bluuebunny/crossref_metadata_2025_split huggingface dataset:
Pathogenic Th17 cell differentiation is negatively regulated by let-7 microRNAs in a mouse model of multiple sclerosis
Authors: Angelou Constance C , Wells Alexandria C , Vijayarhagavan Jyothi , Dougan Carey E , Lawlor Rebecca , Iverson Elizabeth , Lazarevic Vanja , Kimura Motoko Y , Peyton Shelly R , Minter Lisa M , Osborne Barbara A , Pobezinskaya Elena L , Pobezinsky Leonid A 
Published: January 2023

Abstract
Autoimmune disorders such as multiple sclerosis (MS) are caused by proinflammatory events mediated by pathogenic Th17 cells. In MS, these cells arise in response to autoantigen recognition and exposure to the cytokines IL-1β and IL-23, migrate to the central nervous system (CNS) by following gradients of CCR2- and CCR5-cognate chemokines, and secrete GM-CSF. GM-CSF is essential for disease development, as it promotes the activation, differentiation, and recruitment of peripheral inflammatory myeloid cells to the CNS that directly demyelinate neurons and damage axons. Th17 cell pathogenicity in MS has been correlated with microRNA (miRNA) dysregulation, which leads to aberrant post-transcriptional regulation of gene expression and enhanced autoreactive phenotype. We found that the lethal-7 (let-7) miRNA family is abundantly expressed in naive CD4+ T cells, but gets dramatically downregulated over time following antigen encounter, indicating that let-7 may control the differentiation of pathogenic Th17 cells. To investigate a potential regulatory role for let-7 in Th17 cell autoreactivity, we used experimental autoimmune encephalomyelitis (EAE), the animal model of MS. Specifically, we demonstrated that let-7 confers protection from EAE by negatively regulating the proliferation, IL-1β/IL-23-dependent differentiation, and CCR2/CCR5-dependent migration of pathogenic Th17 cells to the CNS. Conversely, absence of let-7 led to enhanced Th17 cell autoreactivity and aggravated disease. Therefore, our results identify a novel regulatory role for let-7 miRNAs in pathogenic Th17 differentiation during EAE development, suggesting a promising therapeutic application for the treatment of MS-related autoimmune diseases.
Eos supports TH1 differentiation through feed-forward propagation of the IL-2/STAT5 signaling pathway
Authors: Read Kaitlin , Sreekumar Bharath , Powell Michael Duane , Jones Devin M , Zafar Shah , Ghoneim Hazem E. , Collins Patrick L. , Oestreich Kenneth J 
Published: January 2023

Abstract
The Ikaros zinc finger (IkZF) transcription factor Eos is a known regulator of CD4+ regulatory T cell (TREG) populations. However, its role in the differentiation of other CD4+ T cell subsets is unknown. Here, we find that Eos deficiency results in both compromised TH1 cell differentiation and reduced production of the TH1 effector cytokine IFN-γ. Historically, IkZF factors have been associated with changes in gene expression via the alteration of chromatin structure. However, ATAC-seq analysis indicated that there were minimal changes in chromatin accessibility between wild type and Eos-deficient cells cultured under TH1-polarizing conditions. Rather, we find that IL-2 signaling promotes the STAT5-dependent induction of Eos expression, and that Eos both interacts with STAT5 and supports tyrosine phosphorylation-mediated STAT5 activation. Consequently, STAT5 enrichment is reduced at TH1 target genes in the absence of Eos. Thus, modulation of STAT5 signaling represents a novel mechanism by which Eos positively regulates the TH1 gene program. Given the important role for IL-2/STAT5 signaling in the differentiation of TH2, TH9, and TREG cell subsets, it will be of interest to determine whether this mechanism is conserved across additional CD4+ T cell populations.
RNA-binding protein HuR promotes Th17 cell differentiation and targeting HuR reduces autoimmune neuroinflammation
Authors: Chen Jing , Yu Shiguang 
Published: January 2023

