute response, we examined peritoneal immune cell composition in SPF and CoH mice before and after sepsis induction. CoH mice have more ‘large’ MHC IIloF4/80hi and ‘small’ MHC IIhiF4/80lo peritoneal macrophages (LPM and SPM) and neutrophils than SPF mice at baseline. Upon the induction of polymicrobial-induced peritonitis, there was increased recruitment of neutrophils, monocytes, T cells, and NK cells into the peritoneum of CoH mice vs. SPF mice. This increased immune cell influx correlated with increased CXCL1 and CCL2 in peritoneal lavage samples from CoH mice. CoH mice devoid of neutrophils or LPM exhibit reduced cytokine storm and improved outcomes after sepsis induction. These data suggest the cells of the innate immune system govern the magnitude of the acute septic response.
The effects of functional platelet-derived extracellular mitochondria on the inflammatory phenotype of neutrophils
Authors: Boudreau Luc H , Foulem Robert D , Léger Jacob L
Published: January 2023

Unregulated inflammation has been associated with chronic inflammatory auto-immune diseases, such as rheumatoid arthritis, multiple sclerosis and atherosclerosis. Polymorphonuclear neutrophils (PMNL) are amongst the immune cells involved in the acute inflammatory response used to fight bacterial infections. Once activated, PMNL release inflammatory mediators in the extracellular milieu to recruit various immune cells required to fight the invading pathogens. Platelets (PLTs) are also implicated in the body’s inflammatory response. Interestingly, activated PLTs can release fully functional mitochondria in the extracellular milieu. Known as the powerhouse of the cell, the mitochondria share similar characteristics with bacteria. Therefore, we hypothesize that PLTs-derived mitochondria present in the extracellular milieu, acting in a similar way as bacteria, induce a sterile inflammatory response that involves the PMNL. The objective of this study was to investigate the sterile inflammatory response of PMNL caused by the exposure of PLTs-derived extracellular mitochondria. Following the co-incubation of PMNL with various physiological doses of PLTs-derived mitochondria, a characterization of the interaction between PMNL and PLTs-derived mitochondria and an investigation of the inflammatory proprieties of PMNL was performed. We have shown that PLTs-derived mitochondria associate with PMNL, resulting in a 2.47-fold mitochondrial-dependent increase in oxygen consumption. Data also shows that PLTs-derived mitochondria significantly induce the release of PMNL microparticles. The knowledge gained from this study provides insight into the mechanism of sterile inflammation in auto-immune diseases.
Glutathione peroxidase 8 negatively regulates caspase-4/11 to protect against colitis
Authors: Hsu Jye-Lin , Chen Tzu-Fan
Published: January 2023

Human caspase-4 and its mouse homolog caspase-11 are receptors for cytoplasmic lipopolysaccharide. Activation of the caspase-4/11-dependent NLRP3 inflammasome is required for innate defense and endotoxic shock, but how caspase-4/11 is modulated remains unclear. Here, we show that mice lacking the oxidative stress sensor glutathione peroxidase 8 (GPx8) are more susceptible to colitis and endotoxic shock, and exhibit reduced richness and diversity of the gut microbiome. C57BL/6 mice that underwent adoptive cell transfer of GPx8-deficient macrophages displayed a similar phenotype of enhanced colitis, indicating a critical role of GPx8 in macrophages. GPx8 bound covalently to caspase-4/11 via disulfide bonding between cysteine 79 of GPx8 and cysteine 118 of caspase-4, which restrained caspase-4/11 activation, while GPx8 deficiency led to caspase-4/11-induced inflammation during colitis and septic shock. Inhibition of caspase-4/11 activation with small molecules reduced the severity of colitis in GPx8-deficient mice. Notably, colonic tissue from patients with ulcerative colitis displayed low levels of Gpx8 and high caspase-4 expression. In conclusion, these results suggest that GPx8 protects against colitis by negatively regulating caspase-4/11 activity.
The adr-2 Mutant Strain of C. elegans and the Effect of Gold Nanoparticles on Life Span and Induction of Immune Responses
Authors: Meiner Kavan V , Elfert Amr , Cotlet Mircea , Idris Sally , Delaney Tetyana
Published: January 2023

Nanoparticles (NP) are widely used for domestic, industrial and medical applications. However, because of their nano-size they can easily penetrate cellular membranes, causing increased damage to the mitochondrial and genomic DNA. NPs also were reported to induce inflammation, cell apoptosis and oxidative stress.

