bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dysregulated CD8+ T cells promote B cell activation and antibody-mediated autoimmune disease (BA2P.123)
Authors: Hoyer Katrina , Gravano David , Sadiqi Sharif , Sanders P. 
Published: January 2023

Abstract
IFNγ-producing CD4 and CD8 T cells accumulate during autoimmune disease in IL-2-KO mice. CD4 T cells are necessary for disease progression, and we have previously shown that IL-2-KO CD4 T cells can transfer autoimmunity. However, the importance of CD8 T cells to initiation and propagation of disease is unclear. Several groups have demonstrated a protective role for CD8 T cells in inflammation and autoimmunity, while others have suggested a pathogenic role. To define the function of CD8 T cells during spontaneous autoimmune disease, we depleted CD8 T cells in IL-2-KO mice and evaluated the pathogenesis of autoimmunity. Elimination of CD8 T cells for only two weeks resulted in a significant augmentation of survival, reduction in the number and activation state of the CD4 T cells and B cells, and a delay in the development of RBC-specific antibodies. In untreated IL-2-KO mice, both CD4+ and CD8 T cells produced IFNγ in response to RBC antigens, indicating that RBC-specific CD8 T cells are present during disease. Our results suggest that autoreactive CD8 T cells contribute to the initiation or propagation of lymphoproliferation and autoimmunity. We are addressing the relative importance of autoreactive CD8 T cells to that of the CD4 T cells, and are evaluating the mechanisms by which CD8 T cells influence the autoimmune disease process. Early data indicate that IL-2-KO CD8 T cells produce elevated levels of perforin, granzyme B, and IFNγ, and induce B cell proliferation.
T follicular helper cells contribute to autoimmunity through the BCMA-BAFF axis (BA2P.117)
Authors: Coquery Christine , Loo William , Wade Nekeithia , Bederman Annelise , Tung Kenneth , Lewis Janet , Hess Henry , Erickson Loren 
Published: January 2023

Abstract
T follicular helper (TFH) cells are critical for the development of protective antibodies via germinal center B-cell responses; however, uncontrolled TFH cell expansion activates autoreactive B-cells to produce antibodies that cause autoimmunity. Although increased circulating level of the cytokine BAFF has been linked to loss of B-cell tolerance in systemic autoimmunity, the potential role for BAFF in TFH cell homeostasis is not known. B-cell maturation antigen (BCMA) is a receptor for BAFF and is critical for the development and survival of bone marrow plasma cells (PCs). Paradoxically, BCMA deficiency exacerbates the formation of autoantibody-secreting PCs in spleens of autoimmune-prone mice. Here, we show that TFH cells uniquely express BCMA and that the absence of BCMA in TFH cells enhances their numbers and responsiveness to BAFF, and in turn, drive autoantibody production. Moreover, circulating CD4+ T cells and TFH-like cells that express another BAFF receptor BR3 (but not BCMA) are elevated in SLE patients and positively correlate with disease severity. These results identify a new BCMA-BAFF axis in controlling TFH cell homeostasis and suggest that the balance between BCMA and BR3 signaling in TFH cells serves as an additional checkpoint of immune tolerance.
Decreased inflammatory dendritic cells in lupus prone ERαKO mice correlate with increased survival (BA4P.214)
Authors: Cunningham Melissa , Wirth Jena , Eudaly Jackie , Gilkeson Gary 
Published: January 2023

Abstract
Lupus is a disease that disproportionately affects females. We previously showed that a functional knockout of estrogen receptor alpha (ERαKO) resulted in significantly reduced renal disease and increased survival in murine lupus. Dendritic cell development, which requires both estrogen and ERα is impacted, as is activation status and cytokine production. Due to altered hormonal feedback loops, ERαKO mice have hypergonadism and partial endocrine sex reversal. Elevated estrogen (E2) and testosterone levels may have immunomodulating effects. Thus, we studied the phenotype of the lupus-prone ERαKO mouse following ovariectomy (OVX) +/- E2 replacement to preserve a physiologic hormonal state. We found that NZM2410 ERαKO mice were protected from lupus disease expression (no early deaths; no proteinuria at 32 weeks) if they were either unmanipulated or if they were both ovariectomized and E2-repleted. These mice also had fewer inflammatory and activated cDCs (CD11c+/CD11b+/MHCII+ cells from Flt3L-cultured bone marrow, or ex vivo spleen or kidney cells), which correlated with increased survival. Interestingly, protection was lost after ovariectomy if no E2 pellet was administered, suggesting that the protective effect requires E2 in the system (despite the lack of a functional ERα). A protective effect was not observed in lupus-prone Ex3a mice (ERα−/−) suggesting that the functional mutant in ERαKO mice, in the presence of estrogen, potentially modulates disease.
Mouse CD300b (CLM-7) regulates the phagocytosis of apoptotic cells via phosphatidylserine recognition. (BA4P.215)
Authors: Coligan John , Tian Linjie , Margulies David , Krzewski Konrad , Murakami Yousuke 
Published: January 2023

