bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of syntaxin11 in cytotoxic T lymphocytes (109.26)
Authors: Halimani Mahantappa , Pattu Varsha , Junker Christian , Marshall Misty , Schwarz Eva , Krause Elmar , Matti Ulf , Hoth Markus , Rettig Jens 
Published: January 2023

Abstract
Cytotoxic T Lymphocytes (CTLs) form immunological synapses (IS) with antigen presenting cells in order to kill them. Killing requires release of lytic granules which occurs via exocytosis, and is dependent on SNARE (Soluble N-ethylmaleimide sensitive factor attachment protein receptor) proteins. Syntaxin11 is a SNARE protein that is predicted to be involved in the fusion of lytic granules in CTLs, since mutations within the coding region of Syntaxin11 lead to Familial Hemophagocytic Lymphohistiocytosis (FHL4), an immune disorder caused by impaired granule exocytosis. We have investigated the contribution of Syntaxin11 to lytic granule release. We used live cell imaging to observe the trafficking of full length Syntaxin11. Using CD3 specific antibodies as a marker for a functional IS, we show that Syntaxin11 accumulates at the IS after CTL polarization and conjugation with the APC has occurred. Syntaxin11 was also associated with Munc18-2, consistent with the role of Munc18-2 as a syntaxin chaperone and as a docking factor. Using a Syntaxin11 antibody we also confirmed the subcellular localization of Syntaxin11 in fixed CTLs with confocal microscopy. Syntaxin11 was associated with Munc18-2 and CD3 at the IS in fixed CTLs, consistent with our observations in live cell imaging of CTLs. These results support the conclusion that Syntaxin11 is involved in lytic granule exocytosis.
Human CD8β isoforms with distinct cytoplasmic tails differ in modulating T cell response and regulated cell surface expression (109.31)
Authors: Coman Maria , Thakral Deepshi , Bandyopadhyay Arunima , Riley James , Kavathas Paula 
Published: January 2023

Abstract
The CD8αβ coreceptor influences CD8 T cell recognition and responses in anti-tumor and -viral immunity. The ancestor to the human and chimpanzee CD8β gene acquired two additional exons absent in the mouse that lead to the expression of multiple isoforms (M1-M4) as a result of alternative splicing. In humans these isoforms differ in their cytoplasmic tails and in their expression pattern. The M-1 isoform is predominant in naïve T cells whereas M-4 is predominant in effector memory T cells. To study functional differences we are co-transducing CD8α, each CD8β isoform, and MHCI restricted NY-ESO-1 specific TCR into human CD4+ T cells and measuring cytokine production after activation. We have found differences in induction of cytokine producing cells such as the MIP-1β chemokine with different isoforms. The M-4 isoform cytoplasmic tail has unique sorting motifs that regulate its cell surface expression and it is modified by phosphorylation after activation. The cytoplasmic tail of M-4 could associate with ubiquitinated substrates in 293T cells and was itself mono-ubiquitinated. The M-4 isoform has unique properties that likely favored its selection in effector memory T cells.
Lck-mediated tyrosine phosphorylation of Discs Large Homolog 1 at position Y222 is required to coordinate alternative p38 activation in T cells (109.33)
Authors: Crocetti Jillian , Silva Oscar , Humphries Lisa , Round June , Miceli M. Carrie 
Published: January 2023

