bluuebunny/crossref_metadata_2025_split huggingface dataset:
PU.1-dependent T cell function limits inflammation in EAE (BA14P.214)
Authors: Awe Olufolakemi , Pham Duy , Kaplan Mark 
Published: January 2023

Abstract
T helper cell subsets mediate autoimmune disease through the secretion of specific cytokines that recruit other immune cells and alter the inflammatory microenvironment. IL-9-secreting Th9 cells can contribute to the development of autoimmune diseases, including Experimental Autoimmune Encephalomyelitis (EAE), although the mechanisms are unclear. Elimination of IL-9 or IL-9R in models of EAE has led to somewhat conflicting results. To attempt a different approach to this question we examined the development of EAE in mice with a T cell-specific deletion of the Sfpi1 gene, which encodes PU.1, a transcription factor important for the production of IL-9. Interestingly, Sfpi1lck-/- mice had significantly worse disease scores compared to WT mice beginning 19 days after initial MOG35-55 immunization. The disease scores for the Sfpi1lck-/- mice persisted through day 30 in contrast to disease resolution in WT mice. Although few IL-9 secreting T cells were found among CD4+ T-cells isolated from the brains of Sfpi1lck-/- mice 25 days after initial immunization, there were significantly higher frequencies of IFN-γ, IL-17a, and IFN-γ+IL-17a+ when assayed by intracellular staining. There was also higher MOG-specific total IgG and IgG1 levels in the Sfpi1lck-/- mice. These results suggest that PU.1 dependent immunity limits autoimmune inflammation, particularly during the resolution phase of disease.
Increased lymph node CD8+ T cell proliferation and memory cell markers upon conditional ablation of Langerhans cells in autoimmune mice (BA14P.215)
Authors: Gutierrez Miguel-Angel , Halder Ramesh , Valera Isela , Singh Ram 
Published: January 2023

Abstract
Systemic autoimmune diseases such as lupus are characterized by inflammation in many organs. The role of local tissue-resident immune cells in specific organ involvement in these diseases remains unclear. We evaluated the role of Langerhans cells (LC) as prototypic tissue-resident dendritic cells (DC) in eliciting local immune defects in the MRL model of lupus. Injection of diphtheria toxin (DT) in Lang-eGFP.DTR knock-in mice results in depletion of langerin+ dermal DC (dDC) and LC. To assess LC’s role in systemic autoimmunity, we introgressed the knock-in mutation from the stock B6 onto the MRL background. Serial DT injections every 7-10 days exacerbate inflammation in the skin (King et al) and cornea in MRL but not in B6 mice. To investigate underlying mechanisms, we assessed the effect of LC depletion on T cells in skin/eye draining cervical lymph nodes (cLN). There was a significantly higher proliferation of CD8+, but not CD4+, T cells in cLN of LC-ablated than control mice; such effect was not seen in spleen CD8+ T cells. Also, LC-ablated MRL, but not B6 mice, exhibited reduced CD62L on cLN T cells. Further, freshly isolated dDC were more efficient than LC in promoting the proliferation of CD8+, but not CD4+, cNL T cells. Such local regulation of CD8+ T cells may play a role in immune tolerance locally. A defect in this mechanism owing to reduced LC migration in MRL mice (Eriksson and Singh, 2008) may confer skin/eye inflammation, without affecting other organs
Foxp3-mediated inhibition of Akt inhibits Glut1 expression in human T regulatory cells (BA14P.203)
Authors: Basu Samik , Bowen Britany , Shevach Ethan 
Published: January 2023