In this research, the effect of gold NPs on growth, reproduction, fecundity, life span as well as induction of immune responses in Caenorhabditis elegans (C. elegans) are studied. The nematode has proven to be the best model to study effects difficult to explore in humans. The adr-2 mutant C. elegans is highly susceptible to environmental toxicity and offer sensitive bioassays to study the impacts of gold NPs on the worm. Two groups of adr-2 mutant C. elegans were grown. One group was exposed to gold NPs and the responses were analyzed and compared to the control group without nanoparticles. The two groups of C. elegans were observed over 2 months, thrashes were counted and the experiment was repeated several times. Findings showed that the thrash count for mutant strains decreases. Thus, at 0 minutes the thrash count was 55, reached 45 at two minutes, and decreased to 30 at four minutes. The other environmental contributors to this effect are under evaluation. The preliminary results showed that both groups had no visible changes. Nematodes were imaged using confocal differential interference contrast (DIC) microscopy. Several different pathways (p38 MAPK (PMK-1) and insulin like receptors (DAF-2)) could be activated to contribute to stress resistance to the NPs. Mutant strains of C. elegans for these receptors need to be tested to get a better understanding of the effect of the gold NPs on the immune responses.
Development of interferon-stimulated gene expression from embryogenesis through adulthood, with and without constitutive MDA5 pathway activation
Authors: Poeschla Eric M. , Bankers Laura
Published: January 2023

Pathogen-associated molecular patterns, e.g. dsRNA, activate expression of interferon-stimulated genes (ISGs), which protects hosts from infection. While transient ISG upregulation is essential for effective innate immunity, constitutive activation typically causes harmful autoimmunity in mice and humans, often including severe developmental abnormalities. We have shown that transgenic mice expressing a picornavirus RNA-dependent RNA polymerase outside the viral context (RdRP mice) exhibit lifelong, MDA5-dependent, and quantitatively dramatic upregulation of many ISGs, which confers broad viral infection resistance. Remarkably, RdRP mice never develop autoinflammation, interferonopathy, or any other discernible abnormalities. Here we used RNA-seq and other methods to analyze ISG expression across five time points from fetal development to adulthood in both wild-type and RdRP mice. In RdRP mice, the proportion of upregulated ISGs increased during development, with the most dramatic induction occurring two weeks postnatally. The immensely amplified ISG profile is then maintained lifelong. We further determined that blocking the type I interferon receptor reverses the amplified ISG transcriptome in adults. Molecular pathways and biological functions associated with innate immune signaling are only activated postnatally, suggesting dampened fetal responsiveness to innate immune stimuli. Similarly, functions supporting viral replication are only inhibited postnatally. In conclusion, the uniquely tolerated amplified ISG state of RdRP mice is mostly triggered early postnatally, is maintained through adulthood, and requires ongoing type I IFN signaling to maintain the augmented innate immune state.
Exercise-stimulated resolvin biosynthesis in adipose tissue is abrogated by obesity-induced adrenergic dysfunction
Authors: Hellmann Jason , Lynch William , Calderin Ernesto Pena , Sansbury Brian E. , Spite Matthew , Hill Bradford G.
Published: January 2023

