bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of Reactive Oxygen Intermediates in B cell activation and differentiation (45.12)
Authors: Crump Katie , Liu Chun , Konopitski Andrew , Poole Leslie , Haas Karen , Grayson Jason 
Published: January 2023

Abstract
Generating a protective humoral immune response requires proper B cell activation. Ligation of the BCR increases the level of reactive oxygen intermediates (ROI). While they have been implicated in oxidative stress, recent studies suggest ROI act as secondary messengers facilitating normal cellular function. Our focus is on the mechanisms by which ROI control B cell function. Following BCR ligation in vitro, the levels of the first oxidation product of cysteine, sulfenic acid, increased in the proteome. Using 5,5-dimethyl-1,3-cyclohexanedione (dimedone), a compound that covalently reacts with sulfenic acid to prevent further oxidation or reduction, we determined reversible cysteine sulfenic acid formation is critical for sustained tyrosine phosphorylation, calcium flux, and proliferation of B cells. To address the role of ROI in vivo, lymphocytic choriomeningitis virus (LCMV) infected mice were treated with Mn(III)tetrakis(4-benzoic acid)porphyrin chloride (MnTBAP), a superoxide dismutase mimetic, from days 0-8 postinfection. On day 38, LCMV specific IgG antibody secreting cells (ASC) were 5-fold lower in the bone marrow. In contrast, the number of LCMV specific memory B cells was unaffected. The ASC decrease was not detected when mice were only treated from days 0-4 or 4-8 postinfection or during secondary viral infection. These data demonstrate ROI and reversible cysteine sulfenic acid formation are critical to B cell function and differentiation.
CD28: a novel negative regulator of plasma cell function and survival. (45.16)
Authors: Njau Modesta , Kim Jin , Chappell Craig , Jacob Joshy 
Published: January 2023

Abstract
CD28 is a cell surface protein that is critical for the activation of naive T cells. Interestingly, CD28 is also expressed on plasma cells. Plasma cells occur in two versions - short lived ones that appear in the secondary lymphoid organs shortly following activation and the long-lived ones that reside in the bone marrow where they produce antibodies for a life time. Here we examined the role of CD28 expression on long-lived plasma cell function and longevity. To generate a host that lacks CD28 expression only on plasma cells, we adoptively transferred CD28-/- or syngeneic wildtype B cells into B cell deficient, μMT mice. We subsequently immunized the recipient hosts and monitored their serum antibody levels, plasma cell frequencies and phenotype for six months. We observed significantly higher antigen-specific IgM and IgG levels in the recipients of CD28-/- B cells in comparison to wildtype counterparts. Consistent with the higher antibody titers, CD28-/- plasma cells produced more immunoglobulin per cell and were present in greater numbers than wildtype plasma cells. Interestingly, CD28-/- plasma cells expressed higher levels of plasma cell survival factors as compared to wildtype plasma cells. Collectively these results suggest that CD28 is a negative regulator of plasma cell function and survival. Insight on the factors that modulate plasma cells can be applied to improve vaccine-induced antibody responses or regulate self-reactive plasma cells in autoimmunity.
CTLA-4 activates the hippo pathway to regulate terminal differentiation of the CD8+ T cell. (46.17)
Authors: Thaventhiran James , Hoffmann Anja , Fearon Douglas 
Published: January 2023

Abstract
Replicating, antigen-specific CD8+ T cells must not commit to terminal differentiation until there has been sufficient clonal expansion. The Hippo pathway of organ size control mediates this requirement by linking expression of the differentiation-inducing transcription factor, Blimp-1, to contact between replicating cells, which would be dependent on their frequency. TCR and IL-2R stimulation assemble the Hippo pathway in the CD8+ T cell by inducing expression of WW45, Mob1, Lats1, and YAP, the transcriptional co-activator that mediates organ growth. Contact between activated CD8+ T cells triggers the Hippo pathway, causing serine phosphorylation and degradation of YAP. This is suppressed by addition of naïve CD8+ T cells, indicating that the ligand-receptor pair triggering the Hippo pathway is expressed only by activated cells. The ligand was identified by suppressing YAP degradation with blocking CD80/86 antibody, and the receptor was defined by inducing YAP degradation by crosslinking CTLA-4. That YAP regulates differentiation was shown by ectopically expressing a non-phosphorylatable, stable form of YAP in activated CD8+ T cells, which suppressed Blimp-1 expression in vitro, and the differentiation/senescence marker, KLRG1, in vivo. This role for CTLA-4 was confirmed by the presence of YAP in T cells from CTLA-4-/-, but not CTLA-4+/-, mice. Therefore, in a process that resembles quorum sensing, the Hippo pathway regulates terminal differentiation of the CD8+ T cell.
Defining the role of mTOR in effector and memory CD8+ T cell differentiation (46.20)
Authors: Pollizzi Kristen , Delgoffe Greg , Waickman Adam , Powell Jonathan 
Published: January 2023

