bluuebunny/crossref_metadata_2025_split huggingface dataset:
Histamine and lipopolysaccharide induce endothelial cell bone morphogenetic protein 2 (60.1)
Authors: Walia Damandeep , Vimala Raveendran Vineesh , Zhou Jianping , Sharma Mukut , Stechschulte Daniel , Dileepan Kottarapat 
Published: January 2023

Abstract
Vascular calcification is one of the complications of chronic inflammatory diseases. It is a complex and multifaceted disease and is recognized as a pathobiological process sharing many features of bone remodeling. BMP2 is a powerful osteogenic morphogen which belongs to TGFβ superfamily and plays a central role in bone remodeling and calcification of atherosclerotic plaques. We have previously shown that histamine (HIS) and LPS independently and synergistically induce endothelial cell inflammatory responses and contribute to vascular inflammation. Our hypothesis is that HIS and LPS stimulate BMP2 expression in endothelial cells. Human vascular endothelial cells were incubated with HIS (10µM), LPS (100 ng/ml) or HIS+LPS for different time periods and the mRNA and protein expressions were determined by qRT-PCR and Western Blot analyses. HIS and LPS induced BMP2 mRNA expression by 4-fold and 3-fold respectively. HIS-induced BMP2 expression was completely abrogated by cetirizine, a histamine receptor 1 (H1R) antagonist, indicating that histamine effect is mediated through H1R. Western blot analysis showed upregulation of BMP2 by HIS, LPS or HIS+LPS as early as 2 hrs post treatment which sustained upto 24 h. These results suggest that HIS and LPS play a role in the regulation BMP2 expression in chronic inflammation and infection.
Sphingosylphosphorylcholine stimulates CCL2 production and ICAM expression from human umbilical vein endothelial cells (60.2)
Authors: Bae Yoe-Sik , Lee Ha Young , Lee Sun Young , Kim Sang Doo , Kim Hak Jung , Jung Young Su , Baek Suk-Hwan 
Published: January 2023

Abstract
We investigated the functional role of sphingosylphosphorylcholine (SPC), a component of high density lipoprotein (HDL) particles, in human umbilical vein endothelial cells (HUVECs). SPC stimulation induced production of the CCL2 chemokine in a PTX-sensitive G-protein-dependent manner. SPC treatment caused the activation of NF-κB and AP-1, which are essential for SPC-induced CCL2 production, and also induced the activation of three MAPKs, ERK, p38 MAPK, and JNK. p38 MAPK or JNK by specific inhibitors caused a dramatic decrease in SPC-induced CCL2 production. The Jak/STAT3 pathway was also activated upon SPC stimulation of HUVECs. Pretreatment with a Jak inhibitor blocked not only SPC-induced p38 MAPK and JNK activation but also NF-κB and AP-1 activation. Our results suggest that SPC stimulates HUVECs, resulting in Jak/STAT3-, NF-κB-, and AP-1-mediated CCL2 production. We also observed that SPC stimulated expression of the adhesion molecule ICAM-1 in HUVECs. Taken together, we suggest that SPC may contribute to atherosclerosis; thus SPC and its as yet unidentified target receptor offer a starting point for the development of a treatment for atherosclerosis.
Pigment epithelium-derived factor as an inflammatory factor — a new role for an old protein (60.4)
Authors: Chavan Sangeeta , Hudson LaQueta , Li Jian Hua , Ochani Mahendar , Katz David , Tracey Kevin 
Published: January 2023

Abstract
Obesity is a major risk factor for insulin resistance, type 2 diabetes mellitus, and atherosclerosis. A chronic inflammatory state plays a pathogenic role in the development and progression of obesity-related disorders. Adipose tissue is infiltrated with macrophages; however the factors mediating the inflammatory response are not yet clear. Here we have identified and characterized a mediator of inflammation that is secreted by adipocytes. Pigment epithelium-derived factor (PEDF), a non-inhibitory serine protease inhibitor, is expressed by differentiated adipocytes. PEDF induces dose dependent increase in expression of proinflammatory cytokines (TNF, IL1 and IL6) in macrophages. The PEDF receptor, adipose triglyceride lipase (ATGL), is expressed in macrophages, and inhibitors of ATGL attenuated PEDF mediated responses indicating importance of PEDF-ATGL interaction in PEDF mediated inflammatory responses. Analysis of downstream effects demonstrated that p38 and ERK1/2 kinases are phosphorylated following PEDF treatment and consistently, inhibitors of p38 and ERK1/2 kinases attenuate PEDF induced cytokine production. PEDF administration induced increased serum levels of TNF and IL6 in mice, while neutralizing PEDF attenuated inflammatory responses. These studies suggest that PEDF secreted by adipocytes contributes to onset and maintenance of chronic inflammation observed in obesity and may serve as a therapeutic target for treatment of obesity related disorders.
Extracellular adenosine as a master regulator of immune cell migration into the central nervous system via induction of CX3CL1 (60.9)
Authors: Mills Jeffrey , Alabanza Leah , Mahamed Deeqa , Bynoe Margaret 
Published: January 2023

