bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bypassing a relatively common deficiency in neurotransmitter synthesis blocks allergic inflammation (97.8)
Authors: Cook-Mills Joan , Mahadevia Riti , McCary Christine , Abdala-Valencia Hiam , Berdnikovs Sergejs 
Published: January 2023

Abstract
An unaddressed triad of symptoms in allergic patients includes: 1) allergic/asthmatic responses, 2) anxiety/depression, and 3) constipation/acid reflux. We proposed that there is a common mediator of these symptoms. It is reported that a deficiency in the neurotransmitter serotonin increases anxiety/depression and reduces peristaltic gastrointestinal movement. In addition, leukocytes synthesize serotonin and serotonin receptors are on leukocytes. Therefore, we proposed that a common mediator is a deficiency in synthesis of the serotonin precursor, 5-hydroxytryptophan (5HTP). A reduction in synthesis of 5HTP occurs due to common polymorphisms in the rate limiting enzyme tryptophan hydroxylase (TPH). TPH converts tryptophan to 5HTP and decarboxylases convert 5HTP to serotonin. To address 5HTP regulation of inflammation, we determined whether dietary supplementation with 5HTP reduces mouse allergic lung inflammation induced by OVA or by house dust mite (DerP1). C57BL/6 mice were used since they have some of the serotonin pathway polymorphisms. 5HTP-containing diets were started two days before antigen. Interestingly, 5HTP blocked lung inflammation 50-70% without affecting body weight, blood eosinophils, Th1/Th2 cytokines, or chemokines. In addition, 5HTP pretreatment of endothelial cells blocked leukocyte transendothelial migration, suggesting that leukocyte recruitment is inhibited. Thus, 5HTP and its metabolites are critical for regulation of allergic inflammation.
Vitamin E isoforms and doses differentially regulate experimental asthma. (97.9)
Authors: McCary Christine , Abdala-Valencia Hiam , Berdnikovs Sergejs , Cook-Mills Joan 
Published: January 2023

Abstract
We have reported that supplemental doses of the alpha and gamma tocopherol isoforms of vitamin E decrease and increase, respectively, lung inflammation in murine experimental asthma. Because asthma clinical trials are currently examining the effect of highly-elevated tocopherol doses in humans, we examined the regulation of murine experimental asthma by highly-elevated doses of tocopherols, defined as 2 mg tocopherol/day (equal to 10 times the supplemental dose). Mice treated with highly-elevated doses of alpha or gamma tocopherol had significantly higher tissue levels of tocopherols, decreased leukocyte infiltration into the lung, and decreased Th2 cytokine and chemokine production. The reversibility of tocopherols’ effects was also determined. At either supplemental or highly-elevated doses, the regulatory effects of alpha and gamma tocopherol were fully reversible, revealing no sustained anti- or pro-inflammatory functions of these isoforms. Overall, gamma tocopherol enhances inflammation at supplemental doses and ablates inflammation at highly-elevated doses. In contrast, alpha tocopherol decreases inflammation at all doses tested. Our results have clinical implications for tocopherol supplementation and alter interpretation of tocopherol studies reported in the literature.
Effects of carotenoids on differential expression of scavenger receptors and cytokines in human monocyte-derived macrophage subpopulations (98.19)
Authors: Wiener Doris , Gong Xiaoming , Marisiddaiah Raju , Rubin Lewis 
Published: January 2023

Abstract
Circulating monocytes differentiate into macrophage (MΦ) subpopulations in response to environmental signals. Carotenoids may reduce MΦ oxidant stress, alter the inflammatory response, and lower risk of atherosclerosis. These observations prompted us to determine the effects of β-carotene, lycopene, astaxanthin and lutein on differentiation of human monocytes into MΦs (M1 and M2) phenotypes. Monocyte-to-MΦ differentiation was driven by treatment for 6 days with GMCSF or MCSF ± individual carotenoids. M1 MΦs (+GMCSF) were activated with LPS+IFNγ; M2 MΦs (+MCSF) were activated with IL-4. We validated that M1 and M2 cells showed distinct morphologies and expression patterns (qRT-PCR) of scavenger receptors (SR-A, LDLR, CD36, SR-B1) and cytokines (IL-10, IL-12). M1 and M2 markers (CD14, CD16, CD36, CD80, CD163) and cytokines (PGE2, IL-6, IL-8, IL-10, IL-12, TNFα, IFNγ) were confirmed by flow cytometry. Expression of CD14, CD16, CD36, CD163, and IL-10 was higher in M2 than M1 MΦs. Lycopene and astaxanthin decreased SRA, CD36 and LDLR mRNA levels in M1 cells but had no significant effect on M2 cells. Lutein also reduced SRA, CD36 and IL-10 expression in M2. Conversely, β-carotene increased M1 and decreased M2 expression of SRA, CD36, IL-10 and IL-12. Results show individual carotenoids can regulate MΦ scavenger receptor and cytokine expression. We speculate that carotenoids influence MΦ polarization, thereby inhibiting cholesterol accumulation in MΦ-derived atherogenic foam cells.
ESTROGEN RECEPTOR SIGNALING PROMOTES MONOCYTE ACTIVATION AND DIFFERENTIATION UPON INFLAMMATION IN VIVO (98.20)
Authors: Bajana Sandra , Turner Sean , Kovats Susan 
Published: January 2023

