bluuebunny/crossref_metadata_2025_split huggingface dataset:
Human IL-7Rαlow memory CD8+ T cells have defect in the calcium signaling pathway in response to TCR triggering, leading to impaired IL-2 production and proliferation. (132.34)
Authors: Lee Ha Na , Yu Mi Na , Lee Dong-Sup , Kang Jae Seung , Hwang Young-il , Lee Wang Jae , Kang Insoo , Kim Hang-Rae 
Published: January 2023

Abstract
We have found two subsets of human CD8+ T cells expressing IL-7Rαhigh and IL-7Rαlow with different cell survival responses to IL-7. These IL-7Rαlow memory CD8+ T cells that produce effector cytokines and perforin have impaired survival in response to IL-7. Although these CD8+ T cell subsets fail to respond to TCR triggering, the mechanism for this is unknown. Thus we investigated the intracellular signaling pathways in differentially regulating proliferation and effector cytokine production in human IL-7Rαlow memory CD8+ T cells. Treating these CD8+ T cells with anti-CD3 Abs did not produce IL-2, but expressed IFN-γ and TNF-α. Although IL-7Rαlow memory CD8+ T cells showed anergic phenotypes in terms of cytokine production and proliferation, activation of MAPK pathway, which is known defects in anergic T cells, was intact. Importantly, human IL-7Rαlow memory CD8+ T cells had reduced calcium flux upon in vitro TCR stimulation, leading to impaired IL-2 production. Moreover, inhibition of IL-2 production was not recovered by PMA/ionomycin treatment. There findings provide new sight into that human IL-7Rαlow memory CD8+ T cells are rendered cellular characteristics of anergic with calcium signaling defects.
Antigen-specific precursor frequencies influence requirement of CD4+ T cell help and CD40-40L costimulation in primary and secondary responses (132.8)
Authors: Sokke Umeshappa Channakeshava , Xiang Jim 
Published: January 2023

Abstract
CD4+ T cells play a decisive role in generation of effective CD8+ T cell-mediated immunity against diseases associated with minor H antigens, including cancer, auto-immunity and allogenic tissue transplantation. The increased precursor frequency in reducing the need of CD4+ T cell help and costimulations in CD8+ T cell primary responses have been demonstrated. However, its similar roles in memory CD8+ T cell responses have not been fully understood. In a non-infectious vaccine model, here we show that, at endogenous precursor levels, CD4+ T cell help, and CD40-40L costimulation are required for both primary and memory responses. At increased precursor frequency, although CD4+ T cell help, and CD40-40L costimulation are not obligatory in primary responses, their requirement is still necessary for functional memory responses. Despite increased precursor frequency is ensured during priming, the memory CD8+ T cells developed from increased precursor frequency in the absence of CD4+ T cell help, and CD40-40L costimulation fail to induce complete protection following tumor challenge. These results further support the previous observations that efficient memory CD8+ T cell responses require CD4+ T cell help and CD40-40L costimulation.
The DRF motif in CXCR6 is important for selectivity among G-proteins in receptor signaling, which is cell-type specific (133.1)
Authors: Singh Satya , Foley John , Zhang Hongwei , Farber Joshua 
Published: January 2023

Abstract
CXCR6 is the receptor for CXCL16. Except for CXCR6, all human chemokine receptors contain the D3.49R3.50Y3.51 sequence, and all but two contain A3.53 at the cytoplasmic end of the third transmembrane helix (H3C). In contrast, the CXCR6 sequence is D3.49R3.50F3.51I3.52V3.53. We investigated the importance of canonical and non-canonical amino acids at these positions in CXCR6 in order to understand the role of this region in chemokine receptor function generally. We produced CXCR6 point mutants, including those where non-canonical amino acids were replaced with canonical amino acids singly and in combination, and analyzed the mutants in HEK-293T and Jurkat E6-1 cells. For the H3C of CXCR6, our data demonstrate that 1) non-canonical residues can function better than canonical residues, 2) sequences are matched, with changes having compensatory effects, 3) residues can affect both ligand binding and receptor signaling independently, 4) residues affect selectivity among G-proteins for receptor signaling, and 5) the affects of these residues on receptor function are cell-type specific. Our findings indicate that the H3C may not only serve as an on-off switch for GPCRs, as has been proposed, but also has broader effects on receptor function, which, surprisingly, depend on cell-type specific factors, and suggest that a GPCR structure may be constrained by a need to use G-proteins promiscuously in all the cells where a given receptors is expressed.
Kinetics and mechanisms of IL-4 suppression of Type I interferons’ responses in murine dendritic cells (134.22)
Authors: Sriram Uma , Bennett Heather , Xu Jun , Shivers Debra , Behrens Edward , Gallucci Stefania 
Published: January 2023

