bluuebunny/crossref_metadata_2025_split huggingface dataset:
Age-related susceptibility to grey matter demyelination and neurodegeneration is associated with meningeal neutrophil accumulation in an animal model of MS
Authors: Le Zuo Michelle Xiao , Fettig Naomi M , Osborne Lisa C , Gommerman Jennifer L , Ramaglia Valeria 
Published: January 2023

Abstract
People living with multiple sclerosis (MS) experience episodic central nervous system (CNS) white matter lesions instigated by autoreactive T cells. With age, MS patients show evidence of grey matter demyelination and experience devastating non-remitting symptomology. What drives progression is unclear and has been hampered by the lack of suitable animal models. Here we show that passive experimental autoimmune encephalomyelitis (EAE) induced by an adoptive transfer of young Th17 cells induces a non-remitting clinical phenotype that is associated with persistent leptomeningeal inflammation and cortical pathology in old, but not young SJL/J mice. While the quantity and quality of T cells did not differ in the brains of old vs young EAE mice, an increase in neutrophils and a decrease in B cells was observed in the brains of old mice. Neutrophils were also found in the leptomeninges of a subset of progressive MS patient brains that showed evidence of leptomeningeal inflammation and subpial cortical demyelination. Taken together, our data show that while Th17 cells initiate CNS inflammation, subsequent clinical symptoms and grey matter pathology are dictated by age and associated with other immune cells such as neutrophils.
Dendritic cells regulate innate cell trafficking into lymph nodes during inflammation
Authors: Gerner Michael Y , Wu Yixuan , Huang Jessica Y 
Published: January 2023

Abstract
Dendritic cells (DCs) have well-established roles in antigen presentation, and we recently showed that within several hours of immunization, lymph node (LN) resident DCs undergo activation and reposition into the T cell zone (TZ) to drive early T cell activation. Concurrently, monocytes are also recruited into LNs via high endothelial venules (HEVs) and cooperate with DCs to enhance the generation of adaptive responses. However, the mechanisms driving monocyte recruitment are unknown. Here, we used mouse models of immunization, quantitative imaging, and RNA sequencing to study early innate responses in draining LNs. We find that during inflammation, monocytes as well as neutrophils rapidly infiltrated the LN parenchyma and the TZ, with the timing of cell influx being concordant with that of DC activation and repositioning. As the DCs migrated towards the TZ, they passed through regions highly enriched in HEVs. DCs also expressed multiple inflammatory chemokines involved in monocyte and neutrophil trafficking, as well as high levels of the integrin ligand, ICAM-1, thus generating chemokine- and adhesion molecule-rich perivascular microenvironments. Depletion of DCs or ablation of ICAM-1 in DCs significantly decreased innate cell trafficking into LNs, with the recruited cells being largely trapped near HEVs and less able to infiltrate the TZ. Disruption of DC repositioning with conditional CCR7 ablation also resulted in the mislocalization of neutrophils and monocytes within the LNs. Thus, in addition to their classical roles as antigen presenting cells, during inflammation, DCs facilitate the recruitment of several innate cell types into LNs and guide them into the TZ to promote the generation of functional immune responses.
Supported by NIH grants R01AI134713, R21AI142667, T32GM007270 (J.H.), and F31AI161316 (J.H.).
The tetraspanin CD53 as a novel regulator for B cell trafficking
Authors: Greenberg Zev Joshua , Li Weikai , Schuettpelz Laura Graves 
Published: January 2023

