bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lack of exacerbated atherosclerotic lesions in apolipoprotein E-inhibitory FcγRIIb deficient mice is associated with increased anti-inflammatory cytokine response (CCR6P.269)
Authors: Nagarajan Shanmugam , Zhu Xinmei , Ng Hangpong 
Published: January 2023

Abstract
Functionally FcγRs can be classified into the activating (CD64, CD16 and FcγRIV) and inhibitory (FcγRIIb) receptors. Previous finding from our lab and others have shown that deletion of activating FcγR in apoE knockout (apoE KO) mice decreased atherosclerosis. This led to our hypothesis that deficiency of inhibitory FcγRII (CD32b) exacerbates atherosclerosis. We tested this hypothesis using apoE-CD32b double knockout mice congenic to C57BL/6 background (apoE-CD32bB6 dKO). On the contrary to our hypothesis, compared to apoE KO mice apoE-CD32bB6 dKO mice did not show exacerbated atherosclerotic lesions. Our finding is in contrast to previous reports showing exacerbated lesions in apoE-CD32b dKO mice in a mixed background (129s/v and C57BL/6, CD32b129/B6). Molecular mechanisms to understand the difference revealed that CD32b129/B6 mice showed more Th1 and Th17 response, while congenic CD32bB6 showed predominant anti-inflammatory Th2 and Treg responses. Moreover, macrophages from CD32b129/B6 mice showed more IL-1β response in response to LPS or IC, while macrophages from CD32bB6 mice showed more IL-10 response. These findings revealed a novel paradigm that inhibitory CD32b does not directly contribute to atherosclerosis. As CD32b polymorphism has been implicated in lupus susceptibility, the role of CD32b, as pro-atherosclerotic mediators in vivo in lupus-associated cardiovascular disease need to be investigated.
IL-6 deletion attenuates psoriasis-like skin inflammation in K5-IL-17C mice. (CCR6P.270)
Authors: Ward Nicole , Klenotic Philip , Diaconu Doina , McCormick Thomas , Johnston Andrew 
Published: January 2023

Abstract
IL-17C is the most abundant IL-17 family member found in lesional psoriasis skin. IL-17C elicits a 4-fold increase in TNFα (P&lt;0.05) from primary human endothelial cells (ECs) and primary human keratinocytes (KCs) stimulated with IL-17C/TNFα produce synergistic increases in psoriasis signature genes. Mice engineered to overexpress IL-17C in KCs (K5-IL-17C) develop a psoriasiform skin phenotype that improves upon TNFα inhibition. IL-17C also elicits a 3-fold increase in IL-6 from ECs (P&lt;0.05), and IL-6 protein levels significantly increase in uninvolved and involved skin of K5-IL-17C mice (42% and 480%; P&lt;0.05). We hypothesized that IL-6 synergizes with IL-17C to elicit chronic skin inflammation, akin to IL-17C/TNFα. To test this, we mated K5-IL-17C mice with IL-6 KO mice. K5-IL-17C-IL-6 KO mice develop less severe skin disease at 8-10 weeks of age vs K5-IL-17C mice (40% decrease in mouse PASI; n=15, P&lt;0.05) and have significant decreases in serum TNFα (64%; n=4; P&lt;0.035). Histological analyses of involved skin show no differences between K5-IL-17C vs K5-IL-17C-IL-6 KO mice, however analyses of uninvolved skin reveal a significantly thinner epidermis (2-fold decrease; P&lt;0.001), decrease in dermal angiogenesis (27%; P&lt;0.04), and decreases in CD4+ and CD8+ T cells and F4/80+ macrophages (30-51%; P&lt;0.05) in K5-IL-17C-IL-6 KO vs K5-IL-17C mice (n=8/group). These data suggest that IL-6 may contribute to proinflammatory IL-17C signaling critical for sustained psoriasis pathogenesis.
A computational model for mechanistic investigation of negative feedback in Inerleukin-17 receptor signaling (CCR6P.274)
Authors: Sheehan Robert , Garg Abhishek , Gaffen Sarah , Faeder James 
Published: January 2023

