bluuebunny/crossref_metadata_2025_split huggingface dataset:
Blockage of of Gi linked G-protein coupled receptor (GPCR) signaling reduces local and inhibits systemic tissue injury in the mesenteric IR model. (B74)
Authors: Moratz Chantal M , Egan Ryan , Zacharia Athina , Tsokos George 
Published: January 2023

Abstract
Ischemia-reperfusion (IR) is a frequent clinical entity encountered in diverse fields such as vascular surgery, trauma, myocardial infarction, hemorrhage, stroke, and multi-organ failure. The acute inflammatory response following an ischemic insult instigates pathology in local and remote organs leading to significant morbidity and mortality. IR induces injury in a biphasic pattern. The acute phase is characterized by oxidative tissue injury resulting in activation of intracellular damage pathway and antibody-mediated complement activation. The sub acute phase is characterized by leukocyte infiltration. It has been shown previously in several models of IR that complement inhibitors reduce tissue injury. In this study the role of G-protein-coupled receptor signaling was examined. Studies in the mesenteric IR model a global Gi inhibitor, pertussis toxin, attenuated local intestinal tissue injury and production of LTB4 and PGE2. In these animals the systemic inflammatory response was also inhibited, assessed by decrease lung tissue injury and blocked cellular infiltrate. Next specific receptor inhibitors such as blocking antibodies to examine the effectiveness of inhibiting chemokine receptors such as CXCR2 or blocking adhesion molecules such as ICAM-1 or P-selectin were utilized. The data indicate specific chemokine receptors are more efficient in inhibiting tissue damage. Comparison of these inhibitors will be useful in developing therapeutics which can control deleterious effects of systemic inflammation after an initial trauma.
USAMRAA W81xWH0610485
Fatal autoimmune disease in scurfy mice is initiated and sustained by conventional effector CD4+ T cells and not by cells that express mutant Foxp3 gene and follow the developmental program of regulatory CD4+ T cells. (B81)
Authors: Kraj Piotr Jan , Kuczma Michal , Mazurek Maciej , Ignatowicz Leszek 
Published: January 2023

Abstract
To determine features of natural regulatory T cells (Treg) controlled by Foxp3 we investigated a subset of CD4+ T cells identified in scurfy mice expressing transcript of mutant, non-functional allele of the Foxp3 gene (Foxp3scurfy+ cells). Foxp3scurfy+ cells poorly responded to TCR stimulation in vitro, produced low level of cytokines including Il-2 and remained dependent on exogenous Il-2 for proliferation. In vivo Foxp3scurfy+ cells did not expand relative to other CD4+ T cells and expressed lower levels of activation markers than conventional CD4+ T cells. Thus, Foxp3scurfy+ cells retained the properties of Treg cells despite losing suppressor function suggesting that Foxp3 is not necessary for the development of Treg cells and that the developmental program followed by these cells is not superseded by inflammatory environment. In contrast, conventional CD4+ T cells expressed highly activated phenotype and produced very high level of Il-2 and inflammatory cytokines. In conclusion, the loss of Foxp3 by Treg cells does not convert them into aggressive, self-reactive T cells. Our results identify conventional CD4+ T cells as the population that initiates and sustains autoimmune disease.
Therapeutic effect of Jagged1 DNA vaccine on collagen-induced arthritis (B86)
Authors: Kijima Mika , Yasutomo Koji 
Published: January 2023

Abstract
Notch signaling plays an important role in functional differentiation of mature T cells. Those findings have allowed us to examine whether modification of Notch signaling has therapeutic effects on immune-associated diseases. We here examined whether DNA vaccine of Jagged1, one of the Notch ligands, is able to prevent collagen-induced arthritis (CIA) in mice. The CIA was induced in male DBA/1J mice by immunizing bovine collagen type II. Those mice were vaccinated with extracellular region of Jagged1 (soluble form: sJagged1) gene three times every one week starting at 2 days before initial immunization. The mice vaccinated with sJagged1 showed reduced clinical score and disease incidence compared with control mice. Histological analysis also showed reduced infiltration of inflammatory cells in the joints by sJagged1-treated mice. The vaccination with sJagged1 did not affect the numbers of regulatory T cell and production of IL-10 from T cells. The coimmunization of OVA and full length Jagged1 reduced the number of OVA specific and granzyme B producing CD8+ T cells. These results indicate that Jagged1 suppresses antigen-specific CD8+ T cell functions resulted in reduced CIA score, suggesting that Jagged1 has the therapeutics effects on autoimmune diseases by suppressing T cell functions.
Anti-desmoglein autoantibody repertoire in pemphigus (B89)
Authors: Qian Ye , Clarke Stephen H , Diaz Luis A 
Published: January 2023