Abstract
Having the ability to modulate lymphocyte entry into the central nervous system (CNS) would benefit patients with neuroinflammatory diseases. We have previously shown that extracellular adenosine regulates CNS entry of lymphocytes during experimental autoimmune encephalomyelitis (EAE), the animal model for the CNS inflammatory disease multiple sclerosis. For instance, while extracellular adenosine levels are vastly increased following inflammatory cellular damage (from the hydrolysis of released cytoplasimic ATP by CD39 and CD73), mice lacking CD73 or given adenosine receptor (AR) antagonists have significantly reduced CNS lymphocyte entry during EAE. We now show through detailed genetic studies that AR signaling regulates lymphocyte migration into the CNS though induction of CX3CL1, a specialized chemokine that acts as both an adhesion molecule and chemoattractant for lymphocytes, monocytes, and NK cells. We show that AR signaling is necessary and sufficient to induce CNS expression of CX3CL1 (as compared to over 40 other chemokines and adhesion molecules). AR regulation of CX3CL1 is critical for EAE progression, as daily anti-CX3CL1 treatments prevent CNS lymphocyte infiltration during EAE. Importantly, AR signaling induces CX3CL1 expression on the choroid plexus, a known CNS entry point for lymphocytes. As AR signaling can promote lymphocyte transmigration across a choroid plexus transwell barrier, we conclude that adenosine is a master regulator of CNS lymphocyte entry.
Blimp1 and two naturally present dinucleotide repeats regulate the expression of mouse CS1 gene. (62.13)
Authors: Dongre Prachi , Mathew Porunelloor 
Published: January 2023

Abstract
CS1 (CD319) is a member of the SLAM family of receptors, expressed on NK cells, T cells, B cells, DCs, etc. It induces NK cell cytotoxicity and is up regulated on B cell activation. CS1 is aberrantly up regulated on B cells in multiple myeloma and a therapeutic humanized anti-CS1 mAb HuLuc63 (Elotuzumab) is in Phase III of clinical trials. Recently, a unique population of CS1high B cells, implicated as major producers of pathogenic auto antibodies was detected in high SLE disease activity index (SLEDAI) patients. Regression analysis indicated direct correlation between the percentage of CS1high B cells and SLEDAI. With the available information on the pathogenic outcome of CS1 up-regulation on B cells, it is vital to study the molecular basis of CS1 expression. Here, we discuss the transcriptional regulation of mouse CS1 gene. According to our data, mouse CS1 promoter binds to Blimp1, mainly a trans-repressor and the master regulator of B cell maturation. Interestingly, mCS1 promoter also bears two unique dinucleotide repeat sequences, (TC) n=48 and (AG) n=42, sensitive to S1 nuclease, indicating presence of two DNA triplexes. Reporter assays indicate a dual regulatory role of Blimp1, as activatory at -1241bp and inhibitory at -704bp on mCS1 promoter and a trans-activating effect of the dinucleotide repeats. This is the first study to demonstrate dual regulation by Blimp1 and the role of unique dinucleotide repeats in transcriptional regulation of a SLAM family member.
Shortening of complementarity determining region 3 of the T cell receptor α and β chains during thymocyte development (62.3)
Authors: Matsutani Takaji , Ogata Masaki , Suzuki Ryuji , Itoh Tsunetoshi , Nishimoto Norihiro 
Published: January 2023