Abstract
Dysregulated Th17 cell differentiation is associated with autoimmune diseases such as multiple sclerosis, which has no curative treatment. Understanding the molecular mechanisms of regulating Th17 cell differentiation will help in identifying a novel therapeutic target for treating Th17 cell-mediated diseases. Here, we investigated the cell-intrinsic processes by which RNA-binding protein HuR orchestrates Th17 cell fate decisions by post-transcriptionally regulating transcription factor Irf4 and Runx1 and receptor Il12rb1 expression, in turn promoting Th17 cell and Th1-like Th17 cell differentiation. Knockout of HuR impaired the transcriptome of Th17 cells characterized by reduced levels of RORγt, IRF4, RUNX1, and T-bet, thereby reducing the number of pathogenic IL-17+IFN-γ+CD4+ T cells during experimental autoimmune encephalomyelitis (EAE). In keeping with the fact that HuR increased the expression of adhesion molecule VLA-4 on Th17 cells, knockout of HuR impaired Th17 cell migration to the central nervous system and abolished the disease. Accordingly, targeting HuR by its inhibitor, DHTS, suppressed Th17 cell differentiation and reduced EAE severity. In sum, we uncovered the molecular mechanism underlying HuR’s regulation of Th17 cell differentiation and functions, underscoring the therapeutic value of HuR as a novel treatment for autoimmune neuroinflammation. (Supported by NIH R01 AI119135)
TCR dependent metabolic programming regulates autocrine IL-4 production resulting in self-tuning of the CD8+ T cell activation setpoint
Authors: Crofts Kali F , Holbrook Beth C , Soto-Pantoja David R , Ornelles David A , Alexander-Miller Martha A 
Published: January 2023

Abstract
The ability of T cells to sense and respond to environmental cues by altering their functional capabilities is critical for a safe and optimally protective immune response. One of the important properties that contributes to this goal is the activation setpoint of the T cell. Here we report a new pathway through which CD8+ T cells can self-tune their activation threshold. We find that in the presence of a strong TCR engagement event, there is a shift in the metabolic programming of the cell where both glycolysis and oxidative phosphorylation are significantly increased. This diverges from the switch to a predominantly glycolytic profile that would be predicted following naïve T cell activation. This altered metabolic program results in the production of autocrine IL-4. Both metabolic pathways are required for this cytokine to be made. IL-4 signaling in the activated CD8+ T cell results in modulation of the sensitivity of the cell, establishing a higher activation setpoint that is maintained over time. Together these data demonstrate a novel mechanism for the regulation of IL-4 production in CD8+ T cells. Further, they reveal a new pathway for the self-tuning of peptide sensitivity. Finally, these studies uncover an unexpected role for oxidative phosphorylation in regulating differentiation in CD8+ T cells.
Invariant NKT cells are unusually refractory to glucocorticoid-induced apoptosis but exhibit impaired functional fitness and fail to trigger antitumor immunity following physiological stress
Authors: Rudak Patrick T , Choi Joshua , Haeryfar S.M. Mansour 
Published: January 2023

Abstract
The nervous system serves critical roles in the regulation of immune responses. Consequently, the neuroimmune functional interface can be disrupted by physiological stress, potentially impeding our ability to combat malignancies. Invariant natural killer T (iNKT) cells are innate-like T cells that, upon activation by lipids such as αGC, participate in antitumor immune surveillance. However, whether mediators of stress impact iNKT cell functions in this context remains unexplored. Here, we subjected mice to prolonged psychological stress due to physical restraint or chronic stress due to heterotypic stressors before administering αGC. We report that stress abrogates TH1-type cytokine production by iNKT cells in vivo, an effect that is reversible by glucocorticoid receptor (GR) blockade and that is diminished in mice selectively lacking GRs in their T cell compartment. Instead, iNKT cell responses are skewed towards an abnormal anti-inflammatory and TH17-type cytokine signature. Gene expression analysis revealed that iNKT cells in stressed mice express reduced levels of critical genes supporting their typical differentiation states. Accordingly, iNKT cells in stressed mice fail to protect against pulmonary metastases of B16-F10 melanoma and are also unable to initiate killing activity against MHC class I-deficient splenocytes in vivo or YAC-1 lymphoma cells in vitro. Finally, the suppressive effects of stress on iNKT cells are not due to their apoptotic death. Unlike conventional T cells, human and mouse iNKT cells are remarkably resistant to glucocorticoid-induced apoptosis. Collectively, our findings define a novel mechanism of stress-induced immunosuppression involving innate-like but not conventional T cells.
Anti-CD3 and anti-CD28 antibodies conjugated with lipid microbubbles (MB-TAC) provide a superior expansion of human naive CD4+ and CD8+ T cells in long term culture
Authors: Lustig Ana , Manor Ty’Keemi , Liu Yu-Tsueng , Shi Guixin , Weng Nan-ping 
Published: January 2023

