bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lunatic Fringe: A key regulator of notch ligand Dll4 mediated Th2 inflammation during viral exacerbation of allergic response. (175.4)
Authors: Mukherjee Sumanta , Berlin Aaron , Maillard Ivan , Kunkel Steven , Lukacs Nicholas 
Published: January 2023

Abstract
Children exposed to environmental cockroach allergen (CRA) have increased susceptibility to develop chronic pulmonary disorders such as asthma later in their life. Moreover when these children encounter a respiratory viral infection it exacerbates their existing condition. The clinical manifestations include increased mucus production and airway hyperreactivity and peribronchial Th2 inflammation, necessitating immediate hospitalization and frequent recurrent hospital visits. In the present study we examined respiratory syncytial virus (RSV) exacerbated allergic responses. Previous studies showed that notch is required for the development of Th2 responses. However the role of notch ligand-Delta like-4 (Dll4) in viral exacerbation is unknown. Here, using a clinically relevant mouse model of RSV exacerbation an abrogation of mucus production and Th2 responses when Dll4 is blocked was observed. Further, our results from CD4-T cell skewing assays suggest that Dll4 augments IL4 production in Th2 cells in an antigen specific manner. Interestingly, both in vitro and in vivo an existing Th2 environment increased lunatic fringe (L-FNG) expression, a glycosyltransferase that enhances Notch activation by ligands of the Delta-like family, in CD4 T cells. Using loss of function assays and an RNAi approach we show that Dll4 augmented IL4 production is L-FNG dependent. These studies highlight a novel regulatory mechanism involved in Dll4 mediated Th2 responses during viral exacerbation.
Structure of Nectin-2 reveals determinants of homophilic and heterophilic interactions that control cell-cell adhesion and immune regulation (176.10)
Authors: Samanta Dibyendu , Ramagopal Udupi , Vigdorovich Vladimir , Rubinstein Rotem , Almo Steven , Nathenson Stanley 
Published: January 2023

Abstract
Nectin comprises a family of four immunoglobulin-like molecules. Homophilic and heterophilic interactions among nectins are implicated in cell-cell adhesion, while their interactions with members of other protein families have diverse biological functions like host-pathogen interaction and immune modulation. In particular, nectin-2, found to be up-regulated on cancer cells is capable of interacting with two receptors, CD226 and TIGIT, expressed on T and NK cells. These interactions lead to the delivery of two opposing signals to both T and NK cells. This situation is reminiscent of the well-studied pathways in T cells, in which the coinhibitory receptor CTLA-4 binds the same ligand (B7) as the coactivating receptor CD28. In order to define the molecular and structural determinants underlying the homophilic and heterophilic recognitions of nectin-2, we examined the biochemical, biophysical and structural properties of human nectin-2. The structure of nectin-2 at 1.3 Å resolution reveals that the architecture of the nectin-2 homophilic dimer resembles other members of the immunoglobulin superfamily and defines the details responsible for recognition and selectivity. Of particular note, the close proximity of charged residues at the interface is a major determinant of binding affinity. Using these biochemical and structural data, we also characterized the heterophilic binding of nectin-2 with TIGIT, which is implicated in T cell and NK cell-mediated immune modulation.
Cell type specific role of B7 for T-dependnet antibody response in vivo (176.14)
Authors: Watanabe Masashi , Hodes Richard 
Published: January 2023

Abstract
B7.1/B7.2 molecules are essential for eliciting immune responses through CD28-mediated T cell activation, as well as for CTLA-4-mediated negative regulation of T cell responses. Although many cell types (DC, MΦ, B cells and T cells) express B7 in vivo, the role of B7 on each cell type for immune response is not well defined. To address these questions, we generated and characterized cell type-specific B7 cKO mice. Mice expressing a B7.1flox BAC Tg on a B7.1/B7.2 DKO background (designated as B7flox) were generated and crossed to several Cre-Tg strains. To evaluate the cell type-specific role of B7 for T-dependent Ab response in vivo, B7 cKO mice were immunized with TNP-KLH/Alum, and TNP-specific Ab was measured. Ag-specific IgG1 production was B7-dependent, as evidenced by the absence of response in B7 DKO mice. Normal levels of IgG1 production were observed in B7flox mice and in B cell-specific B7 cKO (CD19-Cre x B7flox) mice. In contrast, the IgG1 response was essentially absent in DC-specific B7 cKO (CD11c-Cre x B7flox) mice. This result indicates that B7 on DC is critical for induction of IgG1 class switching, whereas B7 on B cells is not required for this response. Further studies are ongoing to elucidate cell type-specific roles of B7 for several critical steps involved in T-dependent antibody responses, including follicular helper T cell differentiation and germinal center formation, using a panel of cell type-specific B7 cKO mice.
Mast cell-macrophage interactions alter inflammatory cytokine production. (177.10)
Authors: Lama Tamang Tenchee , Pullen Nicholas , Ryan John 
Published: January 2023

