bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human gut commensal Prevotella histicola ameliorates disease as efficiently as Copaxone and Interferon-β in a preclinical animal model of multiple sclerosis
Authors: Shahi Shailesh K. , Freedman Samantha N. , Gibson-Corley Katherine , Karandikar Nitin J. , Murray Joseph A. , Mangalam Ashutosh K. 
Published: January 2023

Abstract
Multiple sclerosis (MS) is a demyelinating disease of the central nervous system. We and others have shown that gut microbiota play an important role in disease pathogenesis of MS as there is an enrichment or depletion of specific gut bacteria compared to healthy controls (HC). Thus the specific human gut bacteria found depleted or showing lower abundance in MS patients can be used as potential drug to treat MS. Further, we and others have shown that patients with MS have either lower abundance of Prevotella compared to HC or an increased level of Prevotella in patients receiving disease modifying therapies such as Copaxone and/or Interferon beta (IFN-β). We have previously identified a specific strain Prevotella histicola within Prevotella genus which can suppress disease in experimental autoimmune encephalomyelitis (EAE) animal model of MS. In this present study we compared disease suppressing ability of P. histicola to Copaxone or IFN-β in HLA-DR3.DQ8 transgenic mouse model of MS. We observed that P. histicola suppresses EAE as efficiently as Copaxone and IFN-β. P. histicola treated mice showed higher level of CD4+FoxP3+ regulatory T cell frequencies and decreased levels of pro-inflammatory cytokines IFNγ and IL17 suggesting that P. histicola suppresses disease through immunomodulation. Fecal microbiota analysis of different groups showed that group receiving Prevotella histicola had a distinct gut microbiota than the control group suggesting that P. histicola as might suppresses disease through modulation of gut microbiota. In conclusion, our study suggests that human gut commensal Prevotella histicola can provide an alternate treatment option for MS patients.
SA-FasL-engineered PEG microgels as a novel means of modulating immune response to allogeneic islet grafts
Authors: Shirwan Haval , Woodward Kyle B. , Headen Devon M. , Zhao Hong , Shrestha Pradeep , Tan Min , Bowen William S. , Coronel María M. , Hunckler Michael D. , Weaver Jessica D. , Yolcu Esma S. , García Andrés J. 
Published: January 2023

Abstract
Allogenic islet transplantation is an effective therapy for type 1 diabetes in the clinic. Sustained graft survival requires chronic immunosuppression that has adverse effects. Islet graft rejection is initiated and perpetuated by T effector cells (Teffs). Teffs upregulate Fas death receptor on their surface and undergo apoptosis following engagement with FasL. Therefore, FasL has the potential as an immunomodulator to establish immune tolerance to islet grafts by eliminate alloreactive Teffs. This study used PEG microgels engineered with a novel form of FasL, SA-FasL, for the prevention of allogeneic islet graft rejection. Poly(ethylene glycol) microgels were produced with biotin on their surface. Biotinylated microgels were engineered with SA-FasL (1 μg/103 microgels) by taking advantage of the high affinity interaction between biotin and the streptavidin (SA) domain of the molecule. SA-FasL-PEGs induced apoptosis in Fas expressing A20 lymphoid cells in vitro. Co-transplantation of SA-FasL-PEGs with naïve islets led to prolonged survival of grafts as compared with the control group (islets + unmodified PEGs), with 20% surviving for a 200-day observation period. A short (15 days) treatment with low dose rapamycin further improved the immunomodulatory efficacy of SA-FasL-PEGs, with &gt; 90% of grafts surviving for a 200-day observation period. CD4+CD25+FoxP3+ Tregs were required for graft survival, as depletion of this cell population on day 50 post-transplantation resulted in prompt rejection. These findings provide strong proof-of-efficacy and feasibility for the use of SA-FasL-engineered microgels as an off-the-shelf product for the modulation of alloreactive responses with significant therapeutic potential.
CD45 ligation reduces Tregs’ motility and enhances recognition of activating signals in vitro
Authors: Lee Jounghyun Helen , Lee Joanne Haeun , Rothstein David , Kam Lance C. 
Published: January 2023

