bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Systemic age dependent defect in immune-cell signaling response induced by inflammation (138.2)
Authors: Shen-Orr Shai , Furman David , Kidd Brian , Maecker Holden , Dekker Cornelia , Butte Atul , Davis Mark 
Published: January 2023

Abstract
Immune system function generally degrades with age and is associated with increased risk of infection and disease. Though differences between young and old have been noted in many immune system components, no system wide understanding of how these disparate observation act together exists to date, nor how they relate to genes found to associated with increased longevity, many of which are immune related. Here we characterize the immune system of 29 young and old individuals by concurrently measuring from peripheral blood, immune cell subset frequency, serum cytokines, gene expression and individual cellular responses to cytokine stimuli by pathway specific phospho-protein abundance. We identify age-dependent changes in STAT signaling baseline and in response to stimulation by a panel of 7 different cytokines, particularly prominent in CD8 and CD4 T cells, but also in monocytes and B-cells. The observed differences in cellular responses are not due to adaptation to higher level of cytokine stimuli but rather to an inert inability to mount a full response, many times augmented by a higher base line phosphorylation level in the elderly. We construct an immune network spanning multiple biological layers and identify co-occurring modules which link longevity associated genes with these cellular immune phenotypes. We quantify the contributions of these modules to the observed reduction in cellular response to stimuli in the elderly and suggest a common responsible mechanism.
Long lasting macrophage cytokine suppression by alpha 7 nAChR requires JAK/STAT (138.23)
Authors: Olofsson Peder , Rosas-Ballina Mauricio , Chavan Sangeeta , Parrish William , Hudson LaQueta , Ochani Mahendar , Li Jianhua , Pavlov Valentin , Tracey Kevin 
Published: January 2023

Abstract
The central nervous system regulates innate immunity reflexively, as action potentials transmitted in the vagus nerve suppress cytokine release during endotoxemia. The mechanism of cytokine release by this neural circuit requires signaling through the alpha 7 nicotinic acetylcholine receptor (a7 nAChR), but the duration of this protection is incompletely understood. Electrical vagus nerve stimulation (VNS) in mice significantly inhibited serum TNF levels during endotoxemia, even when endotoxin was given 2, 24 or 48 h after VNS. This inhibition of TNF was abolished in a7 nAChR deficient mice, confirming the mechanism that requires this receptor. One-hour exposure of human macrophages to acetylcholine in vitro reduced endotoxin-induced increases in IKKbeta, IkappaBalpha activity and TNF release for up to 24 h after removal of acetylcholine. Levels of Supressor of Cytokine Signaling 1-3, A20, BCL3, IkappaBNS and IkappaBzeta were not altered in human macrophages 15 min to 8 h after exposure to acetylcholine, but exposure of macrophages to the selective JAK/STAT inhibitor AG490 abolished the cytokine suppressing effect of acetylcholine. Together with previous work indicating that JAK/STAT forms a signaling complex with a7 nAChR, these results suggest that JAK/STAT activation is required for long lasting suppression of macrophage cytokine release after transient exposure to acetylcholine.
Notch signaling regulates human and mouse TH17 development. (139.13)
Authors: Keerthivasan Shilpa , Suleiman Reem , Le Poole Caroline , Nickoloff Brian , Osborne Barbara , Miele Lucio 
Published: January 2023

Abstract
TH17 immune cells, a recently discovered subset of CD4+ T-helper cells, are known to play important role in pathogenesis of several autoimmune diseases, inflammatory disorders and cancers. Although Notch ligand DLL4 was shown to drive TH17 polarization, downstream pathways conducting those signals are unknown. We demonstrated that the Notch signaling pathway has an important role in TH17 polarization. We found that gamma secretase inhibitors (GSI) that inhibit Notch signaling blocked the in-vitro polarization of mouse or human naïve CD4+ T cells towards the TH17 pathway without affecting cell proliferation. Next, we showed that, among various Notch receptors, Notch-1 is upregulated in TH17 polarizing cells and that knocking down Notch-1 by siRNA inhibited TH17 polarization. We further discovered that, in addition to polarizing naïve T cells towards TH17 pathway, Notch-1 is important for maintenance of this phenotype. Furthermore, reporter luciferase assays and chromatin immunoprecipitation (ChIP) assays suggest that the IL-17 as well as RORC promoters are direct transcriptional targets of Notch-1. Finally, in-vivo administration of GSI inhibits TH17 mediated disease progression in mouse experimental autoimmune encephalomyelitis model of multiple sclerosis. Thus our data suggest a key role for Notch signaling, especially Notch-1, in TH17 polarization.
Signaling through the membrane-bound estrogen receptor GPR30 induces IL10 and IL17A expression in Th17 cells (139.2)
Authors: Brunsing Ryan , Hathaway Helen , Prossnitz Eric 
Published: January 2023