Abstract
17-β-estradiol at physiological levels regulates GM-CSF-mediated dendritic cell (DC) differentiation in vitro by promoting the generation of CD11bintLy6C- DC from myeloid progenitors. In this model, estrogen receptor alpha (ER) signaling induces and sustains elevated levels of interferon regulatory factor 4 (IRF4), a transcription factor critical in this pathway. Such events may relate to the de novo DC differentiation that occurs in vivo upon inflammation during infection or autoimmunity, resulting in inflammatory DC that determine the induction of specific Th cell populations. The current work evaluates whether ER signaling regulates new DC differentiation from blood monocytes during inflammation in vivo, and if this is associated with elevated IRF4 expression. ERα+/+ or ERα-/- mice were immunized subcutaneously with complete Freund’s adjuvant. By day 2 post immunization, compared to ERα-/- mice, ERα+/+ mice harbored an increased frequency of CD115+CD11b+ blood monocytes expressing higher levels of IRF4 and MHC class II. On days 2 and 7, relative to ERα-/- mice, lymph nodes of ERα+/+ mice contained greater numbers of monocyte-derived Gr1+CD11b+ CD11clo inflammatory DC. These data show that during inflammation in vivo, ER signaling promotes monocyte activation, IRF4 expression and differentiation, which leads to increased numbers of a de novo inflammatory DC population in draining lymph nodes.
Use of Computer Aided Drug Design (CADD) to identify small molecule inhibitors of TLR2 signaling (98.24)
Authors: Schwarz Ryan , Laird Michelle , Toshchakov Vladimir , Green Shannon , MacKerell Alexander , Vogel Stefanie 
Published: January 2023

Abstract
Toll-like receptors (TLRs) are a family of transmembrane innate immune molecules that play a key role in the detection of microbial and self ligands released from damaged cells. At their C-termini, Toll/Interleukin-1 receptor (TIR) domains dimerize upon ligand binding and initiate a signaling cascade that leads to upregulation of inflammatory cytokines. Overexuberant signaling leads to a “cytokine storm,” causing extensive tissue damage that can be fatal. The ability to block TLR signaling may ameliorate such a scenario. Using CADD, a panel of ~150 low molecular-weight compounds was identified based on their predicted ability to bind a pocket on the TLR2 TIR surface adjacent to BB loop. We performed preliminary screening of these compounds in TLR2-HEK293T stable transfectants and secondary screening of promising candidates in thioglycollate-elicited peritoneal murine macrophages. The impact of compounds on signaling was determined using quantitative RT-PCR of cytokines in response to TLR2-specific agonists (P2C and P3C) and TNF-α, which does not utilize TIR domains for signaling. Compound cytotoxicity was assessed by lactate dehydrogenase (LDH) release. To date, we have identified 10 putative TIR-specific inhibitors, 1 TNF-α-specific inhibitor, and 4 inhibitors of both TIR and TNF-α signaling. This project may lead to the identification of potent small molecule inhibitors of TIR-specific TLR signaling.
Elf3 regulates dendritic cell driven T cell differentiation (99.1)
Authors: Kushwah Rahul , Wu Jing , Oliver Jordan , Hu Jim 
Published: January 2023