Abstract
IL-17 is a pro-inflammatory cytokine that promotes autoimmunity and protects against certain pathogens. Here, we present a computational model of signaling downstream of the IL-17 receptor. To manage the complexity of the system we use a rule-based modeling approach, in which signaling proteins are modeled as structured objects and rules describe their biochemical interactions. The model includes 19 molecules and 46 reaction rules, which expand to a reaction network with 168 distinct chemical species - complexes and phosphoforms of the basic molecules - which are connected through a network of 5,145 unidirectional reactions. The model is defined and simulated using the BioNetGen software. This model encompasses major signaling components downstream of IL-17R, which activate NF-κB and in turn promote the production of pro-inflammatory cytokines. NF-κB also promotes production of A20, a deubiquitinating enzyme that inhibits NF-κB activation in multiple settings. We recently showed that A20 is a negative regulator of the IL-17 signaling pathway. By modeling A20 interactions with TRAF6 and IKK, we are able to recapitulate experimentally-observed oscillations in NF-κB-dependent A20 expression at both the mRNA and protein levels. Further expansion of the model, along with additional experimental work tracking the dynamics of signaling intermediates, will enable us to explore novel hypotheses about the regulatory mechanisms of A20 and other potential regulators of IL-17 signaling.
Noxious pain fibers drive interleukin-23 mediated psoriasiform skin inflammation (CCR6P.275)
Authors: Riol Lorena , von Andrian Ulrich , Ordovas-Montanes Jose 
Published: January 2023

Abstract
To protect against invading pathogens, the skin harbors specialized immune cells, including dermal dendritic cells (dDCs) and T cells, such as IL-17 producing γδT cells (γδT17) whose aberrant activation by IL-23 can provoke psoriasis-like inflammation. The skin is also innervated by peripheral nerves consisting of autonomic and sensory fibers. Interactions between the autonomic nervous system and immune cells in lymphoid organs are known to contribute to systemic immunity, but how peripheral nerves regulate cutaneous immune responses remains unclear. Here, we have exposed the skin of mice to, imiquimod (IMQ), which induces IL-23 dependent psoriasis-like inflammation. We show that a subset of TRPV1+ NaV1.8+ pain fibers is essential to drive this inflammatory response. Imaging of intact skin revealed that a large fraction of dDCs, the principal source of IL-23, is in close contact with pain fibers. Upon selective pharmacological or genetic ablation of nociceptors, dDCs failed to produce IL-23 in IMQ exposed skin. Consequently, the local production of IL-23 dependent inflammatory cytokines by dermal γδT17 cells and the subsequent recruitment of inflammatory cells to the skin were dramatically reduced. Intradermal injection of IL-23 bypassed the requirement for nociceptor communication with dDC and restored the inflammatory response. These findings indicate that nociceptors, by interacting with dDCs, regulate the IL-23/IL-17 pathway and control dermal immune responses.
Critical target cell-specific roles for IL-17 family cytokines/CIKS-mediated signaling in mouse models of skin and lung inflammation (CCR6P.276)
Authors: Siebenlist Ulrich , Claudio Estefania , Ha Hye-Lin 
Published: January 2023

Abstract
The inflammatory cytokine IL-17 has been strongly implicated in human psoriasis and in the imiquimod-induced mouse model of this disease, while the IL-17 family member IL-25 has been associated with allergic asthma in humans and with allergen-induced inflammation in mouse models. Both cytokines signal via the obligatory adaptor protein CIKS (a.k.a. Act1 or Traf3ip2). However what cells are targeted by these cytokines and to what end has not been clearly delineated in these disease contexts. We demonstrate that in imiquimod-induced psoriatic inflammation, IL-17 signals into keratinocytes to dysregulate their growth/differentiation. On the other hand, IL-17 signals into non-keratinocytes to promote cellular infiltration in this psoriasis model and to drive a positive feedback loop, resulting in increased production of IL-17. We further demonstrate that in a chronic house dust mite-induced asthma model, IL-25 uniquely contributes to generation of Th9 cells and to airway remodeling, at least in part by targeting dendritic-like cells. These findings reveal previously unappreciated, cell-type specific functions IL-17 cytokines that contribute to inflammatory diseases at barrier sites.
Chronic activation with a staphylococcal superantigen drives the expansion of CD4, CD8 double negative T cells and promotes multiorgan inflammation mimicking systemic lupus erythematosus in HLA class II transgenic mice. (HUM7P.306)
Authors: Rajagopalan Govindarajan , Tilahun Ashenafi , Chowdhary Vaidehi 
Published: January 2023

