bluuebunny/crossref_metadata_2025_split huggingface dataset:
Differential regulation of human T cell and NK cell function by tobacco product preparations (P4339)
Authors: Arimilli Subhashini , Damratoski Brad , Gaddamanugu Prasad 
Published: January 2023

Abstract
NK cells and T cells play essential roles in innate and adaptive immune responses by protecting against microbial infections and in tumor surveillance. While smoking is known to cause immunosuppression, it is unclear how the use of smokeless tobacco (ST) products affects immune responses. We investigated immunological responses to exposure of reference tobacco product preparations (TPPs) using poly I:C and LPS-stimulated PBMCs. The TPPs included whole smoke-conditioned media (WS-CM) and total particulate matter (TPM) prepared from 3R4F reference cigarettes, 2S3 moist snuff smokeless tobacco extracted in complete artificial saliva (ST/CAS), and nicotine. WS-CM and TPM treatment resulted in dose-dependent reductions in the expression of intracellular IFN-γand TNF-α in NK cells and T cells, secretion of several cytokines by PBMCs and the inhibition of cytolytic activity of effector cells in PBMCs. Although interference from the vehicle effects confounded the interpretation of data from cells exposed to ST/CAS, some immunosuppressive effects were evident only at high doses. Treatment with nicotine caused minimal immunosuppression in this system. These data suggest that the function of human NK cells and T cells is impacted by exposure to TPPs, on an equi-nicotine unit basis, as follows: WS-CM &gt; TPM &gt;ST/CAS&gt; nicotine. These findings support the hypothesis that immune-suppression from smoking is linked to a higher incidence of microbial infections and cancer among smokers.
NKp65 hallmarks a rare subset of human innate lymphocytes (P4347)
Authors: Steinle Alexander , Döring Claudia , Hansmann Martin-Leo , Vogler Isabel 
Published: January 2023

Abstract
We previously provided the first characterization of the human activating C-type lectin-like receptor (CTLR) NKp65. Like its close relative NKp80, NKp65 is a homodimeric CTLR with a cytoplasmic hemITAM-like signaling motif and encoded in the Natural Killer Gene Complex (NKC) juxtaposed to its ligand. While NKp80 ligates AICL expressed by myeloid cells, NKp65 was found to be a high-affinity receptor for KACL, like AICL a member of the human CLEC2 family of CTLR. Strikingly, KACL expression is almost exclusively confined to human skin, i.e. keratinocytes. NKp65/KACL interaction has been shown to activate cytotoxicity and IFNg secretion of an NKp65- expressing NK cell line. However, while NKp80 is expressed by virtually all human NK cells, physiological expression of NKp65 remained enigmatic. In order to identify NKp65 expressing cells we generated NKp65-specific mAb that specifically stain a small subset of human innate lymphocytes isolated from peripheral blood. Transcriptional profiling of NKp65-expressing cells suggested that NKp65 expression is associated with a distinct subset of innate lymphocytes. Phenotypic analyses confirmed exclusive expression of NKp65 on this innate lymphocytes subset, while no NKp65 expression on NK cells was observed. Hence NKp65 hallmarks this subset of innate lymphocytes and may endow this subset with the capacity for dedicated immunosurveillance of human skin.
Vitamin D insufficiency modulates the pro- and anti-inflammatory cytokine balance without altering regulators of circulating 25-hydroxyvitamin D (P6301)
Authors: Barker Tyler , Dixon Brian , Schneider Erik , Weaver Lindell 
Published: January 2023