Abstract
Regulatory T cells (Tregs) play a pivotal role in modulating immune response hence supporting immune tolerance. Recently, CD45 ligation with anti-CD45RB mAb MB23G2 (anti-CD45RB) was reported to enhance graft-survival; evidence shows it was by homeostatic Treg proliferation in vivo. While many in vivo data support the notion that anti-CD45RB supports Tregs expansion, it is still unclear if the ligation ex vivo is sufficient to enhance Tregs’ avidity for activation and further promote their expansion and function. Here, we were to see if anti-CD45RB ligation can enhance Treg’s sensitivity to perceive activating signals in varying strength in vitro. To test this hypothesis, mouse CD4+ T cells (Foxp3-GFP linked B6 mouse, both Foxp3+ and Foxp3−, or Tregs and Tconvs, respectively) were seeded onto surfaces micropatterned with an antibody targeting the CD3 on T cell (anti-CD3) with and without the presence of anti-CD45RB. These surfaces were created by first microcontact printing of 2-μm diameter features of anti-CD3 at various concentrations (i.e., 1, 5, and 20 μg/mL) on glass coverslips and back-filled with ICAM-1. The effect of anti-CD45RB on Treg and Tconv activation was compared by the ratios of phosphorylation of Lck at Tyr 394 to 505 sites, cell registration on anti-CD3 dot array, and motility of each cell type on varying concentration of activation signals. Overall results show that anti-CD45RB specifically slow-down Treg motility and, therefore, enhance the probability to contact to activating signals (cell registration) and improve T cell activation indicated by ratios of phospho-Lck at active over inactive sites. This study assures the importance of further investigation on the role of anti-CD45RB in Treg activation mechanism.
TIPE2, an inflammation transfer switch, regulates the on-and-off immunosuppressive function of MDSCs in tumors
Authors: Yan Dehong , Wang Jinghui , Adeshakin Adeleye O. , Xu Meichen , Chen Youhai H. , Wan Xiaochun 
Published: January 2023

Abstract
Inhibition of the tumor-promoting inflammation of myeloid-derived suppressor cells (MDSCs) is an important determinant of successful clinical immunotherapies. However, the critical checkpoints governing MDSC function in cancer remain poorly characterized. We report here that the critical role of the essential negative regulator of inflammation tumor necrosis factor-α–induced protein 8-like 2 (TIPE2) in the accumulation and immune inhibitory function of MDSCs. TIPE2-deficient MDSCs decreased immune-regulatory activity and enabled the activation of T cell function and the induction antitumor responses. TIPE2 expression in MDSCs was induced by tumor-derived reactive oxygen species (ROS). Importantly, TIPE2 promoted CCAAT/enhancer-binding protein-β (C/EBPβ) by negative feedback inhibiting phospho-AKT/phospho-GSK-3β/phospho-C/EBPβ signaling axis, leading to decreased interleukin-6 (IL-6) and IL-10 production and STAT3 phosphorylation. IL-6 overexpression restored immune-suppressive activity of TIPE2-deficient MDSCs. These findings suggest a critical role of TIPE2 in tumor-induced tolerance and a novel therapeutic strategy of targeting TIPE2 in MDSCs to enhance the potential of cancer immunotherapy.
Profiling the effects of carbonic anhydrase 9/12 inhibition on tumor associated macrophage/microglia polarization in glioma
Authors: Boyd Nathaniel , McFarland Braden , McDonald Paul , Dedhar Shoukat , Hjelmeland Anita 
Published: January 2023

Abstract
Brain tumor microenvironments, including hypoxia and acidic stress, contain a subset of stem-like neoplastic cells termed brain tumor initiating cells (BTICs) that are resistant to chemo- and radiotherapy, providing a reservoir for tumor recurrence and a desirable target for glioma treatments. In prior studies, we have shown that inhibition of carbonic anhydrases 9 and 12 (CA9/12) in combination with chemotherapeutic temozolomide effectively delayed in vivo growth in a BTIC xenograft model. CA9/12 are hypoxia-responsive genes shown to be elevated in tumors that modulate the intra- and extracellular pH in tumor microenvironments. Importantly, downstream effects of hypoxia have shown CSF-1R induction, which can polarize tumor associated macrophages/microglia (TAM) toward an immunosuppressive phenotype and further exacerbate tumor progression. We have profiled CA9/12 inhibitor-treated GBM tumors using integrative omics (kinomics/transcriptomics) and discovered a shift in kinomic activity and gene expression that suggests an immunosuppressive transition in TAMs. Validating this observation, we characterized surface expression of CD11b+ TAMs post-treatment and discovered decreased expression of MHCII (inflammatory) and increased expression of CD206 (immunosuppressive). Together, our data suggest that by understanding the immunophenotypic shift that happens in response to CA9/12 inhibition in glioma, we may be able design a treatment strategy to improve upon the effectiveness of inhibitor therapy.
Mutation of the CD28 costimulatory domain confers increased CAR T cell persistence and decreased exhaustion
Authors: Boucher Justin C. , Li Gongbo , Shrestha Bishwas , Zhang Yongliang , Vishwasrao Paresh , Cabral Maria L. , Guan Lawrence , Davila Marco L. 
Published: January 2023

