bluuebunny/crossref_metadata_2025_split huggingface dataset:
Opposing roles of PGE2 and LXA4 in the regulation of necrosis of Mycobacterium tuberculosis infected macrophages (B175)
Authors: Chen Minjian , Remold Heinz G. 
Published: January 2023

Abstract
The innate immune response against attenuated Mycobacterium tuberculosis (Mtb) can include induction of apoptosis of the host macrophage (Mφ) that limits intracellular growth by sequestering bacilli in apoptotic bodies, facilitates efficient pathogen removal and enhances antigen presentation. In infection with virulent H37Rv a completely different mechanism, necrosis, dominates the outcome of cell death. Necrosis serves as a microbial virulence mechanism allowing dissemination of the pathogen. Initiation of necrosis depends on the induction of mitochondrial inner membrane perturbation causing significant mitochondrial transmembrane potential (Δψm) loss. Here we report that attenuated H37Ra induces in Mφ the production of the prostanoids PGE2, PGD2, PGF2α, and TXA2. PGE2 suppresses Δψm loss in H37Rv-infeted Mφ and rescues the infected Mφ from necrosis. In contrast, virulent H37Rv induces the production of LXA4, which triggers Mφ necrosis by blocking prostanoid synthesis leading to Δψm loss. The production of the above mentioned prostanoids is not induced in H37Rv-infected Mφ. PGE2 acts through the engagement of the PGE2 receptor EP-2, which induces cAMP leading to PI3K and PKA activation. Thus, eicosanoids play a crucial role in the regulation of necrosis of Mtb-infected Mφ and are of critical importance in determining the outcome of an Mtb infection
(Supported by NIH grant AI50216).
Differential role for IL-1 dependent on anthrax infection route (B176)
Authors: Loving Crystal Lynn , Chaplin David D , Iwakura Yoichiro , Merkel Tod J 
Published: January 2023

Abstract
Use of B. anthracis as a bioweapon in 2001 has resulted in an increased emphasis on understanding mechanisms of pathogenesis after spore inhalation. Using a murine aerosol challenge model, our lab is focused on identifying host cells and cytokines involved in early control of disease progression. MyD88−/− mice are susceptible to inhalational B. anthracis, but TLR2, TLR4, TLR9, and TLR2/9 knockout mice are not. As MyD88 is also involved in IL-1R and IL-18R signaling, we investigated the susceptibility of these knockout mice and found that IL-1R−/− mice succumb to aerosol challenge, but IL-18R−/− mice do not. Yet, when challenged by the intraperitoneal (i.p.) route both IL-1R−/− and IL-18R−/− mice exhibit enhanced susceptibility to spore challenge. IL-1α and IL-1β signal via IL-1R, but IL-1β appears to be dispensable for protection, as mice deficient in caspase-1, which cleaves pro-IL-1β into bioactive IL-1β, do not succumb to disease. However, caspase-1−/− mice challenged by the i.p. route succumb to infection. It appears that IL-1α or IL-1β are sufficient for providing protection during inhalational disease, as mice deficient in either molecule alone were resistant to aerosol challenge. Yet, IL-1β is critical for survival after i.p. challenge as IL-1α−/−, but not IL-1β−/−, mice survive i.p. challenge. Overall, these results indicate differential role for IL-1 depending on route of B. anthracis spore challenge.
Mast cells enable heightened humoral immunity during infection (B178)
Authors: Shelburne Christopher P. , John Ashley St. , McLachlan James B. , Staats Herman F. , Abraham Soman N. 
Published: January 2023

Abstract
Dendritic cell (DC) accumulation in T-cell zones of secondary lymph organs is a critical event in the maximization of the primary immune response. Here, we demonstrate that mast cell (MC) control of elevated DC trafficking during infection enables the primary adaptive humoral immune response to proceed with heightened intensity. Elevated DC accumulation in draining lymph nodes (DLNs) was found to be the product of continual and incremental recruitment of DCs into the infected tissue site prior to their egress to DLNs. MCs contribute to this pattern of DC trafficking by the release of TNF, which was found to coordinately activate local blood vessel endothelium and DLNs to increase their expression of CD62E and CCL21, respectively. Blockade of either protein interfered with DC accumulation in DLNs, as well as the intensity of the humoral response to bacterial challenge. Thus, MCs enable the humoral immune response to proceed with enhanced intensity, which may be an important consideration in rationale vaccine design.
Supported by grants DK50814 and AI50021 from the National Institutes of Health.
Immunomodulating Activities of the TLR7 Selective Agonist 851, a Novel Drug Candidate for Treatment of High-risk Human Papillomavirus Infection (B182)
Authors: Tomai Mark Allen , Pederson Linda , Vasilakos John , Gorski Kevin , Reddy Laxma , O’Sullivan Gerry , Frank Rodney , Prestegard Terri , Kieper William 
Published: January 2023