Abstract
Although evident in pathophysiological conditions, it is unknown if regulators of vitamin D are disrupted, if pro-inflammatory cytokines are increased, or if anti-inflammatory cytokines are decreased with vitamin D insufficiency in young adults. The purpose of this study was, therefore, to evaluate circulating regulators of vitamin D, and circulating pro- and anti-inflammatory cytokines in young adults with low serum 25-hydroxyvitamin D (25(OH)D) concentrations. Circulating 25(OH)D, pro- and anti-inflammatory cytokines, parathyroid hormone, calcium, and albumin were measured in young (25 to 42 y), reportedly healthy adults during the winter. Subjects were separated into one of two groups: vitamin D insufficient (25(OH)D &lt; 32 ng/mL, n = 14) or vitamin D sufficient (25(OH)D &gt; 32 ng/mL, n = 14). Age, height, body mass, and body mass index, and circulating parathyroid hormone, calcium, and albumin concentrations were not significantly (all p &gt; 0.05) different between groups. However, pro-inflammatory cytokines (interleukin (IL)-2, IL-1β, tumor necrosis factor-α, and interferon-γ) were significantly (all p &lt; 0.05) greater in the vitamin D insufficient group. The anti-inflammatory cytokines, IL-5 and IL-10, were not significantly (both p &gt; 0.05) different between groups. We conclude that regulators of vitamin D are not disrupted with vitamin D insufficiency, but insufficiency could result in pro-inflammatory stress without modulating anti-inflammatory cytokines in young adults.
Regulation of IL-4-induced STAT6 activation by SUMOylation (P6307)
Authors: Lee Choong-Eun , Tran Tam , Kim Seol-Hee , Sung Ki-Sa , Choi Cheol-Yong 
Published: January 2023

Abstract
As a key signaling molecule in the cytokine-mediated transcriptional activation STATs are shown to undergo PTMs including phosphorylation, acetylation, methylation, ubiquitination and SUMOylation. The SUMO-modification of STAT1 has been implicated in the regulation of protein stability, nuclear translocation, and transcriptional activation, probably through the reduction of the formation and life-span of the active dimer. As a part of regulation of IL-4 signaling mechanism, we have investigated the role of SUMOylation on STAT6 activation. We have identified potential SUMOylation sites on STAT6 by SUMOplot prediction program analysis and observed the IL-4-induced SUMOlyation of STAT6 in HeLa cells. The introduction of mutation at a potential SUMO site K636 and transfection of shRNA of SUMO-conjugating enzyme UBC9 resulted in the inhibition of tyrosine phosphorylation at Y641 induced by IL-4. The results indicate that endogenous SUMOylation reaction of STAT6 is critical for IL-4-induced STAT6 phosphorylation. Conversely, the forced expression of SUMO-1 and STAT6 caused cytosolic relocalization of both STAT6 and phospho-STAT6 induced by IL-4 as demonstrated by Western blot and confocal analysis. Our data suggest that SUMO-modification of STAT6 may play dual roles in the regulation of phosphorylation and nuclear translocation of STAT6 during IL-4 signal transduction.
A critical role of Toll-like receptor 2 in chronic stress-induced immune suppression (P6309)
Authors: Yin Deling , Tan Yang , Feng Yimin , Caudle Yi , Xiang Yanxiang , Hu Dan 
Published: January 2023

Abstract
Stress can enhance or suppress immune functions depending on the duration and severity of stressful situation. Our previous studies observed that Toll-like receptor (TLR) participates in chronic restraint stress-induced immune dysfunction. However, the mechanism by which TLR2 contributes to immune suppression induced by chronic restraint stress remains elusive. Our investigation found that stimulation of TLR2 by peptidoglycan (PGN), a TLR2 ligand, significantly attenuates cell apoptosis in splenocytes induced by chronic restraint stress. Furthermore, we found that administration of PGN markedly blocks chronic restraint stress-induced alterations of anti-apoptotic and apoptotic genes, including Bcl/2 and glycogen synthase kinase-3β (GSK-3β). We also found that activation of TLR2 by PGN inhibits chronic stress-reduced level of c-Jun N-terminal kinase (JNK) phosphorylation. This effect of PGN is abolished in TLR2 deficient mice, suggesting PGN-induced protection of JNK phosphorylation through a TLR2 dependent manner. Moreover, our data have shown that chronic stress caused a dramatic decrease in cytokine IL-2 level but an increase in the cytokines IL-4 and IL-17 in CD4+ T cells. Interestingly, stimulation of TLR2 by PGN could block these alterations of cytokine levels induced by chronic stress. Collectively, our studies thus demonstrate that stimulation of TLR2 attenuates chronic stress-induced immune suppression by modulating apoptosis-related proteins and cytokines.
Comparison of IFN-γ and IFN-γ in antiviral, immune regulatory and apoptotic roles in fish (P6311)
Authors: Nie Pin , Fu Jian , Chang Ming , Huang Bei , Chen Shan 
Published: January 2023