Abstract
The therapeutic promise of chimeric antigen receptor (CAR) T cells was realized when complete remission rates of 90% were reported after treating B cell acute lymphoblastic leukemia (B-ALL) with CD19-targeted CAR T cells. However, with increasing numbers of patients treated, challenges have become evident, especially regarding poor CAR T cell responses, which might be caused by exhaustion. Detailed understanding of the mechanism of CD28-dependent exhaustion in CAR T cells will allow the design of a CAR less prone to exhaustion and reduce relapse rates. We hypothesized that by mutating the CD28 subdomains YMNM and PRRP and leaving only the PYAP CD28 subdomain active we would optimize CAR T cell signaling and reduce exhaustion. We found mutated CAR T cells with only a functional PYAP subdomain secrete 50% less IFNγ, IL6, and TNFα after 24hr stimulation as well as having significantly less TCR signaling as measured by Nur77 compared to non-mutated CD28 CAR T cells. Additionally, mutated CAR T cells injected into mice had decreased expression of PD1 (82% to 62%) in the BM after 2 months. Studies in Rag−/− mice also showed mutated CAR T cells were increased 35% in the BM and 92% in the spleen compared to non-mutated CD28 CAR T cells. After challenge with tumor, exhausted mutant CAR T cells had increased IFNγ (42%), TNFα (62%) and IL2 (73%) secretion compared to exhausted non-mutated CD28 CAR T cells. This suggests that CAR T cells with only the PYAP subdomain are able to persist and be effective longer because they have optimized CD28 signaling. This work allows for development of an enhanced 2nd generation CAR T cell therapy for B cell malignancies by optimizing CAR T cell activation and persistence which may reduce relapse rates and severe toxicities.
Failure to upregulate calmodulin underlies the suppressed KCa3.1 function and enhanced sensitivity to adenosine in CD8+ T cells of head and neck cancer patients
Authors: Chimote Ameet A. , Gawali Vaibhavkumar S. , Madis Erick , Wise-Draper Trisha , Conforti Laura 
Published: January 2023

Abstract
The limited ability of the cytotoxic CD8+ T cells to infiltrate solid tumors presents a major roadblock to developing effective immunotherapy. Adenosine (Ado) accumulates in high concentrations in solid tumors where it contributes to the suppression of T cell function. We have previously shown that Ado reduces the chemotactic ability of peripheral blood CD8+ T cells (PBTs) from head and neck squamous cell carcinoma (HNSCC) patients by suppressing KCa3.1 channel function. Herein, we conducted experiments to elucidate the mechanism of KCa3.1 dysregulation in HNSCC PBTs. KCa3.1 channels are calcium-activated and require binding of calmodulin (CAM). PBTs were isolated from HNSCC patients and healthy donors (HD), and activated with CD3/CD28 antibodies. CAM levels decreased post-activation in HNSCC PBTs (by ~24%, n=7) while they increased in HD PBTs (by ~37%, n=6, p=0.001). To study whether CAM downregulation contributes to KCa3.1 dysfunction, we transfected HD PBTs with siRNA against CAM (siCam). siCam transfection decreased CAM expression and KCa3.1 currents, but not KCa3.1 expression. We then studied whether CAM downregulation affects the T cell chemotactic response to Ado in HD PBTs. Control (scrRNA-transfected) HD PBTs migrated towards CXCL12 in the presence of Ado, but downregulation of CAM abrogated their chemotactic ability in the presence of Ado (~71% inhibition, n=8, p&lt;0.012). Activation of KCa3.1 channels with 1-EBIO restored the ability of siCam-transfected PBTs to migrate towards CXCL12 in the presence of Ado. Our data suggest that downregulation of CAM decreases KCa3.1 activity and suppresses chemotaxis in HNSCC PBTs which may contribute to their limited ability to effectively penetrate Ado-rich tumors.
Expansion and exhaustion of alloantigen-specific cytotoxic T lymphocytes (CTL) in channel catfish, Ictalurus punctatus.
Authors: Spencer David A. , Bengtèn Eva , Wilson Melanie 
Published: January 2023

