bluuebunny/crossref_metadata_2025_split huggingface dataset:
A dynamic approach for treating autoimmune disease.
Authors: Keeler Geoffrey D. , Silverberg Emily L , Hoffman Brad E 
Published: January 2023

Abstract
Cell-based immunotherapies have the potential to reverse autoimmune disease by inducing or restoring immune-tolerance. Extensive preclinical studies have demonstrated that regulatory T cell (Tregs) play a key role in both induction and maintenance of tolerance. Several strategies aimed at harvesting and expanding autologous Tregs and then re-infusing them into patients have been investigated. Unfortunately, development of a large scale in vitro method has not been achieved, especially for human cells. On the other hand, a clinically relevant in vivo approach for inducing Ag-specific tolerance has successfully been demonstrated using ectopic expression of an antigen in the liver. Here, we have developed a long-lasting immunotherapy that induces functionally suppressive, Ag-specific Tregs in the murine model of multiple sclerosis using a liver directed adeno-associated virus (AAV) vector expressing myelin oligodendrocyte glycoprotein (MOG). Modulation of EAE disease in C57BL/6 is typically limited to a single immunodominant epitope. By engineering the AAV vector to express full-length protein (MOG1-247), we have created an immunotherapy that is capable of dynamically inducing Ag-specific tolerance against multiple immunogenic epitopes (e.g MOG35-55, MOG119-132). Mice received a single peripheral injection of AAV.MOG vector, or control. EAE was induced between 2 weeks and 8 months later using various MOG epitopes emulsified in adjuvant. Within 14-days, control mice developed significant neurological deficits, whereas none of the treated mice exhibited any symptoms. This work completely abrogates the deficiencies associated with ex vivo expanded Tregs providing a new paradigm for treating autoimmune diseases.
IL-23 induces regulatory T cell plasticity with implications for inflammatory skin diseases.
Authors: Kannan Arun Kumar , Su Zhi , Gauvin Donna M , Lippert Stephanie E , Marie Honore P , McGaraughty Steve P , Kort Michael E , Scott Victoria E , Gauld Stephen B 
Published: January 2023

Abstract
Foxp3+ regulatory T cells (Tregs) are a major fraction of T cells in the skin. At steady state, Treg cells make little to no inflammatory cytokines and control against inflammation. However, under certain autoimmune inflammatory settings (including psoriasis) Treg cells have been shown to become dysfunctional, making pro-inflammatory cytokines and contributing to disease pathogenesis. To better understand the role of Treg cells in psoriasis we analyzed the homeostasis and function of these cells in a mouse model of psoriasiform dermatitis (IL-23 mediated skin inflammation). We find that IL-23 induced inflammation drove both proliferation and increased accumulation of Foxp3+T cells in the skin. Furthermore, IL-23 induced a population of Foxp3+T cells that expressed RORgt and made IL17A. The induction of this putative pathogenic population was completely abrogated by a RORgt inverse agonist that inhibits RORgt signaling in vivo. IL-23 was also sufficient to drive the generation of Foxp3+RORgt+IL17A+ cells in vitro, suggesting a cell intrinsic role for IL-23. Interestingly, IL-23 did not induce production of other pro-inflammatory cytokines like IFN-g by Treg cells. This is in line with recent studies where both increased accumulation and the ability of Foxp3+ T cells to make IL17A was reported in lesional skin of psoriasis patients. Current efforts are focused on better defining the characteristics of Foxp3+RORgt+IL17A+ cells in lesional skin of psoriatic patients and mice treated with IL-23. Collectively, our data suggests that IL-23 drives Treg cell plasticity by inducing a population of Foxp3+RORgt+IL17A+ cells that is regulated by RORgt signaling and could be relevant to disease pathogenesis.
Families of neurotransmitter receptors are differentially regulated across T cell subsets
Authors: Rosenberg Kenneth M , Singh Nevil J 
Published: January 2023