Abstract
Anti-CD3 and anti-CD28 antibodies are a method of choice for general and physiological activation of human primary T cells in vitro. Here we report a comparison of the growth, cell death, and cytokine production of anti-CD3 and anti-CD28 antibodies conjugated with lipid microbubbles (MB-TCA) and the standard Dynal beads (D-TCA). In 56-day cultures with three repeated stimulation cycles (14 days per stimulation), we found that MB-TCA induced a significantly better expansion (~20-fold increase) of naïve CD4 and CD8 T cells than did D-TCA. We found the main difference between MB-TCA and D-TCA was not the speed of activation-induced initial T cell proliferation but rather the survival of activated T cells. Furthermore, we found that MB-TCA activated T cells had a better telomere length maintenance and produced less TNFα and other cytokines than did D-TCA. Finally, blocking TNFα action via antibody against TNFRSF1A significantly improved expansion of T cells activated by D-TCA in vitro. Together, we demonstrated that MB-TCA offers a better expansion of primary human T cells in vitro and will be a useful method for long term culture of primary human T cells for both basic and clinical applications.
Role of tryptophan metabolism on senescent synovial fibroblasts
Authors: Usategui Alicia , López Abigail , Municio Cristina , Del Rey Manuel J , Pablos José L , Criado Gabriel 
Published: January 2023

Abstract

Background
Cellular senescence is connected to inflammation and altered metabolism. Rheumatoid arthritis (RA) patients show premature senescence and increased Trp metabolism along the kynurenine pathway. We aim to analyze the effect of Trp metabolism on cellular senescence and the expression of the Senescence Associated Secretory Phenotype (SASP) in human synovial fibroblasts (SF).

Methods
For inflammation-induced senescence, SF were cultured for 14 days after exposure to IL6/sIL6R (100 ng/ml for 72h on day 2). On day 14, senescence and SASP were analyzed. Senescent SF were quantified in cultures by detection of senescence-associated beta-galactosidase (SA b-gal). IDO1, IDO2, p16, ATG3, ATG5 and SASP components (IL6, IL8, MMP3, MCP1), were quantified by qRT-PCR and IL6 and IL8 proteins by ELISA. IDO1 was silenced using lentiviral vectors. IDO1 protein was detected by WB. Quantitative data were statistically analyzed by Pearson or Spearman correlation coefficient or Mann-Whitney test where appropriate.

Results
IDO1, but not IDO2, was induced by senescence in SF. Conversely, treatment of SF cultures with the IDO1 product kynurenine significantly reduced senescence, as determined by expression of the senescence marker p16 (p=0.018), although the autophagy response and SASP expression were not affected. IDO1 silencing during senescence induction resulted in a significant increase of IL6 expression without affecting other SASP components.

Conclusion
Upregulation of IDO1 by senescence, together with the reduction of senescence by kynurenine and the increased IL6 production by deficient IDO1 activity suggest that IDO1 and the kynurenine pathway play a negative regulatory role on senescence and SASP expression.
CD4+ T cells are more sensitive to inhibition of TCR signaling under Th17 polarizing conditions
Authors: Zinzow-Kramer Wendy M. , Weiss Arthur , Au-Yeung Byron B 
Published: January 2023

Abstract
Following TCR stimulation, in the presence of polarizing cytokines, naïve CD4+ T cells differentiate into effector cells (e.g. Th1, Th2, and Th17). To study how proximal TCR signaling molecules influence T cell differentiation, we used small molecule inhibitors to reduce signaling through the cytoplasmic tyrosine kinases Zap-70 or Lck. We used PP2 to inhibit Lck signaling and a chemical-genetic system to inhibit Zap-70. In this model system, mouse cells expressing an analog-sensitive mutant of Zap-70 are sensitive to inhibitor while wild-type cells are not. Surprisingly, we found that the generation of IL-17 producing Th17 cells was more sensitive to inhibition than the generation of either IFNg producing Th1 cells or IL-4 producing Th2 cells. These results raised the possibility that cells undergoing Th17 differentiation require TCR signaling of a higher intensity. To test this possibility, we analyzed the TCR-induced expression of IRF4 and Nur77-GFP, which are both inducibly expressed in relative proportion to the strength of TCR signaling. For both IRF4 and Nur77-GFP, the magnitude of expression was decreased under Th17 conditions. Additionally, increasing downstream signaling through NFAT rescued the Th17 sensitivity to inhibition. These results suggest that factors in Th17 cultures suppress TCR signal strength, thus cells stimulated under Th17 conditions experience weaker TCR signals and become more sensitive to further inhibition of TCR signaling.
IL-12 derived from CD8α+ dendritic cells drives the differentiation of innate-like T-bethigh memory-phenotype CD4+ T lymphocytes in steady state
Authors: Kawabe Takeshi , Yi Jaeu , Kawajiri Akihisa , Hilligan Kerry , Fang Difeng , Ishii Naoto , Yamane Hidehiro , Zhu Jinfang (Jeff) , Jankovic Dragana , Kim Kwang Soon , Trinchieri Giorgio , Sher Alan 
Published: January 2023