Abstract
CTL are well studied in mammals, however less is known about the dynamics of memory CTL expansion and exhaustion in teleost fish. Here we provide insights into teleost CTL dynamics using our model alloantigen-specific catfish clonal CTL cell line called TS32.15. Briefly, TS32.15 was cloned from peripheral blood leukocytes isolated from fish immunized with allogenic 3B11 B cells. This clonal line is not synchronized and consists of small resting T cells and larger granular effectors. TS32.15 cells exhibit strict target specificity for 3B11s and they require stimulation with irradiated 3B11 B cells every 7–10 days for their continuous growth. When exposed to irradiated targets, the small T cells initially do not lyse targets, but replicate and differentiate to morphologically distinct cytotoxic effectors. These effectors do not replicate, but efficiently lyse 51Cr-labeled 3B11s. After lysing targets, or with a prolonged absence of targets, effector cells transition to a non-cytolytic exhausted stage and become apoptotic. Late stage apoptotic CTL bind IgM on their surface and are preferentially cleared by myelocyte-enriched head kidney leukocytes. These results indicate teleost CTL likely undergo clonal expansion and differentiation in response to antigen similar to their mammalian counterparts, and establish binding of exogenous IgM is a marker for monitoring exhausted CTL in catfish. Additionally, our studies show that repeated stimulation of a TS32.15 culture mimics a chronic infection and results in CTL exhaustion.
Nlrp12 mediates adverse neutrophil recruitment during influenza virus infection
Authors: Hornick Emma , Banoth Balaji , Miller Ann , Zacharias Zeb Ralph , Jain Nidhi , Wilson Mary E. , Gibson-Corley Katherine , Legge Kevin L. , Bishop Gail A. , Sutterwala Fayyaz S. , Cassel Suzanne 
Published: January 2023

Abstract
Exaggerated inflammatory responses during influenza A virus (IAV) infection are typically associated with severe disease. Neutrophils are among the immune cells that can drive this excessive and detrimental inflammation. In moderation, however, neutrophils are necessary for optimal viral control. In this study, we explore the role of the nucleotide-binding domain leucinerich repeat containing receptor (NLR) family member Nlrp12 in modulating neutrophilic responses during lethal IAV infection. Nlrp12−/− mice are protected from lethality during IAV infection and show decreased vascular permeability, fewer pulmonary neutrophils, and a reduction in levels of neutrophil chemoattractant CXCL1 in their lungs compared to wild-type (WT) mice. Nlrp12−/− neutrophils and dendritic cells (DCs) within the IAV-infected lungs produce less CXCL1 than their WT counterparts. Decreased CXCL1 production by Nlrp12−/− cells was not due to a difference in CXCL1 protein stability, but instead to a decrease in Cxcl1 mRNA stability. Together, these data demonstrate a previously unappreciated role for Nlrp12 in exacerbating the pathogenesis of IAV infection through the regulation of CXCL1 mediated neutrophilic responses.
Protease activated receptor-1 signaling: its potential implication in HIV driven immune activation, inflammation/coagulation
Authors: Chen Hui , Smith Mindy , Herz Jasmin , Gossa Selamawit , Karpova Tatiana , McGavern Dorian , Catalfamo Marta 
Published: January 2023

Abstract
Persistent immune activation remains the hallmark of HIV infection even in the context of successfully suppressed viremia. Its clinical significance is suggested by increased risk to develop non-AIDS defining illnesses such as cardiovascular disease. We previously have shown that human CD4 and CD8 T cells with effector memory and terminal effector memory phenotype expressed the protease activated receptor-1 (PAR1) and that expression was increased in CD8 T cells from HIV infected patients. Thrombin, in a PAR1 dependent fashion, enhanced CD8 T cell function measured by increased secretion of IFN-g and chemokinesis. PAR1 is the main receptor of thrombin and is also expressed on platelets and endothelial cells suggesting a cross-talk between the coagulation system and T cells at the site of vascular injury. To address the role of PAR1 in vivo, we used an infection model with the hepatotropic strain of lymphocytic choriomeningitis virus (LCMV-WE). To evaluate the role of PAR1 signaling in this model, we analyzed the T cells of littermate wild type control and PAR1−/−mice eight days following infection with LCMV-WE (2×105 pfu i.v.). In this model, we observed a similar expansion of total CD8 T cells in the spleens and livers of both WT and PAR1−/−mice at 8 days post infection. Despite similar expansion, PAR1 deficient mice showed impaired viral clearance at day 8 of infection. This lack of viral control was associated with a significant reduction in cytokine secretion by gp-33-specific CD8 T cells in the livers and spleens. These results suggest that PAR1 deficiency leads to an impair viral control that is in part mediated by a decrease ability to secrete cytokine by virus specific CD8 T cells.
Mechanisms of MCMV immune evasion provide insight into MHC-I folding and assembly.
Authors: May Nathan A. , Boyd Lisa , Margulies David H. 
Published: January 2023

