bluuebunny/crossref_metadata_2025_split huggingface dataset:
Cannabinoid receptor modulation of the endothelial cell inflammatory response (P4191)
Authors: Wilhelmsen Kevin , Tran Alphonso , Hellman Judith 
Published: January 2023

Abstract
The cannabinoid receptor (CBR) family is composed of the G-protein coupled receptors, CB1R and CB2R, and potentially GPR18, GPR55 and GPR119, and the transient receptor potential ion channel, TRPV1. Recent reports suggest that several cannabinoids have the ability to modulate the activation of the endothelium in response to inflammatory mediators; however, it is not clear which receptors and what mechanisms are responsible. Our studies aim to determine: 1) the expression repertoire of CBRs in different human endothelial cell (EC) niches; and 2) determine which cannabinoid receptors are responsible for modulating EC activation by toll-like receptor agonists and TNFα. Through qPCR analysis, we have found that all ECs express the same subset of CBRs. Additionally, by utilizing a panel of synthetic, endogenous and plant-derived cannabinoids with differing specificities for each CBR, our data thus far suggest that CB1R and/or TRPV1 activation may be responsible for dampening the inflammatory activation of ECs by TLR2 agonists, as determined by a decrease in IL-6 and IL-8 secretion. Future studies aim to definitively identify the CBR(s) responsible, determine the effect of phytocannabinoids on EC activation in more detail, delineate the cellular mechanisms by which CBRs modulate the EC inflammatory response and more globally, identify the role of endocannabinoids and their receptors in modulation of the inflammatory response to infection.
α1-Acid glycoprotein as an inducer of M2b macrophages (P4139)
Authors: Nakamura Kiwamu , Kobayashi Makiko , Herndon David , Suzuki Fujio 
Published: January 2023

Abstract
Alpha 1 acid glycoprotein (α1-AGP) is an acute-phase protein, and its plasma levels are increased in patients with severe burn injuries and various cancers. It has been reported that α1-AGP has an ability to polarize macrophages (MΦ) from resident MΦ to M2MΦ (IL-10+IL-12− MΦ, an immunosuppressive MΦ with reduced capacity to kill pathogens or cancer cells). So far, three subsets of M2MΦ (M2aMΦ, M2bMΦ and M2cMΦ) have been described. All 3 subsets of M2MΦ are equally immunosuppressive. In this study, resident monocytes (CD14+IL-10−IL-12− cells) cultured with α1-AGP were demonstrated to be specifically converted to M2bMΦ. Healthy donor monocytes cultured for 24 to 72 hrs with 0.0625 to 4.0 mg/ml of α1-AGP produced CCL1 (a biomarker of M2bMΦ) into their culture fluids. The maximum CCL1 production was demonstrated when resident monocytes were stimulated with 4 mg/ml of α1-AGP. Similar to RPMI 1640 medium supplemented with heat-inactivated FBS, CCL1 was produced by healthy donor monocytes stimulated with α1-AGP in serum-free medium cultures. However, CCL17 (a biomarker of M2a monocytes) and CXCL13 (a biomarker of M2c monocytes) were not detected in their culture fluids. Also, these monocytes did not express DC-SIGN (a biomarker of M2a monocytes) and showed weak bactericidal activities against P. aeruginosa and E. faecalis. These results indicate that the MΦ conversion from resident MΦ to M2bMΦ is specifically stimulated by α1-AGP .
Poly (I:C) induces human endothelial barrier dysfunction by disrupting tight junction expression of claudin-5 (P4197)
Authors: Huang Coty , D'Agnillo Felice , Golding Basil 
Published: January 2023

