bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of a specific gene signature in human Th1/17 cells (BA13P.126)
Authors: Duhen Thomas , Ni Chester , Campbell Daniel 
Published: January 2023

Abstract
T cells that co-express the chemokine receptors CCR6 and CXCR3 and produce IL-17, IFN-γ, GM-CSF and low IL-10, also known as Th1/17 cells, have been proposed to be highly pathogenic in several autoimmune disorders. To better characterize these cells and examine their relationship with Th1 and Th17 cells, we compared the whole transcriptome of the aforementioned T cell subsets by RNAseq directly ex vivo. Using this approach, we identified groups of genes shared either with Th1 or Th17 cells. The Th1 signature included genes such as TBX21, IL12RB2, GZMK, PRF1 and CCL5, whereas the Th17 signature contained well-established Th17-associated genes (i.e. RORC and KLRB1) as well as those not previously associated with Th17 cells (i.e. CTSH and PTPN13). Interestingly, we also found genes specifically expressed by Th1/17 cells, among which are IL4i1, LTK and ABCB1. The latter, also known as MDR1, encodes for a transmembrane transporter playing a major role in the efflux of drugs and endogenous metabolites across the cell membrane. We confirmed its unique expression on Th1/17 cells using a rhodamine efflux assay. Moreover, MDR1+ Th1/17 cells displayed a highly pro-inflammatory cytokine profile. Our data reinforce the notion that Th1/17 cells are a heterogeneous population of cells combining Th1 and Th17 phenotypic and functional characteristics but also that these cells have unique properties that could be targeted as new strategies to combat autoimmune diseases.
Regulatory T cells in B cell-deficient mice differ from those in WT mice in function and expression of cell surface markers (BA13P.119)
Authors: Ellis Jason , Braley-Mullen Helen 
Published: January 2023

Abstract
NOD.H-2h4 mice develop spontaneous autoimmune thyroiditis (SAT) with chronic inflammation of thyroids by T and B cells. B cell-deficient (B-/-) mice are resistant to SAT unless Treg are transiently depleted. To determine if Treg from WT and B-/- mice differ in their ability to suppress effectors from a single source, splenocytes from CD28-/-B-/- mice (effector cells and APC) were cultured with or without sorted Treg from Foxp3-GFP WT or B-/- mice before transfer to mice lacking T cells. Recipients of Treg from B-/- mice had significantly lower SAT severity scores compared to those given Treg from WT mice, indicating that Treg in B-/- and WT mice differ functionally. Treg from WT and B-/- mice also differ in expression of TNF receptor superfamily members CD27, TNFR II p75, and GITR. After transient Treg depletion with anti-CD25 or diphtheria toxin, repopulating Treg in B-/- mice had reduced suppressor function, and expression of CD27, p75 and GITR was like that of WT Treg. If B cells were present during development of B-/- Treg in bone marrow chimeras or added to cultures of B-/- Treg and T effectors, expression of CD27, p75 and GITR by Treg from B-/- mice was comparable to that of WT Treg, and suppressive function was reduced. These results establish for the first time that Treg in WT and B-/- mice differ both functionally and phenotypically, and both are altered when B cells are present during Treg development and during interaction of Treg with T effectors.
Leptin promotesTh17 cell response and exacerbates collagen-induced arthritis (BA13P.120)
Authors: Deng Jun , Lu Liwei 
Published: January 2023

Abstract
Leptin regulates CD4+ T helper cell differentiation and functions. IL-17-producing CD4+ T cells (Th17 cells) play a pivotal role in the pathogenesis of various autoimmune diseases. Elevated leptin levels have been detected in the sera and synovial fluid of patients with rheumatoid arthritis (RA). However, whether and how leptin regulates Th17 cell responsiveness and its potential application in autoimmune arthritis remains elusive. Using immunohistochemistry staining and immunofluorescence microscopy, we found that elevated leptin levels and increased Th17 cells were closely related to disease severity and joint damage in mice with collagen-induced arthritis. Leptin treatment significantly promoted Th17 cell differentiation, upregulated ROR-gamma t expression and increased IL-17 production in culture. Intra-articular injection of leptin to the joint led to earlier onset of arthritis, higher clinical score, and severer joint damage compared to PBS-injected mice. Th17 cell frequencies and total cell number in the draining lymph nodes and joint tissue were elevated upon leptin injection. Moreover, increased Th17 cells infiltrated into the synovium with increased IL-17 levels were detected in the synovial cavity in leptin-injected mice. Together, these data have demonstrated that leptin plays an important role in the exacerbation of autoimmune arthritis via promoting Th17 cell differentiation and increases IL-17 production.
Overexpression of Pbx1-d, a novel splice isoform of Pbx1, changes the portion of follicular CD4 T cell populations and impairs regulatory T cell homeostasis (BA14P.201)
Authors: Choi Seung-Chul , Zeumer Leilani , Morel Laurence 
Published: January 2023