Abstract
The cDNA and genomic sequences of grass carp IFN-γ and IFN-γ related (IFN-γrel) genes were cloned, with the absence of an NLS motif in IFN-γrel. IFN-γ and IFN-γrel differed in many aspects. Quantitative analysis revealed that IFN-γ was mainly expressed in spleen and intestine, while IFN-γrel was mainly expressed in gills and intestine, moderately in thymus and muscle. In peripheral blood leucocytes, the expression of IFN-γ and IFN-γrel were both up-regulated after PolyI:C and LPS treatment. Luciferase activities showed that IFN-γ could enhance interferon promoter activity and NF-κB promoter activity. The function of antivirus and antitumor roles was compared between IFN-γ and IFN-γrel in CIK cells. IFN-γ could induce expression of Mx, PKR, IRF1, SOCS1, TNF-α, while IFN-γrel did not have such roles. Nevertheless, IFN-γ and IFN-γrel could not induce the expression of Casp-3, Casp-9 and cathD. Furthermore, IFN-γ but not IFN-γrel inhibited the replication of GCRV, grass carp reovirus in CIK cells and prompted the apoptosis in CIK cells. Surprisingly, whatever IFN-γ was mutated with the deletion of 14 aa in C terminal or using GGGG substitution of RRRR had the similar effect as the intact IFN-γ in inducing the gene expression and the apoptosis. These results thus indicate IFN-γ is a functional homologous gene similar to IFN-γ in other higher vertebrates, while the role of IFN-γrel may need further investigation.
Jun activation domain-binding protein 1 (JAB1)/CSN5 is required for the optimal response to interferon-α. (P6313)
Authors: Muromoto Ryuta , Matsuda Tadashi 
Published: January 2023

Abstract
Degradation of IFNAR1 protein is one of the mechanisms limiting the extent of cellular responses to type I interferon. Tyrosine kinase 2 (TYK2), a JAK family kinase, has been reported to bind and stabilize the basal IFNAR1 protein level, indicating that TYK2 is a fundamental component of IFN receptor complex. In this study, we identified Jun activation domain-binding protein 1 (JAB1; also known as fifth subunit of COP9-signalosome, CSN5) as a new TYK2 binding partner and investigated the role of JAB1 in the regulation of IFN responses. Knock-down of JAB1 by siRNA resulted in the suppression of IFN-α-induced phosphorylation of STAT proteins and their transcriptional activation. Importantly, JAB1 knock-down induced an activation of SCF ubiquitin ligase complex containing Cullin 1 (CUL1), as judged by the enhancement of covalent modification of CUL1 with the ubiquitin-like protein NEDD8, and markedly reduced the basal protein level of IFNAR1. In contrast, NEDD8 knock-down or inhibition of NEDD8-modification by NEDD8-activating enzyme (NAE) inhibitor resulted in an increase of IFNAR1 protein with concomitant reduction of NEDD8-modified CUL1. Furthermore, NAE inhibitor treatment enhanced HeLa cell susceptibility to growth suppression effect by IFN-α. These data suggested that NEDD8-modification pathway is involved in the proteolysis of IFNAR1 and that JAB1 acts as a positive regulator of IFN responses by stabilizing IFNAR1 protein through antagonizing NEDD8 pathway.
Interleukin 22 receptor alpha1 is expressed by both the luminal and glandular epithelia of the day 34 pregnant mare uterus (P6319)
Authors: Brosnahan Margaret , Miller Donald , Adams Mackenzie , Antczak Douglas 
Published: January 2023