Abstract
Thymic-derived CD4+CD25+Foxp3+ T regulatory cells (Tregs) have a diminished capacity to activate the PI3K/Akt pathway. Although blunted Akt activity is necessary to maintain Treg suppressive function, other consequences of decreased Akt signaling are unclear. Glucose transporter 1 (Glut1) is a cell surface receptor responsible for facilitating glucose transport across plasma membranes and expression of Glut1 is tightly coupled to co-stimulatory signals and Akt phosphorylation. Freshly isolated human Tregs were unable to up-regulate Glut1 in response to TCR and co-stimulatory signals when compared to similarly activated effector T cells. Furthermore, stimulated Tregs utilized less glucose than stimulated effector T cells in ex vivo cultures. Introduction of Foxp3 into effector T cells by lentiviral-mediated transduction inhibited not only Akt activation, but also Glut1 expression, indicating that Foxp3 can regulate Glut1. Together, these results illustrate the molecular basis underlying differential glucose metabolism between human Tregs and effector T cells as well as revealing potential pathways for immune modulation.
T cell movement into the pancreas based on insulin receptor expression (BA14P.208)
Authors: Al-Dieri Ali , Esakov Emily , Slotterbeck Brandon , McInerney Marcia 
Published: January 2023

Abstract
In the nonobese diabetic (NOD) mouse, type 1 diabetes is an autoimmune disease characterized by insulitis and T cell-mediated beta cell destruction. The importance of insulin receptor (IR) expression in the pathogenesis of diabetes was examined since IR is a chemotactic molecule capable of directing cell movement in response to a gradient of insulin. Our published data shows that T cells, obtained from NOD diabetic mice, expressing a high density of IR can aggressively transfer insulitis and diabetes while T cells with low to negative amounts of IR, are capable of neither. Therefore, IR may function to deliver T cells to the islet where Ag specific cells within the population can recognize and destroy beta cells. Recently, it has been shown in NOD mice that benign insulitis can occur in the absence of T cell-MHC/Ag interactions. The object of this project was to test whether T cells expressing IR, can move into the pancreas based on insulin chemotaxis alone, in normal strains of mice. A transgenic C57Bl/6 mouse was made in which the mouse IR gene with a 3XFLAG tag was placed behind the CD3 promoter and enhancer to target IR expression on all T cells. In this transgenic model there is IR expression on all T cells and with age the transgenic mice exhibit insulitis, but not diabetes. Distinct areas of the IR mediate metabolic or chemotactic signaling indicating that a specific part of the IR may be selectively targeted for therapeutic intervention to block T cell movement.
Delineating the mechanisms of immunomodulatory effects of sex steroids in autoreactive T cells (BA15P.219)
Authors: Jia Ting , Massilamany Chandirasegaran , Reddy Jay 
Published: January 2023

Abstract
SJL mice immunized with proteolipid protein (PLP) 139-151 show a gender disparity in the development of autoimmune encephalomyelitis in that only females show a chronic relapsing-remitting type of paralysis, but their underlying mechanisms are unknown. We tested the hypothesis that sex steroids differentially regulate the expansion of autoreactive T cells. To this end, we have established an in vitro system to determine the effects of sex steroids on the expansion and functions of antigen-specific T cells based on proliferation assay, major histocompatibility complex class II dextramer staining, cytokine analysis and cell viability assays. Regardless of gender, we noted that the T cell responses to PLP 139-151 were significantly lower in cells exposed to either testosterone or estradiol as compared to vehicle-treated cultures, but the threshold for hormone-sensitivity is two-fold higher in males than females. Importantly, the reduced proliferative responses were found not to be due to hormonal inhibition, rather, the hormones promoted cell death. Conversely, while, the hormones reduced the number of interferon-gamma-producing cells in both the genders, interleukin (IL)-17-producing cells were reduced only in females; whereas other cytokines, such as IL-2, IL-4, IL-10, and IL-22 remained unaltered. Taken together, the data suggest that the immunomodulatory effects of sex steroids involve multiple pathways, and cell death may be the primary underlying mechanism.
Genetic deletion of S100A9 fails to abrogate psoriasiform inflammation in the KC-Tie2 mouse model due to IL-12/23 compensation. (CAM5P.237)
Authors: Ward Nicole , Fritz Yi , Dawes Sean , Diaconu Doina , Golden Jackelyn , Wang Yunmei , Simon Daniel , McCormick Thomas 
Published: January 2023