Abstract
Viral infections are often accompanied by pulmonary microvascular leakage and vascular endothelial dysfunction via mechanisms that are not completely defined. Here, we investigated the effect of the Toll-like receptor 3 ligand Poly (I:C), a synthetic analog of viral double-stranded RNA commonly used to simulate viral infections, on the barrier function and tight junction integrity of primary human lung microvascular cells. Poly (I:C) produced a dose- and time-dependent increase in endothelial monolayer permeability accompanied by a corresponding decrease in the expression of claudin-5, a transmembrane tight junction protein. Immunofluorescence experiments revealed disappearance of membrane-associated claudin-5 and co-localization of internalized claudin-5 with lysosomal-associated membrane protein 1 (LAMP-1), a marker of lysosomes, suggesting enhanced lysosomal degradation of claudin-5. Poly (I:C) also induced a dose- and time-dependent cleavage of TLR3 and TRIF as well as activation of NF-kB and IRF3, but not IRF7 (MyD88-dependent pathway), consistent with the direct stimulation of the TLR3 signaling. Pronounced inflammatory response with the release of IL-6, IL-8, MCP-1 and RANTES and a hallmark TLR3-mediated type 1 IFN production (IFNbeta) was also observed. Together, these findings provide new insight on how virally-activated signaling pathways may disrupt vascular endothelial function and contribute to vascular leakage pathologies.
Column-free isolation of untouched mouse plasmacytoid dendritic cells (P4149)
Authors: Tabatabaei-Zavareh Nooshin , Moor Sameet , Thomas Terry , Fairhurst Maureen 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (pDC) are a DC subset distinct from conventional dendritic cells (cDC) in that they are able to produce large amounts of type I interferon (IFN) after challenge with pathogens. Viruses activate pDC mainly through Toll-like receptors (TLR). Unlike cDC, pDC are poor antigen-presenting cells. In line with the functional differences, pDC express lower levels of CD11c and MHCII as compared to cDC. Moreover, a number of markers expressed by pDC are absent on cDC including: PDCA-1, Siglec-H, B220 and Ly-6C. pDC are found in both lymphoid and non-lymphoid organs. Here, we describe a negative selection method to isolate untouched pDC from mouse spleen. This method uses an immunomagnetic, column-free cell separation technology (EasySepTM). Briefly, single cell suspensions of splenocytes are labeled with biotinylated antibodies against non-pDC. Bi-specific antibody complexes against biotin and dextran are used to cross-link non-pDC with dextran-coated magnetic particles. The unwanted cells are then removed using an EasySepTM magnet. The procedure can be automated using RoboSepTM. Starting with 0.4±0.2% CD11c+PDCA-1+ pDC in spleen, purities of 79±9% (n=21) are achieved. The isolated pDC are functional and produce IFN-α in response to TLR9 ligand, ODN-1585 as measured in ELISA. pDC play an important role in anti-viral immunity and autoimmunity, therefore the isolation of untouched pDC is critical in studies examining their role in these immune responses.
Costimulation through GITR increases follicular helper T cell formation and leads to control of a chronic viral infection (P1021)
Authors: Pascutti Maria Fernanda , Brandao Claudia , van Olffen Ronald , Poplonski Tomasz , van Lier Rene , Nolte Martijn 
Published: January 2023

Abstract
We have previously shown that enhanced costimulation through GITR by overexpression of GITRL increases the formation of both effector and regulatory CD4+ T cells. Here we explored whether it could also affect humoral immunity and T cell help to B cells. Although development of mature B cells was not affected in GITRL transgenic (tg) mice, we found that the number of follicular helper T cells (CXCR5+ PD1+ CD4+ T cells, Tfh) was significantly increased, including the absolute number of Tfh-B cell conjugates, as revealed by ImageStream analysis. Tfh from GITRL tg mice had normal expression levels of ICOS, SLAM and CD44 and slightly lower levels of CD62L and CCR7 compared to wild-type (WT) littermates. Interestingly, Tfh from GITRL tg mice produced more IFN-γ and IL-10, which was accompanied by a biased antibody repertoire (decreased IgG3 and increased IgA, IgG2a and IgG2b). Since Tfh have been implicated in the late control of viral replication, we infected WT and GITRL tg mice with LCMV Clone 13. Surprisingly, at day 30 after infection, we could not detect viral genome in spleen and liver from GITRL tg mice, while WT mice were still infected. Also, PD-1 expression was strongly decreased on virus-specific CD8+ T cells, which correlated with faster viral clearance. All in all, these results indicate that GITRL signaling enhances the control of chronic viral infections, by boosting and modulating Tfh cell responses.
Galectin-9 controls the activity of 4-1BB (CD137) (P1009)
Authors: Madireddi Shravan , Eun So-Young , Mehta Amit , Nemcovicova Ivana , Lee Seung-Woo , Niki Toshiro , Hirashima Mitsuomi , Zajonc Dirk , Croft Michael 
Published: January 2023