Abstract
Pemphigus foliaceus (PF) and pemphigus vulgaris (PV) are autoimmune skin blistering diseases characterized by pathogenic autoantibodies against desmoglein 1 (Dsg1) and desmoglein 3 (Dsg3), respectively. Fogo selvagem (FS) is the endemic form of PF. To study the etiology of PV and FS, hybridomas were generated from the B cells of 5 PV patients and 7 FS patients. The V genes of both H and L chain of the Dsg-specific autoantibodies from these hybridomas were sequenced and analyzed for the H and L chain pairing of the autoantibodies, the diversity of H and L chain V gene usage, and the extent of somatic mutation. The number of somatic mutations in the expressed genes ranged from zero to 42, and the replacement to silent mutation (R/S) ratio was much higher in CDRs than in FWRs of both IgM and IgG hybridomas from FS patients. The R/S ratio was also much higher in CDRs than in FWRs of IgG hybridomas from PV patients. In addition, clonally related sets of anti-Dsg1 or anti-Dsg3 hybridomas were identified from individual FS or PV patients based on VH-D-JH junctional sequence identity that reveals ongoing somatic mutation during clonal expansion. Thus, our evidence suggests that the anti-Dsg autoimmune response in both PV and FS is antigen selected.
The regulatory role of naturally occurring CTLA-4 splice variants:the suppressive function of short splice variant 1/4 WT CTLA-4 (B91)
Authors: Zhang Zheng , Zhu Chen , Xiao Sheng , Chung Denise , Wang Yue , Rainbow Dan , Wicker Linda S , Sharpe Arlene H. , Kuchroo Vijay K. 
Published: January 2023

Abstract
Cytotoxic T lymphocyte Antigen 4 (CTLA-4) is an important cell surface molecule that inhibits T cell activation. Four splice variants of CTLA-4 have been identified, indicating the regulatory complexity of CTLA-4. In this study, we characterized the biology of the shortest splice variant 1/4WT CTLA-4 and its functional correlation to full length (FL CTLA-4). 1/4WT CTLA-4 mRNA was found in quiescent CD4+ T cells and upregulated after immunization with MOG35-55 peptide. The kinetic expression of 1/4WT was opposite to FL CTLA-4. To test functions of 1/4WT CTLA-4, we transduced CD4 T cells with a retroviral vector coexpressing 1/4WT CTLA-4 and the green fluorescent protein (GFP). FL CTLA-4 was downregulated on the cell surface by 1/4WT overexpression, which consequently enhanced antigen specific T cell responses by enhancing T cell proliferation and cytokine production in vitro. This functional correlation between 1/4WT and FL CTLA-4 had profound influence on effector CD4 T cell function. Mice adoptively transferred with MOG35-55 specific CD4 T cells containing ectopically expressed 1/4WT CTLA-4 showed dramatically exacerbated EAE disease, increased CD4 T cell proliferation, and increased IFN gamma and IL-17 production. These data suggest that 1/4WT CTLA-4 plays an important role in regulating T cell activation by affecting expression and/or function of FL CTLA-4.
15-deoxy-12,14-prostaglandin J2 and curcumin regulate the expression of toll-like receptors in autoimmune T lymphocytes (B97)
Authors: Chearwae Wanida , Mo Caiqing , Bright John J 
Published: January 2023

Abstract
Experimental allergic encephalomyelitis (EAE) is a T cell-mediated autoimmune disease model for multiple sclerosis (MS). Toll-like receptors (TLR), expressed primarily on innate immune cells, play critical roles in the pathogenesis of EAE. We have shown recently that 15-deoxy-Δ12,14-prostaglandin J2 (15d-PGJ2) and curcumin ameliorate EAE through modulating inflammatory signaling pathways in T lymphocytes. In this study we have examined the effects of 15d-PGJ2 and curcumin on the expression of TLRs in T cells in EAE. We induced EAE in SJL/J and C57BL/6 mice by immunization with PLPp139-151 and MOGp35-55 antigen respectively. The mice were treated with DMSO, 15dPGJ2 or curcumin on every other day and the spleen cells isolated on day 14 were used fresh or after culture with antigen for 3 days. The cells were stained with TLR4, TLR9, CD4 or CD8 antibodies and analyzed by flow cytometry. Our results show that naïve CD4+ and CD8+ T cells express detectable levels of TLR4 and TLR9 and that increase significantly following induction of EAE. Interestingly, in vivo or in vitro treatment with 15d-PGJ2 or curcumin differentially regulate the frequency and intensity of TLR4 and TLR9 expression in CD4+ and CD8+ T cells in EAE. Although the exact mechanisms are not known, the regulation of TLR expression in T lymphocytes by 15d-PGJ2 and curcumin suggests new therapeutic targets in the treatment of T cell-mediated autoimmune diseases.
The Rate of Absorption of Ragweed Pollen Material from the Nose
Authors: Cohen Milton B. , Ecker E. E. , Breitbart J. R. , Rudolph J. A. 
Published: January 2023