Abstract
Recent studies implicate mTOR in the generation of CD8+ T cell memory. Here, we demonstrate that mTORC1 is required for CD8+ T cell effector function during the activation of naïve CD8+ T cells. We generated mouse T cell-specific knockouts of Rheb, a TORC1 component; TSC2, a TORC1 negative regulator; and Rictor, a TORC2 component. We show both in vivo and in vitro that Rheb-/- CD8+ T cells have diminished effector capabilities, specifically reduced cytokine secretion and proliferation after stimulation or infection. However, these cells persist for over 30 days and retain memory cell characteristics. Conversely, TSC2-/- antigen-specific CD8+ T cells display robust effector function with increased proliferation and cytokine secretion relative to wildtype (WT). The majority of these TSC2-\- cells are unable to transition to a memory phenotype and perish during the contraction phase. Interestingly, TORC2 is not required for effector function as Rictor-/- CD8+T cells are capable of cytokine secretion and proliferation during a primary immune response. Loss of TORC2 may in fact promote memory responses, as we found Rictor-/- CD8+ T cells to have more potent antigen recall in adoptive transfer compared to WT cells. We use this novel approach to selectively probe specific mTOR functions in T cells. Ultimately, our findings highlight the divergent roles of TORC1 and TORC2 in directing CD8+ T cell effector and memory function.
Rapamycin inhibits generation of memory CD8 T cells that localize to mucosal tissues (46.23)
Authors: Sowell Ryan , Rogozinska Magdalena , Marzo Amanda 
Published: January 2023

Abstract
A limitation of current vaccine strategies is the inability to generate effective memory CD8 T cell responses in locations relevant to disease transmission. There is a gap in our knowledge of how memory CD8 T cells are generated and maintained in vivo in different tissues. In response to vaccination, CD8 T cells expand, differentiate into effectors and then undergo contraction. The end result is a population of memory CD8 T cells that survive this process. The mammalian target of rapamycin (mTOR) kinase is a critical regulator of many cell processes including proliferation, differentiation, and survival. The drug rapamycin is a selective inhibitor of mTOR and while it is widely used as an immunosuppressant to inhibit allograft rejection, it has also been shown to increase the number of viral memory CD8 T cells found within secondary lymphoid tissues. The factors that account for the discrepancy observed with rapamycin treatment on pathogen-specific versus graft-specific T cells are unclear. Here we demonstrate that despite enhancing generation of memory CD8 T cells in secondary lymphoid tissues, rapamycin inhibits generation of virus-specific memory CD8 T cells found within non-lymphoid tissues such as the lamina propria of the small intestine and vaginal mucosa. Taken together these data suggest that rapamycin treatment results in an increase in functional memory CD8 T cells residing in secondary lymphoid tissues at the expense of memory CD8 T cells in non-lymphoid sites.
The C3a receptor defines a CD8+ memory T cell subset that produces IFN-γ and TNF-α (46.3)
Authors: Moyron-Quiroz Juan , Ly Minh , Yang Xifeng 
Published: January 2023

Abstract
Complement is an important component of the innate immune system, but is also required for the generation of an optimal adaptive immune response. C3a is a product of the complement activation and is a potent mediator of inflammation. The C3a receptor (C3aR) is involved in chemotaxis, cell activation and granule enzyme release. Previous reports described that C3aR is not expressed on resting T cells, but is inducible on activated T cells. In this study, we describe a subset of C3aR+ T cells present in peripheral blood from normal healthy donors. We found that over 80% of the cells in this population are CD8+ and with an effector memory phenotype (CD45RO+, CD45RA-, CD62L-). Further analysis revealed that the CD8+C3aR+ cells express high levels of CCR4, CCR5, CCR6, CXCR3, and CXCR6, all of which are the receptors of pro-inflammatory chemokines. In addition, this cell population also expresses the adhesion molecules CD11a (LFA-1α), CD18 (β2), CD29 (β1), CD50 (ICAM-3) and CD51/61 (αV/β3), which are involved in costimulation and homing to inflamed sites. Comparing with CD8+C3aR- cells, the CD8+C3aR+ cells produce more IFN-γ and TNF-α. We hypothesize that this memory CD8+ T subset is recruited to inflamed sites, either in response to C3a or chemokines. These cells likely play a pivotal role in protective immune responses.
Genome-wide association study of inbred murine strains discovered nuclear factor of activated T-cells Nfatc4 as a major gene controlling severity of inflammatory arthritis. (47.14)
Authors: Ramez Mohamed , Forde Toni , Chow Chi-Wing , Adarichev Vyacheslav 
Published: January 2023

