bluuebunny/crossref_metadata_2025_split huggingface dataset:
CD73 protects Treg and ILC in the control of Th cell mediated colitis (IRC5P.632)
Authors: Drygiannakis Ioannis , Kurtz Courtney , Klann Jane , Farrow Norma , Thai Rosarlin , Gomez Bobbie , Wilson Jeffrey , Borowitz Molly , Bekiaris Vasileios , Ware Carl , Ernst Peter 
Published: January 2023

Abstract
CD73 is a membrane-bound enzyme catalyzing the final step of the conversion of extracellular ATP to adenosine, which then binds to its receptors and signals via cAMP and non-cAMP-mediated pathways. While CD45RBlow helper T cells from wild-type and CD73 KO mice were both highly enriched in the FoxP3+ regulatory T cell transcription factor, cells from CD73 KO mice produced more IFN-γ and IL-17A and could not inhibit effector T cell proliferation and function in vitro. In vivo CD73 expression by both regulatory (CD45RBlow) and effector (CD45RBhigh) T cells was required for Treg to prevent effector T cell-induced colitis following adoptive transfer into immunodeficient Rag1 KO recipients. The cytokine profile in the intestine was skewed compared to the protective combination of wild-type CD45RBlow and wild-type CD45RBhigh. Donor CD45RBlow recovered from colitic mice had reduced FoxP3 and CD45RBhigh phenotype shifted from Th17 to Th1. If recipients lacked both Rag1 and CD73, wild-type CD45RBlow Th cells did not protect from T cell-induced colitis. CD45RBlow Th cells lost FoxP3, acquired Th1 and Th17 transcription factors, depleted recipient’s innate lymphoid cells and caused disease. Prior adoptive transfer of innate lymphoid cells protected these recipients from disease and maintained FoxP3 expression on cells of the CD45RBlow pool. Thus, CD73 on cells of both adaptive and innate immunity is required for effective regulatory T cell function in intestinal homeostasis.
Fibroblastic reticular cell subsets derived from human lymph node resembles bone marrow stromal cells and interfere with the expression of homing markers in autogous and allogeneic lymphocytes (IRC5P.624)
Authors: Marti Luciana , Alvarenga Heliene , Macedo Antonio , Andrade Maria Claudina , Cury Adriano , Salvalaggio Paolo , Pasqualin Denise , Bromberg Silvio , Rizzo Luiz , Hamaerschlak Nelson 
Published: January 2023

Abstract
The main functions of human bone marrow stromal cell are related to bone marrow homeostasis, even though immune functions already have been described to these cells. A similar cell type named fibroblastic reticular cell was described in animal models secondary lymphoid organs, which the main function was to favor and regulate the immune responses. Therefore, we have obtained informed consent from patients that undergone to organ removal surgery and requested a lymph node donation (Ethics committee approval number - 06592712.4.0000.0071). After treating the lymph node tissue, we were able to isolate stromal cells from several lymph nodes. These cells were cultured for 4 passages, than characterized; we observed that there were two subsets of stromal cells in lymph nodes, one that express gp38 (40%) and meet the already described fibroblastic reticular cell phenotype and another gp38 negative (60%). Either cell subset expressed CD29, CD44, CD73, CD90 or CD105 and do not expressed CD35, CD45, CD19, HLA-DR, CD14, CD34, CD31 or CD106. Both subsets were able to differentiate in adipocytes, chondrocytes and osteoblast, resembling human mesenchymal stem cell bone marrow derived. Regarding their immune function, we observed that either subset was able to interfere with proliferation and CCR6, CXCR4 expression of stimulated allogeneic and autologous T cells.
LPS inhibits ILC2 mediated type 2 immunity in the lung (IRC5P.625)
Authors: Poon Grace , Takei Fumio 
Published: January 2023