Abstract
Migration and homing of B cells involves a myriad of adhesion molecules and chemokines that enable the movement of B cells from one part of the body to another. This tightly regulated process is important to facilitate maturation and optimize B cell function. Accordingly, loss of proper adhesive interactions leads to reduced immunoglobulin production and impaired immune function. As a family, tetraspanins regulate cellular growth, survival, and migration. Specifically, the tetraspanin CD53 has been implicated in early B cell development and immunoglobulin production; yet a role for CD53 in B cell trafficking has not been studied. Using a Cd53−/− mouse, we find that splenic B cells have significantly reduced transmigration when placed on a monolayer of human dermal microvascular endothelial cells, compared to WT B cells (1.7% vs 4.2% transmigration, p&lt;0.0001). We next profiled chemotaxis in response to CXCL12 using a transwell assay and found significantly less migration of Cd53−/− bone marrow-derived pre-B cells (3.2% vs 5.0% of input, p&lt;0.01) and Cd53−/− splenic B cells (14% vs 20% of input, p&lt;0.001), compared to WT littermates. Splenic B cells from Cd53−/− mice also had significantly impaired in vivo homing to both the bone marrow (2.8% vs 7.7%, p&lt;0.01) and spleen (2.6% vs 7.3%, p&lt;0.001) compared to WT splenic B cells. Upon ex vivo stimulation of splenic B cells with CXCL12, loss of CD53 resulted in significantly less phosphorylation of the signaling mediators Akt (7.8% vs 12.2%, p&lt;0.001) and Erk (12% vs 23%, p&lt;0.0001), measured by flow cytometry. Together, these data suggest that the CXCL12 receptor, CXCR4, is a candidate binding partner for CD53 and that loss of CD53 abrogates normal B cell migration and homing.
Supported by institutional funding from Washington University in St. Louis
Delineating the mechanistic pathway and therapeutic potential of Siglec-6 in Chronic Lymphocytic Leukemia
Authors: Nunes Jessica Judith , Ventura Ann , Zapolnik Kevan , Mo Xiaokui , Zhang Liwen , Chen Min , Byrd John , Rader Christoph , Muthusamy Natarajan 
Published: January 2023

Abstract
Sialic acid-binding immunoglobulin-like lectins (Siglec) regulate immune function via glycan recognition. Here, we show for the first time the functional relevance of Siglec-6 and its ligand sialyl Tn (sTn) in cell adhesion and migration in chronic lymphocytic leukemia (CLL). Siglec-6+ primary B-CLL cells have significantly enhanced adhesion and migration towards sTn+ CLL− bone marrow stromal cells. A Siglec-6 targeted antibody inhibited homing of Siglec-6+ MEC1 cells and B-CLL cells to the spleen and bone marrow in NSG mice. Mass spectrometry analysis in MEC1 cells revealed interaction of Siglec-6 with DOCK8, a guanine nucleotide exchange factor. Stimulation of Siglec-6+ MEC1 cells with sTn resulted in Cdc42 activation, WASP recruitment and enhanced actin polymerization, which were compromised in Siglec-6 or DOCK8 KO MEC1 cells. Additionally, cell fractionation experiments revealed that Siglec-6+ MEC1 cells and B-CLL cells had higher levels of DOCK8 at the cell membrane when compared to MEC1 Siglec-6 KO cells and normal donor B cells respectively, indicating that Siglec-6 may be responsible for tethering DOCK8 to the cell membrane and is important for promoting actin polymerization at the leading cell edge. To study the therapeutic potential of Siglec-6, we generated a Siglec-6+ CLL mouse model (Siglec-6 x TCL1). Ex vivo experiments with a Siglec-6/CD3 targeted bispecific antibody showed specific killing of Siglec-6+ leukemic B cells. In summary, our studies have identified a novel role for Siglec-6 in promoting DOCK8 dependent CLL cell migration and opens up new therapeutic avenues to eliminate Siglec-6+ malignant B cells. Ongoing studies are focused on evaluating in vivo efficacy of Siglec-6/CD3 targeted bispecific antibody.
Natarajan Muthusamy: NIH-R21 Grant and Pelotonia Idea grant Jessica Nunes: Pelotonia Graduate Fellowship
Effects of Mycoplasma pneumoniae CARDS Toxin on mitochondrial homeostasis
Authors: Liendo Nathaniel Steven , Benavides Andrea Milenia , Segovia Jesus 
Published: January 2023