Abstract
The proinflammatory proteins S100A8/9 are purported to play a pathogenic role in psoriasis. A recent report showed genetic deletion of S100A9 strongly attenuated ear skin inflammation in the K5-JunB:Jun mouse and imiquimod skin inflammation models. We previously reported increased expression of S100A8/9 (&gt;135-fold; P&lt;0.05) in the KC-Tie2 psoriasis mouse model, with expression downregulated following disease reversal. We sought to confirm the importance of S100A9 in disease pathogenesis and mated KC-Tie2 mice with S100A9 KO mice. In contrast to recent reports, KC-Tie2-S100A9 KO mice (n=11) had similar levels of skin inflammation (acanthosis), T cell and myeloid cell skin-infiltration as KC-Tie2 mice (n=9); despite significantly decreased levels of DefB4, IL-1α, IL-17C and TNFα (all P&lt;0.05). Interestingly, KC-Tie2-S100A9 KO animals had significantly elevated splenic and skin-draining CD11b+Ly-6C+ pro-inflammatory monocytes (27% and 86%, respectively; P&lt;0.05) compared to KC-Tie2 animals, increased expression of skin IL-12/23 and IL-17A/F and sustained levels of skin IFNγ, IL-22, IL-6 and KLK6, potentially explaining the sustained presence of disease. These results suggest that the IL-23-Th17 inflammatory axis may compensate for genetic ablation of S100A9 and sustain psoriasiform skin inflammation in KC-Tie2 animals. Thus, the role of S100A9 in skin inflammation may be model dependent and further investigation is needed to establish S100A9 as a therapeutic target in psoriasis.
SOCS proteins regulate insulin receptor substrate (IRS)-2 activation in response to IL-4 and 13 in monocytes and macrophages (CCR5P.247)
Authors: McCormick Sarah , Gowda Nagaraj , Fang Jessie , Heller Nicola 
Published: January 2023

Abstract
Allergic asthma is a chronic lung disease driven by Th2 cytokines IL-4 and -13. In macrophages, IL-4/13 bind IL-4 receptors, which signal through IRS-2/STAT6, inducing M2 differentiation. We found that IL-4/13 stimulation of human monocytes/mouse macrophages leads to rapid tyrosine phosphorylation (pY) of IRS-2, peaking at 60min and returning to baseline by 150min. Mechanisms that regulate IRS-2 activity and M2 differentiation are not understood. Based on rapid de-phosphorylation of IRS-2, we hypothesized that SOCS proteins regulate IL-4/13 induced IRS-2 activation, thereby limiting M2s. To address this, we tracked the kinetics of SOCS induction following IL-4/13 exposure and found that SOCS1 and CIS are highly induced in human monocytes and mouse macrophages, respectively, while SOCS3 is only moderately induced. By co-immunoprecipitation, we found that SOCS1 and CIS complex with IRS-2 in response to IL-4/13. Furthermore, we found that siRNA knockdown of SOCS1 and CIS prolonged IRS-2 pY and enhanced human and mouse M2 differentiation respectively. Since SOCS proteins complex with ubiquitin ligases to promote proteasomal degradation, we examined the effect of proteasome inhibitors on IL-4/13-induced IRS-2 pY. Proteasome inhibition prolonged IRS-2 pY and enhanced M2 gene expression. These findings identify SOCS1 in human and CIS in mouse macrophages as critical negative regulators of IL-4/13-induced IRS-2 activity/M2 differentiation and are novel therapeutic targets for asthma.
Activator of G-protein signaling 3 (AGS3) binds GRK6 via its G-protein (GPR) domain to modulate CXCR2 mediated cellular responses. (CCR5P.261)
Authors: Singh Vandana , Smith Nikia , Richardson Ricardo 
Published: January 2023