Abstract
TLR agonists have been used in treating diseases including infections, cancer and as vaccine adjuvants. Aldara™ (imiquimod 5% cream) is the first TLR7 agonist approved in the US, and is indicated for the treatment of genital warts caused by human papillomavirus (HPV). Aldara was not well-tolerated when delivered vaginally; therefore a 2nd generation analog, 851, that retained the ability to stimulate cytokine production upon vaginal application was selected for development to treat high-risk cervical HPV infection. In human PBMC and purified plasmacytoid DC cultures, 851 stimulated a &gt;10 fold increase in type I IFN, and indirectly activated human NK cells to express CD69 (8-fold) at concentrations of ≥10μM. In the rat, intravaginal application of 851 gel (0.01% and 0.1% w/v) led to local tissue production of TNF-α and MCP-1 without evidence of systemic cytokine induction. Increases in vaginal levels of TNF-α (9-fold) and MCP-1 (81-fold) were seen with the top strength of 851 tested (0.1%). Vaginal application of 851 results in the influx of NK cells, T cells and macrophages and an efflux of DC from the vaginal tissue. The ability of 851 to induce local cytokine production leading to influx and efflux of various immune cell populations could result in elimination of virally-infected cells from the cervix and vaginal tract including cells infected with high-risk HPV that are known to lead to cervical cancer.
Impaired pro-inflammatory cytokine production and Th1 biased ability of DCs exposed to Taenia antigens (B183)
Authors: Terrazas Luis I , Terrazas César A , Rivera-Montoya Irma , Rodríguez-Sosa Miriam 
Published: January 2023

Abstract
Helminth infectionsinduce Th2-biased immune responses. Mechanisms involved in this phenomenon are notyet clearly defined, but antigen-presenting cells (APC) playan important role in this process. Dendritic Cells (DC) recognize motifs that are conserved between pathogens mainly through Toll-like receptors(TLR) family, which is the main pathway known to be involved in maturating and inducing inflammatory cytokines in DCs. Most of intracellular pathogens drive Th1 type responses largely through these TLRs. By contrast, the result of the interaction between DCs and helminth parasites is less clear. To dissect mechanisms that lead to immune modulation in experimental cysticercosis, we have used DC exposed to Taenia excreted/secreted glycoproteins and examined their role as APCs, as well as in the modulation of DC responses to pro-inflammatory stimuli. We found that Taenia antigens had the ability to downmodulate DC production of TNF-α and IL-12 in response to several inflammatory molecules such as LPS, CpG and Toxoplasma Ags. Taenia-exposed DC displayed increased ability to suppress IFN-γ production but maintained IL-4 production on CD4+ DO11.10 cells in response to OVA in a STAT6-independent manner. Our results reflect the potential ability of T. crassiceps antigens to modulate DC-inflammatory responses to non-related pro-inflammatory molecules.
Work supported by PAPIIT IN208706
Vaccine for Cancer and Infectious Diseases (B194)
Authors: Deisseroth Albert B. , Tang Yucheng , Akbulut Hakan , Maynard Jonathan , Pedersen Line 
Published: January 2023

Abstract
In order to overcome defects in the immune response in older individuals, we have designed an adenoviral vector (Ad-sig-TAA/ecdCD40L) for the in vivo activation and tumor antigen loading of dendritic cells (DCs). This adenoviral vector encodes a fusion protein composed of an aminoterminal tumor associated antigen (TAA) fragment fused to the extracellular domain (ecd) of the CD40 ligand (CD40L) at the carboxyl terminal end. Two sc injections of this vector have broken tolerance to specific tumor associated antigens in two separate TAA.Tg transgenic mouse models (mice transgenic for rat Her-2-Neu (rH2N) and human MUC-1 (hMUC-1)). The immunoprotection extends for over a year and is independent of CD4 cells. We showed that the sc injection of the hMUC-1/ecdCD40L protein following the sc injection of the Ad-sig-TAA/ecdCD40L vector (this vector prime-protein boost is called VPP) increases the levels of both the cellular and humoral immune response over that achievable with just the vector injections alone in the hMUC-1.Tg mice. The antibodies from hMUC-1 VPP vaccinated mice bind to biopsy specimens of human cancers of the breast and prostate. Importantly, the VPP vaccination can induce an immune response even in 18 month old mice which completely suppresses tumor growth in old (18 month old) mice. The Ad-sig-rH2N/ecdCD40L vaccination suppresses the evolution of spontaneous breast cancer in the rH2N.Tg mouse model. The Ad-sig-HA/ecdCD40L vector prime-HA/ecdCD40L protein boost which is directed to the H5N1 avian influenza hemagglutinin (HA) antigen induces neutralizing antibodies against the H5N1 486 avian influenza virus at 1/4000 dilution of the serum.
Novel method for deconstructing humoral responses (B198)
Authors: Barrington Robert A , Feist Frank , Mann Wolfgang , Ketman Ken , Fu Wei , Steen Robert 
Published: January 2023