Abstract
Cytokines cross-talk and modulate immune responses. We have shown before that IL-4 suppresses the response of murine dendritic cells (DCs) to Type I interferons (IFN). In this study we analyzed the kinetics of Type I IFN and IL-4 interactions in myeloid bone marrow-derived DCs (BMDCs) for Type I IFN induced gene and protein expression. Pre-treatment with IL-4 suppressed IFN-a induced gene expression even at the sub-optimal dose of 0.25ng/ml of IL-4; suppressed phosphorylation of STAT1 and STAT2, suggesting that IL-4 acts at the level or upstream of STAT molecules in the Type I IFN signaling pathway. Post-treatment with IL-4 for a range of 2-24h after IFN-a stimulation, also suppressed IFN-b gene expression. IL-4 also suppressed the response to autocrine IFN-b that is induced upon IFN-a stimulation, acting as a positive feedback. Indeed, we found that BMDCs response to exogenous IFN-a is sustained for many hours - washing and replating BMDCs 30 min after IFN-a stimulation leads to a strong reduction of IFN-b expression that is stimulated only by autocrine IFN-b; post-treatment with IL-4 suppressed this second wave of response, suggesting that IL-4 can inhibit the response to paracrine IFNs and the secondary response to autocrine IFN. Cyclohexamide (protein synthesis inhibitor) abrogated the suppressive effects of IL-4 indicating that IL-4 induces the expression of a protein that possibly interacts with Type I IFN signaling and fades out the responses of DCs to Type I IFNs.
Interleukin-19 regulates the immune homeostasis to the airborne environment (134.5)
Authors: Grunig Gabriele , Sisco Maureen , Louten Jennifer , Daley Eleen , Hoffman Carol , Emson Claire , Gordon Terry , de Waal Malefyt Rene 
Published: January 2023

Abstract
Interleukin 19 (IL-19) is thought to have important roles in the immune control of pathogens, and in enhancing chronic inflammatory diseases of the skin (psoriasis), lung (asthma) and intestine (colitis). However, the biological function and the networked control of IL-19 are poorly understood. Naive mice deficient in IL-19 (IL-19-/-) on a 129xBL6 background had decreased alveolar macrophages and increased eosinophils in the broncho-alveolar lavage (BAL) and decreased B cells in the spleen and bone marrow. However, naïve IL-19-/- mice backcrossed to C57BL/6 or BALB/c, and mice housed in different locations exhibited an attenuated phenotype. Therefore, genetic backcross and challenge experiments with environmental stimuli (allergen, urban particulate matter) were conducted. The studies showed that abnormal composition of the cells in the BAL fluid, spleen and bone marrow in naïve IL-19-/- 129xBL6 mice was due to the deficiency in IL-19 combined with the activity of another gene inherited from the 129 strain. IL-19-/- mice on C57BL/6 or BALB/c backgrounds that were intranasally challenged with antigen or with antigen and urban particulate matter had decreased BAL eosinophils, alveolar macrophages and B cells when compared to wild type. Together our studies indicate that endogenous IL-19 is a critical component of immune homeostasis to the airborne environment and that mouse-strain specific genes control the function of IL-19.
Investigating Early Events in Sjogren’s Disease (135.8)
Authors: Shen Long , Wu Jing , Suresh Lakshmanan , Ambrus, Jr. Julian 
Published: January 2023

Abstract
Little is known about the early events in Sjogren’s disease. Previous studies have demonstrated that interleukin 14 alpha transgenic mice (IL14aTG) replicate the clinical and immunological features of Sjogren’s disease (Clin. Immunol. 130:304, 2009). Salivary gland function is lost before lymphocytic infiltration of the salivary glands. The current studies were designed to investigate the early events involved with salivary gland injury. Western blot studies demonstrated lymphotoxin (LTA) in the salivary gland secretions of IL14aTG mice, while salivary gland function was being lost. IL14aTG mice lacking LTA maintained normal salivary gland structure and function, although autoantibodies were deposited in the salivary glands. Because IL-14aTG mice produce high levels of interferon-a (INF-a) and INF-a can induce production of LTA, we investigated IL14aTG mice lacking the α/β interferon receptor. These mice retained normal salivary gland function, although mild lymphocytic infiltration was noted in the lachrymal glands. Because IL14aTG mice have increased numbers of B1 cells, we produced IL14aTG mice lacking btk. Interestingly, these mice maintained normal salivary gland function despite lymphocytic infiltration of the salivary glands. Thus cytokine injury involving LTA and IFN-a are necessary for the development of salivary gland injury in Sjogren’s disease. The roles of particular autoantibodies and lymphocytic infiltration of the salivary glands need to be elucidated.
Hemorrhagic shock primes for IL-1β processing: role of upregulated TLR2 (136.1)
Authors: Fan Jie , Li Yuehua , Xiang Meng , Billiar Timothy , Wilson Mark 
Published: January 2023