Abstract
CD4 and CD8 coreceptors play an important role in the development and maturation of αβ TCR+ T cells. However, T cells lacking CD4 and CD8 (double negative T or DNT cells) are present in the periphery and are significantly expanded in systemic lupus erythematosus (SLE) and Sjogren’s syndrome. The antigens that cause expansion of these DNT cells are unknown. As T cell activation by superantigens does not require coreceptors, we hypothesized that superantigens could activate DNT cells and contribute to autoimmunity. HLA-DQ8 and HLA-DR3 transgenic mice lacking both CD4 and CD8 were generated by intercrossing CD4 and CD8 single knockout mice. The CD4.CD8 double knockout (DKO) mice harbored a diverse repertoire of DNT cells in the periphery, albeit at reduced numbers compared to wild type (WT) mice. DNT cells from DKO mice responded robustly to the superantigen, staphylococcal enterotoxin B, but not to HLA restricted conventional antigens. Implantation of mini-osmotic pumps delivering 10 or 50 μg of SEB continuously over 7 or 28 days induced a lupus-like disease in WT as well as DKO DR3 and DQ8 mice. However, the immunopathology was significantly more pronounced in DKO mice. Immunostaining revealed extensive infiltration of multiple organs including lungs, liver and kidneys with CD3+ T cells in DKO mice. Overall, our study unraveled the role of CD4 and CD8 coreceptors in T cell development/function and the role of superantigens and DNT cells in the pathogenesis of lupus.
Endogenous IL-6 protects exocrine gland integrity and restrains Th1 immune responses in a mouse model of primary Sjögren’s syndrome (HUM7P.309)
Authors: Zhou Jing , Jin Jun-O , Patel Ekta S , Yu Qing 
Published: January 2023

Abstract
IL-6 is a multi-functional cytokine that can either promote or suppress tissue inflammation depending on specific disease context. Previous reports showed that IL-6 is elevated in exocrine glands and sera of patients with Sjögren’s syndrome (SS), a prevalent systemic autoimmune disease primarily affecting tear and saliva secretion. However, the particular function of IL-6 in the SS disease setting is not defined. Here we show that IL-6 levels were increased during the development SS-like disease in C57BL/6.NOD-Aec1Aec2 mice, a well-defined model of primary SS. Interestingly, in vivo neutralization of endogenous IL-6 led to enhanced tissue apoptosis, leukocytic infiltration and Th1 immune responses in salivary and lacrimal glands. The tissue protective effect of IL-6 was also demonstrated in an anti-CD3 antibody-induced multi-organ inflammation model, as neutralization of IL-6 enhanced, whereas administration of IL-6 inhibited tissue apoptosis and caspase-3 activation in both salivary and lacrimal glands. Preliminary molecular analyses showed that the pro-survival effect of IL-6 was associated with altered Bcl-xL and Mcl-1 gene expression in the exocrine glands. Thus, IL-6 exerts tissue-protective and anti-inflammatory effects on exocrine tissues under inflammatory conditions such as primary SS, demonstrating the complex, tissue- and disease-specific function of IL-6 and the possibility of developing new therapies for SS by manipulating IL-6 expression or function.
Peripheral human blood lymphocytes are associated with metabolic risk factors based on glycemic states (HUM7P.285)
Authors: Rosendahl Alexander , Cucak Helena , Vistisen Dorthe , Philipsen Annelotte , Witte Daniel 
Published: January 2023

Abstract
Inflammation is considered a contributing factor to obesity-linked insulin resistance, hyperglycemia and hyperlipidemia leading to type 2 diabetes (T2D). Cytokines are increased in early T2D reflecting a phenotypic switch of the circulating immune cells. To identify cellular inflammatory markers to identify patients at risk to develop T2D, blood from subjects with normal and impaired glucose tolerance was analyzed by flow cytometry. The markers were correlated with known risk factors for T2D (FPG, 2 h PG, HbA1c and BMI). By random forest analysis for each metabolic risk factor the four highest ranked immune cell markers was identified. Foremost, the results showed a reduction of subpopulations of TLR4 expressing T and B cells and T cells expressing the IL-21 receptor with increasing BMI. These data demonstrate that circulating immune cells displayed an altered phenotype already in patients having increase of common diabetic risk factors. These data may guide and provide novel insights to the inflammatory mechanisms involved in diabetic subjects and could be considered as future biomarkers to identify patient groups at risk for disease exacerbation.
Deficiency of Foxp3/Helios co-expressing regulatory T-cells correlates with clinical acitvity in autoimmunity (HUM7P.316)
Authors: Chen Xiaohua , Vander Lugt Mark , Szabolcs Paul 
Published: January 2023