Abstract
T cell receptor (TCR) diversity depends mainly on the hypervariable complementarity determining region 3 (CDR3) α and β loops generated by non-germline-encoded nucleotide insertions and/or deletions. To reveal alteration of CDR3 length during thymocyte development, we analyzed CDR3α and β length distributions in thymic CD4+CD8+ (DP), CD4+CD8- (CD4SP) and CD4-CD8+ (CD8SP) T cells from C57BL/6 (H-2b), C.B10 (BALB/c congenic, H-2b) and BALB/c (H-2d) mice. The CDR3α and β lengths shortened in both CD4SP and CD8SP cells of all strains tested and the extent of shortening varied considerably depending on V segment use. The CDR3α length shortened to the same extent between CD4SP and CD8SP cells whereas the shortening of CDR3β was more considerable in CD4SP than in CD8SP for C57BL/6 and C.B10, but the shortening was only slight for Balb/c. There was no correlation in the extent of CDR3 shortening between CD4SP and CD8SP cells. Interestingly, there was a good correlation in the extent of CDR3β shortening but not of CDR3α in CD8SP among different strains of mice, independent of MHC haplotype. These results suggest that the extent of CDR3 shortening is primarily determined by germline-encoded V segments and partially affected by allelic variation of MHC class I but not of MHC class II. This shows that T cells with shorter CDR3α and β lengths are preferably selected for the functional TCR repertoire, providing an intriguing insight into the interactions of the TCR and p-MHC ligand.
Profiling the repertoire of TCRB usage in induced and natural Treg cells (62.5)
Authors: Carlson Christopher , Livingston Robert , Sherwood Anna , Desmarais Cindy , Warren Edus , Robins Harlan 
Published: January 2023

Abstract
In our recent research, we have found that a subset of all possible T cell receptor beta (TCRB) rearrangements is produced more frequently than expected in unrelated individuals. This subset appears to be defined by rearrangements that are near to germline, with relatively few non-templated nucleotides inserted at the V-D and D-J junctions within complementarity determining region 3. The degree of overlap between TCRB repertoire between individuals does not appear to be dramatically HLA context-dependent. Other groups have described several subsets of regulatory CD4 T cells (Treg), including both thymus-derived natural Tregs (nTregs) and peripherally induced Tregs (iTregs). While both subtypes express the FOXP3 transcription factor that defines them as regulatory T cells, it has been asserted that the expression of a second transcription factor (Helios) can differentiate nTregs (Helios+) from induced Tregs (Helios-). We hypothesize that the nTreg population is likely to be enriched for near-germline TCRB rearrangments, as recurrently produced TCRB that predispose to the nTreg fate are likely to be shared between individuals. We will compare and contrast the TCRB repertoire in three cellular populations within several individuals: CD4+ FoxP3- Helios-, CD4+ FoxP3+ Helios-, and CD4+ FoxP3+ Helios+.
Kinetics of c-Fos induction depends upon strength of TCR stimulation. (63.13)
Authors: Padhan Kartika , Rajat Varma 
Published: January 2023

Abstract
Induction of immediate early genes (like Fos, Jun) in response to T cell receptor stimulation with different MHC-peptide (MHC-p) dose and potency is not well understood. Our aim is to study the kinetics of c-Fos induction in T cells in response to antigen dose and potency. Our model system is AND TCR transgenic mice, whose T cells are specific for moth Cytochrome-C (MCC) peptide. We study the impact of MHC-p potency using altered peptide ligands (APLs). To study c-Fos induction we crossed AND mice to c-Fos-EGFP transgenic mice that express a fusion protein of c-Fos and EGFP driven by the Fos promoter. We observed that upon stimulation of TCR, expression of c-Fos starts after ~30 min, peaks at 3hrs and plateaus for another 2hrs. Strong agonists induce c-Fos even without co-stimulatory molecules where as weak agonists fail to do so. Co-stimulation enhances the percentage of cells responding as well as the level of c-Fos expression. Highering or lowering the peptide dose reduces the expression level of c-Fos. Inhibiton of Calcineurin has no effect on c-Fos expression. Experiment with MAPK inhibitors indicates that all the three MAP Kinases- ERK, p38 and JNK regulate c-Fos induction. ERK primarily regulate c-fos expression in the case of strong agonists (MCC and L98A) where as JNK primarily regulates c-fos expression in case of weak agonists (K99A). We are looking at other regulators of c-Fos induction in response to APLs.
Suppression of Cytokine Signaling 4 (SOCS4) is a novel immunomodulatory molecule in T cells (63.14)
Authors: Luckey Megan , Hong Changwan , Feigenbaum Lionel , Park Hyun 
Published: January 2023