Abstract
The hygiene hypothesis states that the lack of early childhood exposure to infectious agents, which promotes type 1 immunity, results in increased incidents of allergy and asthma, which are predominantly type 2 immune responses. Recently identified group 2 innate lymphoid cells (ILC2s) are essential for the development of protease allergen induced type 2 lung inflammation. To better understand how type 2 immunity is regulated, we tested the effect of LPS-induced type 1 inflammation on ILC2s and type 2 lung inflammation. Intranasal LPS pretreatment of mice suppressed both papain- and IL-33-induced ILC2 activation and eosinophilic lung inflammation. Specifically, the number of activated ILC2s that express IL-5 and IL-13 was decreased with LPS treatment. The LPS effect on ILC2s was dose and time dependent. The in vivo findings were reproduced in vitro. Pretreatment with LPS inhibited the production of type 2 cytokines IL-5 and IL-13 by ILC2s cultured in the presence of IL-33 and TSLP. These results indicate that LPS triggers an ILC2 inhibitory signal, which leads to the suppression of ILC2 activation and type 2 inflammation in the lung upon allergen exposure.
Spontaneous aggregation and novel signalosome organization of BENTA-associated gain-of-function CARD11 mutants in lymphocytes. (IRM5P.641)
Authors: Stinson Jeffrey 
Published: January 2023

Abstract
We recently described BENTA (B cell Expansion with NF-kB and T cell Anergy), a novel lymphoproliferative disorder caused by germline, gain-of-function mutations in the lymphocyte-specific scaffolding protein CARD11. Normally, antigen receptor (AgR) stimulation induces CARD11 phosphorylation to evoke an active conformation that nucleates a signalosome incorporating BCL10, MALT1, and other proteins required for IkB kinase (IKK) activation and NF-κB nuclear translocation. However, ectopic expression of CARD11 mutants triggers constitutive NF-kB activity in B and T cell lines without AgR stimulation. Here we show that these CARD11 mutants form large, cytosolic protein aggregates with a novel substructure, dubbed mCADS (Mutant CARD11 Dependent Shells). Endogenous mCADS are also detected in B cell lymphomas harboring similar CARD11 mutations. In contrast, WT CARD11 cannot form mCADS -/+ AgR stimulation. Only MALT1 and phospho-IKK are stably colocalized with mCADS, indicative of active, sustained signaling. Surprisingly, mCADS formation requires interaction with BCL10 but not NEMO (IKKg). Our results suggest that gain-of-function CARD11 mutants spontaneously assemble into an active, highly-ordered signalosome that is structurally and functionally distinct from WT CARD11, regardless of AgR ligation. The unique structure and composition of mCADS may represent a novel therapeutic target for treating BENTA or lymphoma without disrupting normal AgR-induced NF-kB activation.
The nucleolus contains immune-related RNAs during infection. (IRM5P.644)
Authors: LEE TAEYUN , Lee Sungwook , Park Boyoun 
Published: January 2023

Abstract
The nucleolus is a subnuclear structure that is involved in ribosomal RNA (rRNA) synthesis and ribosome assembly, and it is also the location of viral replication or cellular stress sensing. However, the function of the nucleolus in the immune response is still unclear. Here, we report that the nucleoli are fused in both macrophages and dendritic cells during lipopolysaccharide (LPS) or CpG-DNA stimulation, which activates the Toll-like receptor 4 (TLR4) or Toll-like receptor 9 (TLR9) signaling pathway. Our results from RNA-sequencing analysis reveal that the nucleolus contains diverse RNA transcripts in response to LPS. Furthermore, gene ontology term analysis shows that these RNA species are highly correlated with immune-related functions. More interestingly, intron-containing RNA transcripts are existed in the nucleolus. Our results provide new insight into another mechanism of nucleolus in the immune system.
Regulation of CD4+ T cell activation and effector function by the histone acetyltransferase Myst1 (IRM6P.662)
Authors: Carson William , Mertz David , Cavassani Karen , Mukherjee Sumanta , Harissa Batoul , Kroetz Danielle , Schaller Matthew , Dou Yali , Kunkel Steven 
Published: January 2023

Abstract
Chromatin remodeling events are central molecular mechanisms that serve to govern CD4+ T cell function through regulation of gene accessibility. Myst1 is a histone acetyltransferase responsible for the acetylation of the protein tail of histone 4 at lysine residue 16, and is thought to participate in the activation of gene transcription. Studies of murine and human CD4+ T cells indicate that Myst1 is upregulated upon T cell receptor stimulus. Deletion of the gene encoding Myst1 resulted in a significant proliferation defect in these cells that is unaffected by the addition of exogenous IL-2. Myst1-deficient T cells also exhibit modulations in cytokine production and lineage commitment following skewing to various effector lineages in vitro. Additionally, signal transduction via STAT5 may play a role in Myst1 expression, as downregulation of STAT5a/b expression via siRNA transfection significantly decreased Myst1 expression levels in T cells. These results suggest an important role for the STAT5-Myst1 axis in governing the activation and effector function of CD4+ T cells.
Characterization of human immune phenotypes using a novel transcriptome analysis approach (IRM6P.656)
Authors: Scheid Adam , Van Keulen Virginia , Felts Sara , Zhang Yuji , Markovic Svetomir , Pease Larry 
Published: January 2023