Abstract
IL-1β contributes to post-trauma sepsis, and its release requires processing in inflammasomes. Hemorrhagic shock (HS) primes for IL-1β release in response to LPS via yet unclear mechanisms. We have reported that HS-activated neutrophils (PMN) augment LPS/TLR4 signaling upregulation of TLR2 in alveolar macrophages (AM), thereby sensitizing the AM to TLR2 ligands. Here, we addressed the role of upregulated TLR2 in the HS-primed IL-1β processing. WT, TLR4-mutant and TLR2 KO mice were subjected to HS, followed by intratracheal LPS (30 µg/kg B.W.) and PGN (30 µg/kg B.W.) at 2h and 4h after HS, respectively. Bronchoalveolar lavage fluids (BALF) were collected at 2 h after PGN for detection of IL-1β. In vitro, AM were co-cultured with PMN isolated from HS mice, and treated with LPS, PMN was then removed and PGN was added at 2h. IL-1β and caspase-1 maturation in the AM were assessed. In vivo treatments of LPS and PGN caused a marked increase in IL-1β in BALF in the WT/HS mice, but not in the WT/sham, TLR4-mutant/HS or TLR2 KO/HS mice. HS-activated PMN increased TLR2 expression in AM in response to LPS, and subsequently increased caspase-1 and IL-1β maturation in the AM as well as IL-1β release in response to PGN. Moreover increasing extracellular [K+] failed to prevent the TLR2 signal-induced IL-1β maturation. In HS, amplified TLR2 signaling acts as a second signal in stimulating caspase-1 and IL-1β maturation via mechanisms independent of K+ efflux.
Differential IFN signature for TLR7 and 9 agonists (136.16)
Authors: Puig Montse , Tosh Kevin , Grajkowska Lucja , Verthelyi Daniela 
Published: January 2023

Abstract
Innate immune response modulators that stimulate TLR7 and 9 activate similar signaling pathways but their therapeutic range is distinct. Moreover, 3 structurally different types of stimulatory sequences for TLR9 have been identified (CpG ODN type D/A, B/K and C) that act on the same receptor but activate distinct immunostimulatory pathways and elicit different clinical effects. This study characterizes and compares gene expression in human and macaque cells treated with different TLR7 & 9 agonists. In vitro results showed that overall TLR7 and 9 agonists elicit transcription of a similar repertoire of immune-related genes; however the magnitude and kinetics of the gene expression differed significantly, particularly for IFN-related genes as well as cell mobilization and adhesion genes. Interestingly, while transcription level and kinetics of type I IFNs gene expression in cells treated with CpG ODN D, B and C differed significantly, the relative distribution of IFNα subtypes was almost identical. In contrast, the same cells stimulated with the TLR7 agonist elicited a different pattern of IFNα subtypes and IFNα-inducible genes. In vivo, only rhesus macaque monkeys treated with CpG ODN type D showed a robust IFN type I response both locally and systemically. The data suggest a higher mobilization and activation of the pDC with CpG ODN type D treatment. Understanding the nature of these differences has implications for the therapeutic application of TLR agonists in humans.
NOD2 mediates CNS inflammatory responses of primary murine glia in streptococcal meningitis (136.34)
Authors: Chauhan Vinita , Liu Xinjie , Marriott Ian 
Published: January 2023