Abstract
Invasive trophoblast cells of the equine chorionic girdle (CG) migrate through maternal endometrium to form endometrial cups, dense accumulations of cells that produce equine chorionic gonadotropin from days 40 to 120 of pregnancy. At day 37 CG cells attach to the luminal epithelium, passing through and phagocytosing the epithelial cells. The CG cells then migrate into the stroma to form the mature endometrial cups and the luminal surface of the endometrium re-epithelializes. Penetration of the stroma occurs in association with endometrial glands, but the glandular epithelium is undisturbed. Interleukin 22 (IL22) is involved in the preservation of epithelia. Previously we identified high expression of IL22 in CG cells using a gene expression array on placental tissues from day 34 of gestation. qRT-PCR analysis showed a striking increase in IL22 mRNA in CG cells from days 32 to 35, and absence of IL22 in other conceptus tissues. Our discovery of IL22 in the CG was novel, as this cytokine had been reported only in immune cells. qRT-PCR also detected IL22 receptor mRNA in pregnant endometrium at levels similar to other equine epithelia. Using in situ hybridization we have now visualized IL22RA1 mRNA in both the luminal and glandular epithelia of the pregnant mare uterus. We hypothesize that IL22 produced by the CG binds IL22RA1 on luminal endometrium and glands, serving as a mechanism of fetal-maternal communication and ensuring the integrity of epithelial surfaces.
In vitro cytokine release assays for commercially available biologics for predicting cytokine storm in human patients (P6321)
Authors: Scesney Susanne , Ibraghimov Alexander 
Published: January 2023

Abstract
Therapeutic antibodies undergo numerous safety tests to ensure in vivo safety. One required safety assay looks at the cytokine release from either human whole blood or human peripheral blood mononuclear cells (PBMC) when incubated with the therapeutic antibody. Recently TGN1412, a CD28 super-agonist antibody, while not causing cytokine release from purified PBMC, when administered in vivo, resulted in cytokine storm in all six volunteers. Since that incident, many laboratories have been working to design a cytokine release assay (CRA) that will show cytokine release in response to TGN1412. In this study we compare two CRA formats. In one assay, test antibody is coated to a polypropylene plate, then incubated with freshly purified PBMC for two days. In the second assay, human PBMC are pre-incubated at high density to make the cells more “lymph node like” (LNL) before incubation with soluble antibody. The response of PBMC from multiple donors to TGN1412 and multiple commercially available antibodies were compared. The plastic fixed antibody tends to generate exaggerated cytokine responses probably due to excessive crosslinking. The LNL format has the advantage of using soluble antibody which may better reflect how PBMC in vivo encounter antibody.
The differential regulation of CCR6 biological functions by site-specific phosphorylation of CCR6 C-terminal tail (P6323)
Authors: Lu Mei-Yi , Liao Fang 
Published: January 2023

Abstract
Chemokine receptors are seven-transmembrane G-protein-coupled receptors (GPCRs). For most GPCRs, agonist-induced receptor phosphorylation at C-terminus leads to the recruitment of β-arrestins, resulting in receptor desensitization and internalization. CCR6 is a CC chemokine receptor whose ligand is CCL20. The role of site-specific phosphorylation of CCR6 at C-terminus in CCR6 biological function has not been investigated yet. In this study, we have intended to identify the specific serine/threonine residues at C-terminus of CCR6, and studied their role in the regulation of CCR6 biological function. We have generated a series of site-specific mutants in which specific serine/threonine residues at C-terminal tail of CCR6 were mutated into alanine as well as deletion mutants in which progressive truncation of C-terminus of CCR6 were generated. We have used these CCR6 mutants to characterize the role of site-specific serine/threonine residues in the regulation of CCR6 biology including the association of β-arrestins with CCR6, the internalization and desensitization of CCR6, the intracellular signaling of CCR6, the intracellular trafficking of CCR6, and the CCR6-mediated cell migration. We found that site-specific phosphorylation of threonine/serine residues at C-terminus of CCR6 differentially regulates CCR6 biological functions. The results will be presented and discussed.
A new chemoattractant protein found by PBMC screening platform (P6347)
Authors: Pei Xiaolei , Zhang Yan , Sun Qianying , Xu Enquan , Zheng Yi , Guo Changyuan , Wang Pingzhang , Ma Jing , Peng Xinjian , Chen Dixin , Zhang Yang , Zhang Yingmei , Guo Shuai , Shi Taiping , Ma Dalong , Wang Ying 
Published: January 2023

