bluuebunny/crossref_metadata_2025_split huggingface dataset:
Critical role of STAT5 tetramerization in the pathogenesis of experimental autoimmune encephalomyelitis, a murine model of multiple sclerosis
Authors: Monaghan Kelly Lynn , Farris Breanne Y , Amend Courtney D , Lin Jianxin , Leonard Warren J , Wan Edwin C.K. 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an immune mediated disease that predominantly impacts the central nervous system (CNS). Current MS treatments have demonstrated the ability to reduce the rate of MS relapses; however, they are not effective at preventing disease progression. Elucidating the pathways that govern the pathogenesis of MS may identify molecular mediators, which can act as novel therapeutic targets to treat MS progression. STAT5 is a transcription factor critical for the generation, activation, and function of different immune cell types. Upon activation, STAT5 can form dimers or tetramers, which have distinct functions. Using a Stat5a-Stat5b double knock-in N-domain mutant mouse strain (DKI mice), in which STAT5 dimers are functional, but STAT5 tetramers cannot be formed, we demonstrated that DKI mice developed less severe experimental autoimmune encephalomyelitis (EAE), a model of MS, compared to WT littermates. Moreover, a portion of DKI mice did not develop EAE symptoms. Flow cytometric analysis of peripheral blood revealed that during the early phase of EAE, WT and DKI mice exhibited similar levels of immune cell expansion in the periphery. However, at the peak of EAE, DKI mice had fewer immune cells in the CNS. This suggests that STAT5 tetramers promote immune cell egress to the CNS or/and immune cell expansion within the CNS. In addition, multiplex protein assays established that the expression of a pro-inflammatory cytokine and several chemokines, including IL-1α, CCL3, CCL17, and CCL22, are reduced in DKI mice compared to WT mice, which may account for the reduced disease severity observed in DKI mice. Taken together, the data suggest that STAT5 tetramers plays a multifaceted role in promoting inflammation resulting in EAE.
Distinct waves of CD8 T cell affinity during demyelinating disease progression
Authors: Jacobs Jesica R , Blanchfield Lori , Liu Baoyu , Evavold Brian D 
Published: January 2023

Abstract
The animal model of Multiple Sclerosis, Experimental Autoimmune Encephalomyelitis (EAE), requires CD4 T cells to induce demyelinating disease. However, CD8 T cells are ever present during EAE and have been hypothesized to have both effector and suppressor functions. To understand autoimmune disease progression, we analyzed the affinity and genetic profile of the CD4 and CD8 T cell response over time. Affinity analysis of CD8 T cells during the autoimmune disease course revealed the greatest affinity for myelin at peak disease (d21-23), whereas affinities were significantly lower at onset (d14) and chronic (d36-38) timepoints. Conversely, affinity analysis of CD4 T cells revealed no significant changes at the same time points. Additionally, CD8 T cells displayed a smaller, 10-fold range in affinity, suggestive of a restricted TCR clonal response. CD4 affinities covered a much wider 10,000-fold range, consisting of low and high affinity clones. To further understand the dynamics of the T cell population during disease, we performed single cell RNA-sequencing. Heterogenous populations of CD4 and CD8 T cells were observed at peak EAE disease, including a genetically distinct population of FoxP3+, CD25+ CD8 T cells which accounted for 3.33% the total CD8 population. These genomic data paired with the known associations between affinity and T cell fate suggest phenotypically and functionally distinct waves of CD8 T cells with limited clonality infiltrate the CNS during EAE progression.
Modulation of liver protein sialylation drives spontaneous generation of fatty liver and modulates multiple inflammatory models
Authors: Oswald Douglas M , Jones Mark B , Cobb Brian A 
Published: January 2023

Abstract
Circulatory protein glycosylation has long been recognized as a marker of multiple disease states and has been applied clinically to gauge diseases including liver dysfunction, heart disease, and diabetes. The etiology and mechanism of this relationship, including whether glycosylation changes are causative in or only indicative of inflammatory shifts, has not yet been elucidated. We present here a model in which the α 2,6 sialyltransferase (ST6Gal1) was selectively knocked out of hepatocytes (H-cKO) using a Cre-lox recombination model driven by albumin expression. We confirm the loss of circulatory glycoprotein α 2,6 sialylation and note no obvious dysfunction in young (8–20 week old) H-cKO mice at baseline. Upon aging, the H-cKO mice spontaneously generate fatty liver disease, visible by gross observation and histologic examination, that is indicative of underlying inflammatory dysfunction. Further, H-cKO mice demonstrate exacerbated disease in models of thioglycolate-driven peritoneal inflammation, dextran sodium sulfate-driven colitis, and house dust mite-driven asthma. Our work positions liver protein secretion and glycosylation as not only indicative of inflammatory disease, but also as a causative factor in disease incidence and severity.
Loss of local innervation induces lymph node expansion
Authors: Chen Chien-Sin , Ince Louise , de Juan Alba , Scheiermann Christoph 
Published: January 2023