Abstract
We have previously shown that the differential expression of a novel splice isoform, Pbx1-d, of the Pbx1 gene was associated with the production of autoreactive CD4+ T cells in the Sle1a1 lupus susceptibility locus and lupus susceptibility in the B6.Sle1.Sle2.Sle3 mouse model. Pbx1 regulates the transcription of a large number of genes by controlling chromatin accessibility. Pbx1-d is the only known Pbx1 isoform lacking the DNA binding domain, which suggests a dominant negative function, and its expression is more frequently in the CD4+ T cells from lupus patients than in healthy controls. With the hypothesis that Pbx1-d contributes to lupus development by regulating CD4+ T cell effector functions through novel mechanisms, we have generated B6.CD4-Pbx1-d Tg (Pbx1-d Tg) mice, which specifically express Pbx1-d in CD4+ T cells. These Tg mice reproduce the phenotypes of B6.Sle1a1 mice, with increased inflammatory functions of CD4+ T cells and impaired Treg cell homeostasis. Interestingly, the composition of follicular helper and regulatory T cell (Tfh and Tfr) populations, is different in the Pbx1-d Tg mice as well as the B6.Sle1.Sle2.Sle3 mice as compared with wild-type mice. Currently, we are investigating the mechanisms by which Pbx1 regulates Treg homeostasis and follicular T cell differentiation in the Pbx1-d Tg mice, as well as its role in autoimmune pathology in the B6.Sle1.Sle2.Sle3 mice.
Targeted modification of complex N-glycosylation results in a spontaneous CVID-like immunodeficiency with anti-lymphocyte autoimmunity (BA14P.204)
Authors: Ryan Sean , Abbott Derek , Cobb Brian 
Published: January 2023

Abstract
Common variable immunodeficiency (CVID) is the most prevalent symptomatic primary immunodeficiency affecting humans. Although a clear disease etiology remains elusive, a common characteristic of CVID is deficient IgG antibody production to infection or vaccination. Many patients also exhibit symptoms of abnormal T cell function, including an apparent lack of naïve T cells which correlates with clinical severity. We describe here one of the first animal models of spontaneous CVID that incorporates aspects of both humoral and T cell dysfunction. The CVID mice exhibit characteristic deficiencies in IgG responses to both protein and polysaccharide vaccines. Interestingly, the immunodeficiency is associated with decreased T cell activity due to a persistent autoimmune response depleting naïve T cells, which appears to be mechanistically linked to changes in erythrocyte surface N-glycosylation. The N-glycosylation dependent auto-epitopes that emerge on erythrocytes were found to cross-react with naïve T cells independent of changes to the latter’s glycosylation profile. While the use of Ig replacement therapy and antibiotics has reduced the number of infections in CVID, the major inflammatory complications such as lung disease, cancer, and autoimmunity still develop; thus our findings provide potential insight into human CVID beyond hypogammaglobulinemia by addressing the mechanisms that may contribute to the more broad defects in adaptive immunity linked to clinical severity.
Lectin-targeted dendritic cell immunotherapies against experimental autoimmune encephalitis/multiple sclerosis (CAM5P.242)
Authors: Sagar Divya , Foss Catherine , Khan Zafar , Pomper Martin , Jain Pooja 
Published: January 2023

Abstract
The molecular mechanisms of how circulating dendritic cells (DCs) access the CNS across the blood-brain barrier (BBB) remains to be investigated. During neuroinflammation, DCs transmigrate across the BBB in the presence of chemoattractants, particularly CCL2, and initiate immune response. We showed through near-infrared imaging that DC accumulation into lesions within the CNS correlated with severity of inflammation during experimental autoimmune encephalitis (EAE). Actin polymerization on DCs upon CCL2 exposure indicated a migratory phenotype. Transmigration of DCs was paracellular and more efficient than T cells in vitro. To selectively attenuate DC transmigration, we sought to study expression of DC specific C type lectin receptors- CD205 & CD206 (Type I), CD207, -209, -303, CLEC4A, -9A, -10A & -12A (Type II). Different DC subsets- myeloid (mDC), plasmacytoid (pDC) and monocyte-derived (MDDC) had a unique enrichment of lectin receptors and each subset utilized a different receptor to adhere and transmigrate in the presence of CCL2. Phosphoproteomics revealed that these receptors when triggered, activate molecular events known to be involved in actin dynamics, thus substantiating their role in DC transmigration. Importantly, in EAE mice these lectins were validated targets for inhibiting DC transmigration. Thus, the prospect of selectively regulating DC entry into the CNS will open up new treatments against disease pathogenesis and propagation in multiple sclerosis.
miRNA modulation of M1/M2 macrophage phenotype (CAM5P.231)
Authors: Guerau-de-Arellano Mireia , Jablonski Kyle , Amici Stephanie , Gaodet Andrew , Popovich Philip 
Published: January 2023