Summary
A Prausnitz-Küstner reaction was performed with the serum of a patient highly sensitive to ragweed on 50 non-atopic subjects and 12 patients with asthma, hay fever, or vasomotor rhinitis, nine of whom gave skin reactions by scratch tests. The patients were not naturally sensitive to ragweed. Twenty-four hours later the pollen was blown into both nostrils of the non-atopic and atopic subjects and the sites of injections observed for the appearance of redness and wheal formation. The absorption time of the non-atopic subjects varied from fourteen minutes to thirty-one minutes, while that of the 9 sensitive subjects giving a skin reaction showed a marked delay, ranging from forty-seven minutes to no reaction after two hours. The other three with negative skin tests reacted like the non-atopics.
Signaling by the serine/threonine kinase Drak2 in T cells (35.23)
Authors: Newton Ryan H. , Walsh Craig 
Published: January 2023

Abstract
A role for Drak2 in negative regulation of T cell activation has been well established, yet the manner in which Drak2 mediates downstream events to limit the spurious activation of T cells remains to be elucidated. While T cells from mice deficient in Drak2 exhibit a decreased activation threshold and hyper-respond to sub-optimal stimuli, the proliferative capacity of these cells is diminished as result of a deficiency in survival. A resistance to the induction of EAE in Drak2-deficient mice underscores a dependence on negative regulatory molecules to enforce the requirement of co-stimulation to engage an effective immune response. Biochemical data tracking Drak2 activation has revealed that protein kinase D (PKD) is required for Drak2 signaling. PKD has been shown to respond to the release of calcium from intracellular stores as well as to oxidative stress, two events coupled to T cell activation through the processes of IP3R-mediated ER-calcium depletion and increased mitochondrial respiration. The signaling requirements of Drak2 similarly include both store-operated calcium entry and the production of reactive oxygen species. The data presented indicate that Drak2 and PKD constitute a signaling axis that defines a novel role for PKD in T cell activation and further sheds light on how Drak2 orchestrates decreased responsiveness as a negative regulatory molecule.
The SNARE protein syntaxin 7 is required for immunological synapse formation in cytotoxic T lymphocytes (35.24)
Authors: Pattu Varsha , Qu Bin , Becherer Ute , Matti Ulf , Schwarz Eva , Marshall Misty , Krause Elmar , Hoth Markus , Rettig Jens 
Published: January 2023

Abstract
Cytotoxic T lymphocytes (CTLs) kill target cells by secretion of cytotoxic components such as perforin and granzymes which are contained in lytic granules. Fusion of lytic granules occurs at the contact zone between the target cell and the CTL, the immunological synapse (IS). T cell receptor (TCR) enrichment at the IS is one of the key early events of IS formation. Soluble NSF attachment receptor (SNARE) proteins are required for all fusion events in cells, but the individual SNARE proteins that are important for CTL function are not yet known. We identified syntaxin 7 in CTLs by reverse transcriptase PCR and immunocytochemistry. We found that syntaxin 7 is localised to the IS with a similar distribution to the lytic granule marker - perforin. We then blocked the function of syntaxin 7 by deleting its transmembrane domain and observed a complete loss of TCR accumulation at the IS, indicating that no IS is formed. These results imply that syntaxin 7 is required for IS formation in CTLs. Further studies using specific endosome markers will help us determine the mechanism of syntaxin 7 function in CTLs.
Source of research support - GRK1326
Characterization of SNARE proteins involved in granule exocytosis of cytotoxic T lymphocytes (35.25)
Authors: Marshall Misty R , Pattu Varsha , Qu Bin , Hoth Marcus , Rettig Jens 
Published: January 2023