Adipose tissue (AT) inflammation is associated with obesity-induced insulin resistance. Exercise (Exe) prevents the development of chronic inflammatory diseases and insulin resistance largely through unknown mechanisms. We provide data that specialized proresolving lipid mediators (SPM) contribute to the anti-inflammatory effect of Exe. Compared with sedentary controls (Sed), mice exposed to 4 wk of Exe showed elevated AT expression of SPM biosynthetic enzyme Alox15 (Sed 1.12±0.2 vs Exe 1.68±0.1 relative expression, n=5–7), SPM levels (Sed vs Exe; RvD1 156.7±12.0 vs 223.0±24.7; 17R-RvD1 354.0±35.9 vs 534.4±98.9; RvD4 31.31±6.9 vs 123.1±30.7; and 17R-RvD3 2.92±1.1 vs 8.20±1.7 pg/g, n=5–7), and M2 macrophages (F4/80+CD301+; Sed 17.5±3.2 vs Exe 40.9±4.5 %F4/80+, n=4–6). These changes were diet-dependent as high fat (HFD) abrogated this Exe effect in AT when compared with low fat (LFD) controls. The exercise-stimulated Alox15 expression was restricted to AT macrophages (F4/80+ cells; LFD Sed 0.11±0.03 vs LFD Exe 0.23±0.03; HFD Sed 0.10±0.04 vs HFD Exe 0.08±0.02 relative expression, n=3–6), as adipocytes isolated from Exe mice showed no difference compared with Sed controls. That epinephrine (Epi) is released with exercise and stimulates macrophage Alox15 expression and RvD1 production, we questioned if Epi biosynthesis is affected by HFD. HFD feeding diminished expression of phenylethanolamine N-methyltransferase compared with controls (LFD 1.0±0.25 vs HFD 0.36±0.15, n=4). Similarly, obese-diabetic mice (db/db) also showed reduced catecholamine production and biosynthetic enzyme levels. These results suggest that obesity-induced adrenergic dysfunction abrogates exercise-stimulated resolvin production in AT.
Mutations in the critical domain of human cathelicidin LL-37 affect neutralization of IAV
Authors: De Luna Xavier , White Mitchell , Hsieh I-Ni , Hartshorn Kevan L
Published: January 2023

Influenza A Virus (IAV) is the virus responsible for the flu and affects 3 to 5 million people around the world. Although vaccines exist for common strains of IAV, whenever pandemic strains of IAV arise humans are largely unprotected. Thus, new treatment strategies are required for pandemic strains of IAV as well as serious cases of IAV infection that can result in a bacterial superinfection. Treatments that reduce viral load in an infected host would be optimal to reduce virus spread as well as to potentially alleviate viral burden on the host. Mammals have many host defense peptides such defensins, lectins and cathelicidins. Cathelicidins are short cationic peptides that are antimicrobial in nature, and have been shown to have anti-IAV properties along with other antimicrobial functions. A critical domain of LL-37, the only human cathelicidin, has been shown to have a central helix with a hydrophobic surface. This critical domain is the minimum sequence of LL-37 required to have antimicrobial properties. There is no consensus on the mechanism LL-37 utilizes to neutralize bacteria and viruses. However, we hypothesize that by mutating single hydrophobic residues in the critical domain to a hydrophilic residue we can see changes in LL-37 neutralization of various IAV strains. Using fluorescent focus assays, we can quantify IAV titers after exposure to LL-37 and other cationic peptides. Here we show that reducing the hydrophobic residues within the critical domain of LL-37 reduces seasonal and pandemic IAV titers in vitro. These results lend credence to the theory of the hydrophobic surface of LL-37 is responsible for its anti-IAV effects.
Hyper-ISGylation promotes acute pulmonary pathology during chronic viral infection
Authors: Shaabani Namir , Zak Jaroslav , Teijaro John R
Published: January 2023

ISG15 is one of the strongest type I interferon (IFN-I) stimulated genes (ISGs) which is conjugated to proteins in an enzymatic process termed ISGylation. Protein ISGylation is subsequently reversed by the enzyme Usp18 which acts in a de-ubiqutinase fashion to remove SIG15 from target proteins. Many proteins are ISGylated following IFN-I signaling however, immunological studies have been limited to the anti-viral properties of ISG15 conjugation while its regulation of innate and adaptive immune responses has been poorly studied. Here using Usp18−/− and enzymatically inactive (Usp18C61A/C61A Knock-in) mouse models, we show that hyper-ISGylation during chronic viral infection leads to severe immune pathology characterized by hematological disruptions, cytokine amplification, lung vascular leakage and death. The pathology associated with chronic infection required T cells but not T cell-intrinsic Usp18 expression. We reveal that lack of Usp18 in LysM+ cells reproduced the pathological manifestations observed in Usp18−/− or Usp18C61A/C61A mice. Using single cell sequencing, we observed that hyper-ISGylation during chronic infection altered neutrophil maturation and promoted an inflammatory neutrophil lung infiltrate. Importantly, neutrophil depletion reversed morbidity and mortality in Usp18C61A/C61A mice. In summary, we reveal Usp18’s enzymatic function is crucial for regulating inflammatory neutrophil differentiation and preventing immune hyperactivation during chronic viral infection. Moreover, our results suggest that hyper-ISGylation may drive the inflammatory pathology observed in Usp18-null patients and indicate that regulation of ISG15 expression or conjugation could lessen disease.
CD28 Induces Autophagy in Plasma Cells to Enhance Mitochondrial Respiration and Survival
Authors: Peng Peng , Liu Wensheng , Utley Adam , Chavel Colin , Carlson Louise , Olejniczak Scott H , Lee Kelvin P
Published: January 2023