Abstract
Although primarily expressed in neurons, a surprising array of neurotransmitter receptors (NR) has been found to be expressed in immune cells, and in particular T cells. This suggests that T cell responses may be amenable to direct regulation by the nervous system. However, the absence of adequate information about the subset-specific and activation-dependent variations in NR expression in T cells makes it challenging to decipher how and when such regulation might impact T cell immunity. In this study, we examined the levels of several NR components in different mouse T cell subsets. RNA from FACS-purified CD8 or CD4 (naïve, memory, or regulatory) T cells were analyzed by RT-PCR for 84 known NR transcripts. Preliminary data show that 19 of these are expressed in at least one of the T cell subsets studied. Of these, 3 subunits were stably expressed across all subsets with the remaining 16 transcripts showing at least a 2-fold expression change between groups. 32% of genes showed the greatest expression in naïve CD4 T cells while a further 32% were upregulated most highly among memory CD4 T cells. Intriguingly, the glutamate receptor Gria3 is uniquely expressed by CD8 T cells, while the vasopressin receptor Avpr1a is specific to regulatory CD4 T cells. We also compared our profiles with previously reported metadata including the ImmGen expression database. This study now allows us to develop a functional model for where specific neuronal outputs may impinge on different T cell subsets under physiological contexts.
Elucidating the Role of CD27 in Treg Function during Th17 Responses
Authors: Remedios Kelly Anne , Zirak Bahar , Rosenblum Michael D 
Published: January 2023

Abstract
Tregs and Th17 cells have a very close and complex relationship. Tregs and Th17 cells share a dependence on TGFb for their differentiation and it has recently been shown that Tregs require the Th17-driving transcription factor STAT3 to regulate Th17 responses. Despite sharing a dependence on STAT3, Tregs within healthy tissue tend to produce very low amounts of IL-17. In contrast, in multiple Th17-driven diseases (including Multiple Sclerosis, Psoriasis and Crohn’s) Tregs within the diseased tissue produce increased levels of the IL-17. The costimulatory receptor CD27 and its ligand, CD70, have been shown to play a role in suppressing Th17 responses through the epigenetic suppression of IL-17 and CCR6. We have found that CD27 is highly expressed on Tregs in both human and murine skin, but whether CD27 plays a role in promoting Treg function (and suppressing acquisition of Th17-like traits) remains unknown. Utilizing CD27-deficient mice (CD27KO), we found that Tregs in the skin of CD27KO mice in the steady state produced more IL-17 compared to their wild-type (WT) counterparts. To test the role of CD27 specifically on Tregs during Th17-driven inflammation in vivo, we have developed an adoptive transfer model of cutaneous candida albicans infection where Tregs play an essential role in the development of productive Th17 responses. Using this model, we found that CD27 suppressed Treg expression of IL-17 and CCR6 in a cell-intrinsic manner. We also found that IL-6 could induce the production of IL-17 from Tregs in vitro which was suppressed when co-cultured with CD70 expressing BMDCs. Our studies suggest that the CD27-CD70 pathway promotes Treg function by limiting production of Th17 effector molecules.
Age difference in the immune response to endotoxin (LPS) shapes Th2-mediated airway inflammation and development of asthma.
Authors: Ruiz Beatriz León , Tato Andre Ballesteros 
Published: January 2023

Abstract
In addition to genetic factors, environmental exposures is also of primary importance for the development of asthma/allergy disorders in children, as demonstrated by epidemiological data showing children growing up in traditional farms seem to develop asthma less often than children growing up in urban areas. According to the hygiene hypothesis, this is due to increased exposure to endotoxin (LPS) and other farm-related microorganism-derived compounds. However little is known about how early-life contact to microbial compounds influence the development of asthma. Here we show that house dust mite (HDM) allergen sensitization in adult mice (8 weeks-old) induced T-helper type 2 (Th2)- driven airway inflammation, i.e., influx of activated Th2 cells into the airways, airway eosinophilic inflammation and increased IgE-producing plasmablasts. In contrast, HDM allergen sensitization with low-dose LPS (10 μg) abrogated HDM-driven Th2-mediated allergic airway inflammation without increasing antigen-specific Th1 cell trafficking into the airways. Importantly, unlike adults, infant mice (3 weeks-old) exposed to HDM with low-dose LPS (10 μg) developed Th2 responses allergic airway inflammation and required allergen sensitization with high-dose LPS (100 μg) to effectively suppress allergen-specific Th2 responses and allergic inflammation. These results show that airway exposure levels of endotoxin provide different protection against asthma and allergies in adults and infants and suggest that Infants are more vulnerable to prime type 2 cell dominant responses to inhaled allergens in the presence of low dose endotoxin, which will shape future effector Th2 responses and Th2-mediated airway inflammation throughout adult life.
Adipose type one innate lymphoid cells regulate macrophage homeostasis through targeted cytotoxicity
Authors: Boulenouar Selma , Lynch Lydia 
Published: January 2023