Abstract
Nuclear factor of activated T cells (NFAT) is a family of transcription factors critical in regulating gene expression in development and disease. NFAT is an important target for cyclosporine immunosuppression in transplant rejection. The NFAT pathway is also important in inflammatory and autoimmune diseases such as rheumatoid arthritis. Using genome-wide association study (GWAS) of antibody-mediated arthritis in inbred murine strains, we found NFATc4 within the major arthritis severity peak at 56.35-56.46 Mb of chromosome 14 with genetic association of log(1/p) &gt; 5.5. To confirm the genetic linkage and functional significance of the NFATc4 gene, we performed collagen antibody-induced arthritis (CAIA) and found that NFATc4-deficient mice exhibited significantly reduced inflammation as compared to wild-type counterparts (55% downregulation, p &lt; 0.01). Immunoblotting analysis demonstrated the presence of NFATc4 in articular cartilage and chondrocytes derived from femoral heads of arthritic and naïve mice. Notably, NFATc4 in chondrocytes isolated from arthritic mice was hypo-phosphorylated as compared to naïve cells, indicating activation of NFATc4. NFATc4, as expected, was not found in splenocytes. We also performed genome-wide Affymetrix oligonucleotide array studies in CAIA experimental arthritis model using wild type and NFATc4-deficient mice. Taking together, we uncovered a novel role of NFATc4 in inflammatory arthritis in chondrocytes.
The role of CTSH in narcolepsy susceptibility and autoimmunity (47.16)
Authors: Weiner-Lachmi Karin , Faraco Juliette , R Kornum Birgitte , Lin Ling , Steinman Lawrence , Mignot Emmanuel 
Published: January 2023

Abstract
Narcolepsy, a disease associated with HLA-DQB1*0602, is caused by the selective autoimmune destruction of 70,000 hypocretin neurons in the brain. Recently we identified Cathepsin-H as a novel Narcolepsy susceptibility locus. SNP rs3825932 was associated with Narcolepsy in our set of 807 Caucasian cases vs. 1074 controls (p= 0.002 OR 1.24). The same SNP marker, was previously reported to be associated Type 1 Diabetes in two studies. Cathepsin H is a member of the papain-like family, and participates in intracellular protein degradation. CTSH have also been suggested to be involved in apoptosis and neurodegeneration. CTSH expression levels in peripheral white blood cells are strongly correlated with CTSH genotype at rs1036938 (p&lt;0.0001). Presence of the disease associated C allele is correlated with a lower expression of the CTSH, in a dose dependent manner, in CD8 (T lymphocytes, NK) and CD4 (TH, regulatory, monocytes, macrophages, dendritic) cells, but not in other peripheral blood mononuclear cell types. The low expression variant is also associated with lower CTSH specific activity levels (p=0.029). The genetic association, and results from expression and activity studies strongly suggest a role for CTSH in the development of Narcolepsy. CTSH is implicated in at least two highly selective autoimmune diseases, therefore understanding the role of CTSH in Narcolepsy susceptibility may yield important insights into the development of that and other autoimmune diseases.
Altered IRF5 exon usage and increased expression of IRF7 targets in cells with an allele associated with risk for lupus (47.27)
Authors: Poole Brian , Clark Daniel , Henrie Brandon , Sloan Jason , Baumann Doug , Heap Casey , Bills Trieste , Greenhalgh Kathryn , Read Daniel , Bergsten Sean , Roznik Marinya 
Published: January 2023