Abstract
4-1BB is an inducible stimulatory molecule in the TNFR superfamily expressed on activated T cells, dendritic cells, NK cells, and other lymphoid cell types. Although known to bind the TNF family molecule 4-1BBL, we searched for novel binding partners that might modulate or control the activity of 4-1BB. We found that 4-1BB can bind to Galectin-9, a member of the beta-galactoside binding lectins containing homologous carbohydrate recognition domains (CRDs). Surface plasmon resonance analysis for mouse and human molecules demonstrated that the equilibrium binding constant (KD) of this interaction was very strong at ~11 nM and ~85 nM respectively. Functional studies in vitro showed that the stimulatory activity of 4-1BB was dependent on Galectin-9 in that Galectin-9-/- NK cells, CD4 T cells, or CD8 T cells, were largely unresponsive when 4-1BB was ligated but had normal activity when other stimulatory receptors were engaged. Similarly, 4-1BB activity on dendritic cells was impaired when Galectin-9 was absent. Additionally, in EAE and Asthma models, where an agonist of 4-1BB can suppress disease via induction of CD8β+CD11c+ regulatory cells, we found that this protective effect of anti-4-1BB was lost in Galectin-9-/- mice. Thus, our data demonstrate a novel function of Galectin-9 in facilitating signaling through 4-1BB, and suggest that Galectin-9 may allow 4-1BB to oligomerize and/or cluster in a manner that allows efficient engagement of intracellular signaling pathways.
Cell type specific role of B7 for Ag-specific T and B cell activation and differentiation in T-dependent antibody response in vivo (P1017)
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 and T) express B7 in vivo, the role of B7 on each cell type is not well defined. Utilizing cell type-specific B7cKO mice, cell type specific roles of B7 for T-dependent GC and Ab responses were analyzed. Upon NP-OVA/Alum immunization, NP-specific IgG1 production was profoundly defective in mice lacking B7 expression on DC (DC-B7KO) but was intact in mice lacking B7 on B cells (B-B7KO). Consistent with this, NP-binding GC B cells and Ag-specific CD4+ T cell expansion were not observed in DC-B7KO. Surprisingly, however, GC formation and total numbers of GC B and TFH were undiminished in DC-B7KO; whereas B-B7KO showed reduction of total number of GC B and TFH despite normal number of NP-binding GC B. In addition, NP-specific secondary Ab was absent in DC-B7KO but intact in B-B7KO. These results clearly demonstrate that B7 on DC but not B cells is critical for Ag-specific CD4+ T cell expansion and IgG1 production as well as for secondary Ab responses. In contrast, a distinct pathway dependent upon B7 on B cells but not DC supports apparently Ag-nonspecific GC response. Further studies are ongoing to evaluate cell type-specific roles of B7 for affinity maturation, memory T/B formation, and potentially nonspecific or autoreactive components during T-dependent Ab response.
Discovering the determinants of diversity in the human NK cell repertoire by mass cytometry (P3018)
Authors: Horowitz Amir , Strauss-Albee Dara , Nemat-Gorgani Neda , Dogan Ozge , Dekker Cornelia , Mackey Sally , Swan Gary , Davis Mark , Norman Paul , Guethlein Lisbeth , Parham Peter , Blish Catherine 
Published: January 2023

Abstract
NK cells respond to infected and transformed cells using germline-encoded NK receptors (NKRs). Collectively, these NKRs combine to form distinct repertoires that tune the NK cell response. To provide a framework for understanding how perturbations in expression of NKRs influence disease pathogenesis, we defined the phenotypic heterogeneity of NK cells in 22 healthy individuals (ages 21-62; M=10, F=12), including 5 sets of monozygotic twins. Using mass cytometry, surface expression of 38 markers specific for NKRs and lineage markers to identify B cells, T cells and myeloid cells were examined. KIR and HLA class I genotypes were determined by Luminex, and KIR gene content was further assessed using pyrosequencing. NK cell populations, as defined by inhibitory NKRs, were highly concordant between twins (R2 = 0.90) in comparison to unrelated individuals (R2 = 0.66), indicating that the inhibitory repertoire is highly genetically determined. However, when analyzing all twenty-five NKRs, including activating receptors, the concordance in twins (R2=0.34) and in unrelated individuals (R2=0.11) was greatly decreased, indicating significant environmental influence in determining the overall NK cell repertoire. Overall, these analyses reveal tremendous diversity within the NK repertoire that is both genetically and environmentally determined, and offer the ability to examine the functional capacity of NK cells with far greater resolution.
Loss of CD19 expression alters the CDR-H3 content of marginal zone and follicular B cells, changing the composition of the immunoglobulin antigen receptor repertoire (P3022)
Authors: Khass Mohamed , Freeberg Larry , Zhuang Ynigxin , Carter Robert , Justement Louis , Schroeder Harry 
Published: January 2023

