bluuebunny/crossref_metadata_2025_split huggingface dataset:
A protective role for human IL-10-expressing CD4+ T cells in colitis (121.11)
Authors: Bream Jay , Ramakrishnan Amritha , Wang Fengying , Zhang Hao , Margolick Joseph , Brayton Cory , Ranatunga Dilini 
Published: January 2023

Abstract
Little is known about the relationship between cell- or tissue-specific IL-10 expression and disease susceptibility in humans. Here, we utilized the previously described hIL10BAC transgenic model to examine the role of human IL-10 (hIL-10) in maintaining intestinal homeostasis. Genomically-controlled hIL-10 expression rescued Il10−/− mice from Helicobacter-induced colitis and was associated with control of pro-inflammatory cytokine expression and TH17 cell accumulation in gut tissues. Resistance to colitis was associated with an accumulation of hIL-10-expressing CD4+Foxp3+ T regulatory cells specifically within the lamina propria but not other secondary lymphoid tissues. Co-transfer of CD4+CD45RBlo cells from Il10-/-/hIL10BAC mice rescued Rag1-/- mice from colitis further suggesting that CD4+ T cells represent a protective source of hIL-10 in the colon. In concordance with an enhanced capacity to express IL-10, CD4+CD44+ T cells isolated from the lamina propria exhibited lower levels of the repressive histone mark H3K27Me3 and higher levels of the permissive histone mark AcH3 in both the human and mouse IL10 locus compared to the spleen. These results provide experimental evidence verifying the importance of T cell-derived human IL-10 expression in controlling inflammation within the colonic mucosa. We also provide molecular evidence suggesting the tissue microenvironment influences IL-10 expression patterns and chromatin structure in the human (and mouse) IL10 locus.
Scaffold protein Disc-Large Homolog 1 is required for T cell receptor-induced activation of regulatory T cell function (121.14)
Authors: Zanin-Zhorov Alexandra , Lin Jiqiang , Scher Jose , Kumari Sudha , Blair David , Hippen Keli , Blazar Bruce , Abramson Steven , Lafaille Juan , Dustin Michael 
Published: January 2023

Abstract
Foxp3+CD4+CD25high regulatory T cell (Treg) suppression of inflammation depends on T cell receptor (TCR)-mediated NFAT activation with reduced Akt activity. We investigated the role of the scaffold protein Dlgh1 in linking the TCR to this unique signaling outcome. The Treg immunological synapse (IS) recruited 4-fold more Dlgh1 than conventional CD4+ T cell IS. Tregs isolated from patients with active rheumatoid arthritis, or treated with tumor necrosis factor-α, display reduced function and diminished Dlgh1 recruitment to the IS. Furthermore, Dlgh1 silencing abrogated Treg function, impaired NFATc1 activation, reduced PTEN levels and increased Akt activation. Dlgh1 operates independently of the negative feedback pathway mediated by the related adapter protein Carma-1 and thus presents an array of novel targets to selectively manipulate Treg function.
Quiescence regulator Foxp1 restrains antigen-induced T cell activation and response by mechanisms including a novel negative feedback loop (121.17)
Authors: Wang Haikun , Feng Xiaoming , Takata Hiroshi , Wolf Amaya , Strauman Maura , Tian Lifeng , Erikson Jan , Jiang Ping , Tucker Philip , Hu Hui 
Published: January 2023

Abstract
Previously we have shown that transcription factor Foxp1 is an essential regulator in maintaining the quiescence of naive T cells during homeostasis. In mice, Foxp1 has four isoforms. Now we find that whereas the full-length Foxp1A is constitutively expressed in mature T cells, T cell receptor (TCR) stimulation induces the expression of a short Foxp1 isoform, Foxp1D, which in turn dampens TCR signaling and T cell proliferation, constituting a negative feedback loop. In Foxp1D conditional transgenic mice, Foxp1D transgene in T cells reduces the generation of CD8+ and CD4+ T effectors to viral infections. On the contrary, Foxp1-deficient T cells (lacking all Foxp1 isoforms) elicit a faster and stronger effector response to antigen challenge. These results suggest that quiescence gene Foxp1 plays an important role in restraining antigen-induced T cell activation and response with a novel negative feedback loop by Foxp1D.
cAMP responsive element modulator (CREM)α induces epigenetic remodeling of IL2 in T lymphocytes from SLE patients (121.2)
Authors: Hedrich Christian , Rauen Thomas , Tsokos George 
Published: January 2023