Abstract
Preventing excessive cytokine signaling is critical for a functional immune system. Molecules of the Suppressor Of Cytokine Signaling (SOCS) family play a crucial role in regulating cytokine response, and here we show that SOCS4 is a novel SOCS family member that regulates immune function in T cells. We found that SOCS4 mRNA is highly expressed in the thymus, and specifically in pre-selection CD4, CD8 double positive (DP) thymocytes. TCR signaling significantly downreglated both SOCS4 mRNA and protein expression in DP thymocytes, which also correlated with lower SOCS4 expression in mature thymocytes and in peripheral T cells. Furthermore, T lineage specific overexpression of SOCS4 resulted in impaired T cell development in the thymus, which was presumably induced by a developmental block at the DN stage. Interestingly, SOCS4 overexpression did not result in inhibition of proximal cytokine receptor signaling such as STAT5 or STAT3 phosphorylation as would be expected from other conventional suppressors of cytokine signaling such as SOCS1 or SOCS3. These data suggest that SOCS4 might utilize another mechanism to modulate T cell reactivity. To asses this question, we have generated SOCS4 transgenic mice expressing a transgenic TCR. Preliminary results show further impairment in T cell development in these mice which detailed mechanisms are currently under investigation. Collectively, this study uncovers SOCS4 as a new immunomodulatory molecule in T cells.
The role of deltex1 in regulating T cell tolerance (63.20)
Authors: Hsiao Huey-Wen , Lai Ming-Zong 
Published: January 2023

Abstract
Deltex1 (DTX1) is an E3 ubiquitin ligase and is suggested to be a mediator of Notch signaling. Our previous study found that DTX1 supressses T cell activation and promotes MEKK1 degradation. In this study, we found that T cell activation was partially suppressed by DTX1 with either inactive mutation of RING finger domain or deletion of WWE domain. In addition, we identified another suppressive mechanism of DTX1 by upregulation of Gadd45β, an inhibitor of JNK activation. DTX1 may promote the expression of Gadd45β in the absence of RF domain or WWE domain. Therefore, DTX1 inhibit T cell activation by either E3-dependent or E3-independent mechanisms. To further investigate the physiological function of DTX1 in lymphocyte development and activation, Dtx1-deficient mice were generated. T cell development in thymus and spleen was normal. B cell development in bone marrow was not affected but margine zone B cell population in spleen was reduced in Dtx1-deficient mice. T cell proliferation, cytokine production and MAPK activation were enhanced in Dtx1-deficient T cells. In addition, enhanced autoantibody production, elevated immune complex deposition in kidney and increased leukocyte infiltration in lung and liver were observed in aged Dtx1-deficient mice. These results suggest that DTX1 plays a role in T cell tolerance induction.
CTLA4Ig abrogates allergic airway inflammation through a novel B7-mediated IFNγ/STAT1/NO signaling pathway (63.22)
Authors: Deppong Christine , Bricker Traci , Boomer Jonathan , Green Jonathan 
Published: January 2023

Abstract
Optimal T cell activation requires both TCR signaling and costimulation through CD28. In vivo use of the drug CTLA4Ig (Abatacept) interrupts B7:CD28 binding and has been an effective therapeutic strategy for chronic inflammatory disease. The presumed mechanism of action of CTLA4Ig is that it prevents the initiation of CD28 signaling. However, previous studies in our laboratory have demonstrated that in a murine model of allergic airway inflammation, CTLA4Ig can function by a novel CD28-independent, NOS2-dependent mechanism. In this study we further dissected this novel mechanism of CTLA4Ig by looking at both in vivo and in vitro models. We now demonstrate through the use of bone marrow chimeras and knockout mice that this in vivo mechanism requires IFNγ and STAT-1 mediated signaling. Furthermore, we used an in vitro coculture system of splenocytes and bone marrow derived macrophages to establish that CTLA4Ig binding to B7 on splenocytes induces IFNγ secretion. This cytokine stimulates the bone marrow derived macrophages to activate STAT-1 dependent NO production, which inhibits T cell proliferation. This data establishes a novel mechanism for immunosuppression by CTLA4Ig. In contrast to accepted dogma, CTLA4Ig inhibits ongoing inflammation not solely by preventing CD28-mediated costimulation, but by a novel mechanism whereby B7 ligation leads to the induction of an IFNγ/STAT1/NO mediated signaling pathway and suppression of T lymphocytes.
Multiple receptor-ligand interactions direct activation of epidermal γδ T cells in response to keratinocyte damage (63.7)
Authors: Witherden Deborah , Garijo Olivia , Rieder Stephanie , Komori H. , Havran Wendy 
Published: January 2023