Abstract
In 2005, the orphan G protein-coupled receptor GPR30 was shown to bind estrogen and mediate some downstream events associated with estrogen signaling. While work on GPR30-mediated suppression of autoimmune inflammation has been published, no definitive work on GPR30 in T cells has been reported. To address whether signaling through GPR30 can affect CD4+ T cell populations, CD4+CD62Lhi naïve T cells were stimulated in culture with anti-CD3/CD28 antibodies. We observed a dose-dependent increase in the number of IL10+ and IL10+IL17A+ cells in cultures treated with the GR30-directed agonist G-1, as measured by FACS. This effect is lost in cells from GPR30KO mice. Furthermore, the induction of IL10 and IL17A was limited to the RORγt+Foxp3- (Th17) population, while no effects were observed in Foxp3+ cells. Interestingly, the effects of systemic G-1 in the animal model T-cell mediated colitis varied between the peripheral compartment, where IL17A induction was again observed, and the GI mucosa, where a decrease in the number of IL17A+, IFNγ+, and TNFα+ cells was observed. These findings provide insight into the function of the membrane-bound estrogen receptor GPR30 in CD4+ T cell physiology. In addition, these data may help reconcile an apparent contradiction between work suggesting an immunosuppressive role for estrogen in several autoimmune disease models and clinical reality where women suffer from a substantially higher incidence of autoimmune disease.
Regulatory T cell hybridoma reveal novel Foxp3 inducing factors (139.7)
Authors: Sharma Rahul , Ju Angela , Sung Sun-Sang , Ju Shyr 
Published: January 2023

Abstract
It has been difficult to generate cloned regulatory T cells (Tregs) or Treg hybridoma for the studies on antigen specificity and the regulation of Foxp3 by TCR independent factors. Here, we report the first successful generation of cloned, inducible Treg hybridomas (iTreg-hyb) using the CD4+ T cells of TCR transgenic DO11.10 mice bearing the Foxp3-IRES-eGFP knock-in gene. Although, the iTreg-hyb expressed low Foxp3-GFP, strong expression could be induced by either polyclonal (using anti-CD3 mAb) or antigen specific (using OVA[323-339] peptide presented by syngeneic Rag2 deficient spleen cells) TCR stimulation in the presence of Interleukin (IL) -2 and Transforming Growth Factor-β1. Interestingly, CD11c+ cells isolated from the spleens of Rag2 deficient mice alone could induce Foxp3-GFP expression, albeit with 30-fold lower efficiency. The induction of Foxp3-GFP by the latter mechanism was specific as it did not up-regulate the TCR stimulation dependent Treg associated markers. Using the iTreg-hyb, we have identified several Foxp3 inducing factors which include recombinant IL-6, IL-9, IL-27, and a &lt;5-kd soluble factor produced by Rag2 deficient spleen cells. Our study shows that it is feasible to generate iTreg-hyb, which is useful not only to study the role of self-reactivity but also to uncover TCR independent pathways for the induction of Foxp3 expression by cytokines and novel factors.
Critical role of Ral GTPase as a gatekeeper in controlling neutrophil selective degranulation of secondary granules (140.11)
Authors: Chen Celia , Soto Ileana , Zen Ke , Liu Yuan 
Published: January 2023

Abstract
Neutrophil (PMN) inflammatory responses, including cell adhesion, diapedesis and phagocytosis, are dependent on the mobilization and release of various intracellular granules and vesicles. In this study, we observed that inhibition of the Ras family GTPases by damnacanthal and manumycin A resulted in specifically release of PMN secondary granules, but not primary or tertiary granules, in the absence of stimulation. By screening Ras family members for activity changes during PMN activation and degranulation, we identified Ral GTPase to be a critical regulator that selectively controls secondary granule release. In particular, we found that Ral exists as an active GTPase in resting PMN, while quick inactivation correlates with chemoattractant-induced degranulation. Indeed, we demonstrated that over-expression of a constitutively active Ral mutant (Ral23V) in PMN completely blocked secondary granule release. Subcellular fractionation and immunofluorescence labeling further found that Ral is associated with the plasma membrane and redistributed to secondary granules following chemoattractant stimulation. Blocking clathrin-mediated plasma membrane endocytosis, however, inhibited Ral intracellular translocation and PMN degranulation. Thus, these results provide strong evidence demonstrating Ral to be an essential gatekeeper for secondary granules in PMN. Dysregulation of Ral results in impairment of PMN transmigration in response to inflammatory stimuli.
Interaction between mouse CD99 and CD99L2 proteins influences cellular localization of each protein in the cells and cell-cell interaction. (140.14)
Authors: Ma Min-Jung , Lee Hae-Young , Choi Eun-Young 
Published: January 2023