Abstract
IL-2 functions as an auto- and paracrine growth-factor for T lymphocytes and is essential during their proliferation and activation. The absence of IL-2 has been linked to the development of autoimmune phenotypes in mice and humans. The multi-factorial autoimmune disease Systemic lupus erythematosus (SLE) is characterized by disrupted transcription factor networks and dysregulated cytokine expression, antigen-presentation, and lymphocyte function. The failure to express IL-2 is a hallmark of SLE T lymphocytes and results in reduced numbers of regulatory T lymphocytes that prevent autoimmunity. Impaired IL-2 expression was linked to over-production of the transcription regulatory factor cAMP response element modulator (CREM)α that in turn binds to a CRE site within the IL2 promoter (-180 CRE) in SLE T lymphocytes. Here, we document the involvement of CREMα mediated gene-wide HDAC1-directed deacetylation of histone H3K18, and DNMT3a directed CpG-DNA hypermethylation in the silencing of IL2 in T lymphocytes from SLE patients. We provide evidence that CREMα mediates silencing of IL2 in SLE T cells through histone deacetylation, and CpG-DNA methylation. Our findings add further evidence to the discussion of whether epigenetic modifications display region and tissue-specific patterns in health and disease. Thus, CREMα may serve as a promising target to correct cytokine expression in patients with SLE.
New modes of ERK feedback regulation in T cell receptor signaling revealed by quantitative phosphoproteomic analysis (121.29)
Authors: Helou Ynes , Nguyen Vinh , Beik Samantha , Hung Norris , Salomon Arthur 
Published: January 2023

Abstract
Competing positive and negative signaling feedback pathways play a critical role in tuning the sensitivity of T cell receptor (TCR) activation by selectively enhancing responses to foreign ligands while suppressing signals from self-peptides. In response to TCR agonist engagement, ERK is activated to positively regulate TCR signaling through phosphorylation of Ser59 on Lck. To obtain a wide-scale view of the role of ERK in propagating TCR signaling, a quantitative phosphoproteomic analysis of 417 tyrosine phosphorylation sites by mass spectrometry was performed on Jurkat E6-1 cells in the presence of U0126, a MEK1/2 specific inhibitor of ERK activation. Relative to untreated cells, ERK-inhibited cells showed constitutive decreases in phosphorylation through a TCR stimulation time course on tyrosine residues found on upstream signaling proteins (CD3 chains, Lck, ZAP-70), as well as downstream signaling proteins (LAT, SLP-76, PLCγ1, Itk), consistent with ERK’s proposed role in TCR positive feedback signaling. Interestingly, ZAP-70 Tyr598 and CD3 ζ Tyr123 showed elevated phosphorylation in response to U0126 treatment, suggesting differential regulation of these sites via ERK feedback. Furthermore, elevated phosphorylation on the activation site of Fyn was observed in response to ERK inhibition. These findings shed light on novel signaling events that are regulated by this kinase which may fine tune TCR activation.
Title: PD-1 regulates T cell proliferation in a tissue and subset specific manner during normal mouse pregnancy (122.11)
Authors: Bonney Elizabeth , Shepard Michelle 
Published: January 2023

