bluuebunny/crossref_metadata_2025_split huggingface dataset:
PD-1 controls effectors but not the generation or function of natural or induced regulatory T-cells (P1011)
Authors: Ellestad Kristofor , Thangavelu Govindarajan , Ewen Catherine , Anderson Colin 
Published: January 2023

Abstract
During T cell activation, the balance of co-stimulatory and co-inhibitory interactions plays an important role in shaping the resulting T cell response. Co-inhibitory signals help maintain peripheral immune tolerance, and thus animals with defective co-inhibitory molecules, such as PD-1 KO mice, are predisposed to autoimmunity. Unlike PD-1 KO mice, which develop mild autoimmunity, adult Rag KO recipients of PD-1 KO hematopoietic stem cells (HSC) succumb to rapid and severe multi-organ autoimmune disease soon after T cells first emerge from the thymus, implicating PD-1 as a critical factor in the control of T cell self-reactivity during lymphopenia induced homeostatic proliferation (LIP). While we have found no reduction in the absolute number of FoxP3+ regulatory T cells (Treg) in diseased PD-1 KO vs. wild type (WT) HSC recipients, some studies have indirectly suggested that PD-1 deficiency may inhibit the development of induced Treg (iTreg) cells from conventional T cells in the periphery. We therefore investigated whether PD-1 deficiency results in an intrinsic inability of T cells to convert to iTreg in vivo, which could lead to disease in PD-1 KO HSC recipients. Surprisingly, our findings suggest that PD-1 deficiency does not inhibit the development of iTreg nor does it affect Treg suppressive function, and that in the context of LIP, co-inhibitor deficient effector T cells cause disease primarily by expanding beyond the ability of Treg to control them.
Strong systemic immune stimulation results in profound inhibition on the generation of subsequent primary responses (P3034)
Authors: Sckisel Gail , Bouchlaka Myriam , Mirsoian Annie , Monjazeb Arta , Murphy William 
Published: January 2023

Abstract
The immune system constantly maintains the intricate balance necessary to eliminate pathogens while preserving tolerance to self. We have previously shown that strong immune stimulation using agonistic αCD40/IL-2 immunotherapy (IT) results in the induction of robust, antigen-nonspecific CD8-mediated antitumor responses. In addition to proinflammatory reponses, a number of suppressive pathways become activated as well following IT. We sought to determine the ability of T cells to react to various stimuli immediately following strong immune stimulation. Studies revealed that while CD8 responses remained intact, CD4+ T cells exhibited markedly diminished ability to respond to TCR engagement including lack of CD25 upregulation and proliferation. Paralysis of the naïve CD4 population resulted in the inability to respond to primary antigen in vivo. Further analysis of the naïve CD4 population revealed concomitant upregulation of SOCS3, a negative regulator of JAK/STAT signaling, including STAT5. Consistent with this, STAT5 phosphorylation was diminished in CD4+ T cells restimulated following IT further suggesting a role for SOCS3 in the paralysis. These data have significant ramifications when attempting to apply IT in cancer. It also sheds light into the immune dysfunction documented during periods of strong immune stimulation such as acute infections or sepsis.
TLR-mediated modulation of micro-RNA profiling in human primary bronchial epithelial cells (P3003)
Authors: Vonakis Becky , Sirimalle Srinivas , Plunkett Beverly , Cheadle Christopher , Ceccarelli Michele , D’Angelo Fulvio , Cox Paul , Haddad El-Bdaoui , Stellato Cristiana 
Published: January 2023