Abstract
The nervous system directly innervates both lymphoid and non-lymphoid tissues, but the influence of neural input on regional immune responses remains unclear. Local, unilateral deprivation of nerves to the popliteal lymph node (popLN) and its draining area by surgical denervation led to ipsilateral paw thickening and dramatic nodal expansion of all leukocyte subsets investigated. However, denervation surgeries on nerves bypassing the popLN, such as cutting the femoral nerve, the sciatic nerve below the popliteal fossa or individual branches of the sciatic nerve, failed to increase lymph node cellularity. Expansion was associated with increased expression of Cxcl13, Il1a, Il1b, Il6, Il10, Il17a and Il17f levels in denervated popLNs and paws. Adoptive cell transfer revealed enhanced cellular trafficking of lymphocytes to the lymph node, accounting for increased cellularity as this was ablated in mice receiving antibodies blocking lymph node homing. Additionally, immunofluorescence analyses revealed high levels of B cell proliferation and germinal center formation, which was mirrored by enhanced antibody titers in serum. Blockade of antigen presentation using an MHC class II-neutralizing antibody or surgically ablating drainage of afferent lymphatics to the popLN inhibited denervation-induced nodal expansion. Together, these data provide evidence for an important regulatory role of direct neural innervation in local immune responses.
Induced hyperglycemia promotes sensitization and exacerbates allergic inflammation in a mouse model of asthma
Authors: Queener Ashley , Jeong Brian M , Doan Ton C , Chiarella Sergio E , Walker Matthew T , Coden Mackenzie E , Nagasaka Reina , Berdnikovs Sergejs 
Published: January 2023

Abstract
Glucose is a key source of energy in systemic and cellular metabolism and has known pro-inflammatory properties. It has recently been acknowledged that associations exist between metabolic disorders (diabetes, insulin resistance, obesity), asthma and other allergic diseases. As the prevalence of both metabolic and allergic disorders increases, it is important to understand how metabolism can influence the development of allergy. First, we injected fasted wild type Balbc/J mice intraperitoneally (i.p.) with 2g/kg dextrose (standard hyperglycemic challenge) to test whether hyperglycemia promotes inflammation favoring antigen sensitization. Within one hour of injection, we detected a rapid increase in blood glucose levels (up to 130 mg/dL) followed by an increase in Il1β, Tslp, Cxcl9, and Siglec5 gene expression in the peritoneal tissue compared to vehicle controls. Within 2 hours post injection, IL-1β protein was detected in peritoneal cellular infiltrate. In order to determine whether glucose-induced inflammation promotes allergic response, we sensitized mice by i.p. to chicken egg ovalbumin (OVA) using either alum (standard adjuvant), dextrose or vehicle control. Both alum/OVA and dextrose/OVA mounted lung allergic inflammation in response to inhaled OVA antigen. Dextrose/OVA mice had OVA-specific IgE production similar to alum/OVA group, demonstrating ability to promote sensitization. Remarkably, total cellular infiltrates, bronchoalveolar lavage eosinophils, lung expression of IL-4, IL-13 and IL-33 in dextrose-sensitized mice were equal or surpassing allergic inflammatory responses in mice given standard adjuvant. These results demonstrate a potentially critical role for glucose in promotion of allergy.
ICOS signaling in visceral adipose tissue regulatory T cell homeostasis and function
Authors: Mittelsteadt Kristen L , Campbell Daniel J. 
Published: January 2023