Abstract
Mycoplasma pneumoniae is an atypical bacterial pathogen known to cause mild respiratory infections and pneumonia. M. pneumoniae produces Community-Acquired Respiratory Distress Syndrome toxin (CARDS toxin), a virulence factor that plays a primary role in pathogenesis. CARDS toxin is known to have dramatic vacuolating and inflammatory properties, however these properties have not been fully characterized.
Recent evidence from our lab has supported that the toxin localizes on the mitochondria, affecting homeostasis and triggering inflammation. Our current study is focused on characterizing the effects of CARDS toxin on mitochondrial stress and production of mitochondrial reactive oxygen species (ROS). A549 lung epithelial cells and IMR-90 lung fibroblasts were treated with recombinant CARDS toxin exogenously, and mitochondrial ROS was measured. Additionally, we sought to determine whether ADP-ribosylation or vacuolization were required for CARDS toxin-induced mitochondrial stress by expressing CARDS toxin truncations on a mammalian expression plasmid. Endogenously expressed CARDS toxin truncations were fused to either EGFP or mCherry2. CARDS toxin was detected by western blot and IFA was used to track CARDS toxin localization in the cell.
Our findings support our hypothesis that CARDS toxin disrupts mitochondrial homeostasis and triggers mitochondrial ROS production. Our results provide valuable insight into the effects of CARDS toxin on mitochondrial function, which will help us better characterize the role of CARDS toxin during M. pneumoniae pathogenesis.
Supported by NIH T34GM008073
TLR-9 agonist CpG activates macrophages and modulates expression of microRNAs involved in cancer drug resistance and various signaling pathways
Authors: Kulangara Varsha Kontham , Zhong Yin , Yang Xiaoming , Nagarkatti Prakash S , Nagarkatti Mitzi 
Published: January 2023

Abstract
TLR-9 agonist, CpG induces pro-inflammatory immune response. We studied the immune response elicited in C57Bl/6 female mice by repeated stimulation of TLR-9 through multiple doses of CpG administration. Complete blood count analysis after CpG treatment revealed peripheral pancytopenia and the spleen showed splenomegaly. Flow cytometry analyses of spleen and liver mononuclear cells disclosed macrophage activation, evidenced by substantial increase in F4/80+ cells in the treatment group compared to control. Analysis of serum by ELISA demonstrated enhanced levels of pro-inflammatory cytokines, IL12/23, IL6 and TNFα in the CpG-treated group whereas no significant change was observed in the amount of anti-inflammatory cytokine, IL10. The spleen cells were further subjected to microarray analysis in order to examine the modulation of microRNA expression pattern following CpG treatment. Investigation of miRNA-mRNA relationships using the ingenuity pathway analysis (IPA) revealed differential expression of miRNAs associated with cancer drug resistance, HOTAIR and BAG signaling pathways along with microRNAs involved in immunomodulation. The major microRNAs altered with CpG treatment included mir27a, mir181, mir130a, mir487a, mir29b1, mir130a, mir3473b, mir21a, mir142 and mir144 targeting genes involved in drug resistance, apoptosis, cell cycle, angiogenesis and major histocompatibility complex-related proteins. Together, our studies demonstrate that chronic macrophage activation may lead to uncontrolled activation and proliferation of immune cells. (Supported by NIH grants P01AT003961, P20GM103641, R01AI129788, R01ES030144, R01AI160896 and R01AI123947).
Supported by NIH grants P01AT003961, P20GM103641, R01AI129788, R01ES030144, R01AI160896 and R01AI123947
CXCR4 promotes the stop signal and degranulation of cytotoxic T cells infiltrating influenza-infected lungs
Authors: Mrass Paulus , Byrum Janie , Torres David , Cannon Judy L 
Published: January 2023

Abstract
Cytotoxic T cells can promote protective immunity or exacerbate lung damage during influenza infections. Fine-tuning the functional activity of cytotoxic T cells in situ within influenza-infected lungs is a potential strategy to balance immunity and immunopathology. Using a murine model of influenza, we found that CXCR4 expression strongly correlated with cytotoxic T cell degranulation and that inhibition of CXCR4 reduced the degranulation of cytotoxic T cells in vitro and in vivo. Live tissue imaging revealed that influenza-specific T cells had prolonged dwell times when they were in regions with high levels of influenza antigen. Inhibition of CXCR4 led to increased T cell speed and decreased stop times in influenza-high areas. Moreover, inhibition of CXCR4 expedited the recovery of flu-infected mice. These data identify CXCR4 as a positive regulator of the stop-signal in lung-infiltrating CD8+ T cells and suggest that targeting this pathway could enhance recovery from influenza-induced weight loss.
Type I Diabetes induced by OT-I T-Cells requires ITK-mediated TCR signaling.
Authors: Perrenoud Loni Jean , Conley James , Berg Leslie 
Published: January 2023