Abstract
Global changes in miRNA expression in airway epithelium, a cell type critical in the mucosal response to microbial invasion, allergens and pollutants, have been only partially investigated. Cell stimulation with a TLR3 agonist ± cytokines was used to model disease-specific miRNA changes. PBEC (n=5) were cultured under air-liquid interface and treated with medium or polyI:C (1-10 μg/ml) ±TNFα/IFNγ (10 ng/ml each) for 24 hr. Total RNA was used for miRNA profiling (Agilent) and real-time-PCR analysis of inflammatory genes (control). PBEC supernatants were assayed using antibody arrays. Genome Ontology analysis was performed by Ingenuity pathway analysis. Expression of miR409-3p and miR624 was downregulated &gt;30-fold compared to control following polyI:C treatment. Expression of 16 miRNAs was downregulated following polyI:C/TNFα/IFNγ treatment, with &gt;30-fold reduction for miR617 and miR136 (calculated as Fold Change ≥ 1.5, p&lt; 0.05 after Robust Multichip Average normalization). In cells challenged with the combined treatment, 145 genes (including 16 transcription factors) were predicted with high confidence (by 6/6 programs) as molecular targets for 6 of 16 differentially expressed miRNAs. PolyI:C plus TNFα/IFNγ strongly synergized in upregulating the mRNA and protein expression of CXCL8, GM-CSF, IL-6 and CCL2. The identification of the miRNA signature may identify new mechanisms of posttranscriptional gene control and generate new anti-inflammatory strategies.
Cadmium suppresses IL-1β production and release in monocytes. (P3010)
Authors: Napolitano Jessica , Liu Mingjie , Bao Shengying , Knoell Daren 
Published: January 2023

Abstract
Tobacco smoke is the primary cause of chronic obstructive pulmonary disease (COPD) in the U.S. Immune dysfunction in the lung of chronic smokers increases the risk of respiratory infections. IL-1β is a vital cytokine in host defense and its expression is decreased in the lungs of these patients. Cadmium (Cd) is abundant in cigarette smoke and a major contributor to smoking-related lung disease. The purpose of this investigation is to determine whether Cd suppresses IL-1β in monocytes. THP-1 cells were differentiated into macrophages using PMA and then treated overnight with Cd, followed by combined LPS and ATP stimulation. Primary human monocytes were isolated from blood using CD14+ positive selection and subject to similar exposures. Analysis of THP-1 and monocyte cultures revealed that Cd suppresses IL-1β release following stimulation with LPS and ATP. Additionally, there was a decrease in IL-1β mRNA in Cd-treated LPS stimulated macrophages. We observed a decrease in phospho-p65 translocation into the nucleus of Cd-treated macrophages after LPS stimulation. Consistent with this, using a cell-free system, we observed that Cd inhibits IKKβ kinase activity (IC50 of ~100 nM). From these studies we conclude that Cd acts as an inhibitor of the canonical NF-κB pathway and inhibits IL-1β production. Accordingly, Cd may play a critical role in COPD pathogenesis by impairing monocyte immune function.
In vitro studies of interferon-alpha neurotoxicity (P3016)
Authors: Fritz-French Cari , Bassell Gary , Tyor William 
Published: January 2023

Abstract
Elevated levels of interferon-alpha (IFNα) in the central nervous system (CNS) are linked to cognitive dysfunction in patients with inflammatory CNS diseases such as HIV-associated dementia. Previous studies showed that IFNα treatment of neuronal cultures caused a dose dependent decrease in dendritic branching and length, that was prevented after pre-treatment with IFNα neutralizing antibodies. Antagonists to NDMA receptor were also found to be partially protective against IFNα induced neurotoxicity. To begin to determine the mechanism of IFNα induced neurotoxicity, the cell signaling pathway involving IFNα receptor (IFNAR) was evaluated. We demonstrate increased interferon stimulating gene 15 (ISG15) expression, an indicator for JAK-STAT cascade activation, after IFNα stimulation. Inhibiting IFNAR was found to partially protect neurons from IFNα neurotoxicity, but not as efficiently as neutralizing antibodies to IFNα. Our preliminary studies suggest that IFNα is in part acting through its receptor as well as NMDAR to reduce dendritic arborization in neurons. Further studies are needed to determine what other signaling pathways may be involved in IFNα induced neurotoxicity. Determining the mechanism of IFNα neurotoxicity could lead to therapies for cognitive dysfunction during neuroinflammation.
Nucleotide-binding and oligomerization domain 2 (NOD2) knock-in mice carrying a mutation associated with Blau syndrome show reduced amounts of NOD2 protein and decreased muramyl dipeptide (MDP)-induced inflammatory responses (P1254)
Authors: Dugan Jae , Griffiths Eric , Snow Paige , Rosenzweig Holly , Carr Daniel , Rosenbaum Jim , Davey Michael 
Published: January 2023