Abstract
Obesity and insulin resistance/type 2 diabetes are linked by chronic inflammation in visceral adipose tissue (VAT). Foxp3+ regulatory T cells (TR) are critical for maintaining immune tolerance and are recently appreciated for their role in tissue repair and homeostasis. Indeed, a unique population of TR reside in the VAT of mice and humans, and these TR regulate adipose inflammation and preserve insulin sensitivity and glucose tolerance. ICOS is a costimulatory molecule expressed at high levels by TR. Surprisingly, the abundance of TR in VAT is elevated by nearly 2-fold in IcosKO mice compared to WT controls, and IcosKO TR preferentially reconstitute the VAT of WT:IcosKO mixed bone marrow chimeras. TR in the VAT of IcosKO mice express high levels of VAT-TR markers, like ST2, KLRG1, and IL-10, compared to WT mice, suggesting a potent ability to prevent insulin resistance and diet-induced metabolic syndrome. Ongoing studies aim to determine whether IcosKO mice are more resistant to diet-induced metabolic syndrome by assessing insulin sensitivity/glucose tolerance, indices of inflammation, and VAT immune cell phenotype and function, and to mechanistically elucidate how ICOS signaling controls the accumulation and phenotype of VAT-TR. These studies will yield important information on the molecular pathways involved in regulating the onset of metabolic syndrome and provide new potential therapeutic targets.
Egr2 deficiency significantly impairs the development of pathogenic TH17 cells
Authors: Gao Yuanyuan , Wang Yan , Guan Yukun , Bouladoux Nicolas Jean , Break Timothy , Lionakis Michail S , Belkaid Yasmine , Lazarevic Vanja 
Published: January 2023

Abstract
TH17 cells have critical functions in host defense at mucosal barriers, but they can also be the key drivers of immunopathology. Since their discovery, multiple transcription factor networks have been described that direct Th17 lineage commitment; however, molecular switches that control the development of “pathogenic” versus “non-pathogenic” TH17 cells remain largely unknown. In this study, we have identified the transcription factor Egr2 as a critical regulator of the TH17 pathogenic program. When ectopically expressed under TH17-polarizing conditions, Egr2 significantly enhanced the expression of TH17 signature genes in a RORγt-dependent manner. Although Egr2-deficient CD4+ T cells could be polarized in vitro into TH17 cells, these cells lacked the pathogenic potential in a TH17-dependent disease model, experimental autoimmune encephalomyelitis (EAE). In both active EAE and passive EAE, Egr2-deficeint TH17 cells failed to induce neuroinflammatory response in the CNS. Analysis of CNS infiltrates revealed reduced accumulation of IL-17A-producing CD4+ T cells and impaired recruitment of monocytes and myeloid dendritic cells in mice with T cell-specific loss of Egr2. Most importantly, transcriptional analysis of ex vivo isolated Egr2-deficient CD4+ T cells revealed that Egr2 controlled the expression of pathogenicity-associated genes in TH17 cells in the CNS. We further show that Egr2 deficiency did not affect the effector function of protective TH17 cells at mucosal barriers in Citrobacter rodentium and Candida albicans infections, suggesting that Egr2 is an attractive candidate for the therapeutic targeting of pathogenic TH17 cells while preserving tissue-protective functions of TH17 cells at barrier sites.
Persistent antigen can maintain lung resident memory CD8 T cells long-term
Authors: Uddback Ida Elin Maria , Cartwright Emily , Schøller Amalie Skak , Hayward Sarah L , Lobby Jenny , Takamura Shiki , Thomsen Allan R , Kohlmeier Jacob E , Christsensen Jan Pravsgaard 
Published: January 2023

Abstract
A sub-population of memory T cells, tissue resident memory T cells (TRM), have in recent years gained attention for their indispensable role in heterologous protection against respiratory viruses. For this reason, they are of great interest for vaccine research. However, the number of lung TRM decline within a few months after infection and researchers have been unable to determine how the TRM population, and protection, can be made to last long-term in the lung. We have previously shown that an Adenovirus expressing nucleoprotein (AdNP) from influenza A virus induce CD8 T cell mediated heterosubtypic immunity lasting up to at least 8 months post vaccination when administered locally and systemically, but the underlying reason was not investigated.
Here, we show that the number of CD8 T cell in the lung induced by AdNP is significantly higher compared to cells induced by an influenza infection. Using AdNP, we show that CD8 TRM in the lung can be maintained for at least one year post-vaccination. Our results revealed that lung TRM continued to proliferate in-situ 8 months after AdNP vaccination. Importantly, this required pulmonary vaccination and antigen persistence in the lung, as non-respiratory vaccination routes failed to support lung TRM maintenance. Additionally, parabiosis experiments show that in AdNP vaccinated mice the lung TRM pool is sustained by continual replenishment from circulating memory CD8 T cells that differentiate into lung TRM, a phenomenon not observed in influenza infected parabiont partners. Concluding, these results demonstrates requirements for long-lived cellular immunity to influenza virus, knowledge that could be utilized in future vaccine design.
Aire deficiency during early pregnancy leads to maternal T cell imbalance and embryo loss
Authors: Gillis-Buck Eva Mae , Gardner James M. , Bautista Jhoanne L. , Anderson Mark S. , Erlebacher Adrian , MacKenzie Tippi C. 
Published: January 2023