Abstract
Adipose tissue has a dynamic immune system that adapts to changes in diet and maintains homeostatic tissue remodeling. Recently, adipose type 1 innate lymphoid cells (AT1-ILCs) were reported to promote proinflammatory M1 macrophages in obesity. However, their role at steady state remains unknown. Here we reveal heterogeneous populations of AT1-ILCs in humans and mice. Using parabiotic mice fed HFD, we found that AT1- ILCs had differential trafficking and proliferation features. AT1-ILCs displayed fluctuations in response to short and long term HFD and could revert to regular levels upon diet restriction. Unexpectedly, all AT1-ILCs were able to kill macrophages at steady state, especially tissue repair-associated M2 macrophages. T1-ILCs-depleted and perforin-deficient mice had increased M2:M1 ratios. Diet-induced obesity impaired AT1-ILCs killing ability. Adoptive transfer of AT-1ILCs from lean to obese mice reduced total macrophages but increased M1:M2 ratio. Collectively, we show that AT1-ILCs are essential for regulating macrophage homeostasis through targeted cytotoxicity.
Mechanisms of Glucocorticoid Resistance in Hypereosinophlic Syndromes
Authors: Stokes Kindra N , Yoon Pryscilla , Makiya Michelle , Gebreegziabher Meheret , Holland-Thomas Nicole , Haugen Brandon , Pazdrak Konrad , Klion Amy D. , Khoury Paneez 
Published: January 2023

Abstract
Glucocorticoids (GC) are anti-inflammatory agents commonly used to treat patients with hypereosinophilic syndrome (HES). GC promote eosinophil apoptosis and downregulation of Th2 cytokine production. Although abnormalities of GC receptor (GR) function and elevations in β-GR isoforms have been implicated in GC resistance in asthma, little is known about the mechanisms of GC resistance in HES. To explore the etiology of GC-resistance in HES, samples were analyzed from 19 subjects with HES enrolled on a prospective study of GC-responsiveness and 33 subjects with HES enrolled on a natural history study of eosinophilia for whom response to GC was known. GC responders were defined by a reduction in absolute eosinophil count (AEC) to &lt;1000/mL after 0.5–1mg/kg of GC for ≥1 week. In the prospective trial, GM AEC was significantly decreased in the GC responders (n=14; GM 2430 to 245/mL, p&lt;0.001) but not in GC non-responders (n=5; GM 12800 vs GM 9927, NS). In contrast, an expected rise in absolute neutrophil count was seen in 12/14 GC responders (GM 3.39 to 6.07, p&lt;0.01) and 5/5 non-responders (GM 2.51 vs. 5.27, p&lt;0.01), suggesting that GC resistance in HES is not a global phenomenon. Expression of GR isoforms (α, β, γ and P) was assessed by RT-PCR in RNA from purified eosinophils and showed no differences between responders (n=23) and non-responders (n=12). A panel of serum cytokine levels were measured in 37 subjects. Only IL-5 levels were significantly different between the groups (GM 86.2 pg/mL in non-responders vs. 15.3 in non-responders, p=0.019). IL-5, at levels comparable to those seen in GC non-responders, inhibited GC-induced eosinophil apoptosis in vitro (n=7). These data suggest that increased IL-5 may play a major role in GC resistance in HES.
Dysregulation of Inhibitory Receptor Expression in Systemic Sclerosis
Authors: Fleury Michelle , Belkina Anna C , Gupta Ashna , Zammitti Christopher , Simms Robert , Snyder-Cappione Jennifer , Lafyatis Robert , Dooms Hans 
Published: January 2023

Abstract
Systemic sclerosis (SSc) is an autoimmune connective tissue disorder associated with aberrant fibrosis of the skin and internal organs that can lead to organ failure and death in patients. With few treatment options many patients succumb to fibrosis of the lungs or kidneys, or due to vascular damage culminating in pulmonary disease; the 50 percent survival rate is only about 10 years. We collected PBMCs from 36 SSc patients that were not on immune-suppressants, along with 25 age-matched healthy controls. Using a 15 color flow cytometry panel our lab has observed increases in the inhibitory receptors PD-1, TIGIT, and Tim-3 on PBMCs from SSc patients. This increase in expression is cell type specific with PD-1 and TIGIT shown to be increased on defined T cell types including CD4+ T cells, both Treg and non-Treg subsets, as well as CD8+ T cells. Tim-3 was only statistically significantly increased on a mature subset of CD16+ CD56med natural killer cells. This increase was not seen for all inhibitory receptors. The expression of LAG-3 remained low across all cell types observed. This increase in inhibitory receptor expression was enhanced in patients with a high percent of Tregs within the CD4+ gate. In vitro blockade of these inhibitory receptors provided differential cytokine secretion in patients versus healthy subjects, suggesting a possible role in disease pathogenesis. The observation of increased inhibitory receptors in SSc as well as the ability to modulate cytokines by blocking the interaction of these receptors with their ligands could provide insights into possible immune modulation in patients.
The major roles of monocytes/macrophages and their products in skin inflammation induced by skin deposited lupus IgG
Authors: Deng Guo-Min , Xu Guangqiong , Guo Xuanxuan , Fei Xibin , Li Xiaoyan 
Published: January 2023