Abstract
Stimulation through the T cell receptor (TCR) leads to rapid activation of proximal tyrosine kinases, Lck and ZAP70, which couple to distinct downstream signaling cascades and functional outcomes. We have shown that MAGUK family scaffold protein Discs Large Homolog 1 (Dlgh1) juxtaposes activated Lck and ZAP70 with p38 MAP kinase to selectively activate the alternative p38 pathway and downstream NFAT; but not NFκB. Lck, ZAP70 and p38 are known to complex with Dlgh1 in response TCR engagement. However, Lck and ZAP70 binding sites within Dlgh1 and the mechanism by which Dlgh1 positions and activates Lck and ZAP70 allowing p38 activation remain incompletely understood. Knowing YxxL/I is a preferred motif for Lck tyrosine phosphorylation and that SH2 domains of ZAP70 and Lck prefer to bind Lck-phosphorylated substrates; we identified Dlgh1 Y222 as a potential target of phosphorylation and/or binding. We subsequently assessed the potential role of Dlgh1 Y222 phosphorylation in coordinating the alternative p38 pathway. Preliminary studies using rLck, rZAP70 and GST-Dlgh1 Y222F fusion proteins demonstrate that Y222 is a primary site of Lck phosphorylation required for optimal alternative p38 activation. Experiments are underway to assess the role of Dlgh1 Y222 in Lck and ZAP70 binding and alternative p38 activation in primary T cells. These experiments will elucidate molecular mechanisms behind Dlgh1 coordination of the alternative p38 pathway and TCR signal specificity.
Decreased innate interferon production in type 2 polarized macrophages upon human rhinovirus infection (110.1)
Authors: Stanciu Luminita , Nikonova A. , Traub S. , Khaitov M. , Johnston S. 
Published: January 2023

Abstract
The role of M1 and M2 macrophages (derived in a type 1, respectively in a type 2 cytokine milieu) in human rhinovirus (HRV) infections is not known. M2 macrophage numbers are believed to be increased in asthma and could play a role in the increased susceptibility to HRV-infections. We hypothesized that upon HRV infection M2 produce less innate interferons (IFN-α/β/λ) as compared to M1 macrophages. Methods. Monocyte-derived macrophages (MDM) obtained from PBMC were polarized to M2 (by treatment with IL-4) and to M1 (by treatment with IFN-γ&TNF-α) and infected with HRV16. IFN-α/β/λ gene expression was measured by RT-PCR and protein levels by ELISA. Supernatants of HRV16-infected M1& M2 and MDM were titrated on Hela-Ohio cells to ascertain their 50% tissue culture infective dose TCID50/ml. Results. Upon HRV16 infection we found increased IFN-α1, -β and -λ1 gene expression at 8h and IFN-α1 and -λ1 gene expression at 24 h in M1 as compared to M2 macrophages and MDM. Increased production of IFN-β and -λ1 at 8h and for IFN-α, -β and -λ1 at 24 h, 48 h and 72 h was observed in HRV16-infected M1 as compared to M2 macrophages and MDM. Rhinovirus titer was decreased in M1 macrophages. In conclusion, M1 macrophages produce high levels of antiviral IFN-α/β/λ1 and consequently are less susceptible to virus infection. In contrast M2 macrophages produce low levels of IFN-α/β/λ and this could play a role in virus-induced exacerbations of asthma.
Phospholipases are required for Francisella tularensis induction of macrophage prostaglandin E2 synthesis. (110.15)
Authors: Woolard Matthew , Navratil Aaron , Studt Ashley , Bryan Joshua 
Published: January 2023

Abstract
Francisella tularensis is the causative agent of respiratory tularemia, a serious disease with 30-60% mortality in untreated individuals. F. tularensis is capable of inducing the synthesis of prostaglandin E2 (PGE2) from macrophages. F. tularensis induction of PGE2 synthesis during respiratory tularemia inhibits the development of beneficial T cell responses during respiratory tularemia. Signal transduction events in the macrophage that lead to the synthesis of PGE2 in response to F. tularensis are unclear at this time. Phospholipase (PLases) and lipases are enzymes that cleave phospholipids and lipids repectively leading to the generation of secondary messenger molecules that initiate signal transduction events. PLases are critical to LPS induction of PGE2 production; however the role of PLases and lipases during F. tularensis PGE2 induction is unknown. Utilizing phosholipase and lipase inhibitors we have demonstrated that cPLA2 and DAGL activity are required for F. tularensis to induce PGE2 synthesis in macrophages. This is the first study demonstrating involvement of PLases in the macrophage response to F. tularensis. Subsequent studies will clarify the PLases specific aspects PLases play in the PGE2 synthesis.
CD180 restricts replication of Legionella pneumophila in macropahges and is associated with susceptibility to Legionnaire’s Disease (110.25)
Authors: Misch Elizabeth , Arentz Matthew , Verbon Annelies , Sexson Shannon , Skerrett Shawn , Hawn Thomas 
Published: January 2023