Abstract
As part of its strategy to evade detection by the host immune system, murine cytomegalovirus (MCMV) encodes three proteins that modulate cell surface expression of major histocompatibility complex class I (MHC-I) molecules: the MHC-I homolog m152/gp40 as well as the m02–m16 family members m04/gp34 and m06/gp48. We have explored the interaction between recombinant m06 and both full-length and truncated forms of the MHC-I molecule H2-Ld using in vitro binding assays. Previous work using solution NMR has mapped the interaction footprint of the m06 protein on MHC-I, revealing that the targeted surface area overlaps with that of the MHC-I chaperone molecule Tapasin. Furthermore, we observe enhanced binding between m06 and peptide-free forms of MHC-I, suggesting a possible chaperone-like interaction between m06 and MHC-I. Here, we express recombinant m06 in mammalian cells to explore the interplay between the viral protein and its potential targets through immunofluorescence localization experiments as well as well as binding assays. This study provides insight into the mechanism of the interaction of m06 with MHC-I, suggesting a structural manipulation of the target MHC-I molecule at an early stage of the peptide-loading pathway.
Understanding how AKT phosphorylation of hnRNPA1 modulates T cell fate and function
Authors: White Theresa L , Gable Matthew , Jin Ye , Morel Penelope 
Published: January 2023

Abstract
T regulatory cells (Treg) play a significant role in maintaining self-tolerance and preventing autoimmune diseases. We and others have shown that low dose favors Treg and T helper (Th) 2 cell differentiation, while high Ag dose stimulation activates the PI3K/Akt/mTOR pathway, favoring inflammatory Th1 and Th17 cell differentiation (Tconv). Differences in PI3K/Akt/mTOR signaling not only affects T cell fate but our research shows that Akt phosphorylation of the RNA-binding protein, (RBP) heterogeneous nuclear ribonucleoprotein (hnRNP) A1, is dependent on TCR signal strength. RBPs such as hnRNPA1 are emerging as regulators of RNA processing and stability in immune cells, and the effect of RBP on T cell differentiation is a growing subject of interest. We have shown hnRNPA1 is required for optimal Treg differentiation by performing knockdown experiments, and our present research is focused on identifying a role for Akt phosphorylation in hnRNPA1 function. HnRNPA1 is known to have a single Akt phosphorylation site at S199 and our lab has generated a new mutant mouse model, hnRNPA1-S199A (mA1). This mutation affects the ability of Akt to phosphorylate hnRNPA1 in all immune cells. Using Cytek’s Aurora flow cytometer we characterized the immune cell populations of mA1 at steady state. Preliminary data do not indicate any major changes in innate immune and B cells populations at steady state. We observed modest changes in CD4 T cell population frequencies at steady state. Current work using In vitro skewing suggests that Akt phosphorylation of hnRNPA1 influences Treg fate. This project suggests a novel mechanism by which Akt modulates T cell fate.
AID deficiency greatly improves survival and diminishes renal pathology in the BXSB mouse model of SLE
Authors: Leeth Caroline McPhee , Zhu Jing , Potter Ashley , Hasham Muneer , Richwine Madison , Roopenian Derry C 
Published: January 2023

Abstract
Eight years have passed since the approval of belimumab, a monoclonal antibody directed against B lymphocyte stimulation and the first targeted therapy approved for systemic lupus. erythematous (SLE). While well tolerated, the efficacy of belimumab remains limited and is not recommended for patients suffering severe nephritis, a life threatening complication of SLE. We sought to explore alternative targets of autoreactive B lymphocytes through manipulation of affinity maturation. The BXSB model is a long established model of human SLE developing elevated antinuclear antibodies and immune complex nephritis along with other manifestations of autoimmune disease. We used CRISPR-Cas9 to inactivate Activation Induced Cytidine Deaminase (Aicda, AID) directly in the BXSB background and found greatly improved survival. While mice continued to develop plasma cells, follicular structure was restored and renal pathology was reduced. While antinuclear antibody concentrations were reduced in knockout mice, renal immune complex deposition was grossly the same as wild type while complement deposition was very limited in the knockout mice. Mice develop expanded germinal center B cell populations as in other models of AID deficiency without concurrent expansion of follicular T cells. The prolonged survival in these mice appears to be attributed to the reduced renal pathology warranting further exploration as current therapeutics targeting lupus nephritis are limited and thus in great demand.