Abstract
CD4+ T lymphocytes consist of naïve, antigen-specific memory, and memory-phenotype (MP) cell compartments in steady state. We recently showed that MP cells exert innate-like effector function in host defense. However, it remained to be defined whether MP cell population comprises multiple subsets, and if so, what factors determine their selective differentiation under homeostatic conditions. Here we characterize MP lymphocytes as consisting of T-bethigh, T-betlow, and T-bet− subsets, with innate Th1-like effector activity exclusively associated with T-bethigh cells. We further show that steady-state differentiation of T-bethigh MP cells is promoted by IL-12-producing CD8α+ dendritic cells (DC1). Finally, we present evidence arguing that this tonic IL-12 production requires TLR-MyD88 signaling triggered independently of foreign agonists and is further enhanced by CD40-CD40L interactions with self-dependent MP CD4+ T lymphocytes. Thus, optimal differentiation of T-bethigh MP lymphocytes in steady state requires IL-12 tonically produced by DC1 in an MyD88 and CD40 signaling-dependent fashion.
This work was in part supported by the Intramural Research Program of the NIAID and NCI, NIH.
Non-typeable Haemophilus influenzae P5 binds C4b-binding protein promoting serum resistance
Authors: Thofte Oskar , Mattsson Emma , Su Yu Ching , Thegerström John , Jalalvand Farshid , Blom Anna Marie , Riesbeck Kristian 
Published: January 2023

Abstract
Haemophilus influenzae is a Gram-negative bacterium considered as a commensal in pre-school children, mainly giving rise to infections locally in the upper and lower respiratory tract. Historically invasive disease has been caused by encapsulated strains, mainly H. influenzae serotype b (Hib). However, since introduction of the Hib vaccine, there has been an increased incidence of invasive disease with unencapsulated non-typeable H. influenzae (NTHi). Since NTHi lacks a capsule they must rely on other mechanisms for serum resistance, including binding of complement regulators. We have previously demonstrated that binding of C4b-binding protein (C4BP) is important for NTHi-dependent complement evasion. Here we identified outer membrane protein 5 (P5) as the C4BP-binding protein in NTHi based on an outer membrane proteomic pull down-assay. NTHi P5 has previously been found to bind factor H, an inhibitor of the alternative complement pathway. We found that NTHi grown in nasopharyngeal-simulated conditions has a lower surface expression of P5. This was also corresponding to decreased serum resistance. Importantly, we found that P5-deficient NTHi mutants showed significantly decreased C4BP binding and serum resistance. Additionally, surface expression of P5 on Escherichia coli conferred increased serum resistance and C4BP binding. Our results further highlights P5 as an important protein for protecting NTHi against complement-mediated killing.
Macrophage replacement in Mycobacterium tuberculosis-induced murine lung granulomas
Authors: Herbath Melinda , Marcus Sarah , Spellman Bailey , Adler Samantha , Gilpin Trey , Thanthrige Thiunuwan , Johnson Lexi , Fabry Zsuzsanna , Sandor Matyas 
Published: January 2023

Abstract
Monocytes/macrophages are the dominant cell type in mycobacterial granulomas. Macrophages both provide a survival niche for the bacteria and play a central role in bacterial control. Most of them are recruited from the blood with a Ly6C+ phenotype and they differentiate into F4/80+ macrophages in the lung lesions. It is well demonstrated that monocytes have access to pre-formed granulomatous lesions, but the rate of cell recruitment and the fate of the recruited cells are not well understood.
Previously, testing BCG-induced liver granulomas using a transplantation model, we detected continuous cell replacement in the preformed lesions. Here, using intravital staining and cytofluorimetric detection of CD45+ blood cells in combination with IHC, we show that CD11b+ cells can access the lung parenchyma and Mtb-induced lesions over the course of 24 hours in Mtb H37Rv infected animals. We measured the level of monocyte/macrophage infiltration in both acute and chronic infection in C57BL/6 and C3HeB/FeJ mice. Although monocytes have a short lifespan in the blood, using intravenous cell transfer of tdTomato-expressing sentinel cells we show that they can live longer than a week in the lung lesions of Mtb-infected animals. Interestingly, a population of adoptively transferred CD11b+tdTomato+ cells can be detected for more than 10 days in the blood vessels in the infected lung. Gene expression profiling of recently or earlier recruited (CD11b+F4/80− or CD11b+F4/80+) cells sorted from the lung shows differential inflammatory gene expression between these populations.
The continuous cell replacement that is required to maintain granulomatous lesions offers a novel possibility to manipulate granuloma pathology for a therapeutic advantage.
Does Dysbiosis Drive CVID Enteropathy by Enhancing Bile Acid Cytotoxicity?
Authors: Mohammed Ahmed Dawood , Chatzistamou Ioulia , Roland Mary , Jolly Amy , Schoettmer Lillian , Nagarkatti Mitzi , Nagarkatti Prakash , Kubinak Jason L. 
Published: January 2023