Abstract
Expression of CD19 on B cells modulates B cell receptor signaling threshold. Reduced CD19 expression has been observed in some human patients with common variable immune deficiency (CVID), a disease which is characterized by low production of mature B cells with a paradoxical increase in auto-reactivity. Development of the B cell repertoire in the absence of CD19 expression has not been well characterized. We previously showed that the B cell repertoire is controlled by the sequence characteristics of the complementary determining region heavy chain (CDR-H3). We sought to examine the CDR-H3 repertoire in mice lacking CD19 expression, using our novel bioinformatics system, to test whether its absence can affect the control of B cell repertoire content, yielding altered development and auto reactive antibody production. We analyzed the CDR-H3 based on its length, reading frame usage and average hydrophobicity of its amino acid contents. Follicular B cells lacking CD19 expression were found to express an increase in the use of both highly charged and highly hydrophobic CDR-H3 sequences, sequences that are normally excluded from the follicles. Conversely, when compared to C57BL/6 controls, hydrophobic CDR-H3s were suppressed among marginal zone B cells. These findings indicate a loss of normal repertoire control in the absence of CD19, which may help explain why CD19 deficient patients have altered responses to self and non-self antigens.
The role of endogenous galectin-9 in T cells (P3038)
Authors: Wang Li-Chieh , Li Chi-Shan , Chen Huan-Yuan , Chiang Bor-Luen , Liu Fu-Tong 
Published: January 2023

Abstract
BACKGROUND: Galectins are members of a β-galactoside-binding animal lectin family. Recombinant galectin-9 is a negative regulator of Th1 cells through the Tim3 inhibitory signaling pathway, but the function of endogenous galectin-9 in the immune response is not well studied. Here we examined the role of endogenous galectin-9 in T cells by a mouse model of contact hypersensitivity (CHS), mediated by T cells. RESULTS: CHS response was less severe in galectin-9-deficient (Gal-9 -/-) mice than in Gal-9 +/+ mice by checking the ear thickness change. CD4+ T cells from Gal-9 -/- mice secreted less IFN-γ, IL-17, IL-4 and IL-13 than those from Gal-9 +/+ mice when stimulated by anti-CD3/anti-CD28 mAbs in vitro. CD4+ T cells from Gal-9 -/- mice sensitized with dinitrofluorobenzene topically secreted less IL-17, but not other three cytokines, than those from comparably sensitized Gal-9 +/+ mice. The adoptive transfer with sensitized CD4+ T cells from Gal-9 -/- mice evoked less severe CHS response than those from Gal-9 +/+ mice. Further, CD4+ T cells from Gal-9 -/- mice contained a lower amount of phosphorylated IκBα than those from Gal-9 +/+ mice, when activated by anti-CD3/anti-CD28. CONCLUSIONS: Galectin-9-deficient mice exhibit less CHS response, which might be related to lower IκBα phosphorylation and lower cytokine production in CD4+ T cells. Additional studies of the function of endogenous Gal-9 may provide a new insight on treatment of allergic contact dermatitis.
Critical role for inflammasome-independent IL-1β production in osteomyelitis (P6211)
Authors: Lukens John , Kanneganti Thirumala-Devi 
Published: January 2023

Abstract
The immune system plays an important role in the pathophysiology of many acute and chronic bone disorders, but the specific inflammatory networks that regulate individual bone disorders remain to be elucidated. Here, we characterized the osteoimmunological underpinnings of osteolytic bone disease in Pstpip2cmo mice. These mice carry a homozygous L98P missense mutation in the Pombe Cdc15 homology (PCH) family phosphatase PSTPIP2 that is responsible for the development of a persistent autoinflammatory disease resembling chronic recurrent multifocal osteomyelitis (CRMO) in humans. We found that improper regulation of IL-1β production resulted in secondary induction of inflammatory cytokines, inflammatory cell infiltration in the bone and unremitting bone inflammation. Aberrant Il1β expression precedes the development of osteolytic damage in young Pstpip2cmo mice, and genetic deletion of Il1β, but not Il1α, rescued osteolytic bone disease in mutant mice. Intriguingly, caspase-1 activation in the inflammasome complex was dispensable for Pstpip2cmo-mediated bone disease. Thus, our findings establish a critical role for inflammasome-independent production of IL-1β in osteolytic bone disease.
IL-10-mediated modulation of cochlear inflammation (P6228)
Authors: Moon Sung , Woo Jeong-Im , Oh Sejo , Lee Yoojin , Ahn Okjin , Lim David 
Published: January 2023