Abstract
Legionella pneumophila (Lp) is a gram-negative bacterium that opportunistically infects the human alveoloar macrophage to cause a pneumonic illness known as Legionnaire’s Disease (LD). The role of CD180 (RP105), a non-signalling member of the TLR family that regulates TLR2 and TLR4 function, in Lp pathogenesis is unknown. We hypothesized that CD180 regulates susceptibility to Lp infection in mice and humans. We infected bone-marrow derived macrophages with Lp (Lp02 strain) and observed greater replication (~2-3-fold) in Cd180-/- mice compared to C57Bl/6 controls. This difference was present over a range of MOIs (0.1 to 2.0) at 24, 48, and 72 hours after infection. Enhanced replication of Lp in Cd180-/- macrophages was not associated with increased pyroptotic cell death, as assessed by IL-1β or LDH, or alterations in key innate immune cytokines or effectors, such as IL-6, TNF, or nitric oxide. We next examined whether CD180 polymorphisms were associated with LD with a study of 108 cases and 508 controls in the Netherlands. Four SNPs, rs574464, rs1428469, rs1697143, and rs1803440, were associated with protection from LD, with ODs ranging from 0.27-0.50 (p values: 0.005-0.045, analysis with dominant genetic model). The functional effects of these polymorphisms and the mechanism of CD180 regulation of Lp replication in macrophages are currently being examined. Together, these results suggest that CD180 critically mediates the immune response to Lp in mice and humans.
Naringenin: an immunomodulator of Interleukin (IL)-6, IL-8, and Tumor Necrosis Factor secretion from Chlamydia trachomatis-infected macrophages and epithelial cells (110.28)
Authors: Yilma Abebayehu , Singh Shree , Fairley Stacie , Terrell Kaitlyn , Taha Murtada , Subbarayan Praseetha , Dennis Vida 
Published: January 2023

Abstract
Chlamydia trachomatis, the leading cause of bacterial sexually transmitted infections, is responsible for a myriad of immunopathological changes associated with reproductive health. Chlamydia infects macrophages and epithelial cells, which secrete cytokines such as TNF, IL-6 and IL-8 that are key protagonists of acute inflammation. These cytokines put the patient at risk for major health issues such as pelvic inflammatory disease and infertility if not controlled. Counterbalancing their levels helps mitigate acute inflammation. We have shown that IL-10 regulates inflammatory cytokines elicited by C. trahomatis in innate immune cells. Here we explored the hypothesis that Naringenin, a natural flavonone, with anti-inflammatory properties, when used alone or co-cultured with IL-10 will modulate inflammatory cytokines secreted from C. trachomatis-infected epithelial (HeLa) cells and mouse J774 macrophages. We exposed C. trachomatis-infected cells to Naringenin (100, 10, 1, 0.1, and 0.01 μg/mL) alone or with IL-10 (10 ng/mL). Our results as demonstrated using cytokine ELISA, confocal microscopy and MTT assay, indicate that Naringenin abrogated TNF, IL-8, and IL-6 levels in a dose-dependent fashion. Cytokine levels were reduced further by Naringenin co-cultured with IL-10. The MTT assay showed the anti-inflammatory effect of Naringenin was not due to cell death. The data provides evidence for a novel therapeutic direction in controlling C. trachomatis-induced acute inflammation.
Dendritic cells from burn and septic injured rats modulate CD4+ CD25+ T cells from sham control rats (111.13)
Authors: Fazal Nadeem , Shelip Alla , Siddiqui Erum , Ali Ashraf , Al-Ghoul Walid 
Published: January 2023