Abstract
The CD300 receptor family emerges as group of important molecules that modulate a variety of immune cell processes.We show that mouse CD300b (CLM7/LMIR5), expressed by myeloid cells, recognizes outer membrane-exposed phosphatidylserine (PS) and does not, as previously reported, directly recognize TIM1 or TIM4. CD300b conveys activation signals through its association with adaptor molecules, like DAP12, and accumulates in the phagocytic cups at apoptotic cell contact sites, thereby facilitating the engulfment of such cells. Engulfment requires a functional DAP12 ITAM motif. Importantly, silencing CD300b expression by macrophages diminishes Syk, PI3K and Akt phosphorylation in the presence of apoptotic cells and impairs phagocytosis of these cells. Collectively, our data shows that CD300b recognizes PS as a ligand, and regulates the phagocytosis of apoptotic cells via the DAP12 signaling pathway.
Ste20-like proline/alanine-rich kinase (SPAK) deficiency exacerbates experimental colitis in mouse (CAM1P.239)
Authors: Yan Yutao , Hong Yu , Cao Yifei , Xu Xianrong , Tan Xiaohua , Yang Lei 
Published: January 2023

Abstract
Inflammatory bowel diseases (IBDs) are characterized by epithelial barrier disruption and alterations in immune regulation. Ste20-like proline/alanine-rich kinase (SPAK) plays a role in intestinal inflammation, but the underlying mechanisms need to be defined. Here, SPAK knockout (KO) C57BL/6 mice exhibited significant increase in translocation of intestinal bacteria into mucosa, accompanying with marked elevation in paracellular permeability to FITC-dextran and altered apical side tight junction sodium ion selectivity, compared to wild type (WT) mice. In murine models of colitis induced by dextran sulfate sodium (DSS) and trinitrobenzene sulfuric acid (TNBS), KO mice were more sensitive than WT mice, as demonstrated by colonoscopy features, histological characteristics and MPO activities. Consistent with these findings, KO mice showed decreased IL-10 levels and increased proinflammatory cytokine secretion. Co-immunoprecipitation and immunohistochemical assays demonstrated that SPAK is associated with PKCtheta and CD46, regulators of regulatory T cell differentiation and activation. Flow cytometry analysis found that KO mice had significant lower amount of T regulatory 1 (Tr1) cells in comparison with WT mice. In conclusions, SPAK deficiency compromises intestinal innate immune homeostasis, implying its important role as a potential intervention target against pathological responses in IBD.
19F-MRI for tracking NK Cells after adoptive transfer (CAM1P.240)
Authors: Bouchlaka Myriam , Gordon Jeremy , Ludwig Kai , Niles David , Bednarz Bryan , Fain Sean , Capitini Christian 
Published: January 2023

Abstract
Natural killer (NK) cells have shown anti-tumor responses against solid tumors. However, it is not known if NK cells traffic directly to tumors and mediate their cytotoxicity, or if they indirectly mediate their anti-tumor effects by stimulating other effector cells. Current methods of tracking immune cells by imaging of 1H contrast agents, such as iron oxide particles, are not ideal as a large part of the 1H signal comes from water in the tissues. Because there are trace amounts of fluorine in the body, we have optimized the non-radioactive isotope 19-fluorine (19F) as a means of tracking human NK cells by magnetic resonance imaging (MRI). In vitro, we show that NK cell viability, their ability to secrete granzyme B or IFN-γ and cytotoxicity against K562 leukemia cells are not affected by 19F labeling and are similar to unlabeled cells. There was also no impact on expression of natural cytotoxicity receptors, NKG2D or DNAM-1. MR Spectroscopy has shown that NK cells highly take up 19F, with up to 10^12 atoms/cell. In vivo, 19F-labeled NK cells injected in the footpad of NOD-SCID-gamma (c)-/- mice were nontoxic and detectable by MRI. Overall, perfluorcarbon 19F labeling of NK cells represents a successful method to track and quantify the number of apparent cells in a region of interest without altering their cytotoxic function. Importantly, 19F imaging will enable us to monitor NK cells after their infusion allowing us to determine how they induce their anti-tumor effects.
Regulation of integrin LFA-1 (CD11a/CD18) during T cell migration and activation (CAM1P.227)
Authors: Capece Tara , Abid Anna , Kim Minsoo 
Published: January 2023