Abstract
Skin injury is second common in patients with systemic lupus erythematosus (SLE), but pathogenesis of skin injury remains unclear. Based on skin deposition of IgG in SLE, the role of skin deposited IgG in the pathogenesis of skin injury in SLE was studied. We established skin deposited lupus IgG by intradermal injection of lupus IgG in mice and investigated the role of skin deposited lupus IgG in pathogenesis of skin injury in SLE. We found that skin deposited lupus IgG made by intradermal injection of lupus serum IgG induced skin inflammation, which appeared at 3 h and peaked at 3 d after intradermal injection. This skin inflammation was related to the dose of injected IgG but not to systemic disease activity. This skin inflammation was significantly decreased in mice depleted of monocytes but not lymphocytes. This skin inflammation was markedly decreased in mice with TNF-a deficiency. Lupus IgG promoted monocyte differentiation into dendritic cells (DCs) and enhanced the expression of TNF-α. TNF-α played a major role in the proliferation of keratinocytes. The results indicate that the deposition of IgG in skin exerts an important role in the pathogenesis of lupus skin injury. IgG signaling pathway is a therapeutic target in skin injury of SLE.
Expression of Immune Checkpoint Receptors in Multiple Myeloma Patients
Authors: Kwon Minsuk , Choi Yoon Seok , Park Junsik , Shin Eui-Cheol 
Published: January 2023

Abstract
Multiple myeloma is a malignancy of plasma cells and the second most common hematologic cancer. High-dose chemotherapy plus autologous stem cell transplantation has been performed to achieve remission, but almost all the patients will finally relapse. Alternative therapeutic options are necessary for the multiple myeloma patients. Immune checkpoint inhibitors, such as anti-PD-1 have shown practice-changing therapeutic effectiveness for advanced non-small cell lung cancer, as well as Hodgkin’s lymphoma. To investigate T cell exhaustion in multiple myeloma patients, we assessed the expression of various immune checkpoint receptors, such as PD-1, Tim-3, LAG-3 and TIGIT, in the paired peripheral blood mononuclear cells (PBMC) and bone marrow mononuclear cells (BMMC), by multicolor flow cytometry. In BMMC, PD-1+, and PD-1+TIGIT+ were increased in CD8+ T cells, compared to the paired PBMC. Only a few patients showed Tim-3+ or LAG-3+ T cell population, and the most identified Tim-3+ or LAG-3+ T cell populations were observed in PD-1hiCD8+ T cells in the BMMC. There was significantly increased PD-1+, and PD-1+TIGIT+ expression in CD69+CCR7−CD8+ population compared to CD69− CCR7−CD8+ in BMMC. On direct ex vivo MHC multimer analysis of BMMC, tumor-associated-antigen specific CD8+ T cell showed increased PD-1+, TIGIT+, PD-1+TIGIT+ and CD69+ expression. PD-L1 was also expressed on malignant plasma cell, and the level of expression was correlated with the disease burden. Collectively, our data show that the immune checkpoint receptors-expressing T cells are enriched in bone marrow, where malignant plasma cells reside. Functional restoration of these T cells by immune checkpoint blockers will be investigated in further study.
Small molecule drug inhibition of PD-1 transcription is as effective as anti-PD-1 biologic blockade in cancer therapy
Authors: Rudd Christopher E , Rothstein David , Taylor Alison 
Published: January 2023