Abstract
Dendritic cells (DCs) play a vital role in presenting antigen to CD4+ CD25+ (Reg) T cells. We hypothesize that burn plus sepsis injury adversely affects antigen presenting cells, in particular the DCs, such that these DC are unable to adequately prime Reg T cell; the inadequate stimulation of Reg T cell by DCs may contribute to burn-induced impairment in T cell IL-2 production and proliferation. In this study, we examined the effect of presence of anti-CD3, anti-CD28, and/or IL-2 in the T cell cultures with or without the added APC/DCs. The results show when sham rat Reg T cells were cocultured with DCs or APCs from burn septic rats, IL-2 production and proliferation was decreased, compared to that in sham T cells cocultured with sham DCs or APCs. When exogenous IL-2 was added to sham Reg T cell and burn and sepsis rat APC cocultures, there was an enhancement in Reg T cell proliferation to the level found in cocultures of sham rat T cells with sham rat APCs. Sham rat T cell proliferation was enhanced in the presence of anti-CD3 but not to the level in cocultures with burn plus sepsis rat APCs. Similarly, addition of anti-CD28 produced but a small increase (~20%) in proliferation of sham rat Reg T cell cocultured with rat APCs, compared to the effect of anti-CD28 in cocultures of sham rat T cell with sham rat APCs. These studies indicate a role of burn-derived APC and DCs in modulating a Reg T cell response in burn and sepsis injury.
Bacillus anthracis capsule promotes TNF-α expression and NF-kB activation through activation of TLR2 in phorbol 12-myristate 13-acetate-differentiated THP-1 and human monocyte-derived dendritic cells (111.14)
Authors: Cho Min-Hee , Ahn Hae-Jeong , Shin Youg-Woo , Yoo Cheon-Kwon , Rhie Gi-eun 
Published: January 2023

Abstract
Anthrax is a highly lethal infectious disease caused by the spore-forming Bacillus anthracis. Poly-γ-D-glutamic acid (PGA) capsule is one of the major virulence factors of B. anthracis. Recently, efforts have been made to include PGA as a component of anthrax vaccine; however, the innate immune response of PGA itself has been poorly investigated. In this study, we characterized the innate immune response elicited by PGA in the human monocytic cell line THP-1, which was differentiated into macrophages with phorbol 12-myristate 13-acetate (PMA) and human monocyte-derived dendritic cells (hMoDCs). The treatment of PGA on differentiated THP-1 cells and hMoDCs leads to the specific extracellular release of TNF-alfa in dose-dependent manner as shown by ELISA. Subsequently ELISA results were validated by microarray analysis and RT-PCR. In addition, the increase of TNF-alfa production was inhibited by TLR2-blocking antibody, TL2.1, indicating the enhancement of cytokine productions is TLR2-dependent. Also, we confirmed that PGA-induced TNF-alfa production occurs via activation of transcription factors nuclear factor (NF)-kB through RT-PCR and phosphorylation of IkB by Western blot analysis. These data indicate that PGA of B. anthracis elicite TNF-alfa production through TLR2 pathway via NF-kB activation in PMA-differentiated THP-1 cells and hMoDCs.
Potency of n-3 PUFA anti-inflammatory effects on human and rodent immune cell responses (111.16)
Authors: Kasprowicz Deborah , Ravindran Nair Sangeetha , Gonyon Tom , Gerstein Clare , Fulghum Tim , Carter Phil 
Published: January 2023