Abstract
To test the hypothesis that complex immune phenotypes can be defined by stable gene expression profiles we are using a RNAseq-based transcriptome analysis to relate phenotypes of activated peripheral blood CD4+ cells from healthy subjects and stage IV melanoma patients to their freshly-isolated counterparts. We have defined molecular immune response signatures (MIRS) containing genes that consistently differentiate individuals from one another. Two different phenotypic groups are defined using this approach, both of which are mathematically distinct from randomly-selected groups. First, a MIRS generated using healthy donor T cells stimulated with α-CD3/CD28 beads defines a distinct response phenotype and successfully predicts 17 independent healthy subject and melanoma patient responses, demonstrating the predictive capability of the approach. Second, our comparison of melanoma patient signatures with those of healthy subjects revealed a group of healthy individuals with distinct expression signatures that did not cluster with those of any of the melanoma patients. These results indicate that our RNAseq-based transcriptome analysis has the potential to identify phenotypic groups as well as gene sets that have response-predictive capabilities. Our work has both discovery and translational implications, as our approach can be used to investigate complex genetic contributions to immune responses and predict how individual patients will respond to therapeutic interventions.
Differential long noncoding RNA signatures in activated plasmacytoid dendritic cells (IRM6P.659)
Authors: Joslyn Rochelle , Savan Ram 
Published: January 2023

Abstract
Long noncoding RNAs (lncRNAs) represent a novel class of RNA which act through diverse mechanisms to exert control over a wide range of cellular processes. Through control of transcription, post-transcriptional regulation, or cellular signaling events, lncRNAs can function as potent modulators of innate immune responses, however very few lncRNAs have been investigated. We utilized whole exome sequencing of plasmacytoid dendritic cells (pDC) treated with IFN (interferon)-β, TLR7 agonist (toll like receptor, R848), and R848 with type I IFN blockade to identify 910 noncoding transcripts which are differentially regulated by IFN signaling and TLR7 activation. Through dissection of the stimulatory events controlling lncRNA expression, we have identified subsets of type I IFN- and TLR7-induced or suppressed lncRNAs. We hypothesize that many of these transcripts participate in the orchestration of innate and adaptive antiviral immunity through their modulation of pDC function. Dysregulation of these lncRNAs could significantly alter pDC maturation, IFN production, antigen presentation, turnover or localization, events which would be consequential during viral infection or IFN-driven autoimmune disease.
The intrinsic and extrinsic regulation of γδ-T17 cells through the IL-17-IL-17r axis during homeostasis (LYM2P.729)
Authors: Fleming Christopher , Cai Yihua , Xue Feng , Wei Yu-Ling , Jala Venkatakrishna , Worth Christopher , Liu Na , Chien Yueh-Hsiu , Ding Chuanlin , Yan Jun 
Published: January 2023

Abstract
IL-17 producing γδ T cells (γδ-T17) are major players in promoting several autoimmune inflammatory diseases and cancers. However, γδ-T17 homeostatic regulation has not been fully understood. In naïve, adult IL-17r-/- and IL-17A-/-/F-/- mice we observed higher frequencies of total γδ T cells, predominantly γδ-T17, in the lung, spleen, gut, LN and skin but not in the thymus. We hypothesized that the IL-17-IL-17r axis regulates peripheral γδ-T17 homeostasis. Two different regulatory pathways were identified to be involved in this regulation: extrinsically through microbiota-host interaction and intrinsically through IL-17r-IL-7r signaling interaction. The extrinsic pathway was discovered in vitro using CFSE-labeled IL-17r-/- lymphocytes that had endogenous γδ-T17 proliferation dependent upon DCs, particular CD103+ DCs, and IL-1β. Using mice delivered from IL-17r-/- pregnant females treated either with lymphotoxin beta receptor-Ig to remove LNs or antibiotics (ampicillin, vancomycin, neomycin, and metronidazole) to deplete gut microbiota we found significantly decreased frequency of γδ-T17 in peripheral tissues. Also, intrinsically IL-17r-/- γδ-T17 expressed higher levels of RORγt that suppressed BTLA expression, which is a known inhibitor of IL-7-driven γδ-T17 proliferation. Taken together, our findings suggest that the IL-17-IL-17r axis regulates γδ-T17 via both intrinsic and extrinsic pathways.
The role of miRNAs in the homeostasis of peripheral T cells (LYM2P.732)
Authors: Pobezinsky Leonid 
Published: January 2023