Abstract
Activation of CD8+ T-cells induces changes in T cell trafficking, promoting effector T cell migration into tissues. ITK, a tyrosine kinase activated by TCR signaling, has previously been shown to be required for CD8+ T cell migration into the gastrointestinal tract during viral infection. This finding prompted us to investigate whether ITK signaling was important for effector T cell trafficking into non-infected tissue, as might occur in autoimmune disease. Using the RIP-mOVA mouse model, in which the OVA protein is expressed in the β-islet cells of the pancreas, we examined the ability of WT versus ITK-deficient OT-I T cells to infiltrate the pancreas inducing Type I diabetes. To test this, WT or ITK-deficient OT-I cells were adoptively transferred into RIP-mOVA recipients. Mice were immunized with OVA peptide plus LPS, and monitored for urine glucose. We also examined OT-I cell numbers in the pancreatic draining lymph node and in the pancreas 72hr post-immunization. We found that in the absence of ITK, OT-I cells were impaired in their ability to induce diabetes, with only ~1/3 of the recipients progressing to disease; in contrast 100% of recipients receiving WT OT-I cells developed Type I diabetes. In addition, the numbers of infiltrating ITK-deficient OT-I cells in the pancreas was significantly reduced compared to the numbers of WT OT-I cells detected. These findings indicate that ITK has a key role in generating pathogenic tissue-infiltrating CD8+ T cells capable of migration into β-islets of the mouse pancreas. While exploration of the mechanism underlying this impaired migration needs further investigation, these data provide support for therapeutic potential of an ITK inhibitor in patients with tissue-specific autoimmune diseases.
Supported by NAID TRC SIGNLG
Mechanosensory channel Piezo1 contributes to T lymphocyte migration through direct interaction with integrins
Authors: Laverty Miranda S , Benson Bryan L , Aguilar Alicia , Kingsley Daniel , Myers Jay T , Choi Sung-Hee , Huang Alex Y 
Published: January 2023

Abstract
Response to mechanical signals is an important mechanism by which T lymphocytes drive inflammation. A potential contributor in this process is Piezo1, a mechanosensitive cation channel which is abundantly expressed in T lymphocytes. Previously, our lab has shown that Piezo1 colocalizes with high affinity LFA-1 on activated human T lymphocytes during chemokine-activated crawling, suggesting a role for Piezo1 in the adhesion cascade. However, the mechanism by which Piezo1 contributes to the regulation of the adhesion cascade is unknown. We hypothesize that Piezo1 mediates T lymphocyte migration through direct contact with integrin to regulate its function. We investigate the role of Piezo1 in the expression, binding affinity, and turnover of LFA-1 and VLA-4. Co-immunoprecipitation revealed a preferential association of Piezo1 with the alpha subunit of integrins, specifically LFA-1, MAC-1, and VLA-4 in the active forms. Moreover, microfluidic devices were used to study the contribution of Piezo1 on T lymphocytes under laminar flow-induced shear stress. We found that activated T lymphocytes with CRISPR/Cas9-mediated Piezo1 ablation resulted in a reduced chemokine-induced crawling speed that correlates to the efficiency of Piezo1 gene ablation. Finally, in studies of lymph node homing where fluorescently labeled T lymphocytes were tracked in vivo, Piezo1 deficient cells exhibit increased ingress and decreased egress through the lymph node, suggesting an intricate role of Piezo1 on T lymphocyte migration. Taken together, our data support the hypothesis that Piezo1 modulates inflammatory processes through complex integrin interactions and mediates T lymphocyte migration from the lymph node to the site of inflammation.
Case Postbaccalaureate Research Education Program grant NIH/NIGMS 5R25GM075207-15; St. Baldrick’s Foundation; Hyundai Hope-on-Wheels Program; The I’m Not Done Yet Foundation; and Pediatric Cancer Research Foundation
The CD6-ALCAM pathway selectively modulates pathogenic T cell migration
Authors: Marrocco Valeria , Ampudia Jeanette , Connelly Stephen , Ng Cherie 
Published: January 2023