Abstract
Elf3 belongs to the family of epithelium specific Ets transcription factors and can be upregulated in non-epithelial cells upon cytokine stimulation. Elf3-/- mice have normal T cell development and T cell differentiation into different T helper lineages. Here we show that upon epicutaneous sensitization with ovalbumin(OVA) followed by OVA intranasal challenge, Elf3-/- mice display a defect in Th17 induction, though OVA specific IgE and IgG1 antibody titers are several folds higher and airway inflammation is more extensive compared to Elf3+/+ mice, indicating an exaggerated Th2 response. Furthermore, intraperitoneal sensitization of Elf3-/- mice with OVA followed by OVA intranasal challenge, confirmed that Elf3-/- mice mounted an exaggerated Th2 response. Since, dendritic cells (DCs) are key players in initiation of adaptive immune response, we analyzed DC migration upon OVA challenge and showed that pulmonary DCs underwent hyperactivation in Elf3-/- mice compared to Elf3+/+ mice and rates of DC migration to the airway draining lymph nodes were also significantly higher in Elf3-/- mice, indicating that DCs may mediate the exaggerated Th2 response observed in Elf3-/- mice. Further analysis revealed that Elf3-/- DCs are more prone to maturation than Elf3+/+ DCs and upon maturation, Elf3-/- DCs preferentially secrete chemokines/cytokines that are important in polarizing Th2 differentiation. Taken together, these findings reveal a role of Elf3 in DC driven T cell differentiation.
RNA-binding protein HuR coordinately regulates genes essential for Th2 polarization and function at the posttranscriptional level (99.7)
Authors: Gubin Matthew , Stellato Cristiana , Casolaro Vincenzo , Tartar Danielle , Magee Joseph , Chen Jing , Calaluce Robert , Atasoy Ulus 
Published: January 2023

Abstract
Posttranscriptional gene regulation controls the expression of genes implicated in a variety of processes, including those of the immune system. The RNA-binding protein HuR regulates many early response genes as well as cytokines such as IL-4 and IL-13. Therefore, HuR has been implicated in regulating Th2 polarization and cytokine production. GATA-3, a transcription factor that is essential for Th2 polarization contains a putative HuR binding site in its 3’UTR. Immunoprecipitation (IP) of ribonucleoprotein complexes with an anti-HuR Ab revealed significant enrichment for GATA-3 mRNA. HuR association with GATA-3 3’UTR was confirmed by biotin pull-downs. To understand the role of HuR in Th2 polarization in vivo we generated a transgenic HuR CD4+ T-cell mouse model. Th2-skewed cells over-expressing HuR displayed increases in both numbers of GATA3-expressing cells, and in overall GATA-3 expression. The frequency of Th2-polarized cells, but not of Th1, increased in CD4+ T cell HuR transgenic cells and the levels of secreted IL-4 and IL-13, but not IFN-γ, also increased significantly in both activated splenocytes and polarized cells. Furthermore, a knockdown of HuR in Jurkat cells decreased protein levels of GATA-3. Hence, HuR may be functioning to coordinately posttranscriptionally regulate genes essential for Th2 polarization and function. Better understanding of posttranscriptional regulation may elucidate control mechanisms of naïve CD4+ Th2 polarization.
Cellular sentinels for Heat Shock Protein-peptide complexes in vivo. (100.1)
Authors: Messmer Michelle , Binder Robert 
Published: January 2023

Abstract
Heat shock proteins (HSPs) released into the extra cellular environment via cell death or through vaccination are taken up by antigen presenting cells of the immune system. HSP-chaperoned peptides are then cross-presented for priming of specific T cell responses. Our goal is to identify the subset(s) of cells that sample HSPs in vivo and then initiate T cell priming. By introducing a fluorescent labeled HSP into mice, cells in the draining lymph node that engage extracellular HSP have been identified. Our data demonstrate that CD4+CD11c+ dendritic cells (DC) are the predominant cells that internalize the HSP, while CD8+CD11c+ DCs did not significantly internalize the HSP. The localization of these cells within the lymph node and their ability to prime T cells are examined. These studies illustrate how HSPs act to alert the immune system of cellular damage and will be useful for optimizing HSP-based vaccination regimes in clinical studies of cancer patients.
Listeria-derived recombinant proteins are processed by a unique MHC class I processing pathway (100.10)
Authors: Wolf Benjamin , Princiotta Michael 
Published: January 2023

Abstract
Studies examining antigen processing in infected cells suggest that the efficiency of peptide-MHC class I (pMHC) production can differ by as much as 100 to 1000-fold depending on the source of the protein from which the peptide determinant is derived. For recombinant proteins expressed from vaccinia virus (rVV), ~3000 protein molecules must be degraded to generate one surface pMHC complex. By contrast, the measured efficiency of pMHC production from native Listeria monocytogenes proteins is ~1 surface pMHC complex per 3.5 to 35 protein molecules degraded. What accounts for this 100 to 1000-fold difference in efficiency remains unresolved. One possible explanation is that processing efficiency is a function of the peptide and/or protein studied. It is also possible that exogenously derived proteins, such as those secreted by Listeria, enter a different processing pathway than host-cell derived proteins. To better understand these differences in processing efficiency, we engineered Listeria strains to express recombinant proteins similar to those used for the rVV efficiency studies. We found that surface pMHC production was 20-fold more efficient from recombinant proteins expressed from Listeria than from a rVV-derived recombinant protein. These data suggest proteins expressed from Listeria follow a distinct and highly efficient processing pathway relative to host cell derived proteins.
Combinatorial peptide library scans of multiple HLA A*0201 restricted CTL reveal high levels of degeneracy at MHCI anchor positions (100.12)
Authors: Wooldridge Linda , Edwards Emily , Ekeruche Julia , Miles John , van den Berg Hugo , Clement Mathew , Sewell Andrew 
Published: January 2023