Abstract
Blau syndrome is an autosomal dominant disorder caused by mutations in NOD2 and characterized by arthritis, dermatitis and uveitis. NOD2 binds MDP and activates NF-kB and MAPK signaling cascades. Prior in vitro studies reported that NOD2 containing Blau mutations caused enhanced activation of NF-kB, suggesting a gain of function in mutated NOD2 caused Blau syndrome. We tested this hypothesis in vivo by creating a knock-in mouse where a point mutation resulted in a change of arginine [R] to glutamine [Q] at position 314 (R314Q) of NOD2 (position 314 in mice corresponds to 334 in humans). R314Q heterozygous (+/m) and homozygous (m/m) mice did not spontaneously develop arthritis or dermatitis. Bone marrow derived macrophages (BMDM) from R314Q mice showed a reduction in NOD2 protein levels compared to wild type (WT) mice despite comparable amounts of NOD2 mRNA. MDP treatment of BMDM showed reduced activation of NF-kB and p38 MAPK in +/m and m/m compared to WT mice that correlated with the copy number of mutated NOD2, with the greatest reduction in m/m mice. In response to ip MDP, reduced levels of IL-6 and KC were detected in the serum of +/m and m/m mice, also in manner correlating with the copy number of the mutation. These data indicate that R314Q-NOD2 mice do not demonstrate a gain of function of the NOD2 pathway. Rather, R314Q causes a deficiency of NOD2 and raises the possibility that Blau syndrome may fall within the spectrum of an immunodeficiency disease.
ATPase-driven oligomerization of RIG-I on an RNA ligand allows optimal induction of type I interferon (P1260)
Authors: Patel Jenish , Jain Ankur , Chou Yi-ying , Baum Alina , Ha Taekjip , Garcia-Sastre Adolfo 
Published: January 2023

Abstract
RNA with an exposed 5’-triphosphate (5’-ppp) and terminal double-stranded structures as found in cells during infection with many viruses, constitutes a strong activating ligand for the cytosolic pathogen sensor RIG-I. RNA binding and subsequent ATP hydrolysis by RIG-I are believed to be critical for downstream signaling. During its ligand-mediated activation, RIG-I has been suggested to form multimers but there is insufficient evidence on whether RIG-I is able to form higher-order oligomers on RNA and how multimerization affects signaling. In addition, RIG-I has been shown to translocate on 5’-ppp-dsRNA in an ATP-dependent manner and a precise role for the ATPase and associated RNA translocase activities in RIG-I activation has not been established. Using biochemical analyses of native RIG-I-RNA complexes, we show that RIG-I oligomerizes on this 5’-ppp dsRNA in an ATP hydrolysis-dependent manner. Importantly, there is a drastic reduction in ATP-dependent oligomerization of RIG-I on DI RNA with shorter dsRNA stems, correlating with the level of IFN-I activation by these RNAs. These results demonstrate a clear role for RIG-I ATPase in dsRNA length-dependent oligomer formation and provide a rationale for the ATP-driven translocase activity of RIG-I in stimulation of the IFN response.
Transcription factor ELF4 promotes formation of CD62LLOW effector and memory CD8+ T cells in response to infection (P1422)
Authors: Mamonkin Maksim , Puppi Monica , Lacorazza Daniel 
Published: January 2023

Abstract
At the peak of response majority of pathogen-specific CD8+ T cells are short-lived CD62Llow effectors which eventually die off, revealing CD62Lhigh central and CD62Llow effector memory populations. Intrinsic regulation of CD8+ T cell differentiation to effector and memory lineages still remains poorly understood. We have previously demonstrated that the transcription factor ELF4 regulates homeostatic and antigen-induced proliferation of CD8+ T cells. In this work, we show that ELF4 plays a critical and specific role in the generation of CD62Llow effector and memory CD8+ T cells following Listeria monocytogenes infection. Adoptively transferred Elf4—/— CD8+ T cells displayed impaired formation of CD62Llow effectors, lower magnitude of expansion and reduced numbers of effector memory without affecting development of central memory. Consistent with this finding, we observed decreased levels of Elf4—/— effector and memory CD8+ T cells in non-lymphoid tissues and blood but not in lymph nodes or bone marrow. Impaired expansion and formation of memory was due in part to enhanced cell death of Elf4—/— effector cells and suboptimal priming, evidenced by lower numbers of IL-2Rα positive cells and reduced proliferation 2.5 days after infection. Finally, loss of ELF4 compromised generation of secondary effector cells from central memory in a recall response. Thus, ELF4 emerges as a novel transcriptional regulator of CD8+ T cell differentiation in response to infection.
CD8 T cell metabolism and differentiation in an acute immune response (P1440)
Authors: Verbist Katherine , Wang Ruoning , Green Douglas 
Published: January 2023