Abstract
T cell entry into a lymph node is dependent upon several events, including activation of chemokine receptor CCR7 and adhesion of integrin LFA1. T cells become highly motile to scan dendritic cells (DCs) for cognate antigen (Ag), utilizing active LFA1 at the leading edge. Ag challenge induces stable T-DC interactions and immunological synapse (IS) formation, where active LFA1 relocates. LFA1 is an adhesive force in migration and IS formation, but the outcome of chemokine and T cell receptor (TCR) signals are different, as the former induce “go” and the latter mediate “stop” signals. We hypothesize the magnitude of chemokine and TCR signals will determine the outcome of LFA1 mediated interactions and thus T cell activation. We generated fluorescent knock-in (KI) mice to visualize LFA1 distribution and activation patterns. We mated these KI mice with OTI TCR transgenic mice to study LFA1 distribution in T cells activated by varying TCR signal strength with altered peptide ligands (APL). We have thus far shown LFA-1 clusters faster and remains clustered longer at the contact site in interactions with cognate Ag compared to those with weaker APLs. We are repeating these experiments in our LFA1 FRET mice, which allow us to visualize the LFA1 activation, to determine if LFA1 activation induces the LFA1 clustering seen in interactions with cognate Ag. We have also incorporated a photoactivatable chemokine receptor, PA-CCR7, to assess the effect of chemokine signal strength on LFA1.
Essential role of elmo1 in dock2-dependent lymphocyte migration (CAM1P.228)
Authors: de la Rosa Gonzalo , Stevenson Catherine , Anderson Christopher , Murphy Patrick , Capece Tara , Kim Minsoo , Elliott Michael 
Published: January 2023

Abstract
Elmo1 and Elmo2 are highly homologous cytoplasmic adapter proteins that interact with Dock family guanine nucleotide exchange factors to promote activation of the small GTPase Rac. In T lymphocytes, Dock2 is essential for CCR7- and CXCR4-dependent Rac activation and chemotaxis, but the role of Elmo proteins in regulating Dock2 function in primary T cells is not known. Here we show that endogenous Elmo1 but not Elmo2 interacts constitutively with Dock2 in mouse and human primary T cells. CD4+ T cells from Elmo1-/- mice were profoundly impaired in polarization, Rac activation and chemotaxis in response to CCR7 and CXCR4 stimulation. Transfection of full-length Elmo1, but not Elmo2 or a Dock2-binding mutant of Elmo1, rescued defective migration of Elmo1-/- T cells. Lymphocytes from Elmo1-/- mice showed a four-fold reduction in total Dock2 protein levels despite normal levels of Dock2 mRNA, suggesting Elmo1 is critical for the maintenance of Dock2 levels in primary lymphocytes. Indeed, Dock2 polyubiquitination was increased in Elmo1-/- T cells, and treatment with proteasome inhibitors partially restored Dock2 levels in Elmo1-/- T cells. Taken together, these findings reveal a previously unknown, non-redundant role for Elmo1 in controlling Dock2 levels and Dock2-dependent T cell migration. This work provides valuable insights into the molecular regulation of Dock2 by Elmo1 that can be used to design improved Dock2-based therapeutic approaches.
CD103 downregulation marks memory-phenotype CD8 T cells and excludes them from the gut (CAM1P.231)
Authors: Ligons Davinna , Sato Noriko , Park Jung-Hyun 
Published: January 2023

Abstract
CD103 is a cell surface integrin whose primary role is the retention of lymphocytes in epithelial tissues where its ligand E-cadherin is expressed. CD103 is found on all newly generated CD8 single positive thymocytes but not on CD4 lineage cells. Because of such lineage specific expression, CD103 is currently considered a lineage specific marker whose expression is proposed to be controlled by Runx3, a runt-family transcription factor involved in CD8 lineage specification. Here we report the surprising finding that Runx3 was neither necessary nor sufficient for CD103 expression on CD8 T cells. Notably, we identified a subpopulation of CD8 T cells that had downregulated CD103 expression, despite expressing high levels of Runx3. Upon correlating CD103 expression with activation and memory markers, we found that CD103-negative CD8 T cells corresponded to memory phenotype cells and that homeostatic cytokines signals terminated CD103 expression on CD8 T cells. Importantly, CD103-negative CD8 T cells were found in every lymphoid organ and other peripheral tissues with the exception of the gut. Indeed, all CD8+ IELs were CD103-positive, and adoptive transfer of CD103-negative CD8 T cells resulted in their recovery from the LN but not the intestinal mucosal tissues. Collectively, these results suggest that CD8 memory-phenotype acquisition is accompanied by CD103 downregulation to control their tissue distribution and to exclude them from the gut.
The development and survival but not function of follicular B cells is dependent on IL-7Rα Tyr449 signaling. (CCR1P.242)
Authors: Abraham Ninan , Patton Daniel , Plumb Adam , Redpath Stephen , Osborne Lisa , Perona-Wright Georgia 
Published: January 2023