Abstract
Helios is expressed by thymus-derived natural CD4+Foxp3+ Tregs. There is limited knowledge of the role of Helios+ Tregs in human autoimmune disease (AD). In this study, we monitored the phenotype and activation status of these cells in 3 patients with AD pre/post autologous-CD34 selected hematopoietic stem cell transplant (auto-HSCT) and 7 controls on an IRB-approved protocol. Treg related phenotypes were tested with conjugated antibodies against CD4, CD25, CD127, Foxp3, Helios, CD45RO, CD62L. The gating strategy is shown in Fig1. In patients, the mean percentage of single Foxp3+ or single Helios+ cells among CD4+CD25+CD127low Treg was comparable or somewhat higher than in controls. However, the proportion of Foxp3/Helios-co-expressing Tregs (FH-Treg) were significantly lower in patients (21 vs 46%; p=0.01) (Fig2a). Amongst Foxp3+/Helios+ Tregs, the central memory (CD45RO+CD62L+) population was significantly lower in AD compared to control (5 vs 15%; p=0.03) (Fig2b). In one patient, the percentage of FH-Treg normalized post-auto-HSCT and decreased to pre-transplant value after relapse (Fig3c). To our knowledge, we are the first to report deficiency of FH-Treg in human AD. FH-Treg, rather than Foxp3 or Helios single positive Treg, may play an important role in regulating autoimmunity. Co-expression of Foxp3+Helios+ may represent a critical functional stage of Treg. This will be studied in a large cohort.
Maturation defects in NOD macrophages impacts beta cell regeneration (HUM7P.286)
Authors: Coudriet Gina , Criscimanna Angela , Esni Farzad , Piganelli Jon 
Published: January 2023

Abstract
In Type 1 Diabetes (T1D), CD4+ T cells drive autoimmune destruction of pancreatic β cells by activating M1 macrophages which hinders β cell repair. Endogenous β cell growth can occur if inflammation is tempered. A more controlled inflammatory response is necessary to induce the regenerative process, characterized by a transition of macrophage phenotypes from M1 to M2. Macrophages from NOD mice have several maturation defects. We hypothesize that upon β cell destruction in autoimmunity defective macrophages fail to transition from the M1 to M2 phenotype, rendering β cells unable to initiate the repair program. We adoptively transferred NOD.scid mice with BDC-2.5-TCR-Tg splenocytes to cause T1D. Mice receiving a CD4+ T cell-depleting antibody (GK1.5) had a restricted inflammatory response and delayed T1D onset. We observed low levels of M1 macrophage genes in pancreatic tissue after GK1.5, while M2-macrophage gene expression was maintained. Control mice demonstrated a persistent M1 phenotype overwhelming the M2 macrophage lineage. β cell maintenance is achieved though β cell expressed growth factors, facilitated through M2 macrophages. In pancreatic injury we observed that autoimmune mice have decreased growth factor expression, e.g. HGF, compared to non-autoimmune mice. These macrophage defects may render β cells unable to initiate replicative programs. We propose that addition of HGF to the islet will provide the necessary cues from macrophages for β cell growth and function.
Failure of human maternal immune tolerance at the uteroplacental interface in pre-eclampsia (HUM7P.322)
Authors: Loewendorf-Snead Andrea , Nguyen Tina , Yesayan Maria , Kahn Daniel 
Published: January 2023

Abstract
Background: Pre-eclampsia is an elusive disease specific to human pregnancy. Risk factors suggest an immunologic cause, but specific data is lacking. Methods: A case control study of normal to pre-eclamptic mothers was undertaken (UCLA IRB # 11-002962). Cells were harvested at the uteroplacental interface at the time of cesarean section using a surgical sponge and, together with peripheral blood lymphocytes, subjected to FACS analysis. Results: Comprehensive evaluation of the peripheral and local maternal cellular milieu demonstrates maternal immune activation during pre-eclampsia. Significant reduction of peripheral naïve CD4 T cells concomitant with increased central memory CD4 T cells is accompanied by a significant increase of HLA-DR+ Activated Treg in pre-eclampsia. At the uteroplacental interface, CD8 HLA-DR+ effector T cells and Fas+ CD56hi NK cells are significantly enriched. Discussion: Systemic and local maternal immune activation is a feature of pre-eclampsia suggesting a breakdown in maternal immune tolerance. The observed peripheral increase in Activated Tregs is not mirrored at the uteroplacental interface suggesting a potential deficit in Treg trafficking. In pre-eclampsia, maternal cellular immune activation is invoked at the uteroplacental interface involving both the T cell and NK cell compartments. The kinetics of peripheral blood CD4 immune activation suggests long-standing underlying inflammation potentially providing a diagnostic- and prognostic tool.
Monocyte-derived dendritic cells from cirrhotic patients retain similar capacity for maturation/activation and antigen presentation as those from healthy subjects (HUM7P.290)
Authors: Kaplan David , Tanoue Shiroh 
Published: January 2023