Abstract
To find new human gene encoding protein which has potential to be chemokine like or chemokine, our laboratory has established the peripheral blood mononuclear cell (PBMC) chemoattractant platform, which includes four steps. First, we screened out 212 genes potential to be cytokines from the genes library in which the genes’ cytokine-related function was unknown or unreported. And then, we constructed the eukaryotic vectors expressing the candidate genes. And third, the proteins encoded by the candidate genes, which could be secreted, were chosen. At last, the secretive proteins were applied on the PBMC chemoattractant platform to find the protein which had the chemoattractant ability. The PBMC chemoattractant platform was based on the Boyden Chamber Assay. Among the proteins which were potential to be chemokine or chemokine like, one protein was found having the ability to chemoattract peripheral blood monocyte and lymphocyte, which was low expressed in normal tissue and detected in human breast cancer and prostate cancer. What’s more, we have found that CCR2B was the protein’s chemoattractant receptor in vitro. In consideration of CCR2B is normally expressed in lots of cancer sites, the protein which we have found may play some kind role in the procession of cancer development.
FAM19A4: a novel cytokine promotes phagocytosis of zymosan but negatively regulates TNF-α secretion associated with NF-κB pathway in macrophages. (P6349)
Authors: Wang Wenyan , Wang Pingzhang , Li Ting , Wang Xiaolin , Zhang Heyu , Yuan Wanqiong , Zhang Yingmei , Ma Dalong , Han Wenling 
Published: January 2023

Abstract
Cytokines play a key role in the immune system. When pathogen infection occurs, macrophages undergo phagocytosis first, accompanied with inflammatory cytokines secretion. FAM19A4 is a novel cytokine identified by the strategy of immunogenomics. It is a classical secreted protein, low expressed in most normal tissues, but is up-regulated in inflammatory stimulated monocytes-macrophages. FAM19A4 can enhance phagocytosis of murine macrophages against zymosan both in vitro and in vivo by noticeably increasing the phosphorylation of Akt, but reduce TNF-α secretion in zymosan-challenged macrophages by impacting activation of p38 and ERK1/2 in NF-κB pathway, due to the degradation of p65 both in nucleus and cytoplasm. These findings provide a novel cytokine FAM19A4 expressed by stimulated macrophages, which can promote phagocytosis of macrophages by activating Akt, but impact TNF-α increasing with challenge by inhibiting NF-κB pathway.
Suppressor of cytokine signaling proteins are induced by IL-7 and regulate CD127 expression in human CD8 T cells (P6291)
Authors: Al-Ghazawi Feras , Parmar Parmvir , Faller Elliott , El Salfiti Abdulkareem , MacPherson Paul 
Published: January 2023

Abstract
Interleukin (IL)-7 is an essential non-redundant cytokine influencing cell survival and proliferation throughout the life-span of a T cell. IL-7 is currently being investigated as a potential therapy in HIV+ individuals with poor immunological response to antiretroviral therapy. In order to optimize the use of IL-7 in therapeutic settings, it is crucial to understand how expression of the IL-7 receptor-α (CD127) is regulated. We elucidate in this study the mechanism by which IL-7 targets surface CD127 protein for degradation and describe the role of SOCS proteins, a family of eight proteins (CIS and SOCS1-7), in this process. We show here that in primary human CD8 T cells, upon binding IL-7, surface CD127 is rapidly internalized and phosphorylated at Y449 within 20 minutes of IL-7 stimulation, while activation of the JAK/STAT5 pathway stimulates expression of CIS transcripts and proteins. Following IL-7 stimulation, CIS binds directly to CD127 as demonstrated by Co-IP and co-localizes with both CD127 and the early endosomal marker EEA1, then with the proteasomal marker MCP20, as demonstrated by confocal microscopy. Subsequent proteasomal degradation of CD127 and CIS is dependent on an E3 ligase and can be blocked with the E3 ligase inhibitor SMER3. Through siRNA-mediated knockdowns we confirm roles of CIS in the IL-7 mediated degradation of the CD127 protein.
Regulation of IL-4-induced insulin receptor substrate-2 activation in allergic inflammation (P6294)
Authors: Gowda Nagaraj , Fang Jessie , Heller Nicola 
Published: January 2023