Abstract
Background. The single nucleotide polymorphism rs2004640 within interferon regulatory factor 5 (IRF5) is a well established genetic risk locus for systemic lupus erythematosus (SLE). The overall effects on the cell, as well as the mechanisms through which this polymorphism enhances the risk for lupus remain elusive. Since an alternate first exon is created by this polymorphism, we hypothesized that the risk haplotype causes changes in splicing patterns. We further hypothesized that the risk allele may alter the expression levels of the transcriptional targets of either IRF5 or IRF7, which is modulated by IRF5. Methods. Peripheral blood mononuclear cells were isolated from lupus unaffected individuals after informed consent. DNA was extracted and genotyped for the allele at rs2004640. IRF 5 splice variant usage in EBV-transformed cells from nine race, sex, and age-matched pairs differing in rw2004640 genotype were compared using quantitative RT-PCR. Expression of genes regulated by IRF5 and IRF7 was also compared using quantitative RT-PCR. Statistical differences were evaluated using the Wilcoxon signed rank sum test. Results. Cells with the risk haplotype had more IRF5 mRNA (1.5-fold, p=0.05), but less exon 1C (1.9-fold, p=0.017) and less exon 1D (3.5-fold, p=0.012) than those with the protective type. Additionally, transcription of three targets of IRF7 was increased in risk cells, (average 1.5-fold increase, p&lt;0.05).
Children with autism spectrum disorders experiencing neuropsychiatric symptoms, cognitive skills, and gastrointestinal symptoms exhibit distinct innate immune abnormalities and transcriptional profiles in peripheral blood monocytes (47.7)
Authors: Jyonouchi Harumi , Geng Lee , Streck Deanna , Toruner Gokce 
Published: January 2023

Abstract
ASD is considered a behavioral syndrome associated with multiple genetic and environmental factors. This study focuses on a subset of ASD children characterized by fluctuating behavioral symptoms and cognitive skills, triggered by immune insults, in addition to chronic GI symptoms (ASD-immune subtype; ASD-IS). The study included ASD-IS (N=19), control ASD (N=29), and normal control (N=26) children. We evaluated production of pro-inflammatory and counter-regulatory cytokines by peripheral blood mononuclear cells (PBMCs) in response to agonists of toll like receptors (TLRs), polyclonal T cell stimulants, and recall antigens (Ags), and transcriptional profiles of PB monocytes. ASD-IS PBMCs produced less pro-inflammatory (IL-1β, IL-6, IL-12, and IL-23) and counter-regulatory (IL-10 and sTNFRII) cytokines with TLR agonists and less Th1 cytokines (IFN-γ and TNF-α) with recall Ags than control cells. ASD-IS PBMC also produced higher amounts of CCL2 than controls. Transcript profiles of ASD-IS monocytes were distinguished by altered mRNA expression (&gt;2x) in &gt;900 genes compared with either ASD or normal controls. Among these, 392 genes overlapped in the two control groups, including genes regulating TLR signaling, inflammatory responses, cell migration/adhesion, chemokines (CCL2 and CCL7), and genes associated with risks for autism, sychizophrenia, and depression. Our findings indicate a role of innate immunity in the puzzling clinical features observed in ASD-IS children.
Tumor-specific CD4+ T cells enhance the therapeutic efficacy of adoptive immunotherapy with CD8+ T cells in the lymphopenic setting and establish a long-term memory response (48.10)
Authors: Church Sarah , Jensen Shawn , Fox Bernard 
Published: January 2023

Abstract
TCR-gene-modified CD8 T cells are used for cancer treatment. We examined if tumor-specific CD4 T cells would enhance persistence and function of adoptively transferred CD8 T cells. RAG1-/- mice bearing 3-day B16-D5 melanoma pulmonary metastases received both naïve tumor-specific Trp-1 TCR CD4 T cells and effector gp100-specific TCR CD8 transgenic T cells or CD8 T cells alone. Significantly fewer mice had pulmonary metastases when treated with both CD4 and CD8 T cells compared to CD8 T cells alone at day 10 (3/20 vs. 14/14 p&lt;0.001) and 20 (7/20 vs. 13/14 p=0.001). CD4 T cells alone were ineffective. More effector CD8 T cells were observed in the blood of mice treated with both CD4 and CD8 T cells at day 20 (1.7-fold p&lt;0.001). These cells had a tumor-specific type 1 cytokine profile (IFNγ, TNFα, IL-12 and GM-CSF). CD4 and CD8 T cell treated short-term (50 days) and long-term (120 days) survivors rejected subsequent tumor challenge (0/5 and 0/15 respectively, p&lt;0.001). T cells in these long-term survivors maintained tumor-specific cytokine expression (IFNγ, Grz B and IL-2). Contrary to non-transgenic models, depletion of either CD4 or CD8 cells prior to challenge at day 120 did not reduce ability to reject tumor. While no mice grew tumor at the challenge site, 30% of mice developed amelanotic metastases. This tumor escape was associated with loss of T cell function. These data provide a strategy for augmenting the therapeutic efficacy of TCR gene therapy of patients with cancer.
Forced NF-κB in T cells leads to tumor rejection (48.24)
Authors: Evaristo Cesar , Gajewski Thomas , Alegre Maria-Luisa 
Published: January 2023