Abstract
CD99 and CD99 like 2 protein (CD99L2) are membrane glycoprotein leukocyte antigens that do not belong to any known protein family. They are expressed at the endothelial cell junction and are known to participate in the diapedesis process. Using mouse CD99 and CD99L2 gene, we show that CD99 interact with CD99L2. To examine molecular interaction between mouse CD99 and mouse CD99L2 and to localize the site of the interaction, we used a bimolecular fluorescence complementation (BiFC) assay via flow cytometry and confocal analysis. The interaction between mouse CD99 and mouse CD99L2 was verified by isothermal tiltration calorimetry (ITC) and coimmunoprecipitation. The interaction between CD99 and CD99L2 proteins is mediated by cytoplasmic domain of CD99. Their interaction enhanced the localization of each protein to the cell surface membrane and also increased cell-cell interaction via homophilic way. The results from this study suggest that the interaction of CD99 and CD99L2 may control the location and functions of CD99 and CD99L2 as cell adhesion molecules. *This study is supported by a fund from national research foundation (2007-2003619), republic of Korea.
Monocyte trafficking to hepatic sites of bacterial infection is chemokine-independent and directed by focal ICAM-1 expression (140.2)
Authors: Shi Chao , Velázquez Peter , Hohl Tobias , Leiner Ingrid , Dustin Michael , Pamer Eric 
Published: January 2023

Abstract
Recruitment of CCR2+Ly6C+ monocytes to sites of infection is essential for efficient clearance of microbial pathogens. While CCR2-mediated signals promote monocyte emigration from bone marrow, the contribution of CCR2 to later stages of monocyte recruitment remains unresolved. Herein, we show that CCR2 deficiency markedly worsens hepatic L. monocytogenes infection because Ly6C+ monocytes are retained in the bone marrow. Intravenously transferred, CCR2-deficient Ly6C+ monocytes traffic normally to hepatic foci of infection and contribute to bacterial clearance. Pertussis toxin treatment of adoptively transferred monocytes does not impair their intrahepatic trafficking, suggesting that chemokine signaling, once CCR2+Ly6C+ monocytes emigrate from the bone marrow, is not required for monocyte localization to sites of bacterial infection in the liver. Expression of ICAM-1 is induced in close proximity to foci of bacterial infection in the liver and blockade of CD11b and CD44 diminishes monocyte localization to these hepatic foci. Our studies demonstrate that Ly6C+ monocyte recruitment from the bloodstream to the L. monocytogenes-infected liver does not require chemokine receptor mediated signals but instead is principally dependent on integrin- and extracellular matrix-mediated monocyte adhesion.
Long-term immune characteristics of cow's milk allergy in a mouse model (141.12)
Authors: Gangur Venu , Gonipeta Babu , Parvataneni Sitaram 
Published: January 2023

Abstract
Specific changes in immune memory underlying the ability to outgrow cow’s milk allergy (CMA) are incompletely understood. We studied the long-term immune characteristics of CMA in an adjuvant-free mouse model we have recently published (Gonipeta et al 2009. J. Dairy Sci. 92:4738-4744). Thus, BALB/c mice were sensitized to cow’s milk whey protein using a transdermal sensitization protocol. Subsequently, exposure to whey protein was withdrawn for 3, 5 or 8 months. Fate of circulating IgE and IgG2a antibodies was monitored. Subsequently, mice were given booster exposures and examined for memory IgE, IgG2a, and spleen cell IL-4 and IFN-γ responses. Following allergen withdrawal, IgE antibody levels began to drop. Whereas, significant memory IgE response to booster exposures was evident after 3 months of allergen withdrawal, a weak or no memory IgE response was noted after 5 or 8 months of allergen withdrawal. Despite booster exposures, spleen cell memory IL-4 responses to whey protein was absent after 3, 5 or 8 months of allergen withdrawal. Milk sensitized mice, upon 8 months of allergen withdrawal, exhibited elevated IFN-γ and IgG2a responses suggesting a shift to a TH1 immune profile. In summary, these data suggest that: (i) upon withdrawal of allergen exposure, whey protein specific memory IgE, and memory IL-4 do not persist for long periods; and (ii) there occurs a shift in immune memory towards elevation of TH1 responses as mice age. Funding: US EPA STAR Grant#R833133
Role of T-cells in diesel particle exacerbation of allergen-induced pulmonary inflammation (141.2)
Authors: Brandt Eric , Lee Gerald , Gibson Aaron , Bass Stacey , Khurana-Hershey Gurjit 
Published: January 2023