Abstract
Despite the importance of the co-receptor PD-1 in T cell immunity, the upstream signaling pathway(s) that regulate PD-1 expression has not been defined. Glycogen synthase kinase 3 (GSK-3, isoform α and β) is a serine-threonine kinase implicated in cellular processes. We have shown that GSK-3 is a key upstream kinase that regulates PD-1 expression in CD8+ T cells. GSK-3 inactivation increased Tbx21 transcription for enhanced Tbet expression, and its suppression of Pdcd1 transcription in CD8+ cytolytic T-cells (CTLs) (Taylor et al 2016 Immunity 44, 274–86). Here, we show that GSK-3 inhibitor blockade of pcdc1 (PD-1) transcription with a small molecule inhibitor (i.e. SB415286) is as effective as anti-PD-1 and PDL-1 blocking antibodies in the control of pulmonary metastasis of B16 melanoma, intra-dermally injected B16 and EL4 lymphoma solid tumors, and in the ex vivo pre-treatment of T-cells before adoptive transfer into mice carrying tumors. In an analysis of knock out mice, GSK-3α/β−/− mice, which had greatly reduced PD-1 expression on T-cells, showed the same reduction in B16 pulmonary metastasis as Pdcd1−/− mice. Further, the ex vivo treatment of T-cells with SB415286, anti-PD-1, or in combination followed by adoptive transfer, had identical effects in inhibiting EL4 lymphoma growth. In all tumor models, GSK-3 inactivation markedly inhibited Pdcd1 transcription and PD-1 expression on tumor infiltrating T-cells (TILs), while increasing Tbx21 (Tbet) transcription and the presence of CD8+ TILs expressing CD107a+ (LAMP1) and granzyme B (GZMB). Our findings define for the first time that a next generation approach using small molecule inhibition of PD-1 expression is as effective as anti-PD-1/PL1 biologics in cancer therapy.
Tumor infiltrating T cells acquire exhaustion-associated epigenetic programs
Authors: Youngblood Ben A , Ghoniem Hazem , Moustaki Ardiana , Dogra Pranay , Abdelsamed Hossam Aly , Fan Yiping 
Published: January 2023

Abstract
Antigen-specific CD8 T cells play a critical role in controlling chronic infections and cancer, but progressively lose their effector functions during prolonged antigen exposure. Repression of CD8 T cell effector functions, commonly referred to as T cell exhaustion, limits the ability of the immune system to purge the chronic pathogen from the host. It has recently become recognized that CD8 T cell exhaustion programs can be reinforced and heritably maintained. Therefore in order to develop and/or improve current therapeutic approaches that utilize host antigen-specific T cells to treat chronic infections or cancer a major challenge for the field is to identify mechanisms that stabilize T cell exhaustion. Using the LCMV model system of chronic viral infection we determined that Dnmt3a mediated de novo DNA methylation plays a causal role in establishing CD8 T cell exhaustion. We then performed whole-genome methylation profiling of WT and Dnmt3a cKO CD8 T cells from chronically infected animals and identified Dnmt3a-dependent DNA methylation programs in genes, including interferon gamma and Tcf7, that are coupled to the progressive decline in effector function and developmental plasticity of the antigen specific cell. Lastly, we extended these findings to the tumor setting using a syngeneic mouse tumor model. We found that tumor-infiltrating PD-1hi CD8 T cells acquire exhaustion-associated de novo DNA methylation programs. These results have significant implications for therapeutic strategies that utilize reactivation of host pathogen-specific CD8 T cells to control chronic viral infections or cancer and provide a nucleotide-resolution map of epigenetic programs progressively acquired during T cell exhaustion.
Assessing the infection rates and pathogenesis of Borrelia burgdorferi in wild rodents and ticks in Western Maryland
Authors: Kuchinsky Sarah Catherine , Elliott Hannah , Brown Drew , Granger Mallary , Lloyd Nicole , Nyachieo Fred , Weakland Amy , Wygal William , Taylor Rebekah 
Published: January 2023

Abstract
Lyme disease is the most prevalent vector-borne disease in the United States. The causative agent is the spirochete Borrelia burgdorferi. Many rodents are reservoir species that carry B. burgdorferi, through which immature ticks (Ixodes sp.) readily transmit the bacteria. Human cases of Lyme disease have dramatically increased over the past 10 years due to tick range expansion. Incidence has expanded from its New England origin into the Midwest and Mid-Atlantic. We hypothesized that Western Maryland is a prime candidate for B. burgdorferi expansion. To prepare, we sought to (1) determine the incidence of B. burgdorferi infection in wild rodent and tick populations in Western Maryland, (2) identify simple and quick methods for diagnosing B. burgdorferi infection in the field, and (3) gain insight into the pathogenesis of B. burgdorferi in wild rodent populations. DNA was extracted from 67 rodent blood samples and 202 ticks, collected from sites in Western Maryland. Approximately 35.14% of these samples tested positive for B. burgdorferi by PCR. Blood samples were tested for the presence of anti-B. burgdorferi antibody using SNAP 3Dx tests, a rapid ELISA-based veterinary diagnostic. Interestingly, the SNAP 3Dx tests were successful in identifying B. burgdorferi infection in both wild mice and chipmunks, suggesting that the capture antibody used in these canine tests is cross-reactive. Finally, whole B. burgdorferi bacteria were identified on frozen sections of wild mouse spleen by immunofluorescence. Bacteria were found throughout the red pulp and in splenic germinal centers of infected mice. Studies are underway to further characterize the infected cells in these samples.