Abstract
IL-7 is a critical cytokine for lymphocyte development but its role in B cells is less well characterized. Using a knock-in mouse with a Tyr to Phe mutation at position 449 (IL-7Rα449F/449F mice) within the cytoplasmic domain of IL-7Rα, we evaluated the role of this YxxM motif in spleen B cells. IL-7Rα449F/449F mice had reduced numbers and increased death of follicular B cells compared to WT, but had significantly more follicular cells than IL-7Rα-/-. The death of IL-7Rα449F/449F follicular cells was not due to a failure to respond to BAFF or lower levels of BAFF. Marginal zone B cells were unaffected in IL-7Rα449F/449F mice. A role for TSLP was ruled out, as TSLPR-/- mice had an identical B cell phenotype to wild-type mice. Bone marrow chimeras and the absence of IL-7Rα on B cells suggested that IL-7 did not directly regulate mature B cells, but that an IL-7-responsive cell was influencing B cells. IL-7 was also critical at the checkpoint between the T1 and T2 stages in the spleen. We tested the functional responses of IL-7Rα449F/449F mice and found no difference in antibody responses to T-dependent or T-independent antigens, or to Influenza/A. IL-7 was important for generation of antibody responses to the intestinal worm H. polygyrus and for naive levels of IgA. Our data shows that IL-7 regulates follicular B cell numbers and survival in a cell-extrinsic manner, via a bone-marrow derived cell, but is not critical for antibody production outside the gut.
CCR2-dependent dendritic cell recruitment to CNS tissues during EAE promotes disease progression and inflammatory cytokine production by CNS T cells (CCR1P.243)
Authors: Clarkson Ben , Rayasam Aditya , Walker Alec , Harris Melissa , Sandor Matyas , Fabry Zsuzsanna 
Published: January 2023

Abstract
Dendritic cells (DCs)—while absent from the healthy central nervous system (CNS)—rapidly accumulate within brain and spinal cord tissue during neuroinflammation associated with experimental autoimmune encephalomyelitis (EAE, a mouse model of multiple sclerosis). Yet, while DCs have been appreciated for their role in initiating adaptive immune responses in peripheral lymphoid organ tissues, how DCs infiltrate the CNS and contribute to ongoing neuroinflammation in situ is poorly understood. Therefore we sought to identify key chemokines and chemokine receptors on DCs that govern DC migration across the blood-brain barrier and within the CNS. Here we report that 1) CD11c+ bone marrow-derived (BM)DCs and CNS-infiltrating DCs express CCR2, 2) compared to CCR2+/+ cells, adoptively transferred CCR2-/- BMDCs or DC precursors do not accumulate in the CNS during EAE, despite abundance in blood, 3) CCR2-/- DCs show less accumulation in the inflamed CNS in mixed bone marrow chimeras, when compared to CCR2+/+ DCs, and 4) ablation of CCR2+/+ DCs during EAE clinical onset delays progression and attenuates T cell cytokine production. These data suggest that CCR2-dependent DC recruitment to the CNS during ongoing neuroinflammation plays a crucial role in effector T cell cytokine production and disease progression, and signify that CNS-DCs and circulating DC precursors might be key therapeutic targets for modifying ongoing neuroinflammation in CNS autoimmune diseases.
Demonstrating the immune response of D. rerio using keyhole limpet hemocyanin (EDU1P.249)
Authors: Young Matthew , Criscitiello Michael 
Published: January 2023