Abstract
After spinal cord injury (SCI), “M1” (pro-inflammatory) and “M2” (anti-inflammatory) macrophages populate the site of injury. M1 macrophages are neurotoxic and hamper neuroregeneration while M2 macrophages have a pro-regenerative, anti-inflammatory phenotype. Therefore, therapeutic strategies that suppress M1 and enhance M2 macrophages after SCI are actively sought. Micro RNAs (miRNA) are small, non-coding RNAs, approximately 19-24 nucleotides in length, which regulate gene expression at the transcriptional level. miRNA are known to regulate inflammation in autoimmune disease and inflammatory disorders. We therefore hypothesized that miRNA play a key role in differentiation process of macrophages into the pro-inflammatory M1 phenotype. The relative expression of several miRNA was analyzed in vitro in bone marrow-derived macrophages undergoing differentiation into M1 versus M2 phenotypes. We found that M1 macrophages expressed significantly higher levels of miR-155 than M2 macrophages. In addition, miR-155 expression was increased in the injured spinal cord mirroring the increase in M1 macrophages at the site of injury. miR-155 deficiency or silencing resulted in blunted iNOS, IL-1β and TNFα up-regulation in M1 macrophages, among others. In contrast, Arginase-1 expression remained stable in M2 macrophages. These results indicate that miR-155 plays an important role in the differentiation of macrophages into an M1 phenotype.
Visfatin increases the activity of aggrecanases-1 and -2 in human chondrocytes (CCR5P.264)
Authors: Yaykasli Kursat , Dogan Mustafa , Hatipoglu Omer , Yaykasli Emine , Kaya Ertugrul , Ozsahin Mustafa , Uslu Mustafa 
Published: January 2023

Abstract
Visfatin, characterized in 1994 is an adipokine (adipocytokines). It was secreted by not only visceral adipose tissue but also by several tissues. First, visfatin with insulin-mimetic activity has been implicated in the insulin resistance. Finally, it was understand that visfatin involves in several pathological processes like inflammation. The degradation of aggrecan by inflammation induced aggrecanases is a key process for diseases such as rheumatoid arthritis (RA) and osteoarthritis (OA). Aggrecanases-1 and -2, called ADAMTS-4 and -5 respectively are members of a disintegrin and metalloproteinase with thrombospondin motifs enzyme family. The aim of this study was to investigate the potential role of visfatin in aggrecan destruction. Human articular chondrocytes were stimulated with visfatin at 10, 25, 50, 100, 250 and 500 ng/ml doses. Total RNA was isolated and reverse-transcribed by random primer. The aggrecanases-1 and -2 genes expression were analyzed and standardized by β-actin gene expression using real-time polymerase chain reaction. The highest upregulation for aggrecanases-1 and -2 was determined at 500 ng/ml. Obesity is associated with RA by secreting adipocytokines. In addition, the increased serum visfatin level in patients with RA was reported before. Therefore, visfatin become a potential target for aggrecanases upregulation. In conclusion, the high level of visfatin accompanied by obesity has catabolic effect on the aggrecan degradation.
Multi-color fluorescence microscopy detection of chemokine receptors using novel Brilliant Violet™-conjugated monoclonal antibodies (CCR5P.267)
Authors: Zhang Hong , Lundsten Kelly , Ransom John , Yang Xifeng , Nguyen Dzung 
Published: January 2023