Abstract
The regulation of T cell homeostasis during pregnancy has important implications for maternal tolerance and immunity. Evidence suggests that Programmed Death-1 (PD-1) participates in regulation of T cell homeostasis and peripheral tolerance. To examine the contribution of PD-1 signaling on T cell homeostasis during normal mouse pregnancy, we examined T cell number or proportion, PD-1 expression, proliferation, and apoptosis by flow cytometry, BrdU incorporation, and TUNEL assay in pregnant mice given anti-PD-1 blocking antibody or control on days 10, 12, and 14 of gestation. We observed tissue, treatment, and T cell-specific differences in PD-1 expression. Both pregnancy and PD-1 blockade increased T cell proliferation in the spleen while this effect was limited to CD4 T cells in the uterine- draining nodes. In the uterus, PD-1 blockade markedly altered the composition of the T cell pool. These studies support the idea that pregnancy is a state of dynamic T cell homeostasis and suggest that this state is partially supported by PD-1 signaling.
Imaging the initial cellular events in EAE. (123.26)
Authors: Barkauskas Deborah , Myers Jay , Othman Youmna , Huang Alex 
Published: January 2023

Abstract
Limitations in current techniques to capture rare immune cell events have hindered the characterization of the initiation and progression of experimental autoimmune encephalomyelitis (EAE). With the cranial window and intravital two-photon microscopy techniques, we followed the initial dynamic and sequential cellular processes preceding EAE development. Disease progression was imaged daily in the first 12 days post EAE induction in either CX3CR1-GFP or CD11c-GFP transgenic mice containing adoptively transferred naïve ubiquitin-CFP myelin oligodendrocyte glycoprotein (MOG) specific T cells. Blood-brain barrier (BBB) permeability was assessed by i.v. injection of fluorescent dextran or quantum dots. We hypothesize that lymphocyte accumulation follows an initial transient BBB leak caused by pertussis toxin-initiated inflammation, with subsequent accumulation and activation of microglia and perivascular antigen presenting cells (APCs) facilitating the tissue-specific infiltration of MOG-specific T cells into the brain parenchyma via intra-luminal contact. Indeed, our sequential imaging experiments demonstrated transient leaks in BBB at the pia surface within the first 4 days post EAE induction, local APC accumulation from days 2 to 6, followed by T cell infiltration adjacent to local CNS vessel leak. Future work involves further defining the cellular and molecular regulation of CNS peri-vascular microenvironment responsible for cross-BBB attraction of MOG-specific cells.
Intrinsic Regulation of Autoimmune CD4+ Memory T Cells by Subtle Variations in CTLA4 levels (123.38)
Authors: Devarajan Priyadharshini , Miska Jason , Torres Juan , Chen Zhibin 
Published: January 2023

Abstract
Cytotoxic T lymphocyte antigen 4 (CTLA4) is a negative regulator of immune function. Genetic polymorphisms in the CTLA4 locus leading to subtle variations in CTLA4 expression have been linked to various autoimmune diseases including type 1 diabetes. We hypothesize that CTLA4 regulates the immune response in these disease settings through memory T cells. To test this hypothesis, a unique transgenic model of CTLA4 RNAi has been used to mimic subtle quantitative variations of CTLA4 in humans. BDC2.5 transgenic mice with the CTLA4 RNAi had a higher incidence of diabetes. An analysis of different T cell populations in the lymph nodes and pancreatic tissue revealed an increased number of CD4+ effector memory T cells. To investigate if the increase in CD4+ effector memory T cells is a direct effect of CTLA4 reduction on the conventional T cell compartment (cell-intrinsic) or if it is mediated through reduced CTLA4 on regulatory T cells (cell-extrinsic), a series of adoptive transfer experiments were done with CD4+ naïve, effector, effector memory and regulatory T cells. These experiments suggested a cell-intrinsic mechanism where reduced CTLA4 in the CD4+ effector memory T cell itself causes increased activation and survival/proliferation. These results indicate that subtle reductions in CTLA4 levels have a selective impact on CD4+ effector memory T cells, which may play a critical role in CTLA4-determined risk of autoimmune diabetes.
Metabolic regulation of CD4+ T cells by Estrogen Related Receptor-α modulates autoimmune and allergic disease (123.48)
Authors: Michalek Ryan , Gerriets Valerie , Nichols Amanda , Inoue Makoto , Theriot Barbara , Giguere Vincent , McDonnell Donald , Shinohara Mari , Walker Julia , Rathmell Jeffrey 
Published: January 2023