Abstract
The lifelong presence of mature T cells in the organism is vital for immune responses. Despite being extensively studied, the molecular mechanisms governing T cell homeostasis are still not fully understood. Here we investigated how microRNAs, acting through post-transcriptional regulation, contribute to the maintenance of mature naïve T cells in the periphery. We found that T cell-specific Dicer ablation caused lymphopenia, due to survival defects in mature T cells. Using miRNA knockout mice, we identified specific miRNAs which, when deleted, resulted in T cell death, and overexpressing these miRNAs in the T cells of transgenic mice provided complete protection. Finally, we characterized the downstream molecular mechanisms responsible for miRNA-induced suppression of cell death. Altogether our findings represent a novel miRNA-mediated mechanism that controls homeostasis of peripheral naïve T cells in vivo.
GATA-3 regulates the development of ILC3 subsets at multiple stages (LYM2P.721)
Authors: Zhong Chao , Hu Gangqing , Zhao Keji 
Published: January 2023

Abstract
Innate lymphoid cells (ILCs) are emerging lymphocytes that maintain the homeostasis between the host and commensals, and play essential roles during secondary lymphoid organogenesis and innate immune responses. Mature ILCs, similar to their adaptive counterparts of CD4 T cells, can be divided into three major subsets based on their signature cytokine production and selective expression of master transcriptional regulators, T-bet, GATA-3 and RORγt. However, how the development and functions of these subsets are precisely regulated, especially by transcription factors including GATA-3 that are expressed at various levels, is still elusive. We have previously reported that in addition to its critical role in the development and maintenance of ILC2s, GATA-3 is also required for the development of other IL-7Rα+ ILCs including ILC3s. Here we show that GATA-3 is continuously expressed by all ILC3 subsets and has crucial regulatory functions at multiple stages during the development of ILC3 subsets. Deletion of Gata3 gene after RORγt expression by Rorc-Cre resulted in a significant reduction of ILC3s, especially for the NKR+ ILC3 subset. Genome wide analysis of gene expression in the wild type and Gata3-deficient ILC3s revealed that the expression of Il7r and a number of genes associated with the functions of ILC3s was dramatically reduced in the absence of GATA-3. Thus, our results shed light on the regulatory function of GATA-3 at multiple developmental stages of ILC3 subsets.
Defining the mechanisms and cues that influence cytotoxic T cell homeostasis during malnourishment (LYM2P.727)
Authors: Gubbels Bupp Melanie , Scott Samantha , Murphy Sarah , Edwards Whitney , Litvin Seth 
Published: January 2023

Abstract
T cell homeostasis is clearly disrupted by malnutrition, but the precise cues and mechanisms through which it occurs are unknown. Short-term malnutrition reduces the size of the peripheral CD8+ T population in mice. Perhaps paradoxically, malnourished CD8+ T cells are less sensitive to death-by-neglect and express higher levels of CD127, a component of the IL-7 receptor. We aimed to identify whether vitamin and mineral deficiency or reduced caloric intake triggers these changes and to better understand the molecular mechanisms through which such changes occur. While vitamin and mineral deficiency is not sufficient to reduce total CD8+ T cell numbers or trigger CD127 up-regulation, it is sufficient to reduce CD8+ T cell sensitivity to death-by-neglect. Mechanistically, we have determined that the reduced sensitivity to death-by-neglect observed in malnourished CD8+ T cells is Foxo1-dependent. Given that CD127 is a transcriptional target of Foxo1, additional studies were conducted to determine if CD127 is up-regulated or if CD127low CD8+ T cells are simply out-competed in a malnourished environment. Our results suggest that during malnourishment, Foxo1 induces the up-regulation of CD127 on CD8+ T cells, which may relate to the cells’ reduced sensitivity to growth factor withdrawal. While different mechanisms may be triggered by calorie versus vitamin and mineral deficiency, both deficiencies reduce CD8+ T cell sensitivity to death-by-neglect.
Decreasing initial stimulation increases generation of early effector cells that survive the contraction phase in malaria infection (MPF2P.754)
Authors: Stephens Robin , Opata Michael , Carpio Victor , Ibitokou Samad , Dillon Brian , Obiero Joshua 
Published: January 2023