Abstract
BioLegend has generated a large library of high affinity antibody clones against mouse and human chemokine receptors that are validated in flow cytometry applications. They are offered as conjugates with the violet-excited Brilliant VioletTM (BVTM) fluorophores. Use of the BVTM fluorophores has greatly increased phenotypic multiplexing capabilities in flow cytometry by increasing the utility of the violet laser. Violet-excited BV421TM antibody conjugates are at least as bright as yellow/green-excited phycoerythrin (PE) and red-excited Alexa Fluor® 647 conjugates. The brightness of violet-excited BV510TM is more on par with the green-excited Alexa Fluor® 488. BV421TM and BV510TM are far brighter and more photostable than the violet-excited Alexa Fluor® 405 and Pacific BlueTM. Here, using a variety of discrete immunologic cell populations bearing different chemokine receptors, BV421TM and BV510TM are shown to also increase multiplexing capabilities and increase resolution in fluorescence microscopy applications. Elsewhere, both BV421TM and BV510TM were excited with a multiphoton laser, indicating their potential utility in deep tissue and live cell imaging applications. The results show that BV421TM- and BV510TM-conjugated monoclonal antibodies are valuable new tools for the scientist wanting to resolve multiple antigens simultaneously with fluorescence microscopy.
CCR6 expression by small intestine lamina propria dendritic cells is critical for surveillance of the luminal contents at the villous epithelial surface (CCR5P.251)
Authors: McDonald Keely , Wheeler Leroy , McDole Jeremiah , Miller Mark , Newberry Rodney 
Published: January 2023

Abstract
The immune system underlying the villous epithelium is continually monitoring its complex environment to appropriately respond with the induction of tolerance or immunity. However little is known about the factors promoting immune surveillance of the luminal contents at this surface. Mice deficient in lymphotoxin beta receptor (LTBR) have a decreased population of dendritic cells (DCs) associated with the villous epithelium. The chemokine CCL20 is expressed by the villous epithelium in a LTBR dependent manner, and CCR6, the only receptor for CCL20, is expressed by the LP-DCs associating with the villous epithelium. Using an in vivo imaging approach, we observed that LP-DC from CCR6 deficient mice were further from the intestinal lumen and flow cytometric analysis revealed they had a decreased population of epithelia associated LP-DCs. Mixed bone marrow transfers confirmed that CCR6 expression by DCs was required for association with the epithelium. In vivo imaging and flow cytometry revealed that LP-DCs from CCR6-/- mice were not impaired at migrating in the lymphatics or populating the mesenteric lymph node. However LP-DCs from CCR6-/- mice were deficient at acquiring epithelial cell proteins and intraluminal ovalbumin, and were deficient in priming immune responses to luminal antigen in the mesenteric lymph nodes. These findings identify an essential role for CCR6 expression by LP-DCs in surveillance of the luminal environment at the villous epithelial surface.
Murine aortic vascular cells produce IL-17C and support inflammation in atherosclerosis. (CCR6P.271)
Authors: Butcher Matthew , Galkina Elena 
Published: January 2023

Abstract
The progression of atherosclerotic lesions involves regular communication between arterial hematopoetic cells and vascular cells through cytokine and chemokine networks. While recent evidence has demonstrated the presence of Interleukin-17A (IL-17A)-producing T cells within atherosclerotic Apolipoprotein E-deficient (Apoe-/-) aortas, the roles of the other IL-17 family members have not been explored. As several recent studies have demonstrated a pro-inflammatory role for epithelial and keratinocyte-derived IL-17C, we sought to examine aortic Il17c expression within atherosclerotic western diet-fed Apoe-/- and Il17a-/-Apoe-/- mice. Il17c mRNA was detected within the aortas of both Apoe-/- and Il17a-/-Apoe-/- mice and several major sources of aortic IL-17C were identified by flow cytometry; including smooth muscle cells (SMCs, 5±0.8%) and CD45-αSMA- cells (5.8±0.8%). Interestingly, elevated aortic IL-17C production was detected within Il17a-/-Apoe-/- mice in comparison to Apoe-/- littermates (6.16±0.24% vs 3.6±0.1%), suggesting an inverse relationship between T cell derived-IL-17A and vascular-IL-17C. To examine the effects of IL-17C and TNFα on aortic chemokine and cytokine production, we explanted whole aortas with IL-17C or IL-17C and TNFα. In this system, IL-17C broadly supported the expression of Ccl2, Ccl7, Cxcl1, Cxcl2, Cxcl5, and Il6. Together, these data suggest that SMC-derived IL-17C supports atherogenic inflammation via cytokine and chemokine production.
Accentuated dichotomy between activation and resting state results in lower basal IL-2R signaling potential in CD4 memory T cells of T1D subjects (HUM7P.284)
Authors: Long S. Alice , Schwedhelm Katharine , Tatum Megan , Cerosaletti Karen , Greenbaum Carla , Buckner Jane 
Published: January 2023