Abstract
Increased levels of mast cells and macrophages are seen at the tumor microenvironment. Mast cells are associated with tumor progression, and macrophages have been observed to induce cancer cell motility and angiogenesis. Like mast cells, macrophages produce inflammatory cytokines such as TNF, IL-13, IL-6 and MIP-1α. The individual role of these cells are well-recognized, but very little is known about their potential interactions in this microenvironment. Our preliminary studies demonstrate that co-culturing mast cells with macrophages elevates inflammatory cytokine production. Cultures of IgE-sensitized, antigen stimulated mast cells mixed with macrophages produced higher concentration of inflammatory cytokines than either cell population alone. These data demonstrate the possibility that communication between mast cells and macrophages may alter one another’s functions, especially cytokine secretion. These studies will help us gain a better understanding of the role of mast cell-macrophage interactions and how they may contribute to both normal and pathological immunity.
Autocrine secretion of hemokinin-1 promotes the innate immunostimulatory function of mast cells. (177.21)
Authors: Sumpter Tina , Ho Chin , Tkacheva Olga , Larregina Adriana 
Published: January 2023

Abstract
Mast cells (MC), with pro- and anti-inflammatory functions, control innate and adaptive immune responses. Mechanisms causing such disparate functions are only starting to be elucidated. Recent advances in neuroimmune regulation suggest that MC function may be regulated, in part, by neuropeptides. MC express the neurokinin-1 receptor (NK1R), the preferred receptor for the inflammatory neuropeptides, Substance P (SP) and Hemokinin-1 (HK1). We hypothesized that agonistic signaling through the NK1R regulates MC function. We compared the ability of C57BL/6 wild type (WT) or NK1RKO bone marrow-derived MC stimulated via FcϵRI or TLR4 to secrete pro-inflammatory IL-1β, IL-6, IL-33, and TNFα, or anti-inflammatory, IL-10. NK1RKO MC secreted significantly less TNFα and IL-33 but equal levels of IL-10, IL-6 and IL-1β compared to WT-MC, results that were further confirmed with WT-MC incubated with the NK1R antagonist L760,330. As our experimental conditions lack nerve fibers and exogenous SP or HK1 did not increase cytokine secretion we postulated that the observed effects were caused by autocrine secretion of NK1R agonists by MC. Using quantitative RT-PCR, we found high expression of the tac4 gene encoding for HK1 which was further increased following MC stimulation while the levels of SP mRNA remained significantly lower in spite of MC simulation. Together our results demonstrate that autocrine HK1 and NK1R signaling are potent stimulators of the pro-inflammatory function of MC.
ADAP Regulates the Stability and Movement of SLP-76 Microclusters (178.10)
Authors: Lewis Juliana , Nguyen Ken , Bunnell Stephen 
Published: January 2023

Abstract
The adaptor molecule ADAP and its paralog, PRAM-1, are expressed in mononuclear cells, lymphocytes, and granulocytes. ADAP plays a crucial role in T cell receptor-induced integrin activation, conjugate formation, NF-kB activation, IL-2 production, and T cell proliferation. However, it is not clear how ADAP exerts its profound effects on T cell activation. Here, we demonstrate that endogenous ADAP is an integral component of TCR-induced SLP-76 microclusters, and is essential for their persistence and movement. Our live cell imaging studies show that when we break the ADAP-SLP-76 interaction ADAP is not recruited into SLP-76 MC and SLP-76 MC are immobile and non-persistent. Consequently, T cell adhesion via the TCR is severely impaired, indicating a novel role for ADAP in adhesion to TCR ligands. Phosphorylation of highly conserved, SLP-76-interacting tyrosines within the ADAP C-terminus is necessary for ADAP to enter SLP-76 MC, and associations mediated by the N-terminus are required to stabilize ADAP within SLP-76 MC. These data demonstrate that ADAP could participate in TCR-mediated adhesion to antigen in vivo. Consequently, ADAP and PRAM-1 could be therapeutically relevant targets for modulating TCR- and FcR-dependent signals in clinical settings.
High-throughput sequencing of Memory and Naïve T-cell receptor repertoires at the RNA and DNA levels reveals differences in relative expression of expanded TCR clones (178.12)
Authors: Desmarais Cindy , Robins Harlan 
Published: January 2023