Abstract
Inflammatory response is commonly involved in the pathogenesis of various cochlear diseases such as acoustic trauma and cisplatin ototoxicity. Previously, we have demonstrated that the spiral ligament fibrocytes play a pivotal role in cochlear inflammation through secretion of chemokines such as CCL2 and CXCL2 in response to pro-inflammatory signals. In this study, we aim to determine molecular mechanism involved in negative modulation of cochlear inflammation. We found that IL-10 inhibits SLF’s CCL2 up-regulation and migration of THP-1 cells in response to SLF-derived molecules. The SLFs appeared to up-regulate HMOX1 expression via NRF-2 activation. The inhibitory effect of IL-10 was mimicked by CoPP and CORM-3 but was blocked by silencing of IL-10RA and HMOX1. IL-10 was found to inhibit binding of p65 NFκB to the enhancer region of the CCL2 gene. IL-10 deficiency appeared to enhance cochlear inflammation secondary to nontypeable H. influenzae-induced otitis media. Furthermore, we demonstrated that the cochlear macrophages serve as a local source of IL-10, which appeared to be primarily distributed in the stria vascularis. Our results suggest that cochlear macrophage-derived IL-10 modulates cochlear inflammation by inhibition of SLF’s CCL2 up-regulation through HMOX1-mediated CO production. This study is anticipated providing us with a novel strategy for the management of inflammation-associated cochlear diseases.
Antigen-specific CD4+ T cell derived IL-10 is uniquely pro-inflammatory in the context of acute influenza (P6234)
Authors: Dutta Avijit , Chen Tse-Ching , Lin Chun-Yen , Lin Yung-Chang , Chang Chia-Shiang , He Yueh-Chia , Huang Ching-Tai 
Published: January 2023

Abstract
IL-10 is regarded as an anti-inflammatory cytokine which prevents damage from immunopathology with exacerbated immune responses. However, severe lung inflammation in fatal human influenza is accompanied with increased levels of IL-10 as well as other pro-inflammatory cytokines. This information raises questions regarding the role IL-10 plays in inflammation associated with acute influenza. We found that IL-10 was produced by CD4+ T cells in synchronization with pro-inflammatory cytokines during an acute influenza virus infection in mice. IL-10 supplementation during the acute phase increased inflammation leading to death, while IL-10 knockout mice displayed decreased lung inflammation and survival benefits. In the absence of IL-10, influenza-specific CD4+ T cells divided less and produced less pro-inflammatory cytokines despite unaffected Th1 polarity, no increase of regulatory T cell populations, and no alteration in the kinetics of viral eradication. Influenza-specific CD4+ T cell derived IL-10 was more important than IL-10 from other sources for the detrimental effect. In this context of acute influenza in mice, IL-10 also impeded the release of mature TGF-β with decreased neuraminidase-mediated desialylation of host immune cells. The abundance of neuraminidase may contribute to the pro-inflammatory role of IL-10 we demonstrated in this context of acute influenza virus infection.
Catechin modulates inflammation in TNBS induced colitis. (P6238)
Authors: Hajj Hussein Inaya , Zeenny Marie-Noel , Shams Ali , Eid Assaad , Jurjus Abdo 
Published: January 2023

Abstract
This study assessed the effect of catechin in modulating the inflammation of colitis. Thirty rats were randomized into three groups: Group I: chemically induced with colitis and received no treatment, Group II: chemically induced with colitis and received daily injection of EGCG, Group III: control group provided with water only. In groups I and II, colitis was induced by TNBS solution. Rats were sacrificed on days 3, 10 and 17 and biopsies of distal colons were assessed by histology and molecular techniques for ROS and changes in cytokines like IL-6 and IL1-β. Colitis induced by TNBS and non-treated with EGCG group, showed high inflammatory scores resulting in colonic mucosa and submucosa damage, necrosis, absence of goblet cells, infiltration of inflammatory cells, crypt abscesses and granulomas. Infiltration of inflammatory cells and tissue damage peaked at day 17 after the induction. At day 3 and day 10, the treated rats with EGCG showed amelioration in the severity of inflammation, but at day 17, the state of the colonic crypts and goblet cells was close to normal. The number of mast cells in the colon of treated rats decreased by more than 30% at day 17 compared to the non- treated rats. In addition, the expression of ROS decreased significantly. IL-1B and IL-6 expressions decrease with time by 50% in the treated group. Our data confirmed the anti-inflammatory effect of EGCG in TNBS-induced colitis and provided a clue to a potential therapeutic agent.