Durable humoral immunity relies on the persistence of long-lived plasma cells (LLPC) that continuously produce protective antibodies. The prototypical T cell costimulatory molecule CD28, that is also expressed on LLPC, is critical to their survival but the mechanisms involved remain unclear. We found CD28 activation enhances autophagy in LLPC/multiple myeloma as evidenced by increased level of autophagy marker LC3II, elevated autophagosome numbers and decreased level of autophagic cargo receptor P62. Inhibition of autophagy by 3MA or BafA abolished CD28’s pro-survival effects. Mechanistically, CD28 activation increased the protein level of ATG5, a critical autophagy regulator, even with translation blockade, but this was abrogated by proteasome inhibition. Knocking down atg5 eliminated CD28’s pro-survival effects, suggesting CD28 signaling regulates ATG5 degradation to sustain pro-survival autophagy. CD28 activation significantly increased mitochondrial respiration and blocking autophagy prevented this. Free fatty acids (FFA) can be degraded from cellular lipid droplets (LD) by lipophage, a selective form of autophagy, to fuel mitochondrial respiration. We found CD28 activation decreased LD staining and blocking autophagy prevented this. Blocking lipophagic lipase by lalistat eliminated CD28-induced oxidative phosphorylation and pro-survival effects. Addition of FFA (oleic and palmitic acids) boosted oxidative phosphorylation and restored survival with autophagy inhibition but not with fatty acid oxidation blockade. These data suggest CD28 signaling regulates ATG5 protein degradation to enhance autophagy and this in return produces FFA to sustain mitochondrial respiration and support LLPC survival.
Friend or foe? A systematic evaluation of NKT and NK cells as prognostic markers in colorectal carcinoma
Authors: Feng Hao , Tong Huan , Thasler Wolfgang , Xia Qiang
Published: January 2023

BACKGROUND

Natural killer t(NKT) cells and Natural killer (NK) cells orchestrate immune responses in tumor progression. NKT and NK cells have been suggested as prognostic markers for cancer patients. However, the results were controversial. In the present study, we present a systematic review and meta-analysis of studies assessing the prognostic value of NKT and NK cells in colorectal cancers, respectively.

METHODS

We performed a systematic literature search and included studies using immunofluorescence or flow cytometry and survival analysis to assess the prognostic value of NKT and NK cells in colorectal cancer. We performed a meta-analysis of studies providing necessary statistical data and investigated prognostic value of NKT and NK cells.

RESULTS

Of the 275 articles identified, 169 were eligible for this systematic review, and eight were included in the meta-analysis. Both NKT and NK cells were the most promising predictors of disease-free survival (DFS) and overall survival (OS) in colorectal cancer patients. Both NKT-high and NK-high were positively correlated to better OS, respectively (NKT: p=0.002, HR 2.91, 95%CI 1.46~5.80; NK: p<0.0001, HR 3.87, 95%CI 1.97–7.62). Intriguingly, NK-high was also associated with better DFS (NK: p<0.0001, HR 3.42, 95%CI 2.05–5.69); however, NKT-high presented the opposite trend for DFS(NKT: p<0.0001, HR 5.28, 95%CI 2.35–11.84).

CONCLUSION

Proper reporting of the association between NKT/NK allows the reader to evaluate the quality and reliability of the previously reported data. The promising results might boost the interest in tumor immunology such as CAR-NK and CAR-NKT. The prognostic value of NKT and NK cells should be validated in large, standardized studies.
Novel role for gelatinases in modulating metabolic reprogramming associated with T cell receptor stimulation in CD4+ T cell
Authors: Onwuha-Ekpete Lillian , Tokmina-Roszyk Dorota , Fields Gregg
Published: January 2023

The gelatinases, MMP-2 and MMP-9, are proteolytic enzymes with diverse physiological roles. They are linked with various aspects of the immune response, such as leukocyte trafficking and cytokine activation. However, their dysregulation is associated with various pathologies such as metastasis in cancer progression and blood-brain barrier disruption in multiple sclerosis. Much of what is known about the gelatinases is in context of their extracellular activities. However, recent studies including this study suggest an intracellular role in immune cell activation.