Abstract
Allergic asthma is a chronic airway disease driven by Th2 cytokines, IL-4, -5 and -13. IL-4 receptors activate IRS-2/STAT6 signaling to induce differentiation of M2 macrophages, cells associated with allergic lung inflammation in mice and humans. IL-4, but not IL-13, strongly induced tyrosine phosphorylation (pY) of IRS-2 and M2 gene expression. Understanding regulation of IRS-2 activation is essential for designing novel therapeutics. We hypothesized that both induction of SOCS proteins and serine phosphorylation (pS) of IRS-2 downregulates IRS-2 signaling. To test this, we stimulated both human monocytes (U937 cells) and mouse primary macrophages with IL-4 and observed robust pY-IRS2 that peaked at 60 min and returned to baseline by 150 min. Kinetics of induction of mRNA for SOCS1, SOCS3 and CIS by IL-4 matched the decrease in pY-IRS-2. Using a siRNA knockdown approach, we found that SOCS1 is responsible for downregulating pY-IRS2 but not CIS and SOCS3. SOCS proteins bind to ubiquitin ligase complexes and target proteins for proteasomal degradation. Proteasome inhibitor-treated, IL-4-stimulated U937 cells showed extended pY-IRS2 signaling. In U937 cells, IL-4-induced pS-IRS-2 had an opposite pattern to pY-IRS-2 and when serine phosphatases were inhibited, pY-IRS-2 dramatically decreased. Using serine kinase inhibitors, we found that inhibition of mTORC1 prolonged pY-IRS-2 signaling in BMM and U937 cells, thus uncovering novel, targetable regulators of allergic inflammation.
Engineering DNA nanoparticles as immunomodulatory reagents (P1076)
Authors: Li Lingqian , Huang Lei , Lemos Henrique , Chandler Phillip , Pacholczyk Gabriela , Baban Babak , Mellor Andrew 
Published: January 2023

Abstract
Cytoplasmic DNA sensing triggers anti-microbial immunity by inducing interferon type I (IFNαβ) via STING-TBK1 activation but sustained STING activation incites autoimmunity. Systemic DNA nanoparticle (DNP) treatment triggered rapid and potent tolerogenic responses in mice via selective activation of the STING-IFNαβ pathway in myeloid dendritic cells (DCs) to induce indoleamine 2, 3 dioxygenase (IDO) activity. DNPs associated rapidly with splenic marginal zone (MZ) macrophages (MΦs) and some DCs, and IFNαβ and IL-1β were induced soon after DNP injection. IFNβ1 was primarily induced in myeloid (CD11b+) DCs and this response was STING dependent, while IL-1β was induced primarily in granulocytes and was STING independent. IFNαβ-mediated IDO induction caused DCs and regulatory T cells (Tregs) to acquire potent tolerogenic phenotypes. Physiologic depletion of (CD11c+) DCs and STING ablation abolished IDO induction and tolerogenic responses to DNPs, but MZ (CD169+) MΦ depletion did not. Moreover, systemic treatment with the cyclic dinucleotide (c-diGMP) to activate STING directly induced IDO and suppressed antigen-specific Th1 responses. Thus myeloid DCs sense circulating cargo DNA and c-diGMP to induce selective IFNαβ release that promotes dominant tolerogenic responses via IDO. Targeting this distinctive tolerogenic pathway may be an effective strategy to suppress hyper-immune syndromes, including autoimmune diseases incited by excessive IFNαβ release.