Abstract
Healthy pregnancy relies on maternal tolerance to a unique set of pregnancy associated self-antigens (PAAs), which are encoded in the maternal genome, but have not been produced since the mother herself was a fetus with a placenta. We hypothesize that Aire-mediated expression of PAAs promotes deletion of PAA-reactive T cells and induction of PAA-specific Tregs, and that Aire-deficiency may play a role in pregnancy complications. We used an Aire-diphtheria toxin receptor (DTR) transgenic mouse model to ablate maternal Aire-expressing cells only during the first nine days of pregnancy. AireDTR+DT plugged females were 6.5 times as likely to show complete embryo resorption by E9.5, compared to WT+DT dams (RR 6.5; 95% CI 1.61–26.1; p&lt;0.01), and twice as likely to show no signs of implantation at E9.5 compared to WT+DT dams (RR 2.0; 95% CI 1.05–3.94; p&lt;0.05). AireDTR+DT dams had significantly fewer FoxP3+ Tregs (p&lt;0.0001) and more CD4+ T cells (p&lt;0.0001) in the maternal thymus, and increased frequency of CD3+ CD4− CD8− T cells in the uterus (p&lt;0.05). AireKO mice are known to develop ovarian insufficiency and subsequent progesterone (P4) deficiency. However, we found no significant differences in serum P4 levels of AireDTR+DT compared to WT+DT dams. P4 supplementation failed to prevent embryo resorption in AireDTR+DT dams or to change the alteration in thymic Tconv:Treg. Thus, Aire deficiency during early pregnancy leads to maternal T cell imbalance and embryo loss without inducing ovarian insufficiency, suggesting a novel mechanism for autoimmune mediated infertility.
Absence of conventional dendritic cells type 1 (cDC1) impairs intestinal homeostasis and permeability and leads to the development of obesity
Authors: Hamade Hussein , Stamps Jamsine T , Thomas Lisa S , Stamps Dalton , Shimodaira Yosuke , Michelsen Kathrin S 
Published: January 2023

Abstract
The maintenance of intestinal homeostasis depends on a complex interaction between microbiota, intestinal epithelial barrier and immune system. Alteration in of one of these components could lead to the development of chronic inflammatory diseases such as inflammatory bowel diseases (IBD). Recent studies have associated obesity with more severe IBD. In obesity, pro-inflammatory macrophages accumulate in adipose tissue and their numbers correlate with inflammation and insulin resistance. BATF3 is a transcription factor implicated in the development of conventional dendritic cells type 1 (cDC1). Here, we show that aged Batf3−/− mice developed obesity as characterized by increased body weight, fasting glucose levels (Batf3−/−: 213, WT: 115 mg/dl, p &lt; 0.005), size of abdominal white adipocyte, and development of hepatosteatosis. We also observed increased intestinal permeability in Batf3−/− mice (FITC-dextran gavage; p &lt; 0.01) suggesting impaired epithelial barrier function. Feeding high-fat diet for 2 months further enhanced the metabolic phenotype in Batf3−/− mice, leading to increased fasting glucose levels (300 vs. 246 mg/dl, p &lt; 0.05), and increased adipocyte size. Moreover, during high-fat diet Batf3−/− mice were more susceptible to dextran sulfate sodium (DSS) colitis with more severe cecal inflammation, increased gut and mesenteric fat M1 macrophages (p &lt; 0.05). Our data suggest that deficiency of the transcription factor BATF3 and lack of cDC1 alters the intestinal epithelial barrier, promotes the development of obesity, and increases susceptibility to DSS. In conclusion, BATF3-dependent cDC1 play an important role in maintaining the intestinal epithelial barrier and are protective for the development of obesity.
Isolation of mouse CD45 positive leukocytes from tissues
Authors: Antignano Frann , Poon Grace , Facca Valerie , Separovic Lea , Valdez Yanet , Kokaji Andy I , Woodside Steven M. , Eaves Allen C. , Thomas Terry E. 
Published: January 2023