Abstract
A popular concept in CD8 T cell biology is that manipulation of T cell metabolism represents a method by which CD8 T cell differentiation, and thus the size and quality of the resultant memory population, is controlled. Indeed, it has long been known that different metabolic programs (i.e. utilization of distinct metabolic pathways to meet the cell’s energetic demands) are associated with different phases of CD8 T cell differentiation during an immune response. Specifically, T cell activation via TCR stimulation results in dramatic increases in the uptake and consumption of glucose and glutamine by effector T cells, whereas naïve and memory CD8 T cells rely primarily on fatty acid oxidation to meet energetic and biosynthetic demands. Interestingly, it has been shown that the initial metabolic programming upon activation is dependent on expression of the transcription factor c-Myc. Here, we utilize c-Myc reporter mice and conditional c-Myc knockout animals to examine the metabolic re-programming that occurs as CD8 T cells transition from the effector to memory phase, and we explore the possibility that c-Myc expression can predict the metabolic profile and eventual fate of CD8 T cells participating in an acute immune response.
Mucosal infection drives rapid development of protective resident memory CD8 T cells (P1444)
Authors: Sheridan Brian , Pham Quynh-Mai , Lee Young-Tae , Cauley Linda , Lefrancois Leo 
Published: January 2023

Abstract
The intestinal mucosa contains unique sites designed for promoting specialized T cell responses. Oral infection with a murinized Listeria monocytogenes (Lm) induced a robust mucosal CD8 T cell response which rapidly converted to a memory population during the peak of the intestinal T cell response. Concomitant with this event, the mice were nearly completely protected against re-infection. Blocking migration with anti-α4β7 from 7 dpi prevented continued migration into the intestinal compartment but did not alter the rapid conversion of early migrating cells into memory suggesting in situ events regulated early memory formation. These early memory cells, but not other effector subsets, preferentially upregulated CD103 identifying them as precursors to mucosal resident memory CD8 T cells. Surprisingly, CD103 expression on infiltrating CD8 T cells was required for early T cell entry into the epithelium but not for long term retention. Moreover, CD103 upregulation on Lm-specific CD8 T cells within the epithelium was predominately independent of TGFβ signals. Thus, the generation of distinct CD8 T cell subsets uniquely tailored to respond to intestinal pathogens may be a hallmark of oral challenge. Indeed, CD8 T cells that infiltrate the intestinal mucosa following intranasal influenza virus infection failed to generate MPECs and were not maintained long-term, demonstrating the importance of intestinal priming for generating robust mucosal memory CD8 T cells.
Temporal dynamics of the primary human T cell response to yellow fever virus 17D as it matures from an effector- to a memory-type response (P1406)
Authors: Blom Kim , Braun Monika , Ivarsson Martin , Gonzalez Veronica , Falconer Karolin , Moll Markus , Ljunggren Hans-Gustaf , Michaelsson Jakob , Sandberg Johan 
Published: January 2023

Abstract
The live attenuated yellow fever virus 17D vaccine provides a well-controlled model to study T cell responses to an acute viral infection in humans. Here, we have studied the temporal dynamics, composition and character of the primary human T cell response to YFV. The acute YFV-specific effector CD8 T cell response was broad and complex, composed both of dominant responses that persisted into the memory population, as well as of transient subdominant responses that were not detected at the memory stage. Furthermore, HLA-A2- and HLA-B7-restricted YFV epitope-specific effector cells predominantly displayed a CD45RA-CCR7-PD- 1+CD27high phenotype, which transitioned into a CD45RA+CCR7-PD-1-CD27low memory population phenotype. The functional profile of the YFV-specific CD8 T cell response changed in composition as it matured from an effector- to a memory-type response, and tended to become less polyfunctional in the course of this transition. Interestingly, activation of CD4 T cells as well as FoxP3+ T regulatory cells in response to YFV vaccination preceded the kinetics of the CD8 T cell response. The present results contribute to our understanding of how immunodominance patterns develop, as well as the phenotypic and functional characteristics of the primary human T cell response to a viral infection as it evolves and matures into memory.
A GATA-3 function distinct from Th2 determination controls basic T cell function (P1410)
Authors: Wan Yisong , Wang Yunqi , Gu Aidi , Curitis Thomas 
Published: January 2023