Abstract
Upon activation, naïve CD4+ T cells differentiate into effector (Teff) subsets that provide protective immunity or inducible regulatory (Treg) subsets which balance inappropriate inflammation and autoimmunity. Consistent with distinct immunological roles, Teff and Treg utilize specialized metabolic programs for specification and function with Treg increasing lipid oxidation and Teff dependent on glucose metabolism. We have identified the nuclear receptor estrogen related receptor-α (ERRα) as a critical regulator of metabolic gene expression required for Teff. Here, we demonstrate through the use of an experimental autoimmune encephalomyelitis model that ERRα contributes to the severity of autoimmune disease as limited glucose metabolism in the absence of ERRα selectively diminished Th17, but not Treg generation. ERRα contributed to inappropriate inflammation as Teff glucose metabolism, expansion, and function were reduced in ERRα-/- mice and in siRNA treated human CD4+ T cells. Additionally, elevated ERRα expression and glycolytic metabolism were also observed in a murine model of asthma and from lavage fluid of asthmatic patients. Importantly, pharmacological inhibition of ERRα following an aerosol response alleviated airway inflammation, demonstrating a metabolic requirement for ERRα in Teff maintenance. Thus, ERRα is a selective transcriptional regulator of Teff metabolism that may provide a metabolic means to modulate immunity.
Oral tolerance in patients with food protein induced enterocolitis syndrome (FPIES); Evidence of impaired induction of oral tolerance in FPIES patients with persistent reaction to multiple foods (123.9)
Authors: Jyonouchi Harumi , Geng Lee , Streck Deanna , Toruner Gokce 
Published: January 2023

Abstract
Treatment for FPIES is the avoidance of offending food, i.e., a restricted diet (RD). While most FPIES children are good responders to the RD (FPIES-GR), some are not (FPIES-poor responders: FPIES-PR) with persistent non-IgE mediated reactivity to multiple foods. This study assessed production of counter-regulatory cytokines crucial for oral tolerance by peripheral blood mononuclear cells (PBMCs) in FPIES-PR (N=18, 0.6 -4.8 yr), FPIES-GR (N=22, 1.1-5.9 yr), and normal control (N=19, 1.0-5.7 yr) children after FPIES children were on the RD. PBMCs were stimulated with agonists of toll like receptors (TLRs) and representative luminal antigens (Ags) (soy, cow’s milk, and wheat proteins, and candida Ag). Microarray analysis of PB moncytes was also performed on samples from some of these children. FPIES-PR children revealed more severe clinical features including failure to thrive, intolerance to probiotics, and anaphylaxis like reaction to rice than FPIES-GR children (p&lt;0.005). FPIES-PR PBMCs produced less sTNFRII and IL-10 both with TLR agonists and without a stimulus than the control groups. FPIRS-PR cells also produced less IL-6 with a TLR4 agonist and without a stimulus than normal controls (p&lt;0.05). Such changes were not observed in FPIES-GR cells. FPIES-PR PBMCs produced less TGF-β with and without luminal Ag than PFIES-GR PBMCs. Taken together, FPIES-PR children may have impaired production of counter-regulatory cytokines that are crucial for oral tolerance.
Optimized buffer system for detection of transcription factors and surface proteins in T-cells validated in high throughput screening of activated T-regulatory cells. (124.10)
Authors: Hingorani Ravi , Vidal Jason , Le Hai , Waterman Paul , Rohrer Jurg , Boyce Christopher 
Published: January 2023