Abstract
CD8+ cytotoxic T lymphocytes (CTLs) hold the key to the successful prevention and eradication of infectious and neoplastic disease. T-cells recognize ‘foreign’ peptide fragments often referred to as epitopes, in the context of ‘self’ major histocompatibility complex class I (MHCI) molecules. Epitope identification is essential to define targets for the effective control of disease. Peptides bind to MHCI molecules by anchoring at position 2 and the C-terminus of the peptide ligand. Techniques such as pool sequencing have been hugely instrumental in revealing HLA allele-specific peptide binding motifs. However, this approach only reveals the most dominant MHC anchor residues. The preferred binding motif identified by pool sequencing for the most common MHCI, HLA A*0201, is L/M at position 2 and V/I/L/A at the C-terminus. Here, we use an extremely sensitive combinatorial library screening approach to show that in the context of HLA A*0201, degeneracy at anchor residue positions is much higher than previously appreciated. We also examined the recognition of peptides with substitutions of all 20 amino acids at both anchor positions in three different clonal systems which confirm these observations. Indeed, in one clonal system, 9-mer peptides containing A, C, F, I, K, L, M, Q, S, T, V in position 2 and A, C, F, I, L, M, T, V at position 9 were recognized efficiently. These observations have implications for T-cell epitope prediction and future vaccine design.
ERAAP-deficiency differentially affects peptide presentation across multiple MHC I (100.15)
Authors: Lind Kristin , Escobar Hernando , Reyes-Vargas Eduardo , Delgado Julio , Shastri Nilabh 
Published: January 2023

Abstract
Genome-wide association studies in humans have linked polymorphisms in the ER aminopeptidase associated with antigen processing (ERAAP) as well as particular MHC class I molecules to autoimmune diseases. How ERAAP polymorphisms affect function of different MHC I molecules is not known. ERAAP normally trims antigenic precursor peptides to generate potential peptide-MHC I ligands that can be recognized by CD8 T cells. To detect changes in the peptide repertoires displayed by different MHC I molecules, we immunized wild-type B10.D2 (H-2d) mice with ERAAP-deficient (ERAAP-KO) cells and analyzed the specificity of the CD8 T cell response. In wild-type mice, anti-ERAAP KO T cells responded primarily to peptides presented by Kd, and not the other two MHC I molecules, Dd and Ld of the H-2d haplotype. We also assessed the MHC I bound peptide repertoire by mass spectrometry. This analysis revealed that Dd and Kd molecules were bound to large number of shared as well as unique peptides, some with N-terminal extensions. Remarkably, Ld bound peptides were not detected in ERAAP-deficient mice. Thus, changes in ERAAP expression can have a major influence on the peptide repertoire presented by different MHC I molecules as well as on CD8 T cell responses.
Beryllium-specific T cells adopt an unusual binding topology for antigen recognition (100.25)
Authors: Bowerman Natalie , Falta Michael , Mack Douglas , Kappler John , Fontenot Andrew 
Published: January 2023

Abstract
Unconventional antigens, such as metals, stimulate T cells in a very specific manner. To delineate the binding landscape for metal-specific T cell recognition, alanine screens were performed on a set of beryllium (Be)-specific T cell receptors (TCR) derived from the lung of a chronic beryllium disease patient. These TCRs were HLA-DP2-restricted and expressed nearly identical Vβ5.1+ chains coupled with different Vα chains. Site-specific mutagenesis of all amino acids comprising the complementarity determining regions of TCR Vα and Vβ chains showed a dominant role for Vβ5.1 residues in Be recognition, with little contribution from the Vα domain, suggesting an abnormal binding topology. To confirm these findings, solvent-exposed residues along the α-helices of the α- and β-chains of HLA-DP2 (i.e., potential TCR contact sites) were mutated to alanine. Two β-chain residues, located near the proposed Be binding site of HLA-DP2, played a dominant role in T cell recognition, while the α-chain mutations had no effect on T cell activation. These findings strongly suggest that Be-specific T cells recognize antigen using an unconventional binding topology, with the majority of interactions contributed by residues in the TCR Vβ5.1 domain and HLA-DP2 β1-chain. We propose that unusual docking topologies are not used exclusively by T cells recognizing self-antigens, but also for the recognition of metal antigens, such as beryllium. Supported by NIH grants: HL062410 and HL092997.
Deficiency in Ab response to insulin in gamma-IFN-inducible lysosomal thiol reductase (GILT) knock-out mice (100.29)
Authors: He Xiao , Wang Xiaomin , Jensen Peter 
Published: January 2023