Abstract
Recent advances in high throughput sequencing technologies have enabled the profiling of TCR repertoires at an unprecedented level of resolution. Adaptive Biotechnologies has developed sequencing methods that are able to sequence TCR rearrangements by sequencing either using cDNA isolated from RNA, or using genomic DNA directly. We used Adaptive’s immunoSEQ technology to profile CD8+ Memory, CD8+ Naïve, and unsorted PBMC repertoires from 2 individuals using cDNA and genomic DNA isolated from the same starting population of cells. Using this method, it is possible to identify instances of either differential over- or under-expression of particular rearrangements, and to study the overall impact of starting material on sequencing results. Understanding the implications of assaying TCR repertoires at the genomic versus RNA level is important for the interpretation of experimental results, as well as for informing the design of immune profiling experiments that use high-throughput sequencing methodologies.
The B7-CD28 paradigm revisited--- a systemic receptor array approach (178.19)
Authors: Yao Sheng , Zhu Yuwen , Zhu Gefeng , Augustine Mathew , Goode Diana , Wang Jun , Zheng Linghua , Han Xue , Luo Liqun , Broadwater Megan , Ruff William , Chen Lieping 
Published: January 2023

Abstract
T cell cosignaling pathways regulate T cell activation and control the fate of T cell responses. They consist of complex receptor-ligand interactions on cell surface between T cell and antigen presenting cell. The effort to identify co-signaling interactions has been hampered by low expression levels and weak affinities. We generated a comprehensive receptor array system in which over 2,500 human transmembrane proteins of hematopoietic system are expressed individually at high level and could be screened in a high throughput format to determine the binding partner of a fluorescence-labeled target protein. We report here the identification and characterization of two new interactions in the B7-CD28 family. Firstly, B7-H2, formerly known as the ligand for ICOS, is identified as the third ligand for CD28 and CTLA-4 in human. B7-H2 interacts with CD28 through a non-overlapping domain as B7-1 and B7-2. B7-H2-CD28 interaction is essential for the costimulation of human T cells’ primary responses to allogeneic antigens and memory recall responses. B7-H2-CD28 costimulation upregulates survival factor Bcl-xL, suppresses cell cycle inhibitor p27kip1 and induces signaling cascade of ERK and AKT kinases-dependent pathways. Secondly, B7-1 is identified as an inhibitory counter-receptor on T cell for B7-H1. Our study reveals new cosignaling interactions as potential targets for immune-modulation and provides a platform to rapidly discover receptor-ligand interactions in a large scale.
T lymphocytes are sensitive to LIF and possess (i) a LIF/IL6 axis and (ii) a LIF/NANOG axis (178.2)
Authors: Metcalfe Su 
Published: January 2023

Abstract
Subtractive genome-wide analysis of transplantation tolerance has revealed a novel stem cell gene - MARCH-7 - as one of only 8 genes specifically associated with tolerance in vivo. Further studies revealed (i) MARCH-7 as an E3 ligase required for degradation of leukaemia inhibitory factor receptor (LIF-R); and (ii) LIF itself is linked to T cell lineage determination and tolerance. Further, LIF proved to be functionally linked to IL6, each generating opposing feed forward loops during T cell differentiation and leading respectively to either Treg (LIF), or TH17 (IL6). Modelling this LIF/IL6 axis would place MARCH-7 as pivotal: indeed, experimental evidence supports this notion with low MARCH-7 correlating with high LIF-R expression and high responsiveness to LIF. Conversely, high MARCH-7 reduced LIF responsiveness. In stem cells LIF induces NANOG, critical for pluripotency. Given the various links between (i) MARCH-7 and stem cells; (ii) MARCH-7 and regulation of endogenous LIF signalling; and (iii) LIF and NANOG expression in stem cells, we asked, in MARCH-7 null T cells, (a) is NANOG expressed ? (b) is NANOG expressed in response to exogenous LIF ? (c) does MARCH-7 gene-dose influence expression of short non-coding RNAs (miR) ? Each question was positively confirmed, unmasking a novel LIF/NANOG axis linked to MARCH-7 in the T cell lineage. A model is proposed where Treg and TH17 are downstream of a common precursor lineage sensitive to “stemness”-type signals.
IGSF4 is a novel TCR ζ-chain-interacting protein that enhances TCR-mediated signaling (178.5)
Authors: Kim Hye-Ran , Jun Chang-Duk 
Published: January 2023