Abstract
Measurement of antibody production is a critical feature in vaccine design. However, the production and persistence of memory B (Bmem) cells may be more informative for long-term humoral protection against pathogens. In addition to the frequency of available Bmem cells, the heterogeneity of Bmem cells may influence protective responses. To determine heterogeneity, production and persistence of memory B cells with time after encountering pathogens, we implemented the AmpliGrid platform developed by Advalytix AG in concert with 6-parameter polychromatic flow cytometry to deconstruct B cell subpopulations during ongoing immune responses to a model protein antigen. Flow cytometry analysis and cell sorting was performed on a 3-laser FACSAria system. Antigen-binding B cells participating in germinal center (GC) and post-GC responses were sorted directly onto AmpliGrid chips. Subsequent single cell analysis using on-chip 1 ul RT-PCR and sequencing reactions revealed that responding B cells expressing diverse IgH genes can be subdivided into those being selected versus those being eliminated. After the initial response, we find evidence of memory B cells persisting during the subsequent 4–5 months following antigen encounter. This preliminary work suggests it will be of value in the evaluation of experimental vaccines.
Immunodeficiency and accumulation of B1-like B cells in transgenic mice overexpressing catalytically inactive RAG1 (B2)
Authors: Swanson Patrick C. , Anderson Dirk K. , Hassaballa Ashraf E. , Parks Kevin W. , Perry Greg A. 
Published: January 2023

Abstract
Antigen receptor genes are assembled by V(D)J recombination, which is mediated by two lymphoid cell-specific proteins called RAG1 and RAG2. In B cells, primary V(D)J rearrangements yield B cell receptors displaying diverse antigenic specificity. Recent evidence suggests that certain populations of peripheral CD5+ B cells may express the RAG proteins in order to initiate secondary V(D)J rearrangements to “edit” receptor specificity away from autoreactivity or “revise” receptor specificity to further expand an antigen-specific B cell repertoire. In principle, if a means were identified to block this process, B cell maturation and/or function may be impaired. To explore this idea, we generated transgenic mice overexpressing catalytically inactive full-length RAG1 in the periphery. In two founder lines, transgenic mice exhibit a polyclonal expansion of B1-like B cells (B220loCD19+CD5+), pronounced hypogammaglobulinemia, and an impaired humoral immune response. Interestingly, B1-like B cells fail to accumulate in mice expressing both catalytically inactive RAG1 and a heavy chain transgene specific for dsDNA (3H9H56R.KI), but bi-transgenic mice still exhibit hypogammaglobulinemia and may also be susceptible to bacterial infection.
This work was supported by the Health Future Foundation, and the State of Nebraska LB506 and LB692 Research Programs.
Profiling the Mammalian Cell Surface Glycome (B201)
Authors: Li Yu , Tao Sheng-Ce , Qian Jiang , Schnaar Ron , Goldstein Irwin , Schneck Jonathan , Zhu Heng 
Published: January 2023

Abstract
Glycosylation is one of the most complex posttranslational modifications with an extremely high level of diversity that has made it refractory to detailed analysis. To facilitate studies in glycosylation, we have developed a comprehensive lectin microarray for use as a high-throughput analysis of investigating cell surface glycosylation in mammalian cells. Using this approach, we have established a "GlycoCode" and hierarchical organization of 23 mammalian cell lines based on the profile of their surface glycan structures. We also show the potential use of this approach as a sensitive analysis of cell development and differentiation by probing immune system cells. Lectin microarrays represent a novel approach to high-throughput analysis of cell heterogeneity; the results of which can be used to studying diverse cell processes including cell development and differentiation, cell-cell communication, pathogen-host recognition, cell surface biomarker identification, and to aid in development of novel therapeutics.
Real-time PCR array for simultaneous evaluation of multiple cytokine mRNA expression (B204)
Authors: Arikawa Emi , You Min , Wang Jie , Lo Jing-yi , Yu Sean , Yang Jingping 
Published: January 2023