Abstract
Resident cells of the central nervous system (CNS) such as microglia and astrocytes initiate and/or augment inflammation following trauma or infection. We have previously demonstrated that glia constitutively express nucleotide-binding oligomerization domain-2 (NOD2), a member of the novel nucleotide-binding domain leucine-rich repeat region-containing family of proteins (NLR) that functions as an intracellular receptor for a minimal motif present in all bacterial peptidoglycans. We have also shown that this NLR is essential for glial responses to Gram-negative pathogens and the subsequent in vivo CNS inflammation. In the present study, we have established that intact Streptococcus pneumoniae, the major causative agent for Gram-positive bacterial meningitis in adults, is a potent stimulus for the activation of the pivotal inflammatory transcription factor NF-kB and production of inflammatory cytokines in primary murine microglia and astrocytes. We demonstrate that NOD2 is essential for the maximal responses of these cells to intact S. pneumoniae but not cellular lysates. Finally, we have shown that NOD2 is required for the elevated inflammatory mediator levels, astrogliosis, and demyelination, following in vivo administration of this Gram-positive CNS pathogen. As such, we suggest that NOD2 plays a critical role in the establishment of the lethal inflammation associated with streptococcal meningitis.
Cross-talk between innate and adaptive immunity through pentraxins and Fc receptors (136.42)
Authors: Lu Jinghua , Marjon Kristopher , Marnell Lorraine , Mold Carolyn , Du Clos Terry , Sun Peter 
Published: January 2023

Abstract
The classical pentraxins include serum amyloid P component (SAP) and C-reactive protein (CRP), which are two of the acute-phase proteins and able to opsonize microbial pathogens. In 2008, we reported the structural mechanism for pentraxin’s binding to FcγRIIa and its functional activation of FcγR-mediated phagocytosis and cytokine secretion. We further extend these findings in two aspects: 1) High-affinity IgG receptor, FcγRI, is also recognized by pentraxins. The crystal structure of human FcγRI reveals a unique architecture of the three-Ig domains of FcγRI, which is reminiscent of the head of a seahorse. Despite the additional third Ig domain (D3), the first two Ig domains (D1 and D2) of FcγRI adopt a similar structure to that of FcγRIIa and also confer the high affinity for the binding of IgG. Therefore, like FcγRIIa, FcγRI binds to two diagonally-spanned subunits of each CRP or SAP pentamer with both D1 and D2 domains of the receptor contacting the ridge helices from two SAP subunits. 2) We show that pentraxins also recognizes FcαRI, both in solution and on cells. FcαRI bound to the effector face of CRP and SAP in a region close to but not identical with that of FcγRs. The binding of CRP to FcαRI transfected RBL cells induced degranulation and the phosphorylation of Syk kinase. Taken together, these results establish antibody-like functions for pentraxins in the Fc receptor pathway and suggest a new crosstalk between the innate and adaptive immune systems.
Proteomics decode RLR and TLR signaling circuitry (136.5)
Authors: Li Shitao , Wang Lingyan , Dorf Martin 
Published: January 2023

Abstract
RIG-I-like receptors (RLRs) and Toll-Like Receptors (TLRs) play a critical role in sensing microorganisms and activating the early innate immune response. The signaling circuit of innate immunity remains incompletely understood. To systematically investigate the network, we initiated a global proteomic analysis of protein complexes found in the innate immunity pathway. Fifty-five genes involved in TLR and RLR pathways were tagged with FLAG epitopes and stably transfected into 293 cells engineered to express TLR3 or TLR4. After treatment with or without poly(I:C), LPS, or poly(dA:dT), cells were collected to purify the tagged protein complexes. Proteins in each complex were identified by mass spectrometry and data were statistically analyzed. Protein interaction networks were established using Cytoscape software. Sixty-five known interactions were confirmed and seventy-nine new interactions were detected.
Crucial role of effector T cell-derived IL-10 in regulating lung inflammation and pulmonary function during primary RSV infection (137.13)
Authors: Sun Jie , Cardani Amber , Braciale Thomas 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) infection is the leading viral cause of severe lower respiratory tract illness in young infants. Clinical studies have documented that genetic polymorphism of a regulatory cytokine IL-10 is associated with the development of severe bronchiotis in RSV infected infants. However the tempo, source and function of IL-10 during the RSV infection remain unclear. Here, we have examined the role of IL-10 in a murine model of primary RSV infection. We found that high levels of IL-10 are produced in the respiratory tract at the onset of adaptive immune responses. The main cellular sources of the IL-10 are infiltrating anti-viral effector CD4+ and CD8+ T cells (Te). We showed that the function of the Te-derived IL-10 in vivo is mainly to limit the pro-inflammatory activity of the anti-viral Te themselves and thus to control the exaggerated pulmonary inflammation induced by the infiltrating Te. Furthermore, we demonstrated that the inhibition of the exaggerated inflammation by IL-10 is essential in maintaining normal pulmonary function of the host. Our data not only discovered a novel function of IL-10 (i.e., autocrine inhibition of the inflammatory activity of Te) during infection but provided potential explanation and cellular mechanism for the early clinical studies demonstrating a link between IL-10 gene polymorphism and the development of pulmonary diseases during RSV infection.
HCV+ hepatocytes induce human CD4+ regulatory T cells through the production of TGF-β (137.19)
Authors: Hall Caroline , Hahn Young 
Published: January 2023