Abstract
Immunoglobulin superfamily member 4 (IGSF4) is a known ligand of CRTAM, a receptor expressed in activated NKT and CD8+ T cells, but its function in T cell immunity has not been elucidated. Here, we show that IGSF4 directly interacts with the TCR ζ-chain and enhances TCR signaling by enhancing ζ-chain phosphorylation. Ectopic overexpression of IGSF4 enhances TCR-mediated T cell activation. In contrast, IGSF4 knockdown shows a dramatic decrease in markers associated with T cell activation compared with those in control siRNA. The transmembrane domain is essential for TCR ζ-chain association and clustering to the immunological synapse, and the ectodomain is associated with T cell interaction with APCs. IGSF4-deficient mice have impaired TCR-mediated thymocyte selection and maturation. Further, these mice reveal attenuated effector T cell functions accompanied by defective TCR signaling. Collectively, the results indicate that IGSF4 plays a central role in T cell functioning by dual independent mechanisms, control of TCR signaling and control of T cell-APC interaction.
A mixed Th1/Th2 response induces optimal resistance to Leishmania braziliensis (43.21)
Authors: Novais Fernanda , Lee James , de Oliveira Camila , Scott Phillip 
Published: January 2023

Abstract
Leishmania braziliensis is a protozoan that causes cutaneous lesions that heal spontaneously or induces a disease where the parasites metastasize to the nasopharyngeal mucosa. The factors responsible for the severe forms of the disease remain poorly understood, but an exaggerated Th1 response is associated with the immunopathology observed in these patients. In the absence of IFN-γ or iNOS mice are extremely susceptible to L. braziliensis, indicating that Th1 responses are essential to control these parasites. However, we hypothesized that a more moderated response might be associated with less pathology. Since we previously found that CD8 T cells inhibit a Th2 response in leishmaniasis, we tested whether depletion of CD8 T cells might promote a mixed Th1/Th2 response and less pathology. In fact, BALB/c mice infected with L. braziliensis and depleted of CD8 T cells developed smaller lesions, and unexpectedly were better able to control the parasites. This enhanced protection was accompanied by an increase in IL-4 and IL-5 levels, and an infiltration of neutrophils and eosinophils into the lesions. Eosinophils were essential for the increased resistance, since the protective effect of anti-CD8 treatment was reversed when mice were depleted of eosinophils by anti-CCR3 or anti-IL-5 monoclonal antibodies. Taken together, these data demonstrate that a mixed Th1/Th2 response induces optimal control of a L. braziliensis infection, and eosinophils are required for this outcome.
Polyfunctional T cell responses to ESAT-6 and PPD after treatment in multidrugresistant tuberculosis patients. (43.26)
Authors: Sada Isabel , Carrillo-Alduenda José Luis , Chávez-Galán Leslie , Sköld Markus , Salazar-Lezama Miguel Angel 
Published: January 2023

Abstract
Background: Treatment failure against Mycobacterium tuberculosis (Mtb) infection has contributed to the development of multidrug resistant (MDR) tuberculosis. Polyfunctional (PF)-T cells secrete IFN-γ+IL-2+TNF-α+ and mediate protective immunity during chronic infections. Objective: characterize and compared the frequency of PF T cells responding to ESAT-6 and PPD antigens. Methods: We followed 10 MDR TB patients longitudinally 3 months after anti-MDR TB treatment. All samples were obtained in accordance with the Institutional Review Board’s protocol. Peripheral blood mononuclear cells (PBMCs) were stimulated with ESAT-6 or PPD and prepared for FACS analysis of CD3, CD4, CD8, IFN-γ, TNF-α and IL-2. Results: Basal samples from all patients showed a positive response to ESAT-6 and PPD antigens; however, the frequency of CD4+ and CD8+ T cells producing any combination of cytokines was not different. The proportion of IFN-γ+TNF-α+IL-2+, IFN-γ+ and TNF-α ESAT-6 CD4+ T cells increased significantly (p=0.02) when compared with pretreatment values. When cells were stimulated with PPD, IFN-γ+TNF-α+, IFN-γ+ and TNF-α+ CD4+ T cells also increased. The frequency of triple positive ESAT-6 CD8+ T cells was not different. For both, ESAT-6 and PPD antigens IFN-γ+TNF-α+, IFN-γ+ and TNF-α+ CD8+ T cells increased significantly (p&lt;0.001). Conclusions: Our results support that PF T cell responses are actually recovered during the first 3 months of therapy.
IL-18-primed ‘helper’ natural killer cells mediate the attraction and activation of dendritic cells, promoting the accumulation of type-1-polarized T cells at effector sites (45.6)
Authors: Wong Jeffrey , Muthuswamy Ravikumar , Kalinski Pawel 
Published: January 2023