Abstract
The mechanisms underlying the adjuvant effect of diesel exhaust particles (DEP) on allergen-induced airways responses are not well understood. We aim to define how T-cells are affected by exposures to DEP and house dust mite (HDM). Balb/c mice were exposed intra-tracheally to DEP, HDM, or both, three times a week for three weeks. Airway inflammation and FACS analysis of lung and draining lymph node T-cells were assessed 24h or 1 week after the last exposure. Exposure to DEP alone generated airway neutrophilia whereas exposure to HDM was characterized by eosinophilia. Coexposure to HDM and DEP resulted in a significant increase in BALF cells over HDM alone. The percentage of CD4+ T-cells expressing the activation marker CD69 were increased in all three groups over saline exposed mice. However, significantly more activated CD4+ T-cells were observed in the lungs and mediastinal lymph nodes of mice coexposed to HDM+DEP. Alterations in Foxp3+ regulatory T-cells were limited following coexposure to HDM and DEP resulting in a favorable ratio of activated T-cells over Tregs. One week after the last exposure, pulmonary inflammation and DEP containing macrophages could still be observed. Interestingly, levels of activated T-cells remain elevated in all three groups compared to saline exposed control mice, but the further increase observed after coexposure was no longer significant. Taken together, these results suggest that DEP exacerbates the early acute allergic responses.
Resveratrol induces myeloid-derived suppressor cells (MDSCs) and down-regulates CXCR3+ T cell expression and NF-kB activity to abrogate chronic colitis in IL-10-/- mice (142.10)
Authors: Singh Udai , Singh Narendra , Singh Balwan , Hofseth Lorne , Price Robert , Nagarkatti Mitzi , Nagarkatti Prakash 
Published: January 2023

Abstract
Inflammatory bowel disease, a chronic mucosal inflammation, is caused by hyperactivation of effector immune cells, which produce pro-inflammatory cytokines. Macrophages, along with Gr-1+ CD11b+ cells known as myeloid-derived suppressor cells (MDSCs), are known for their immunosuppressive function. Resveratrol is one of the most promising natural molecules for the treatment of chronic inflammatory diseases. In the present study, we investigated the resveratrol-mediated cellular and molecular mechanisms of chronic colitis inhibition in IL-10-/- mice. Resveratrol effectively attenuated various pathological markers of colitis by down-regulating Th1 responses. Resveratrol reversed colitis-associated decreases in body weight and increased levels of serum amyloid A and colon TNF-α, IL-6, RANTES, IL-12, and IL-1β. After resveratrol treatment, the percentage of CD4+ T cells that expressed CXCR3+ was decreased in the spleen, mesenteric lymph nodes (MLNs), and intestinal lamina propria (LP). However, the percentages of MDSCs in LP lymphocytes and spleens were increased after resveratrol treatment as compared with controls. Interestingly, resveratrol treatment after chronic colitis significantly reversed NF-κB activity in the colon. This study demonstrates for the first time that resveratrol protects against chronic colitis in IL-10-/- mice by inducing MDSCs, down-regulating mucosal and systemic effector CD4+T cells that express CXCR3+, and reducing NF-κB activity in the colon LP.
The splicing factor ASF/SF2 regulates the expression of the human T cell receptor CD3 zeta chain (143.11)
Authors: Moulton Vaishali , Tsokos George 
Published: January 2023