Abstract
Pharmaceutical parenteral emulsions consisting of n-6 poly-unsaturated fatty acids (PUFAs) alone (Intralipid®), n-6 PUFAs and n-9 mono-unsaturated fatty acids (MUFAs; e.g. Clinoleic®), n-3 PUFAs alone (Omegaven®), or a combination of n-3, n-6, n-9 and medium chain triglycerides (SMOF®) provide nutritional benefit to critical care patients, as well as influencing immune cell function. The efficacy of these compounds in modulating cell function varies depending on the formulation composition, route of administration, dose, and immune status of the individual. In general while n-9 MUFAs are neutral, n-6 PUFAs promote inflammatory responses. In contrast, n-3 PUFAs have immunosuppressive properties including blocking cell proliferation, cytokine production, and surface protein expression. However, there is limited data assessing the effects of n-3-containing pharmaceutical emulsions on immune cells across different species. In the current work, we determined the anti-inflammatory effects of n-3 PUFA-containing emulsions on an LPS-induced cytokine response in mouse, rat, and human mononuclear cells. We found that while n-3 PUFAs reduced the LPS-induced cytokine response in all species tested, these compounds were less potent in rodent cells compared to human cells. These findings provide insight into differences in the potency of n-3 PUFA anti-inflammatory effects across species and may be relevant for selecting the appropriate model system for assessing n-3 PUFAs efficacy in vivo.
Phenotypic and functional profiling of GM-CSF- and M-CSF-driven macrophages (111.18)
Authors: Wiener Doris 
Published: January 2023

Abstract
Monocyte-derived macrophages(MΦs) are a heterogeneous population of cells. MΦ function is dependent on polarization to M1 (proinflammatory) or M2 (anti-inflammatory) phenotypes. Described inconsistencies in M1 and M1 vs. M2 function may be due to different differentiation protocols. We performed analysis of GM-CSF-driven and M-CSF-driven M1 and M2 cells. Monocyte-to-MΦ differentiation was driven by exposure to GM-CSF or M-CSF for 6 days. GM-CSF-driven MΦs were polarized toward M1(GM-M1) with LPS+IFNγ; alternatively, M-CSF-driven MΦs were polarized toward M1(MC-M1) with LPS+INFγ or toward M2 with IL-4. MΦ phenotypes were determined by flow cytometry, ELISA and qRT-PCR. GM-M1, MC-M1 and M2 cells have distinct morphologies as well as different expression patterns of (1) scavenger receptors (SR-A, LDLR, CD36, SR-B1); (2) surface markers (CD14, CD16, CD36, CD80, CD163); and (3) cytokines (IL-1β, IL-6, IL-8, IL-10, IL-12, TNFα, IFNγ). CD14, CD16, CD36, and CD163 expression was significantly higher in M2 than in MC-M1 or GM-M1 MΦs. CD80 expression was higher in both M1 populations compared to M2 cells. IL-10 expression was higher in MC-M1 than in GM-M1 or M2 cells; IL-12 expression was detected only in GM-M1 cells. Finally, MC-M1 cells had more phagocytic activity than GM-M1 cells. Our results characterize the profiles of GM-CSF and MC-SF driven MΦs. A rigorous analysis of MΦ polarization may facilitate the search for novel therapeutic strategies in disease prevention and treatment.
Alternative macrophage activation and lung host defense against Pneumocystis murina (111.20)
Authors: Nelson Michael , Christmann Benjamin , Metz Allison , Werner Jessica , Steele Chad 
Published: January 2023