Abstract
The chemokine-driven interaction of immune cells is essential for effective immunity against infection and cancer. We have previously shown that natural killer (NK) cells can be primed by IL-18 for unique ‘helper’ activity, promoting dendritic cell (DC) activation and DC-mediated induction of type-1 immune responses. Here, we further demonstrate that such IL-18-treated ‘helper’ NK cells are selectively primed for high expression of immature DC (iDC)-attracting chemokines, including CCL3 and CCL4, upon subsequent exposure to type I interferons, tumor cells, or other secondary signals. These ‘helper’ NK cells potently attract iDCs in a CCR5-dependent mechanism, situating them for efficient NK cell-mediated activation. We further describe the unique ability of these ‘helper’ NK cells to induce high DC production of CXCR3 and CCR5 ligands, including CXCL9, CXCL10, and CCL5, facilitating the recruitment of type-1 effector T (Teff) cells to sites of infection or tumor. Furthermore, using cells from the ascites of patients with advanced ovarian cancer, we demonstrate the therapeutic potential for using helper NK cell-inducing stimulatory factors to enhance Teff cell-attracting chemokines directly within the human tumor environment. This study demonstrates for the first time the unique chemokine regulation of ‘helper’ NK cell differentiation, and supports the therapeutic use of properly-activated NK cells in promoting type-1 immune responses against infection and cancer.
Role of Dectin-2 in the host defense against Streptococcus pneumoniae infection (45.8)
Authors: Ishii Keiko , Akahori Yukiko , Toyama Masahiko , Aoyagi Tetsuji , Yamamoto Natsuo , Saijo Shinobu , Iwakura Yoichiro , Yamasaki Sho , Kawakami Kazuyoshi 
Published: January 2023

Abstract
Streptococcus pneumoniae possesses a thick polysaccharide capsule. Host defense to this bacterium is mediated by neutrophilic inflammatory responses, which is critically regulated by innate immune mechanism. S. pneumoniae has been reported to be sensed by pattern-recognition receptors such as TLR2, TLR4 and SIGN-R1. In this study, we examined the role of Dectin-2 in the host defense to S. pneumoniae infection. When bone marrow-derived dendritic cells from wild-type (WT) and Dectin-2KO mice were stimulated with culture supernatants (Sup) of a strain of S. pneumoniae (URF918) or its ConA-bound fraction, IL-12p40 production was abrogated in Dectin-2KO mice. In a NFAT-GFP reporter assay, Sup provided a positive signal in Dectin-2 gene-transfected reporter cells. These results suggest that Dectin-2 senses mannose-containing polysaccharides in S. pneumoniae capsule. Dectin-2KO mice infected intratracheally with S. pneumoniae showed shorter survival and larger bacterial burden and lower IFN-γ production in lungs than WT mice. Although the neutrophilic infiltration in lungs was equivalent in both mice, the ratio of engulfed/un-engulfed neutrophils was lower in KO mice than in WT mice. The levels of anti-pneumococcal polysaccharide Ab in bronchoalveolar lavage fluid and serum were lower in KO mice than WT mice. These results suggest that Dectin-2 plays critical roles in the host defense to S. pneumoniae infection through promoting the Ab-mediated phagocytosis by neutrophils.
MHC II retrograde signaling in MDSC (46.15)
Authors: Nagaraj Srinivas , Nelson Allison , Nair Sajitha , Mcmillan Christina 
Published: January 2023

Abstract
It is now evident that inadequate function of the host immune system is due to suppressive factors such as MDSC. A number of studies demonstrated that MDSC induced antigen-specific tolerance of CD8+, but not CD4+ T cells. In our recent study we found that the ability of MDSC to induce antigen-specific CD4+ T-cell tolerance in vivo was dependent on the expression of MHC class II. We also found that MDSC in tumor-bearing mice can induce this suppression, as long as MDSC express a sufficient level of MHC class II molecules. Antigen-specific CD4+ T cells, but not CD8+ T cells, were able to convert MDSC to non-specific suppressor cells in vitro and in vivo and this effect was dependent on MHC class II. Our data have demonstrated that this effect required MHC class II cross-linking, which leads to up-regulation of cox2 and PGE2, which were previously implicated in MDSC mediated immune suppression. These functions of MHC class II was linked to increase in intracellular Ca2 mobilization, and activation of PKC and the mitogen activated kinases p38 and Erk. Cross-linking of IAb resulted in up-regulation of Ets-1 in MDSC. Down-regulation of Ets-1 abrogated increased cox2 expression and PGE2 production caused by IAb ligation. Our data suggests that CD4+ T cell / ETS-1/ COX2 play a major role in retrograde MHC class II signaling in MDSC and results in PGE2 synthesis. Our study may provide an understanding of the initiation and maintenance of immune suppression in cancers.