Abstract
CD6 is a costimulatory receptor on T cells that, through binding to its ligand ALCAM, modulates both the activity and trafficking of effector T cells (Teff). ALCAM has been seen to be overexpressed on various tissues during inflammation and is associated with increased infiltration of pathogenic CD6high CD4+ T cells in patients with autoimmune and inflammatory diseases. However, the role of the CD6-ALCAM pathway in the chemotaxis of pathogenic T cells into inflamed tissues is not yet clear.
Here, we investigate the contribution of CD6 in mediating chemokine-induced migration of Teff cells through the endothelial barrier by using itolizumab, a humanized anti-CD6 monoclonal antibody that inhibits the CD6-ALCAM interaction and reduces CD6 levels on T cells in the presence of monocytes. The migration of Teff cells from PBMCs or in vitro polarized TH17 cells expressing high levels of CD6, was tested under static conditions using a Boyden chamber assay incorporating human umbilical vein endothelial cells (HUVEC) which express high levels of ALCAM.
Analysis of migrating CD4+ T cells from total PBMCs showed that cells expressing higher levels of CD6 preferentially migrated through the HUVEC layer in response to CXCL12. Blocking the CD6-ALCAM pathway reduces the migration of CD4+ CCR7− CD45RA− TEM cells by ~60%. Furthermore, itolizumab treatment reduced the migration of pathogenic TH17 when co-cultured with monocytes.
This data demonstrates that the CD6-ALCAM pathway contributes to the chemokine-driven migration of Teff cells into inflamed organs through the endothelial layers. Consequently, blockade of this pathway will not only inhibit the activity of Teff but may also selectively affect their infiltration into inflamed organs.
Improving the Effectiveness of Combining Voice-Over PowerPoint Presentations and Review Games for Delivering Immunology Content.
Authors: Cusick John K , Goel Pranshul , Tang Justin Shiann , Abram Alyssa , Shyu Yennie , Gerriets Valerie 
Published: January 2023

Abstract
This study aims to improve the combined delivery of voice-over PowerPoint lectures and review games to increase student engagement and learning. Medical students were assigned immunology voice-over lectures prior to in-class or online learning sessions, in which students participated in teams to entertain sample exam questions in review games. Students provided quantitative feedback using a seven-point Likert scale and provided suggestions on how to improve the delivery of this pedagogy. The participating students were divided into groups that had differing access to either the voice-overs and/or review games before taking a formative quiz on the subject material. We are currently unable to conclude whether voice-overs or review games are more effective at increasing student performance on formative quizzes, yet future studies are planned to address this. More importantly, substantial feedback received from students through this ongoing study has been incorporated to improve the delivery of combining pre-assigned voice-overs with in-class gaming. Students overwhelmingly favored practice questions embedded within voice-overs and preferred multiple voice-overs of shorter length over long voice-overs for a given subject. Additionally, students greatly appreciated a brief question-and-answer review of the main points of the voice-over before beginning the in-class review game. Collectively, the students preferred this active learning pedagogy over traditional lectures. This study has provided valuable feedback to enhance the delivery of an active learning pedagogy that is relevant to both undergraduate and graduate-level immunology courses and is readily adaptable for either in-class or online learning sessions.
Seed grant from California Northstate University Institute of Teaching and Learning Excellence
The zombie pirate invasion: an introduction to immunology concepts.
Authors: Csencsits-Smith Keri , Robson Rachel 
Published: January 2023

Abstract

Introduction
The terminology that is used to describe the parts of the immune system can be very intimidating to student learners. We have noticed that some students disengage from the learning process early on in the semester because they are overwhelmed by “jargon” (e.g. CD markers, cytokines). The classroom exercise described here serves to introduce students to the concept of immunology as the body’s defense system as well as to provide a simplified framework for students to refer to as they learn more advanced concepts.

Methods
First-year medical students at Saba University School of Medicine, located on the small Dutch Caribbean island of Saba, are engaged in this group-based exercise at the beginning of the immunology content block. We present them with the following scenario: “Our island is being invaded by zombie pirates! How can we defend against this attack?” Each learning group is assigned a specific task, such as figuring out how to communicate across the island, determining how to differentiate zombies from residents, or planning how to train the populace in defense. Students then share their answers with the class. Instructors facilitate the linking of student answers with applicable concepts in immunology, e.g. communication = cytokine signaling, zombie identification = MHC presentation, training centers = lymph nodes.

Results/Conclusion
We have found that students remember this discussion in the following weeks, and that we can use the ideas they presented as a framework for reference as we introduce immunology concepts. This activity could be adapted to a number of different educational settings, contingent on the unique geographic or structural features of the campus on which other students study.