Abstract
AGS3, a receptor-independent activator of G-protein signaling, comprises of seven tetratricopeptide (TPR) motifs and four GPR motifs joined by a linker sequence. Previous studies have demonstrated that TPR domain of AGS3 is important for determining the positioning of AGS3 within the cell, whereas, GPR domain serves as a docking site for Gαi-GDP leading to AGS3-Gαi signaling. We have recently shown that GRK6 complexes with AGS3-Gαi to regulate CXCR2-mediated cellular functions including Ca2+ mobilization, mitogen-activated protein kinase (MAPK) activation, receptor desensitization and down-regulation. In the current study mutants from the GPR and TPR domains were used to determine which motif(s) of AGS3 is (are) involved in binding with GRK6. Co-imunoprecipitation studies in RBL cells co-transfected with wild type and mutants AGS3 showed that, upon stimulation with interleukin 8 (IL-8/CXCL8), GRK6 interacts with the GPR domain of AGS3. Overexpression of AGS3 and the GPRs motif (ΔTPR7-link), but not the TPRs motif (TPR), in RBL-CXCR2 inhibited CXCL8-induced intracellular Ca2+ mobilization and MAPK activation. Taken together, these results indicated that AGS3 binds to GRK6 via the GPR domain to modulate CXCR2-mediated signal transduction.
FcγRIIB-mediated dissociation of BCR-CD19 microclusters and hyperreactivity of lupus B cells (HUM7P.303)
Authors: Liu Wanli , Xu LiLing 
Published: January 2023

Abstract
B cell activation is regulated through the interplay between the B cell receptor (BCR) and the inhibitory co-receptor FcγRIIB or activating co-receptor CD19. Recent studies suggest that the BCR microcluster is converted to a signaling active state upon its colocalization with CD19 microclusters to activate B cells. Here using TIRF microscopy-based live cell imaging techniques, we show that FcγRIIB can inhibit B cell activation by a previously uncharacterized mechanism of blocking the spatial-temporal colocalization of BCR and CD19 microclusters within the immunological synapse (IS). This inhibitory function of FcγRIIB mainly relies on its transmembrane domain (TM). Indeed, human primary B cells from SLE patients homozygous for the gene encoding FcγRIIB-I232T lose the ability to block the synaptic colocalization of BCR with CD19, leading to the dis-regulated recruitment of pCD19, pSyk and pPI3K to membrane proximal signalosome. Furthermore, FcγRIIB-I232T homozygous SLE patients are more likely to develop severe clinical symptoms characterized by high anti-ds DNA antibody titers and the involvement of kidney, central nervous system, gastrointestinal tract, lung and heart compared to patients carrying either one or both WT copies of FcγRIIB. These observations may explain the hyper-reactive features of SLE patient B cells and provide a potential target through FcγRIIB for the prophylactic diagnosis, evaluation of disease progression and potential therapies.
Inflammatory responses in myopia progression (HUM7P.317)
Authors: Wan Lei , Lin Hui-Ju , Hsu Yu-An 
Published: January 2023

Abstract
Myopia is an important and common eye disease as it can induce many other associated eye diseases and even lead to legal blindness. The prevention and treatment of myopia is an important public problem worldwide. We found that the expression levels of several inflammatory molecules are altered in the retina and sclera of FDM hamsters. To affirm the possible correlation between inflammatory reaction and myopia progression, cyclosporine A was used to treat FDM rats. We found that inhibiting inflammatory responses in the eye slowed myopia progression, whereas increasing inflammation by LPS and PGN enhanced myopia advancement. The expression levels of TGF-β, MMP2, CHRM1, CHRM3, Id1, c-Fos, NF-κB, IL6, and TNF-α were up-regulated in occluded eyes and down-regulated when treated with atropine and cyclosporine A. Id1, c-Fos, NF-κB, IL6, and TNF-α are known to have important roles in inflammatory reactions. Our results showed that immune and inflammatory reactions would alter the progression of myopia through modulation of inflammation responses.
An iNKT cell expansion causes premature delivery in mice (HUM7P.323)
Authors: Stlouis Derek , Sanchez-Rodriguez Elly , Milovic Tatjana , Romero Roberto , Gomez-Lopez Nardhy 
Published: January 2023