Abstract
NF-κB activity has been reported to be reduced in T cells from tumor-bearing hosts. Our previous results indicate that reduced NF-κB activation results in impaired T cell survival, decreased Th1 and Th17 differentiation and increased iTreg differentiation. Mice with reduced T cell-NF-κB activity fail to reject cardiac and pancreatic islet allografts in absence of any pharmacological treatment. We hypothesize that forced NF-κB activation in T cells may have the opposite effect and promote T cell survival, facilitate Th1/Th17 differentiation and prevent iTreg differentiation, which would be beneficial to reject tumors. We generated mice with constitutively active (CA) IKKβ in T cells (henceforth CA-IKKβ mice). Ectopic CA-IKKβ expression resulted in NF-κB phosphorylation. Transgene expression was limited to CD4, CD8 and NKT cells and T cells showed increased NF-κB activation and nuclear translocation. T cell numbers were comparable to littermate controls, but CA-IKKβ mice had fewer Tregs and increased frequency of activated T cells that produced IFNγ upon re-stimulation. When B16-SIY melanoma cells were injected subcutaneously, tumors grew progressively in control littermates, but were rejected by mice expressing CA-IKKβ in T cells. Furthermore, CA-IKKβ expressing CD4+ and CD8+ T cells were necessary and sufficient for tumor control. Our results demonstrate NF-κB to be at the cross-roads of major T cell fate decisions that should uniquely synergize for control of tumor growth.
Survivin TCR transgenic mice develop T cell lymphoblastic lymphoma (48.25)
Authors: Cui Yongzhi , Garber Haven , Mackall Crystal 
Published: January 2023

Abstract
Survivin has been considered a potential tumor antigen due to high expression in most cancers and limited expression in normal tissues. To explore the potential for survivin reactive TCRs to mediate antitumor effects in mice, we generated several founders of TCR transgenic mice with specificity for the H-2b restricted immunodominant epitope of survivin. In transgenic mice, survivin reactive T cells were predominantly CD8+ and mediated specific immune reactivity toward survivin peptide pulsed targets. Some antitumor reactivity was observed, but it was not potent, and the survivin reactive transgenic T cells were unable to mediate objective tumor regression of survivin bearing tumors in vivo. Spontaneous T cell acute lymphoblastic leukemia (T-ALL) was observed at 4-6 months of age in offspring of all 3 founders and was found in both survivin TCR+Rag+/- and survivin TCR+Rag-/- mice. Flow cytometry demonstrated that these lymphoblastic cells were CD3+ and survivin TCR+, and were variably CD8+ or CD4-/CD8-. Analysis of alpha gene rearrangements revealed oligoclonality but the cell were malignant, grew continuously in vitro and induced tumors in C57BL/6 recipients. The occurrence of T-ALL in 3 founders suggests that the transgene itself, rather than insertional mutagenesis, is causative. We postulate that self-reactive TCR serves as an oncogene if expressed early during thymopoiesis. TCR signaling may cooperate with NOTCH activation and/or other events to generate the malignancy.
Over-expression of uridine cytidine kinase like-1 protects tumor cells from NK-mediated lysis in vitro and in vivo (48.26)
Authors: Ambrose Elise , Gullickson Gail , Kornbluth Jacki 
Published: January 2023

Abstract
NK lytic-associated molecule (NKLAM) plays a critical role in NK-mediated tumor cytotoxicity. Our hypothesis is that NKLAM and other granule proteins are released from NK cells upon interaction with tumor targets. Acting as a ubiquitin ligase, NKLAM ubiquitinates uridine cytidine kinase like-1 protein (UCKL-1) in tumor cells, leading to its degradation. Downregulation of UCKL-1 by RNA interference was previously found to increase tumor susceptibility to NK killing. Here we show that UCKL-1 over-expression protects tumor cells from NK killing and from apoptosis mediated by chemotherapeutic agents. Gene array analyses and protein expression studies indicate an involvement of UCKL-1 with the NF-κB and death receptor (DR) 3 pathways. Studies were performed to evaluate the role of UCKL-1 in tumor survival in vivo. Wild type and NKLAM-deficient mice were injected intravenously with RMA-S tumor cells expressing various levels of UCKL-1. Results to date indicate that RMA-S cells expressing the highest levels of UCKL-1 have the greatest survival in wild type mice. Tumor survival is potentiated in NKLAM-deficient mice, suggesting that the NKLAM/UCKL-1 interaction contributes to eradication of tumor cells in vivo. These findings indicate an involvement of UCKL-1 in tumor cell survival. Findings suggest an important role for UCKL-1 and its interaction with NKLAM in cancer control mediated by NK cells.