Abstract
Common variable immunodeficiency (CVID) is a primary immunodeficiency characterized by defects in the humoral immune response. CVID patients commonly present with a gastrointestinal enteropathy characterized by intestinal malabsorption. CVID is linked with abnormal gut microbiota composition (i.e. dysbiosis), which may be a contributing factor to CVID enteropathy. Recently, we demonstrated that CD19−/− mice represent a spontaneous animal model of CVID enteropathy that develop an associated dysbiosis. We now expand upon this work to identify how microbial dysbiosis may contribute to CVID enteropathy. We first demonstrate a significant association between dysbiosis in CD19−/− mice with defects in bile acid (BA) homeostasis (increased luminal BA titers and decreased abundance of conjugated BA species). Enteropathy in CD19−/− mice is associated with enhanced apoptosis of intestinal epithelial cells (IECs). Adoptive transfer of WT B cells (but not CD19−/− B cells) reduces luminal BA concentrations, protects IECs from apoptosis, and resolves enteropathy, indicating that functional B cells are important regulators of BA homeostasis and gastrointestinal health. Additionally, dietary supplementation with a taurine conjugated BA also protects animals from disease demonstrating that enhanced BA deconjugation enhances disease severity. Finally, direct ablation of the ability of a model commensal microbe to deconjugate BAs also rescues animals from disease. Collectively, these data suggest that enhanced bacterial deconjugation of BAs in the gut contributes to CVID enteropathy by enhancing BA cytotoxicity to IECs. Thus, microbial BA deconjugation may represent a novel therapeutic target for the treat of CVID enteropathy.
Deficiency of TAM receptor family increases γδT cell population by promoting chemokine-induced gut homing
Authors: Yi Jawoon , Choi Harim , Kim Su-Man , Kang Hyung-Sik 
Published: January 2023

Abstract
Intestinal intraepithelial lymphocytes (IELs) are a various community of lymphoid cells located in between the intestinal epithelial cells (IECs) that configure the intestinal mucosal barrier. The epithelial γδT cells act as a major T cell population in epithelia, which is support in tissue homeostasis and repair. However, we found that the γδT cell population was strikingly increased in IELs in TAM receptor-deficient mice. Based on these data, we tested whether this receptor acts as a crucial regulator, which may generate better homing to the intestinal epithelium. We also tested that adoptive transfer of γδT cells to show γδT cell homing to the intestinal epithelium. Our study shows that a key factor in γδT cell homing into the intestinal epithelium, to maintain homeostasis.
Role for B-cell lymphoma 6 in intestinal innate lymphoid cell homeostasis and inflammatory disease
Authors: Ohayon David E , Seelamneni Harsha , Chaimowitz Natalia S , Cazares Tareian , Waddell Amanda , Huber Alex , Kotey Amos , Rosen Michael J , Miraldi Emily R , Waggoner Stephen N 
Published: January 2023

Abstract
Group 3 innate lymphoid cells (ILC3) are critical for intestinal homeostasis. The functional activity of ILC3 must be tightly regulated as cytokines (e.g. IL-17 and IL-22) produced by these cells are critical for antimicrobial immune defense but can also contribute to intestinal disease when produced in excess. This regulation is achieved via incompletely defined transcriptional networks that include B-cell lymphoma 6 (BCL6), which we recently demonstrated is critical for maintaining ILC1- and ILC3-specific gene expression programs during homeostasis (Pokrovskii et al 2019). We now show that Bcl6-deficient small intestine lamina propria ILC3 exhibit elevated expression of the transcription factors retinoic acid receptor-related orphan nuclear receptor alpha (RORα) and RORγt. These factors are known to support ILC3 cytokine expression, potentially explaining elevated Il17a and Il17f expression by Bcl6-deficient ILC3. Of note, expression of Il22ra2 (IL-22 binding) was also markedly increased in these ILC3, likely limiting bioavailability of IL-22. Accordingly, these mice exhibit marked reduction in IL-22-dependent expression of epithelium-associated anti-microbial peptides in the terminal ileum. Of note, Bcl6ΔILC mice exhibited augmented susceptibility to dextran sodium sulfate-induced inflammatory bowel disease. Thus, BCL6 expression in ILC3-intrinsic BCL6 expression is important for mucosal immune defense and limiting intestinal inflammation.