Abstract
Intracellular detection of antigens at the single cell level by flow cytometry requires optimized buffers and methods for detection of a given specificity and the compartment probed. Using a new buffer system tightly controlled for low non-specific staining, increased resolution, and significantly lower cell loss, we have developed an optimized system for detection of intracellular proteins by flow cytometry. This system works across multiple tissues from human and mouse models offering a one-method system for reliable antibody cross-reactivity tests. The optimized system detects proteins within the nucleus, endoplasmic reticulum, outer mitochondrial membrane, and the Golgi complex, with focus on optimal resolution of DNA-binding transcription factors. Intracellular targets detected included; FoxP3, T-bet, RORγT, GATA-3, Satb1, Bcl6, Bcl2, pH2AX, Ki-67, Themis, Nanog, Sox1, Sox2, Oct 3/4, Pax-6, Nestin, and various cytokines. In addition, fluorescent antibodies to cell surface antigens for multi-color flow were also found to be compatible with the optimized buffer system. Further verification of the buffer was done by high throughput screening of human cell surface markers with the BD LyoplateTM on activated human T-regulatory cells (Treg). GARP positive and negative Treg subsets were screened for various markers and the results are discussed in the poster.
Type I interferon inhibits acute IL-5 and IL-13 expression in human memory CD4+ T cells (125.12)
Authors: Gonzales Sarah , Huber Jonathan , Farrar J. David 
Published: January 2023

Abstract
Type I Interferon (IFN-α/β) is an innate cytokine important for fighting infections and is used to treat hepatitis C infection and multiple sclerosis, among other diseases. Although the innate role of this cytokine has been studied extensively, our group has shown that IFN-α/β plays a role in shaping the adaptive immune response in humans. Recently, we demonstrated that IFN-α/β inhibits Th2 development by regulating mRNA and protein levels of GATA3. Regulation of GATA3 in turn suppresses Th2 cytokine production, without effecting IFN-γ production. In addition to blocking Th2 development, we now demonstrate an acute inhibition of human memory T cell function upon treatment with IFN-α/β. Human memory CD4+ T cells acutely activated with anti-CD3 in the presence of IFN-α/β resulted in reduced mRNA levels of Th2 cytokine genes, IL5 and IL13, but did not effect IL4 expression. GATA3 levels were not affected, suggesting a GATA3-independent mechanism. Further experiments have determined that this regulatory mechanism does not require new protein synthesis. This mechanism seems to be specific to IFN-α/β, as neither IFN-γ nor IFN-λ treatment were able to repress Th2 cytokine mRNA expression to the extent of IFN-α/β. This study has uncovered an additional level of regulation that blocks early TCR signaling events that affect Th2 function. This novel pathway suggests that treatment of atopic asthmatic individuals with IFN-α/β can regulate both Th2 development and memory Th2 function.
Inhibition of Tyrosine Kinase Reduces Pulmonary Inflammation in A Mouse Model of Asthma (125.14)
Authors: Kim Jiyoun , Yang Beom seok , Remick Daniel 
Published: January 2023

Abstract
Protein tyrosine kinases play a critical role in activation of inflammatory cells and expression of airways inflammatory mediators. We investigated if a discoidin domain receptor family tyrosine kinase inhibitor, LCB03-0110 will reduce pulmonary inflammation in a mouse model of asthma induced by cockroach allergen (CRA). BALB/c mice were exposed to 3 pulmonary challenges via hypopharyngeal administration of CRA on day 0, 14 and 21. Two hours after the last challenge, airway hyperresponsiveness (AHR) was measured and the mice sacrificed for bronchoalveolar lavage (BAL) fluid and lung tissue. Groups of mice were treated with LCB03-0110 or vehicle one hour prior to the 2nd and 3rd CRA challenge on day 14 and 21. AHR was significantly reduced in the LCB -treated mice when compared to vehicle -treated group (454 vs. 235% Penh increase of baseline). Inhibition of tyrosine kinase activity significantly decreased levels of BAL CXC chemokines, KC (2322 ±169 vs. 1096 ±117 pg/ml) and MIP-2 (174 ±13 vs. 64 ±8 pg/ml) and pulmonary recruitment of neutrophils (903±31 vs. 364 ±27 x1000 cell/mouse). Further, mucin production in airways was significantly reduced. Taken together, treatment with a tyrosine kinase inhibitor significantly reduces AHR and pulmonary inflammation in a mouse model of asthma induced by cockroach allergens. This suggests that inhibition of tyrosine kinase may be a novel therapeutic target in the treatment of asthma and other inflammatory airway diseases.
Rapid polyclonal desensitization with monoclonal antibodies (125.15)
Authors: Khodoun Marat , Strait Richard , Kucuk Zeynep , Krishnamurthy Durga , Finkelman Fred 
Published: January 2023