Abstract
The γ-IFN-inducible lysosomal thiol reductase (GILT) is a unique thiol reductase with optimal activity at low pH, and is expressed in Ag presenting cells (APC). GILT is critical to processing protein Ag by catalyzing disulfide bond reduction, thus facilitating the unfolding of native Ag containing disulfide bonds for further processing. GILT-/- mice are defective in processing exogenous protein Ag. However the role of GILT in Ab response is not clear. Here we report that after immunization, GILT-/- mice failed to produce Ab to exogenous bovine insulin (bIns), which has A and B chains and contains both inter and intra-chain disulfide bonds, while Ab response to OVA was normal in both GILT-/- and wt B6 mice. GILT-/- APC was able to present bIns to an insulin B chain peptide-specific T hybridoma, although it produced less amounts of IL2, compared to when using B6 APC. IL2 production was MHC-II mediated (could be blocked by α-mouse, but not α-human, MHC-II Ab) and the reduced IL2 production of GILT-/- cells was bIns-specific since GILT-/- APC could not present HEL, another Ag containing disulfide bonds, to an HEL-specific T hybridoma to produce IL2. This data indicates that the T cell response to bIns is only partially affected in GILT-/- mice, which was further supported by the fact that CFSE labeled, in vivo bIns primed, GILT-/- CD4 cells proliferated equally well as wt B6 CD4 cells upon restimulation by bIns. Our data indicates a novel role of GILT in Ab response to insulin.
Microparticles released by Listeria monocytogenes-infected macrophages contribute to the generation of protective immunity: insight into interaction between macrophages and dendritic cells (100.33)
Authors: Huang Bo , Zhang Yi , Liu Jing 
Published: January 2023

Abstract
Mutual interaction among immune cells is of paramount importance to an efficient protective immunity against bacterial infection. However, the interplay between macrophages and dendritic cells (DC) in the processing and presentation of bacterial antigens for T cell immune responses remains poorly understood. In this report, using Listeria monocytogenes (Lm) infection model, we demonstrate that DCs require the support of macrophages to elicit the protective immunity against Lm infection. We found that DCs themselves were inefficient at taking up Lm but capable of taking up microparticles released by Lm-infected macrophages. These microparticles transferred Lm antigens to DCs, allowing DCs to present Lm antigen to effector T cells. Such microparticles-mediated Lm antigen transfer required MHC class I participation, since MHC class I deficiency in macrophages resulted in a significant reduction of T cell activation. Moreover, the vaccination of mice with microparticles from Lm-infected macrophages produced strong protective immunity against the challenge of Lm. Thus, we have identified an intrinsic antigen transfer program between macrophages and DCs during Lm infection, and emphasized that macrophages also play an essential role in DC-elicited Lm-specific T cell responses.
Obese concentrations of leptin enhances dendritic cell indution of T cell activation (100.37)
Authors: Radovich Abraham , Garza Kristine , Gonzales Amanda , Delgado Christine 
Published: January 2023

Abstract
The treatment with leptin has shown an increase in the functionality in DCs as they produce longer dendrites, however, the interaction with T-cells and the migration in vivo have not been studied. Therefore the relevance of the investigation conveys immunological responses. The increase in T-cell interaction with the dendritic cells, will lead to a quicker migration in vivo of the DCs to the high concentration areas. The decrease in time of activation will then optimize the immunological response. The treatments of the DC are to be performed by concentration seen in obese conditions, that way the interaction of leptin can be assured. The usage of KODAC imager will demonstrate the increase of T-cell/DC interaction. The experiments demonstrated an increase of T-cell interaction with the DC. Also, the leptin increase the functionality of the DC lines, as could be seen to have longer and more prominent dendrites. The profileration of DC was also seen as they were treated in leptin and LPS. The treatment of DC with leptin, was tested to enhance the T-cell/DC activation and the results correlated the fact that dendritic cells migrate faster in vivo as they are treated with leptin. The fact that it can be demonstrated that the DCs, involved in immunological responses, can be enhance will open the path to bettering the immunological responses in order to be more effective in less time.