Abstract
T cells from patients with systemic lupus erythematosus (SLE) express decreased levels of the T cell receptor associated (TCR)-associated CD3 zeta (ζ) chain, a feature directly linked to their aberrant function. The decrease in CD3ζ protein expression is in part due to decreased levels of the functional wild-type (WT) isoform of the 3` untranslated region (UTR) of CD3ζ mRNA with concomitant increased levels of an unstable alternatively spliced (AS) isoform. In order to identify factors involved in the post transcriptional regulation of CD3ζ, we performed mass spectrometric analysis of Jurkat T cell nuclear proteins “pulled down” by a CD3ζ 3`UTR defined oligonucleotide which identified the splicing protein Alternative Splicing factor / Splicing factor 2 (ASF/SF2). We show for the first time that ASF/SF2 binds specifically to the 3`UTR of CD3ζ and regulates expression of CD3ζ protein by limiting the production of the alternatively spliced isoform. During activation of human T cells, an increase in the WT CD3ζ mRNA is associated with increased expression of ASF/SF2. Finally, we show a significant correlation between ASF/SF2 and CD3ζ protein levels in T cells from SLE patients. These results identify ASF/SF2 as a novel factor in the regulation of CD3ζ expression in human T cells, and a potential molecular mechanism underlying the altered CD3ζ chain expression in SLE.
Non-MHC genes increase susceptibility to autoimmune myocarditis by driving T cells towards a Th17 response (143.14)
Authors: Chen Ping , Ligons Davinna , Baldiviano G. , Talor Monica , Zheng Dongfeng , Barin Jobert , Rose Noel , Cihakova Daniela 
Published: January 2023

Abstract
MHC genes do not fully explain susceptibility to experimental autoimmune myocarditis (EAM) since A.SW and B10.S mice share the same MHC (H-2S) haplotype, yet A.SW is susceptible to EAM while B10.S is resistant. These two inbred mice strains provide an opportunity to study myocarditis susceptibility factors while excluding the effects of MHC-associated genetic components. We induced EAM in A.SW and B10.S mice by immunizing with murine cardiac myosin on day 0 and 7. A.SW displayed severe myocarditis compared to B10.S (p=0.008). The number of CD4+ T cells from heart and myosin specific T cells proliferation were greater in A.SW (p&lt;0.001). A.SW and B10.S had different proportions of T helper (Th) subsets in their hearts after immunization; A.SW had more Th17 cells (7.05±1.25 vs. 2.24±1.05, p=0.003) and B10.S more Th1 cells (8.94±1.26 vs. 2.9±1.06, p=0.001). In support of these in vivo data we found that CD4+ T cells from naïve A.SW have higher potential to differentiate towards the Th17 pathway than CD4+ T cells from naïve B10.S (11.93±3.72 vs. 5.5±1.14, p=0.04). In contrast, B10.S T cells differentiated towards a Th1 phenotype compared with that from naïve A.SW (12.73±0.61 vs. 8.14±0.85 p=0.002). Additionally, A.SW have a lower proportion of regulatory T cell in their hearts (p=0.002) and spleens (p&lt;0.001) compared to B10.S. Our data suggest that the tendency of A.SW differentiate towards a Th17 phenotype contributes to their heightened susceptibility of autoimmune myocarditis.
Role of IDO in B Cell-Mediated Immune Responses (143.2)
Authors: Mandik-Nayak Laura , Pigott Elizabeth , Galanti Rebecca , Ridge Natalie 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is a chronic and debilitating inflammatory autoimmune disease of unknown etiology. As with other autoimmune disorders, elevated tryptophan catabolism has been detected in RA patients. This is indicative of activation of the immunomodulatory enzyme indoleamine 2,3-dioxygenase (IDO); however, the role that IDO plays in the disease process is not well understood. The current paradigm for IDO function is immunosuppressive, providing an important inhibitory role to limit effector T cell responses. In contrast, the role of IDO in B cell-mediated responses has not been addressed. The widely held belief that IDO acts solely in a T cell immunosuppressive role has led to the general assumption that inhibition of IDO activity should exacerbate autoimmune disorders. However, using the K/BxN mouse model of RA and 1-methyl-tryptophan (1MT), an inhibitor of IDO, we found that inhibiting IDO activity had the unexpected consequence of ameliorating, rather than exacerbating arthritis symptoms. This alleviation of joint inflammation was not due to an altered T cell response, but rather resulted from a diminished autoreactive B cell response. We have preliminary data demonstrating that 1MT is also able to inhibit B cell responses to immunization with model antigens. These data suggest for the first time an important role for IDO in directing B cell responses to both self and foreign antigens.