Our findings suggest, that gelatinase activity can modulate T cell bioenergetics, affecting both oxidative phosphorylation and the glycolytic pathways with a greater influence on the latter. This results in a mitigated CD4+ T cell response to TCR stimulation. Upon stimulation, CD4+ T cells lacking gelatinase activity, demonstrate a reduced capacity to be activated as determined by the expression of early activation markers, proliferation, and the production of effector cytokines. In addition, RNA-seq analysis of control and gelatinase inhibited CD4+ T cells highlight key proteins associated with both the immune system and metabolic processes as predicted by STRING. We confirm our pharmacological studies with the genetic ablation of the gelatinases and further analysis of these shRNA treated T cells using the Nanostring metabolic panel. In addition, we demonstrate in-vivo relevancy using a murine model for multiple sclerosis known to be initiated by CD4+ T cells; gelatinase inhibition resulted in reduced clinical severity. These results suggest an intracellular role for gelatinases in CD4+ T cell mediated diseases and their potential as therapeutic targets.
Perinatal androgen exposure impacts mast cell development and mediator storage
Authors: Bali Vedrana , Mackey Emily , Moeser Adam
Published: January 2023

Mast cell (MC)-associated diseases often exhibit a female sex bias in both prepubertal children and adults. We have shown that female MCs store and release greater amounts of histamine which correlates with heightened anaphylaxis in both prepubertal and adult mice. Further, perinatal androgen exposure, a major organizing event in sex biology, reduced MC histamine storage and release, and the severity of anaphylaxis in prepubertal and adult female mice, thus demonstrating that perinatal androgens are critical early life drivers of MC sex differences, but the mechanisms remain poorly understood. The objective of this study was to determine how perinatal androgens impact MC development. RNA sequencing analysis of BMMCs derived from adult female and male mice that were previously exposed to perinatal androgens revealed significant down-regulation of a number of genes related to granule biogenesis and mediator storage of which included lactotransferrin (by −2.6 fold), proteoglycan 2 (by −3.9 fold), lipocalin 2 (by −1.4-fold), elastase (by −2.5 fold) and myeloperoxidase (by 1.4 fold). Current studies aim to confirm RNA seq data with qPCR and protein expression analysis and to define the critical period of androgen programming on MC development and function. Understanding the mechanisms by which early life androgens drive sex differences in MCs will be critical in understanding the origins of sex differences in MC and its role in health and disease.
Each feature of Th plasticity is regulated individually, balancing pathology and protection in malaria
Authors: Stephens Robin , Aussenac Florentin , Clark Lucinda Puebla , Wilson Kyle D. , Dent Alexander L. , Carpio Victor H.
Published: January 2023

Hybrid Th1/Tfh cells (IFN-γ+IL-21+CXCR5+) predominate in response to persistent infections. However, it is not clear if the function of these cells is plastic and might be modulated. In infection with Plasmodium spp, an IFN-γ+ T helper-1 (Th1) response controls initial parasitemia, while antibody and IL-21+CXCR5+ T follicular helper (Tfh) function effect final clearance. Supporting plasticity, we found that CD4-intrinsic Bcl6, Blimp-1 and STAT3 all regulate T-bet expression, which controls IFN-γ expression. While Bcl6 and Blimp-1 regulate the level of CXCR5, only T-bet, IL-27Rα and STAT3 strongly affected the cytokine bias of the Th1/Tfh phenotype. Infected mice with STAT3-deficient T cells produced less antibody, and more Th1-like IFN-γ+IL-21−CXCR5lo T cells, significantly increasing protection from re-infection. Conversely, reduced Th1 bias in re-infected T-bet KO was reflected in prolonged secondary parasitemia. Signs of pathology were somewhat prolonged in Th1-biased animals supporting an evolutionary tradeoff between control of parasite and tissue damage. In summary, each feature of hybrid Th1/Tfh population in Plasmodium infection is uniquely regulated and the cytokine bias of memory T cells can be modified to enhance the effectiveness of the response.