Abstract
Rapid desensitization, a procedure in which individuals allergic to an antigen are treated at short intervals with increasing doses of that antigen until they tolerate a large dose, is an effective, albeit risky way to induce temporary tolerance. We have adapted this procedure to suppress IgE- or IgG-mediated immediate hypersensitivity in mice by injecting them with activating mAbs to IgE, FcϵRIα, or FcγRII/RIII, starting with doses too small to induce detectable reactions and doubling doses every 90 minutes. Rapid desensitization with anti-IgE mAb suppressed IgE-mediated immediate hypersensitivity; however, some mice developed mild anaphylaxis during desensitization. Rapid desensitization with an anti-FcϵRIα mAb that only binds FcϵRI that is not occupied by IgE suppressed both active and passive IgE-mediated anaphylaxis and induced no detectable reactions, but took several days to become fully effective by eliminating nearly all mast cell FcϵRI. Rapid desensitization with this mAb also rapidly, but temporarily, desensitized mast cells by decreasing their responsiveness to FcϵRI crosslinking. Rapid desensitization with anti-FcγRII/RIII mAb induced no detectable adverse reactions and rapidly suppressed IgG-mediated anaphylaxis. Similar approaches with mAbs to human IgE or IgG receptors may be useful as prophylaxis for human immediate hypersensitivity disorders, including anaphylaxis, food allergy, atopic dermatitis and asthma.
Diversity of epitope recognition of IgG4 antibodies induced by birch-pollen specific immunotherapy (125.3)
Authors: Subbarayal Brinda , Schiller Dirk , Möbs Christian , Pfützner Wolfgang , Vieths Stephan , Bohle Barbara 
Published: January 2023

Abstract
Birch pollen (BP)-allergic patients sensitized to the major BP allergen, Bet v 1, often show IgE-cross-reactivity with Mal d 1 in apple and Cor a 1 in hazelnut. We recently found that specific immunotherapy (SIT) with BP induces high titers of Bet v 1-specific IgG4. In parallel, many BP-SIT-treated patients developed food-reactive IgG4. However, around 25% of patients showed no detectable food-specific IgG4 response. The diversity of epitope recognition of IgG4 antibodies (abs) in BP-SIT-treated patients developing or lacking food-reactive IgG4 were compared. Sera containing Bet v 1-, Mal d 1- and Cor a 1-specific IgE and Bet v 1-specific IgG4 with and without food-reactive IgG4 abs were included. Blocking capacity of the sera was analysed in facilitated antigen-binding and basophil activation assays. Potential epitopes recognized by allergen-specific IgE and IgG4 abs were screened by competitive immunoscreening with a random phage-displayed 12-mer peptide library. Patients showing food reactive-IgG4 with blocking capacity recognized a higher number of putative epitopes distributed over the surface of allergens. In contrast, a restricted number of putative epitopes was recognized by sera from patients who had failed to develop food-reactive IgG4. The induction of polyclonal IgG4 responses during BP-SIT may be a prerequisite for the development of abs that cross-react with Bet v 1-related food allergens and consequently, block food-reactive IgE abs.