Abstract
γδ T cells represent a major T cell population in epithelial tissues where they function in maintenance of the epithelium and provide a crucial first line defence against epithelial insults. Despite their importance, little is known about the molecular mechanisms involved in their activation and function. γδ T cells resident in the epidermis are a prototypic epithelial γδ T cell population and are an essential component of the wound healing process in the skin. We have exploited this population to identify molecules involved in the complex cell-cell interactions between epithelial resident γδ T cells and the neighboring cells they support. In the skin, keratinocyte damage is a primary trigger of γδ T cell activation, however the molecular interactions regulating this activation are unknown. Here we confirm a vital role for TCR-ligand interactions in the initiation of the γδ T cell response to keratinocyte damage. In addition, we identify two receptor-ligand pairs crucial for an effective γδ T cell response both in vitro and in vivo. Interactions between JAML and CAR provide costimulatory signals for γδ T cell activation and signals through CD100 and plexin B2 are responsible for γδ T cell morphology changes in response to wounding. Disruption of these interactions delays the γδ T cell-mediated wound healing response. Our results provide insight into the molecular mechanisms of γδ T cell activation and describe two novel receptor-ligand pairs vital to this process.
How we achieve tolerance for our mothers: evidence for a layered immune system in humans (64.1)
Authors: Mold Jeffrey , Venkatasubrahmanyam Shivkumar , Burt Trevor , Michaelsson Jakob , Weinberg Kenneth , Nixon Douglas , Stoddart Cheryl , McCune Joseph 
Published: January 2023

Abstract
The developing fetal and neonatal immune system is known to be exceedingly prone to tolerance induction following exposure to foreign antigens. In mice this has been partially attributed to the absence of a functional adaptive immune system prior to birth. By contrast, the adaptive immune system is highly developed in human beings as early as the second trimester of fetal gestation. Nonetheless, evidence for intrauterine tolerance to non-inherited maternal alloantigens has been observed in clinical studies suggesting that the human fetal immune system is also prone to tolerance induction. In these studies we provide the first evidence for a mechanism underlying the development of immunological tolerance in human beings involving the generation of fetal CD4+CD25+ regulatory T cells (TReg) in the fetal peripheral lymphoid tissues. Additionally we provide evidence that the tolerogenic properties of the fetal adaptive immune system are in part attributed to the presence of a unique population of fetal T cells distinct from those found after birth. Our findings suggest that fetal hematopoietic stem cells (HSC) give rise to phenotypically and functionally distinct T cells than those derived from adult HSC. These findings have significant implications for many different clinical settings including intrauterine infections, transplantation, and autoimmunity.
The cKrox transgene affects the development and function of multiple T cell types. (64.10)
Authors: Twu Yuh-Ching , Tong Jade , Teh Hung-Sia 
Published: January 2023

Abstract
The zinc finger transcription factor cKrox is crucial for CD4 T cell development. Mice lacking a functional cKrox gene lack CD4 T cells. Furthermore, in cKrox-deficient mice, developing T cells expressing a MHC class II-restricted TCR are redirected into the CD8 T cell lineage. We investigated the effect of the cKrox transgene on the development and function of two other types of T cells, namely self-specific CD8 T cells and CD4 T regulatory cells. In TCR transgenic mice that express a male-specific TCR (H-Y TCR), the only CD8 T cells that developed in male TCR transgenic mice are self-specific CD8 T cells. These H-Y TCR+ self-specific CD8 T cells are characterized by high expression of CD44, CD122, Ly6C, 1B11, NK cell markers and proliferative response in response to IL-2 or IL-15. We found that the cKrox transgene converted these self-specific CD8 T cells into CD4 T cells. Furthermore, the converted CD4 T cells lost the characteristics of these self-specific CD8 T cells described above. Notably, the converted self-specific CD8 T cells express genes that are characteristic of CD4 T cells (GATA-3 and IL-2). The numbers of CD4 T regulatory cells were significantly higher in cKrox transgenic mice and these cells expressed higher suppressor activity compared to CD4 T regulatory cells from control mice. These studies indicate that the cKrox transgene affects the development and function of multiple T cell types. This work is supported by the Canadian Cancer Society.