Abstract
The IL-2/IL-2R pathway is clearly implicated in type 1 diabetes (T1D). However, not all of the underlying mechanisms are understood. Known genetic risk alleles in humans account for some, but not all, of the reduction in IL-2R signaling. To determine additional factors, we interrogated signaling in healthy controls and T1D subjects held constant for IL-2R associated risk alleles. Glucose and HbA1c did not correlate with IL-2 response. Nor was reduced IL-2R signaling observed in T2D subjects or CD4 memory cells rested in fresh media. Instead, IL-2 signaling in CD4 memory cells of T1D subjects remained significantly lower than controls even after in vitro activation and proliferation. Together, these data suggest that intrinsic factors contribute to reduced IL-2R signaling in T1D subjects. IL-2R signaling was comparable regardless of CCR7, CD27 or CD62L expression suggesting that differentiation state is not a factor. However, there was an accentuated dichotomy between IL-2 response in activated versus quiescent cells of T1D subjects resulting in a more pronounced reduction in resting cell IL-2 signaling. Thus, in addition to known T1D-associated risk alleles, intrinsic properties related to activation state may modulate levels of basal IL-2R signaling. This is consistent with our finding that reduced IL-2R signaling occurs as early as autoantibody conversion. Therapeutically targeting multiple components of the IL-2 pathway may increase treatment options for T1D.
Serum Melatonin levels in Sjögren patients (HUM7P.310)
Authors: Keskin Goksal , Keskin Dilek , Inal Ali , Ilikci Rahsan 
Published: January 2023

Abstract
OBJECTIVES: Primary sjögren syndrome is a systemic autoimmune disease characterized by immune system abnormalities. It’s unclear how melatonin influences the immune responses. In this study we investigated the levels of serum melatonin in sjögren patients and melatonin’s diurnal rhythm. METHOD: In this study, 47 sjögren patients (35 female, 12 male, mean age 39,7 ± 11,5 years, mean disease duration 8,2 ± 2,7 years) and 28 healthy controls ( 21 female, 7 male, mean age 41,3± 7,7 years) were included. Serum melatonin levels were measured by ELISA, in the morning and night. RESULTS: In the sjögren patients group; the mean serum melatonin levels were 49, 2 ± 12,3 at night and 52,6 ± 5,1 in the morning. In the healthy control group; the mean serum melatonin levels were 77,1 ± 12,9 at night and 27,2 ± 6,1 in the morning. There was significant change in the diurnal rhythm of the melatonin in the sjögren patient group. Melatonin secretion at night was found to be lower, in the SSc patient group than the healthy control group, but in the morning was found to be higher. Those were statically significant (p&lt;0.001, p&lt;0.001 respectively). In the patient group, there was no statically significant difference between the serum levels in the morning and night. CONCLUSION: In this study, we found that diurnal rhythm of the melatonin changed in sjögren patients. These findings can be due to the effects of melatonin on the inflammatory responses.
Immune exhaustion is associated with development of cardiopulmonary disease in a non-human primate model of HIV-infection (HUM8P.340)
Authors: Kling Heather , Shipley Timothy , Tarantelli Rebecca , Champion Hunter , Morris Alison , Gladwin Mark , Norris Karen 
Published: January 2023

Abstract
Cardiopulmonary disease remains a leading cause of morbidity and mortality in HIV infection, despite widespread use of anti-retroviral therapies. Chronic inflammation due to ongoing viral replication may lead to immune exhaustion, a potential mechanism for the development of chronic obstructive pulmonary disease (COPD) and/or pulmonary arterial hypertension (PAH). Previously, we have shown that simian immunodeficiency virus (SIV) infection of macaques results in PAH and COPD. In the current study, we examined whether lymphocyte exhaustion and/or activation is associated with cardiopulmonary disease in SIV infection. Macaques were intravenously infected with SIVΔB670 (n=6, PAH study) or SHIV89.6P (n=11, COPD study). Blood and bronchoalveolar lavage (BAL) fluid cells from S(H)IV-infected animals were analyzed by flow cytometry. Serial pulmonary function testing and right heart catheterization were performed to assess pulmonary obstruction and mean pulmonary arterial pressure (mPAP), respectively. Increased effector memory CD8+ T (Tem) cells (CD95+CD28-) (p=0.008) and expression of programmed death-1 (PD-1), a marker of exhaustion (p&lt;0.001), in the lung correlated with increased mPAP in SIV-infected macaques. Increased peripheral blood CD8+ Tem cells correlated with worse pulmonary function (p&lt;0.001) in SHIV-infected macaques. These results support the hypothesis that HIV-related cardiopulmonary disease is associated with chronic immune activation and exhaustion.