Abstract
Cytotoxic T lymphocytes (CTLs) exert their cytotoxic activity through the polarized secretion of cytotoxic granules at the immunological synapse (IS). Because soluble NSF attachment receptor (SNARE) proteins mediate numerous intra- and intercellular fusion events and are essential effectors of exocytosis of synaptic granules in neurons, we examined the contribution of specific SNARE proteins to the formation of the IS. To determine the functional significance of specific SNAREs, our lab has examined the expression profile of more than 40 SNARE proteins by RT-PCR and quantitated various protein levels of these SNAREs in resting and activated CTLs. We have found that the expression profile of some SNARE proteins is up or down-regulated upon CTL maturation. In conjunction with these studies, we have also analyzed the sub-cellular localization and mobilization following IS formation of all SNAREs that are expressed in CD8+ T-cells for which specific antibodies are available. Some but not all SNARE proteins are translocated towards the immunological synapse and some co-localized with cytotoxic lytic granules at the immunological synapse. We found that syntaxin 4, syntaxin 6, VAMP 3, and VAMP 8, among other proteins, seem to polarize and colocalize with cytotoxic granule markers at the immune synapse suggesting they play important roles in cytotoxic granule secretion.
Grant/ other support: DFG RE 1092/6-1
HS1 and Itk cooperate to regulate actin remodeling and Ca2+ signaling in T cells (35.28)
Authors: Carrizosa Esteban , Gomez Timothy S. , Labno Christine M. , Chang Briana L. , Klos Dehring Deborah A. , Zhu Peimin , Freedman Bruce D. , Billadeau Daniel D. , Burkhardt Janis K. 
Published: January 2023

Abstract
Productive T cell activation requires efficient reorganization of the actin cytoskeleton. We have shown that the actin regulatory protein HS1 is required for the stabilization of F-actin and Vav1 at the immunological synapse (IS) and for efficient Ca2+ responses. The Tec family kinase Itk also regulates actin responses, in part by facilitating the localization of Vav1 at the IS, suggesting that these proteins act in the same pathway. Using video microscopy, we found that T cells lacking Itk or HS1 behaved similarly, extending unstable actin-rich protrusions upon TCR stimulation. HS1 and Itk could be coimmunoprecipitated from T cell lysates, and GST-pulldown studies showed that Itk's SH2 domain binds directly to two phosphotyrosines in HS1. In the absence of Itk, HS1 failed to localize to the IS. Since Itk is required for PLCγ1 phosphorylation and Ca2+ store release, we examined the Ca2+ signaling pathway in HS1 knockout T cells in greater detail. T cells lacking HS1 exhibited abnormal Ca2+ store release. These cells showed normal phosphorylation but altered localization of PLCγ1. We conclude that Itk-dependent recruitment of HS1 is required for actin stabilization at the IS, and directs the localization of PLCγ1.
Supported by NIH F31-AI071385 to EC and NIH R01-A1065644 to JKB
Inhibition of T cell function in vitro and in vivo using small molecule antagonists of PIM kinases (35.33)
Authors: Jackson Leila J , Wright Dale , Gross Stefan , Robinson John , Marmsater Fredrik P , Allen Shelley , Munson Mark , Carter Laura L 
Published: January 2023

Abstract
The PIM family consists of three highly conserved serine/threonine kinases whose activity is regulated at the level of expression. PIM kinase expression has been shown to be stimulated by a variety of cytokines, hormones, and mitogens and it functions in the regulation of cellular proliferation, differentiation, apoptosis and tumorigenesis. The role of PIM-induced oncogenic transformation has been extensively studied in hematopoietically derived tumors; however the function of PIM in normal immune cells has yet to be fully defined. We sought to elucidate the role of PIM in activation of primary murine T cells by utilizing a highly selective, small molecule PIM inhibitor. We found that inhibition of PIM activity resulted in decreased anti-CD3 and antigen-induced CD4+ T cell proliferation. These effects were observed under Th0, Th1, Th2 and Th17 polarizing conditions, and were not due to increased cell death, but appear to be mediated by a G0/G1 cell cycle arrest. Inhibition of PIM during polarization prevented the production of Th1, Th2, and Th17 effector cytokines suggesting that PIM expression is required for naïve T cell differentiation. Consistent with the in vitro data, mice treated with the PIM inhibitor prior to immunization had a significant reduction in ex vivo recall responses. These results support a key role for PIM in the activation and differentiation of murine CD4+ T cells.
Biochemical evidence for bivalent T cell antigen receptor expression in primary T cells (35.42)
Authors: Schrum Adam G. , Turka Laurence A. , Palmer Ed 
Published: January 2023