Abstract
T cells are pivotal to controlling the immune response to fight infection, eradicate tumor and elicit autoimmunity. CD8 T cells are the “cytotoxic” T cells to mediate the killing of target cells. And CD4 T cells differentiate into “helper” T (Th) cell subsets to modulate immune function. To achieve desired T cell response, series of events including cell activation, expansion and differentiation will have to be accomplished following antigenic and mitogenic stimulation. Therefore, identifying the factors critical for T cell activation, expansion and differentiation becomes the central question for T cell immunology. GATA-3 is a master regulator for Th2 differentiation. It is however poorly understood if GATA-3 controls mature T cell function beyond Th2 determination. We found that TCR-signal and cytokine-stimulation promoted GATA-3 expression in CD8 T cells with enhanced proliferation but not Th2-like differentiation. In addition, while GATA-3-deficient CD8 T cells developed normally, their peripheral maintenance was impaired. More strikingly, TCR- and cytokine-promoted expansion of CD8 and CD4 T cells was abrogated in the absence of GATA-3. This study thus reveals an unprecedented GATA-3 function central to basic T cell functions beyond Th2 differentiation.
Notch signalling shapes the CD8 T cell response following Listeria infection (P1416)
Authors: Mathieu Mélissa , Duval Frederic , Labrecque Nathalie 
Published: January 2023

Abstract
Following an infection, naïve CD8 T cells expand and differentiate into effectors able to eliminate the pathogen. At the peak of the response, two populations of effectors are distinguishable: short-lived effector cells (SLECs) meant to die by apoptosis and memory precursor effector cells (MPECs) destined to survive as memory cells that will confer long-term protection. Thus, following activation, the CD8 T cell faces a binary cell fate decision. We postulate that the Notch signalling pathway, known for its role in cellular differentiation and binary cell fate choice, acts as a key player in the MPEC/SLEC choice. To elucidate the role of Notch signalling in CD8 T cell response, we infected mice lacking or not expression of Notch1 and Notch2 in mature CD8 T cells with Listeria monocytogenes expressing OVA. Notch deficiency led to the generation of more OVA-specific effector CD8 T cells but less of these effectors had a SLEC phenotype (CD127loKLRG1hi) at the peak of the response. Surprisingly, Notch did not impair CD8 T cell memory generation following infection. Thus, Notch signalling influences the expansion and the acquisition of a SLEC phenotype following Listeria infection but not the generation of memory cells. Understanding the molecular pathways leading to memory generation is crucial as this knowledge will ultimately contribute to new vaccine development.
Human anti-HIV-1 envelope monoclonal antibodies with a low percentage of mutations have the ability to neutralize sensitive Tier 1 viruses. (P6189)
Authors: Gorny Miroslaw , Li Liuzhe , Wang Xiao-Hong , Williams Constance , Volsky Barbara , Seaman Michael , Zolla-Pazner Susan 
Published: January 2023

Abstract
Antibodies (Abs) specific to the HIV-1 envelope regions, the CD4 binding site (CD4bs), V3 and V2 loops display neutralizing activity and may have protective functions. These Abs are present in almost all HIV-infected subjects and can also be induced by vaccines, as shown in clinical trials, but exhibit rather low neutralizing activities. To determine a threshold of mutations which may represent the maturity needed for cross-neutralizing antibodies, we have studied the percentage of mutations in the variable fragment of the heavy and light chain genes of 63 human monoclonal Abs (mAbs) which are specific to CD4bs, V3 and V2 domains, and tested their neutralizing activities against a panel of 41 pseudoviruses (psVs). All 63 mAbs neutralized, with various frequencies, a panel of sensitive Tier 1 psVs while the more resistant Tier 2 psVs were neutralized sporadically and only by V3 mAbs. Mutation frequencies ranged from 2.4% to 16.7% of the nucleotide sequences were observed for both the heavy and light chain genes of all mAbs. The percentage of mutations was comparable for CD4bs, V3 and V2 mAbs and had no correlation with the breadth of neutralization. The results indicate that mAbs with at least a few percent of mutations can neutralize sensitive Tier 1 viruses, which probably can be achieved during HIV vaccination.