Abstract
Objective: Maternal-fetal tolerance involves a tight balance between the innate and adaptive immunity, which avoids rejection of the semi-allogeneic embryo. The innate and adaptive limbs of the immune response are bridged by iNKT cells; therefore, we hypothesized that an alteration in their number and/or activation breaks maternal-fetal tolerance, leading to premature delivery. Methods: Pregnant mice were injected on E16.5 with either α-galactosylceramide (an iNKT activator; 2 μg) or PBS as a control (n=19 each). Gestational length and the rates of stillbirth and long-term pup survival were recorded. Peripheral blood, decidua, liver, spleen and lymph nodes were collected just prior to delivery from additional treated and control mice (n=8-9 each). In tissue cell suspensions, iNKT phenotype and activation were analyzed by flow cytometry. Results: 1) iNKT activator treatment promoted premature delivery (E18.09±0.60; controls delivered on E19.51±0.78); 2) iNKT activator treatment increased the rate of stillbirth (63.63 vs. 21.94%); 3) iNKT activator treatment did not reduce long-term pup survival; 4) iNKT activator treatment caused an expansion of iNKT cells (CD1d tetramer+NK1.1+CD49b+) in peripheral blood, liver and decidua, but not in spleen and lymph nodes; 5) iNKT cell activation was unchanged by iNKT activator treatment. Conclusion: Treatment with α-galactosylceramide causes an expansion of systemic and local iNKT cells leading to premature delivery in mice.
Examining a potential link between antigen-presentation and metabolic dysregulation (HUM7P.289)
Authors: Bhagwandin Candida , Whalen Michael , Ishido Satoshi , Szajman Adam , Truong Sarah , Tsao Tsu-Shuen , Lybarger Lonnie 
Published: January 2023

Abstract
Antigen-presentation is induced following interaction of an antigen-presenting cell (APC) with stimuli, including microbial ligands and cytokines, whereupon Major Histocompatibility Complex II (MHC-II) and co-stimulatory molecule, CD86, surface expression is significantly upregulated. At the surface of the APC, the immune modulators provide signals critical for T cell activation and induction of the appropriate adaptive immune response. As key regulators of the T cell-APC conversation, it is imperative to gain a thorough understanding of the underlying mechanisms regulating expression and function of antigen-presenting molecules, and the biological consequences of dysregulation. Indeed, our lab has demonstrated that Membrane-Associated RING-CH1 (MARCH1), a highly-conserved E3 ubiquitin ligase identified as a potent regulator of both MHCII and CD86 in APCs, is stringently regulated at multiple steps, indicating a need for precise control of antigen-presentation. Interestingly, MARCH1-deficient and hemizygous mice show signs of mild hyperglycemia, an increase in the visceral adipose tissue (VAT), and increased inflammation of the VAT, all characteristic of insulin-resistance and type II diabetes progression. Collectively, the data support the hypothesis that antigen-presentation is tightly regulated to ensure a rapid adaptive immune response, and suggests a novel role for components of this pathway in the maintenance of insulin sensitivity in healthy adipose tissues.
In vitro hyporesponsiveness of CD4+ and CD8+ T cells in septic patients with faecal peritonitis (HUM8P.326)
Authors: Gore David , Preciado-Llanes Lorena , Mills Gary , Heath Andrew , Read Robert 
Published: January 2023