Abstract
In this project, immunization of a model aquatic organism is used in the classroom to facilitate understanding of molecular biology, evolution and the vertebrate adaptive immune system. Molecular immunoglobulin and T cell receptor cloning, sequencing and repertoire analysis techniques that I learned in the Criscitiello Comparative Immunogenetics Lab at Texas A&M this summer were applied to analysis of the lymphocyte antigen receptor repertoire in zebrafish. Students at A&M Consolidated High School performed experiments to quantify and qualify various aspects of the immune response of the model organism, D. rerio. Hemocytometry was used to measure leukocyte counts before and after immunization with keyhole limpet hemocyanin. Serum IgM was analyzed using ELISA. Repertoire analysis was performed using PCR and DNA sequencing. The data from these experiments was used in the classroom to discuss the evolution of vertebrate adaptive immunity, immunization techniques, and vaccination design. Furthermore, the repertoire sequence data contributed to an important phylogenetic perspective for ongoing NSF funded studies at A&M on the plasticity of B and T cell receptor immunogenetics. One goal is for high school students to earn co-authorship on journal publications from the Criscitiello lab. This was the first stage in an iterative system of teaching and research that will continue to include reciprocal pedagogical exchange and hands-on experience in comparative molecular immunology.
Integrating immunology into a clinical presentation-based medical school curriculum: lessons learned (EDU1P.251)
Authors: Piskurich Janet , Baatar Dolgor 
Published: January 2023

Abstract
Clearly there is no one best way to study the complex adaptive system that comprises our ability to respond to a wide variety of microorganisms, many of which have developed medically relevant means to evade the immune system. As a teacher or learning facilitator it can be difficult to even decide where to start. Different text books adopt a variety of approaches. A further problem unique to the educational process of medical students is how to provide a relevant framework to enhance their retention of the concepts of basic immunology and their ability to apply these concepts later as they begin to practice medicine. Paul L. Foster School of Medicine, a new medical school located on the Texas Mexico border, utilizes a clinical presentation-based curriculum model. This model has also been recently adopted by several other new medical schools across the country. In this model, students learn clinically-relevant basic science in the context of common patient presentations. Pros and cons of the integration of the basic science of immunology into a clinical presentation-based curriculum as well as lessons learned over the first five years will be addressed.
Pre-clinical studies of inflammation: automating the most common methods of animal randomization and distribution (EDU1P.254)
Authors: Ibsen Eric , Kumer Jeff 
Published: January 2023

Abstract
To more closely emulate clinical research methodology and improve clinical relevance, researchers are increasingly utilizing more clinically-similar group randomization and distribution methods for Inflammation studies in animals. A common pre-clinical approach is paradoxically the least clinically-relevant; a random number generator in a spreadsheet, which is subject to manual and transcription errors and provides no easy way to exclude animals from distribution based on other parameters. Pre-clinical trial software is available which automates the most recommended randomization and distribution methods based on tumor volume or other parameters: deterministic and pair-matched distribution, pure randomization, stratified sampling randomization and randomization across multiple numeric parameters. Subjects may be excluded initially based on recorded clinical observations and comments and then by acceptable value ranges for any numeric parameter. Animals can be randomized iteratively in a rolling or staggered way and can be re-randomized into new groups as required on drug-resistance studies. Proposed randomization results by ANOVA can be displayed instantly to ensure that the group means are acceptably similar. Standardizing and automating methods of randomization and distribution which are similar to this used clinically can potentially improve the ease, clinical relevance, quality and outcome of pre-clinical Inflammation studies relative to spreadsheet-based methods.
CXCR4 antagonist AMD3100 mobilizes leukocytes from bone marrow and thymus to blood in mice (HEM2P.265)
Authors: Liu Qian , Li Zhanzhuo , Gao Ji-Liang , Wan Wuzhou , McDermott David , Murphy Philip 
Published: January 2023

Abstract
AMD3100 is approved for mobilizing hematopoietic stem cells to blood for transplantation in cancer by blocking chemokine receptor CXCR4. It also mobilizes mature leukocytes to blood; however, there is conflicting published evidence regarding the source of mobilized neutrophils (bone marrow vs lung), and the source(s) of other mobilized leukocytes is unknown. Here we used surgical, pharmacologic and histologic approaches to address these questions in mice. AMD3100 10 mg/kg sq mobilized the same leukocyte subsets to blood in mice as in humans, and increased neutrophil and monocyte content in lung and lymph node, while reducing neutrophil, lymphocyte and monocyte content in bone marrow as well as T cell content in thymus. Direct intrathymic labeling showed that AMD3100 mobilized naïve CD4+ and CD8+ T cells to blood. In contrast, splenectomy and experiments with FTY720, which blocks T cell egress from lymph node, suggested that spleen and lymph node were not major sources of AMD3100-mobilized cells. Importantly, AMD3100 did not inhibit neutrophil trafficking to thioglycollate-inflamed peritoneum. These data provide evidence that CXCR4 is a selective compartmentalization factor for both bone marrow (lymphocyte, monocyte, neutrophil) and thymus (naïve T cells), but is not critical for homeostatic trafficking of leukocytes to uninflamed tissues or for innate immune responses to inflamed sites, a profile that is favorable for chronic administration of AMD3100 in patients.