Abstract
Hepatitis C Virus (HCV) is remarkably efficient at establishing persistent infection and impaired T cell responses are reported to be responsible for facilitating and maintaining this infection. CD4+ regulatory T cells detected in chronic HCV patients have been implicated in dampening antiviral T cell responses to HCV infection. However, the underlying mechanism for induction and expansion of regulatory T cell responses has yet to be determined. To this end, we examined whether HCV-expressing hepatocytes, as the primary target for HCV infection and replication, are able to induce regulatory T cells. These studies were accomplished by establishing the co-culture of Huh7.5 containing the full-length genome of HCV genotype 1a (Huh7.5-FL) with activated CD4+ T cells, using Huh7.5 with no HCV (Huh7.5) and a subgenomic portion (Huh7.5-SG) as controls. The production of IFN-γ and proliferative response of antigen-independent CD4+ T cells was diminished following co-culture with Huh7.5-FL compared to controls. Notably, these CD4+ T cells in contact with Huh7.5-FL express an increased level of regulatory T cell markers (FoxP3, CTLA-4, LAP) and are able to suppress the proliferation of effector T cells. Intriguingly, Huh7.5-FL upregulate the production of TGF-β and blockade of TGF-β abrogates FoxP3 expression and IFN-γ production. These results suggest that hepatocytes may play a pivotal role in impairing antiviral T cell response by the induction of regulatory T cells.
TAT-CD137 Ligand cytoplasmic domain as the effective activator for reverse signaling (138.19)
Authors: Kim Jung , Lee Eun , Quang Nguyen , Kwon Byungsuk 
Published: January 2023

Abstract
The CD137 (also called 4-1BB), a T-cell co-stimulatory receptor induced by TCR activation, is a member of tumor necrosis factor (TNF) receptor superfamily. The ligand of CD137 (4-1BBL) , also a TNF superfamily member, is known to be expressed on activated antigen-presenting cells such as B cells, macrophages and dendritic cells (DCs). Bi-directional transduction of signals exists for the CD137 receptor/ligand system. Here we developed the TAT fused CD137 Ligand cytoplasmic domain (TAT-CD137Lcy), which has cell-penetrating ability, to regulate the reverse signaling. The TAT-CD137Lcy protein induced the production of IL6 and TNF-a on peritoneal macrophages and also increased the level of mRNA transcripts of IL-6 and TNF-a. The TAT-CD137Lcy protein increased phosphorylation of p38, JNK and Erk but not the degradation of the inhibitor of NF-kB (IkBa). The TAT-CD137Lcy protein potentially functions as activator for reverse signaling. To confirm the positive effect of TAT-CD137Lcy protein, B cells proliferation assay was performed. The TAT-CD137Lcy protein enhanced the proliferation of splenic B cells stimulated with anti-CD40 monoclonal antibody (mAb) and anti-IgM mAb. Our results indicated that the TAT-CD137Lcy protein is the effective activator for reverse signaling. For activation of reverse signaling, The TAT-CD137Lcy protein is more effective than 4-1BBFc and TKS-1 mAb because the TAT-CD137Lcy protein does not interrupt the interaction between receptor and ligand.
Validation of markers for phenotyping naïve and memory T-cells by multicolor flow cytometry. (138.22)
Authors: Hingorani Ravi , Cheung Jocelyn , Ernst David , Lane Cynthia , Rohrer Jurg 
Published: January 2023

Abstract
The antigen experienced T-cell population can be broadly divided into the effector and memory cell compartments, both of which are needed for sustaining a responsive immune system. Simplistically, the effector T lymphocytes are described as cells needing active antigenic stimulation to eliminate pathogens. Memory T lymphocytes, on the other hand are described as cells which remain present in the absence of antigenic stimulation and have the capacity to expand rapidly upon secondary challenge. Recently, with the identification of central and effector memory T-cell subsets, tremendous effort has been devoted to characterize markers on the surface of these cells. Though various markers have been used to identify the subsets, no single marker that segregates one subset from the other has been described. Thus, multiple markers are needed to subset the cells in order to characterize them. Here we report the verification of a 9-color panel that can successfully identify 5 distinct CD4 and CD8 T cell populations within the naïve and effector cell subsets. In addition, functionality of the memory subsets was validated by stimulating the cells and using the production of IFN-γ as a read-out. Avoiding potential pitfalls when setting up 9 color panels for use in flow cytometry will be discussed.