Abstract
We have recently reported that Src TKO mice, which demonstrate augmented lung clearance of P. murina (PM) (Infect Immun, May 2009), have enhanced expression of alternative macrophage activation (M2a) markers such as RELM-α and Arg-1 as well as the M2a-associated chemokines CCL17 and CCL22 (J Immunol, February 2011). We have subsequently discovered that M2a development is a specific response to PM lung infection as AMs isolated from PM-infected mice have increased expression of RELM-α and Arg-1 and produce CCL17 and CCL22 ex vivo. Moreover, M2a development during PM lung infection requires CD4 T cells, as their depletion leads to an inability to maintain optimal lung expression of M2a markers. A putative effector function of the M2a response is CCL17/CCL22-mediated recruitment of eosinophils. Analysis of eosinophil peroxidase mRNA expression revealed increased levels in Src TKO mice and decreased levels in CD4-depleted mice, which directly correlated to the magnitude of M2a development in these two models. In preliminary studies, mice deficient in eosinophils demonstrated impaired PM lung clearance. In addition, administration of simvastatin to mice increased AM M2a marker expression and reduced PM lung burden. Collectively, our data provides evidence that M2a development is the natural immune response to PM, a response regulated by SFKs and requires CD4 T cells for maintenance. Moreover, our work supports a role for eosinophils in lung host defense against Pneumocystis.
Activation of NOD2 induces an IL-32 regulated dendritic cell program in leprosy (111.22)
Authors: Schenk Mirjam , Modlin Robert 
Published: January 2023

Abstract
The ability of the innate immune system to recognize and respond to microbial pathogens is determined by the interaction of microbial ligands with host receptors located in specific cellular compartments. For Mycobacterium leprae, tricaylated lipoproteins are recognized by cell surface TLR2/1 and muramyl dipeptide (MDP) by intracellular NOD2. The association of NOD2 polymorphisms with susceptibility to leprosy led us to hypothesize that activation of NOD2 regulates key innate immune host defense pathways in leprosy. We first examined gene expression profiles of monocytes stimulated with NOD2 ligand (MDP) or TLR2/1 ligand (19kD) and compared them to those found in lesions of tuberculoid (T-lep) vs. lepromatous (L-lep) leprosy. NOD2 activation and T-lep lesions are both characterized by DC gene programs. NOD2 activation induces differentiation into CD1b+ DC that are potent antigen presenting cells compared to TLR2/1L-triggered DC. Gene expression analysis and gene knockdown experiments revealed that NOD2-induced DC differentiation is regulated by IL-32. Furthermore, NOD2L activation of monocytes from L-lep patients show reduced induction of IL-32 and CD1b+ DC; however, CD1b expression is rescued by the addition of exogenous IL-32. In conclusion, we provide evidence that NOD2L induces a IL-32-regulated DC program in monocytes, with relevance to the pathogenesis and therapeutic intervention in infectious diseases such as leprosy.
Regulation of FoxO1 by mTORC2 negatively controls Toll-like receptor responses (111.24)
Authors: Wang Huizhi , Brown Jonathan , Suttles Jill , Kinane Denis , Graves Dana , Martin Michael 
Published: January 2023

Abstract
Activation of the Akt pathway plays a pivotal role in regulating the inflammatory response. Rictor is a component of the mTORC2 complex, and its loss has been shown to abrogate the phosphorylation of Akt at serine 473. However, the biological importance of mTORC2 in innate immunity is currently unknown. Here we demonstrate that rictor plays a critical role in TLR-signaling via its ability to regulate FoxO1. Genetic deletion or siRNA-mediated knockdown of rictor enhanced the production of pro-inflammatory cytokines upon LPS-stimulation, whereas the levels of the anti-inflammatory cytokine IL-10 were suppressed. The hyper-inflammatory phenotype was due to a defective Akt signaling axis, since rictor-deficient cells or rictor knockdown DC exhibited attenuated Akt phosphorylation and kinase activity, whereas complementation with a constitutively active Akt ablated the enhanced inflammatory response. Analysis of downstream targets showed phosphorylation of FoxO1 was impaired in rictor-deficient or rictor knockdown cells. The loss of rictor resulted in elevated levels of nuclear FoxO1 and an inability to export FoxO1 into the cytoplasm upon TLR-stimulation, an event that was restored upon complementation with a constitutively active Akt. FoxO1 deletion in DC resulted in an attenuated TLR mediated inflammatory response in which rictor knockdown was unable to rescue. These findings identify a novel signaling pathway by which mTORC2 regulates the TLR-mediated inflammatory response