Abstract
The immunological processes behind sepsis are ill defined. Predicting or diagnosing sepsis, based on cellular immune parameters is a research priority. This study quantified differences in peripheral blood mononuclear cell (PBMC) and cytokine levels between septic patients with fecal peritonitis, matched surgical patients (no sepsis) and healthy controls. To assess function, PBMC were stimulated in vitro with anti-CD3±anti-CD28 antibodies. Four days later T and B cell proliferation were measured by CFSE and activation by expression of CD25 and CD69 on T cells and CD86 on B cells. Viability was assessed by flow cytometry. Concentrations of a range of cytokines were determined from plasma and supernatants (post-stimulation) by multiplex cytometric bead array. On day of phlebotomy there were reduced CD4+ and increased CD19+ cells in septic patients. Plasma IL-6 and IL-8 levels were higher in septic patients, IFN-γ and IL-13 were lower. There was reduced proliferation and activation of T cells from septic patients in response to anti-CD3+anti-CD28. Supernatant levels of IFN-γ, TNF-α and interleukins 1β, 5, 6, 13 and 17A were lower in septic patient samples. There were no initial differences in PBMC viability, but septic patient PBMC had reduced survival on incubation. In summary, we found reduced T cell numbers and a functional impairment of remaining T cells in septic patients.To our knowledge this is the first report of an immune function assay across these three groups.
Intensive immune suppression of Bolivian squirrel monkeys activates latent polyomavirus (HUM8P.337)
Authors: Nehete Pramod , Ruiz Julio , Nehete Bharti , Vanchiere John 
Published: January 2023

Abstract
The squirrel monkey polyomavirus (SMPyV) is commonly found in Bolivian squirrel monkeys, and, as such, the squirrel monkey may be a useful model for the study of polyomavirus-associated pathogenesis and experimental treatment and prevention strategies. Squirrel monkeys were given single or multiple doses of an anti-B cell antibody (Rituximab) and/or an anti-CD8 monoclonal antibody (7pt-3F9) which resulted in a complete CD20+lymphocyte depletion and transient CD8+lymphocyte depletion as compared to control animals. The animals remained clinically well, suggesting that the intensity and/or duration of immune suppression were inadequate to trigger pathogenic reactivation of the latent polyoma and herpes viruses. Quantitative PCR assays revealed excretion of the SMPyV in the urine within one week after the 7pt-3F9 infusion, continuing until the end of the study. At necropsy, spleen, liver, lymph nodes, cerebrum, cerebellum and kidney were positive for SMPyV DNA. Histologic and immunohistochemistry studies revealed moderately increased numbers of meningeal cells and inflammatory mononuclear cells in the kidney, renal medulla and foci of increased Ki67 staining in distal tubular epithelial cells in antibody treated monkeys compared to untreated monkey. One animal had multifocal encephalitis with prominent CD3+lymphocyte collections throughout the cortex. These results suggest that the transient depletion of CD8+lymphocytes may be a useful trigger for SMPyV shedding.
Multi-drug resistant Th17 cells: new players in autoimmune and steroid-resistant inflammation (HUM8P.351)
Authors: Sundrud Mark , Ramesh Radha , Kozhaya Lina , McKevitt Kelly , Djuretic Ivana , Carlson Thaddeus , Quintero Maria , McCauley Jacob , Abreu Maria 
Published: January 2023

Abstract
Th17 cells are generally regarded as key effectors of autoimmune disease. However, not all Th17 cells are pro-inflammatory. Furthermore, the defining features of "pathogenic" Th17 cells in humans are unknown. We have found that pro-inflammatory human Th17 cells are restricted to a subset of CCR6+ CXCR3(hi) CCR4(lo) CCR10- CD161+ T cells that expresses the drug transporter multi-drug resistance-1. Unlike MDR1- human memory T cell subsets, MDR1+ Th17 cells produce both Th17 and Th1 cytokines upon stimulation; do not express IL-10 or other anti-inflammatory molecules; express high levels of IL23R as well as other genes selectively expressed by pathogenic mouse Th17 cells; and display heightened responsiveness to IL-23 stimulation. In vivo, MDR1+ Th17 cells accumulate in the gut of patients with active Crohn's Disease, where they display a marked pro-inflammatory transcriptional signature relative to local MDR1- effector T cells. In addition, MDR1+ Th17 cells are refractory to the immunosuppressive effects of several steroids used to treat autoimmune disease in the clinic. Thus, MDR1+ Th17 cells may be important mediators of autoimmune inflammation, particularly in clinical settings of steroid-resistant disease.