Abstract
The αβ T cell antigen receptor (TCR)/CD3 complex is currently viewed as a monovalent receptor with the subunit composition αβγδε2ζ2. This model has emerged from among at least five different potential stoichiometries that were proposed in previous years. Recently, the case for the accepted stoichiometry has been supported by experiments involving co-immunoprecipitation (IP) followed by Western blot, and by blue native (BN)-PAGE. We have revisited the issue of TCR/CD3 stoichiometry by examining immunoprecipitated complexes by flow cytometry (IP-FCM), a technique which allows a quantitative estimate of the subunit constituents in native multi-protein complexes. Together with size-exclusion chromatography, the data supports the conventional stoichiometry, but provides evidence that the copy number may be two, making the TCR/CD3 a precisely bivalent complex, [αβγδε2ζ2] x 2. The expression of a bivalent TCR/CD3 complex has implications regarding possible mechanisms by which the interaction with antigen may trigger signaling.
Research support: Adam G. Schrum, start-up funds from the Mayo Foundation; Laurence A. Turka, grants AI41521 and AI 43620 from NIH; Ed Palmer, grants from the Swiss National Science Foundation.
A Monoclonal Antibody that inhibits IgE binding to the High affinity IgE receptor recognizes a glycolipid (36.11)
Authors: Guiraldelli Michel F , Berenstein Elsa H , Siraganian Reuben P 
Published: January 2023

Abstract
The mAb BD6 is a monoclonal IgM that binds to the surface of RBL-2H3 cells and inhibits the binding of IgE to FcεRI, without reacting with this receptor. By cloning, mAb BD6 binding was resulted by the expression of the alpha1,3-galactosyltransferase gene. In binding assays on RBL-2H3 and alpha1,3-galactosyltransferase transfected PEAK cells there was partial competition between mAb BD6 and IB4, a galactose binding lectin. Both galactose and melibiose decreased the binding of mAb BD6 in a dose dependent manner on RBL-2H3 cells and abolished its binding on alpha1,3-galactosyltransferase transfected PEAK cells. MAb BD6 recognized a low molecular weight band by immunoblotting suggesting that it may be binding a lipid. This was confirmed by blotting lipids from RBL-2H3 cells separated by thin layer chromatography. By sucrose gradient analysis mAb BD6 bound to the lipid rafts fractions. RBL-2H3 variants were isolated completely deficient in mAb BD6 binding; these cells were also deficient in the ganglioside GD1b and had low expression of ganglioside GM1. However, these cells deficient in mAb BD6 binding and GD1b ganglioside still had normal FceRI induced degranulation. These results demonstrate that mAb BD6 inhibits IgE binding and reacts with a glycolipid present in lipid rafts; furthermore the ganglioside GD1b is not essential for degranulation.
Supported by the Intramural Research Program, NIDCR, NIH.
Novel dendritic cell (DC)-stimulating antibody, B7-DCXAb, effectively downregulates allergen-specific IgE, IgG1, Th2 cytokines, and allergic inflammation (36.13)
Authors: Pease Larry R , Checkel James L , Iijima Koji , Radhakrishnan Suresh , Van Keulen Virginia P , McClimon Brad J , Kobayashi Takao , Plager Douglas A , Kita Hirohito 
Published: January 2023

Abstract
We investigated whether human IgM antibody, B7-DCXAb, which binds to B7-DC and enhances DCs' antigen uptake and retention, migration, and cytokine production, improves the efficacy of allergen immunotherapy. Mice were sensitized by intraperitoneal injection of OVA or house dust mite (HDM) antigens plus alum and then treated by intravenous B7-DCXAb injection intranasal or sublingual administration of antigen. OVA-sensitized mice developed anti-OVA IgE and IgG1, Th2-type splenocyte responses to OVA; marked airway eosinophilic inflammation was observed after intranasal OVA challenge. When these mice were treated by intravenous B7-DCXAb plus intranasal OVA, anti-OVA IgE and IgG1 decreased by 75~95% within 1 week; this decrease persisted for &gt;8 weeks. Anti-OVA IgG2a decreased transiently. Splenocyte production of IL-4, IL-5, and IL-13 in response to OVA was abolished; no increases in IFN-g or IL-17 were noted. In OVA-challenged mice, airway eosinophilia and lung levels of IL-4, IL-5 and IL-13 were reduced by &gt;90%. Immunotherapeutic effects were also observed with intravenous B7-DCXAb plus sublingual OVA, but not with intravenous B7-DCXAb alone. Furthermore, HDM-sensitized mice were treated successfully by intravenous B7-DCXAb plus intranasal HDM. B7-DCXAb injection plus mucosal administration of antigen effectively treats allergic responses and may provide novel strategy for allergen immunotherapy.