Abstract
Immune cell function is often tissue-specific, therefore isolating cells from their tissue microenvironment is necessary to better understand their role in health and disease. This can be challenging in the presence of non-immune, tissue-derived cells and cellular debris from tissue dissociation. These factors reduce the leukocyte start frequency, which can prolong the isolation process and limit the identification of small but critical leukocyte subsets. To overcome these obstacles, we developed a simple and rapid selection method to enrich for CD45+ leukocytes from mouse tissues. Starting with a single cell suspension, cells are labelled with an antibody complex that links CD45+ cells to magnetic particles and following magnetic separation, the isolated CD45+ cells are ready for use. Using healthy mouse lung tissue as an example, CD45+ leukocytes were enriched from 63.5 ± 9.4% to 97.1 ± 1.2% purity, and the recovery of viable CD45+ cells was 37.7 ± 14.5% (mean ± SD, n=37). Preliminary testing on tumors from a mouse 4T1 breast tumor model resulted in efficient enrichment of tumor-infiltrating leukocytes. Importantly, the composition of immune subsets from both healthy lung tissues and tumor samples was maintained following CD45 selection. To demonstrate functionality, T cells within the CD45 selected population from spleen were able to upregulate CD25 and CD69, and proliferate upon stimulation. In as little as 20 minutes, the EasySep™ Mouse CD45 Isolation Kit allows researchers to easily enrich for functional leukocytes from tissue samples. Examining immune cells directly from healthy and tumor tissues will enhance our understanding of how these cells function and aid in developing new approaches to combat disease progression.
Identifying predictive cytokine biomarkers in asthma and chronic obstructive pulmonary disease using the Invitrogen ProQuantum high-sensitivity immunoassay platform
Authors: Bourdon David M , Ekoff Helena , Sjölander Anders , Fujisawa Takao , Nagao Mizuho , Molin Magnus 
Published: January 2023

Abstract
Asthma and chronic obstructive pulmonary disease (COPD) affect hundreds of millions of people worldwide and are a significant cause of morbidity and mortality. These heterogenous and complex inflammatory diseases have few good biomarkers for disease etiology, diagnosis, and monitoring of treatment efficacy. Characterization has previously been performed at genomic and proteomic levels, as well as airway histology, lung function, and quality of life metrics. Since inflammation typically involves one or more cytokine cascades, or pathways responsible for triggering and maintaining such inflammation, cytokine profiles are critical to advancing our understanding of the disease. However, a powerful immunoassay platform with a simplified workflow that is capable of early cytokine detection and minimum sample consumption is lacking in the asthma/COPD research space.
The goal of the following study was to determine whether the Invitrogen ProQuantum high-sensitivity immunoassay platform (www.thermofisher.com/proquantum) could prove useful in uncovering biomarker profiles in asthmatic populations. Additional benefits included running the proteomic assay on ubiquitous qPCR instrumentation and the small 2 μL sample requirement. In this study, 168 human serum samples were analyzed (IL-5, IL-6, IL-8, IL-13, IFN-γ and eotaxin-1) across wheezing and asthmatic phenotypes, as well as healthy age-matched controls among both cohorts. Data presented include group-wise analyses that indicate that the ProQuantum high-sensitivity immunoassay has helped uncover predictive biomarkers in samples from wheezing and asthmatic children, that were not previously distinguishable using more traditional immunoassay testing platforms.
Immunoregulatory effects of Mesenchymal Stem Cells-Derived Exsomes in a Rabbit Model of Autoimmune Dacryoadenitis
Authors: Nian Hong , Li Na , Wei Yankai , Zhao Lu , Chen Sisi , Wei Ruihua 
Published: January 2023

Abstract
We previously demonstrated that mesenchymal stem cells (MSCs) ameliorated autoimmune dacryoadenitis in rabbits. Recently, MSC-derived exsomes (MSC-Exo) were considered to carry functions of MSCs. In this study, we investigated the effect of local administration of human MSC-Exo on rabbit autoimmune dacryoadenitis. Diseased rabbits were treated with MSC-Exo or PBS via subconjunctival injection on days 1, 3, 5, 7 and 9 after infusions of activated autologous lymphocytes, and clinical scores were determined by assessing tear production, break-up time and fluorescein. The in vitro effects of MSC-Exo on activated peripheral blood mononuclear cells (PBMCs) were determined by BrdU proliferation assay and Q-PCR assessing the mRNA expression levels of T cell or macrophage related cytokine genes and subpopulation-related marker genes. We found that subconjunctivally administered MSC-Exo greatly reduced the severity of autoimmune dacryoadenitis as their parent cells by reducing the infiltration of T cell subsets and other inflammatory-related factors in lacrimal glands. In vitro, the proliferation of autoreactive T cells was inhibited by MSC-Exo in a dose-dependent manner. MSC-Exo treatment significantly down-regulated the expression of M1 macrophage-related gene, while promoted the expression of anti-inflammatory M2 macrophage related genes. These results suggests that MSC-Exo effectively ameliorated autoimmune dacryoadenitis partially by inducing the generation of M2 macrophage, indicating a potential novel therapy of MSC-Exo for autoimmune dacryoadenitis.