Abstract
Defects in dendritic cell function have been implicated in impaired T-cell responses against chronic hepatitis viruses but few studies have investigated the impact of liver cirrhosis on DC function. The purpose of this study was to compare the activation and APC capacity of MoDC from cirrhotic relative to healthy donors. CD14+ monocytes isolated from PBMC from cirrhotic or healthy donors were matured with GM-CSF/IL-4 x 24h, then activated with TNF-α/IL-1β/IL-6/PGE2/GM-CSF/IL-4 x24h with pre- and post-activation phenotyping. MoDCs were irradiated to 30Gy and pulsed with 15mer peptide pools for two liver tumor antigens alphafetoprotein (AFP) and glypican-3 (GPC3) as well as 9-10mer peptides for CMV/EBV/influenza (CEF). Autologous T-cells purified from PBMC were plated with irradiated/pulsed DC (5:1) and expanded with IL-2, IL-15 and IL-21, refeeding on d13 and d20. Expanded T-cells were evaluated by IFNγ ELISPOT and intracellular cytokine staining on d27. 13 patients with cirrhosis (CIR) and 5 healthy age-matched donors (HD) were studied. No significant difference were identified in MoDC maturation/co-stimulatory phenotype between HD and CIR examining CD40, CD70, CD83, CD86, CD137, OX40L, or HLA-DR. No significant differences were observed in MoDC expansion of AFP, GPC3 or CEF-specific IFNγ+ T-cells by Elispot or ICCS. Conclusion: MoDCs isolated from cirrhotic individuals retain similar capacity for activation, maturation and antigen-presentation as those from healthy donors.
Activated PD-1highCD127highCD27highCD45RAlowCD4 T cell phenotype is associated with increased susceptibility to HIV infection rather than exhaustion. (HUM8P.339)
Authors: Petrovas Constantinos , Paris Robert , Koup Richard 
Published: January 2023

Abstract
The role of PD-1 as regulator of CD4 T cells in HIV infection is not well understood. We found a differential expression of PD-1 on CD4 and CD8 T cells with the PD-1highCTLA-4lowCD127highCD27highCD45RAlowCD4 T cell population (PD-1highCD4TCM) declining with disease progression. Induction of PD-1highCD4TCM cells after HAART was accompanied by reduction of PD-1highCTLA-4highCD27highCD45RAlowCD4 T cells, a population enriched with HIV-specific CD4 T cells. Increased HIV-1 gag DNA was found in PD-1high compared to PD-1lowCD127highCD27highCD45RAlowCD4 T cells. This profile was associated with increased activation and expression of the HIV coreceptor CCR5 on PD-1highCD4TCM cells. We found increased telomerase activity indicating that PD-1 upregulation is due to a direct rather than a bystander activation of CD127highCD27highCD45RAlowCD4 T cells. In vitro stimulation of sorted populations revealed an increased production of IFN-g, MIP1-a, IL-4, IL-10, and IL-17 by PD-1high compared to PD-1lowCD127high CD27high CD45RAlow CD4 T cells. In line with our previous studies, the majority of CCR7highCXCR5highCCR6highCD4 T cells, a population with increased capacity for delivering in vitro help to autologous B cells, was found in the PD-1highCD4TCM cell compartment. Our data show that expression of PD-1 on early-differentiated CD4 T cells may represent a higher susceptibility to HIV infection associated with a recent activation and differentiation rather than exhaustion of these cells.
Investigating the role of inducible T cell kinase in CD8+ T-lymphocyte cytolytic effector function (HUM8P.343)
Authors: Kapnick Senta , Schwartzberg Pamela 
Published: January 2023

Abstract
CD8+ cytotoxic T lymphocytes (CTLs) are critical for eliminating virally infected cells, and defects in CTL responses are implicated in lymphoproliferative diseases. Patients with mutations in inducible T cell kinase (Itk), a tyrosine kinase that serves as an amplifier of T cell receptor (TCR) signaling, develop lymphoproliferative disease associated with ineffective responses against EBV, a virus that infects B cells. This phenotype is similar to X-linked lymphoproliferative syndrome, where mutations in the adaptor SAP lead to fatal EBV infections associated with specific defects in B cell cytolysis. Unlike SAP-deficient CTLs however, we found CTLs from Itk-deficient mice exhibit impaired killing of multiple targets, suggesting Itk-deficiency leads to global defects in cytolysis. Killing by CTLs begins when TCR signaling triggers adherence to targets, centrosome polarization, and release of secretory granules inducing cytolysis of targets. Although early events such as adhesion, actin ring formation, and polarization of lytic machinery were intact in Itk-deficient murine CTLs, we found defects in granule secretion suggesting Itk may play an unappreciated role in the final stages of killing. Exploration of the role of Itk in CTL function may provide new insight into the contribution of early TCR signaling to late stages of cytolysis, and why mutations in Itk lead to lymphoproliferative diseases.