Abstract
CD4 T cells orchestrate immunity against blood-stage malaria. However, a major challenge in designing malaria vaccines is understanding generation of memory from effector T cells (Teff) in chronic infection. We use a transgenic mouse model with T cells specific for the Merozoite Surface Protein (MSP)-1 of Plasmodium chabaudi to show that activated T cells generate three Teff (CD127-) subsets representing progressive activation. The earliest observed subset (CD62LhiCD27+) divides first, and maintains memory gene expression. An intermediate (CD62LloCD27+) effector subset has more multi-cytokine producing T cells and a terminal phenotype, including expression of PD-1 and Fas. Fully activated (CD62LloCD27-) late effector cells contain fewer cytokine+ and more AnnexinVhi apoptotic cells. Using an adoptive transfer system, we show the ability of CD62Lhi Teff to generate the terminal Teff subsets but also to survive the contraction phase better than the CD62Llo subsets. Furthermore, we show that progression towards terminal differentiation increases with duration of infection, but is inhibited by IL-2, which promotes expansion but not progression. Therefore, we have defined markers of progressive activation of CD4 Teff at the peak of malaria infection, and shown that CD127-CD62Lhi Teff survive the contraction phase. Therefore, antigen and cytokine levels during CD4 T cell expansion influence the proportion of activated cells that can survive T cell contraction in malaria infection.
Schistosoma Mansoni infection induces anti-atherogenic transcriptional changes in hepatic macrophages (MPF2P.758)
Authors: Fairfax Keke , Elvington Andrew , Pearce Edward , Randolph Gwendalyn 
Published: January 2023

Abstract
Hepatic macrophages play an essential role in the granulomatous response to infection with the parasitic helminth Schistosoma Mansoni, but the transcriptional changes that underlie this participation are poorly understood. To explore this, we sorted the two previously recognized hepatic macrophage populations (perivascular and Kupffer) from naïve and S. mansoni infected mice and performed microarray analysis as part of the Immunological Genome Project. Consistent with the pattern of great diversity identified in other organ macrophages, the two hepatic macrophage populations displayed signatures distinct from all other macrophages, with the two populations exhibiting remarkable differences between them. However, this diversity was greatly reduced following infection with S. mansoni, and in fact, many of the transcripts identified as uniquely perivascular or Kupffer cell specific were lost following infection, raising the possibility that both populations may be replenished by monocytes following infection. Our analysis showed a profound alteration in phospholipid and cholesterol metabolic pathways, including prostaglandin signaling, in addition to the expected M2 markers. These changes suggested a possible mechanism for the previously reported atheroprotective effects of S. mansoni infection. Indeed we find that ApoE null mice fed a high fat diet in combination with S. mansoni infection have reduced body mass in addition to reduced plaque area as compared to control mice.
Hepatic recruitment of the inflammatory cell subsets upon liver injury duo to Echinococcus multilocularis (MPF2P.761)
Authors: Zhang Chuanshan , Wen Hao , Lin Renyong 
Published: January 2023

Abstract
Alveolar echinococcosis (AE) is a chronic disease characterized by slowly expanding liver lesions.In addition to liver-resident Kuffer cells, infiltrating immune cells have recently been linked to the development of liver injury.The role of these inflammatory cell subsets and functional contributions of their differential recruitment into the injured liver are not defined.Here we studied in vivo the phenotype and function of infiltrating immune cells around the lesion in AE patients and mice model by means of immunohistochemistry and flow cytometry.AE patients with parasitic lesions displayed a significant increase of activation of predominantly CD163+ kuffer cells and CD14+ monocyte cells around lesion compared to the diatal liver tissue.Infected mice showed that inflammatory CD11bhi monocytes are massively recruited into the liver upon E.m-induced injury in CD11b++F4/80+ intrahepatic macrophages,and the proportion of resident macrophages and eosinophils increased after 6 weeks infection. In addition, CD8+ effector T cells displayed a less exhausted phenotype characterized by decreased expression of programmed cell death 1 (PD-1).These findings indicate that inflammatory monocytes recruited into the injured liver and the percentage of kuffer cells, eosinophils and CD8+PD-1lo T cells increased around lesion, promote the progression of liver injury. These data suggest that they could be related to resistance or sensitivity to infection by E. multilocularis in humans.