Abstract
Cytokine quantification is an important element in studies of inflammation and immune responses. Quantitative RT-PCR, a rapid and sensitive assay, is the preferred method to quantify cytokine mRNA levels because they are often expressed at low levels. The RT2Profiler™ PCR Array combines the reliable performance of SYBR® Green based RT-qPCR with multi-gene profiling capabilities to simultaneously analyze the expression of a panel of genes from the same pathway. Using PCR Arrays, we have monitored the mRNA levels of 84 different cytokines in human peripheral blood mononuclear cells (PBMC) in response to treatment with 50ng/mL PMA and 1μg/mL ionomycin for up to 24 h. The results identify 23 up-regulated and 6 down-regulated genes (with &gt;5 fold-change & p&lt;0.005) in the stimulated cells when compared to the resting cells at 6 h. At 24 h, the effects of PMA-ionomycin on genes such as BMPs, CSFs, IFN-γ, IL-1β, IL-6, IL-11, TGF-β and TNF are continuously observed, while the effects on other genes such as IL-2, 3, 5, 9, 10, 13, 17 and 22 diminish 24 h after stimulation. To validate these results, the protein level of eight selected cytokines secreted by the PBMC was measured using a multiplex ELISA array. Our data show that changes in cytokine mRNA levels detected by PCR Arrays accurately predict changes in protein levels measured by ELISA. Hence, the PCR Array offers a simple, reliable and sensitive tool for multiple cytokine profiling.
Antitumor effect of resveratrol and potential for combination with immunotherapy (B205)
Authors: Soto Brenda Liz , Hank Jaquelyn A , Polans Arthur S , Sondel Paul M 
Published: January 2023

Abstract
Resveratrol (3, 4′, 5 – trihydroxy-trans-stilbene) is a natural compound currently being investigated for possible health promoting activities. It has antitumor effects on various types of cancer cells in vitro and in some animal models. Clement et al (Blood, 92:3, 996–1002, 1998) demonstrated that trans-resveratrol (t-RV) induced apoptotic cell death in HL60 leukemia cells and T47D breast cancer cells in vitro at doses which were minimally toxic to peripheral blood lymphocytes (PBMC). Additional studies on the effect of t-RV on the immune system have found t-RV to be immunosuppressive in vitro in a time and dose-dependent manner, but not suppressive in vivo. We are currently examining the ability of t-RV to be combined with immunotherapy to treat cancer. Using proliferative and cytotoxicity assays we are defining a dose in vitro and a regimen in vivo that does not suppress the immune system and has additive or synergistic anti-tumor effects when combined with immunotherapy. Our data suggest that t-RV inhibits the proliferation of K562, a natural killer sensitive target, and M21, a human melanoma cell line, in a dose dependent fashion. In addition, t-RV inhibits the ability of PHA to stimulate proliferation of PBMC. Furthermore, t-RV suppresses proliferation of cells already activated with PHA. These responses were dose dependent. We are examining the effects of t-RV on target and effector cells in cell mediated and antibody dependent cytotoxicity assays. These investigations may shed light on ways of combining t-RV with immunotherapy to treat cancer.
Assessing the regulation of T cell function by distinct cytokine combinations using GuavaFlo, a cytometry-based system for multidimensional cellular analysis. (B206)
Authors: Gillis Katherine Anne , Fishwild Dianne M , Lefebvre Ray , Zieske Lynn R , Snyder-Cappione Jennifer E. , Cappione Amedeo J 
Published: January 2023

Abstract
Precise regulation of effector function is critical for mounting a potent, yet specific, immune response to a given antigenic challenge. Distinct cytokine combinations present within a given microenvironment can induce changes in the phenotype and effector functions of immune cells that differ dramatically from the effects exerted by each cytokine alone. Using a multiparametric analysis approach, we investigated the phenotypic and functional changes of all combinations of 6 cytokines (IL-2, IL-4, IL-7, IL-12, TNF-α, and TGF-β) on human T-cell isolates derived. Cultures were assessed for changes in maturation/activation state, as well as determination of their individual proliferative capacity and viability. Cultures were subject to cytokine production profiling through intracellular staining. Samples were acquired using Guava’s EasyCyte Plus system and integrated GuavaFlo software, an analysis platform designed specifically to simplify and expedite the assessment of plate level flow cytometry data sets. We found that unique combinations of cytokines had profound and highly varied effects on lymphocyte fate and function in vitro. Demonstrations of synergy, dominance, and substitution amongst multiple cytokines in our experimental system may reflect actual paradigms underlying such processes as the differentiation of T cell subsets into effector and resting memory phenotypes in vivo.
Live Cell Imaging of Human Mast Cells on a Pico-well LiveCell Array at Single Cell Resolution (B207)
Authors: Maliakal James , Tagen Michael , Kempuraj Duraisamy 
Published: January 2023

Abstract
Mast cells are important effecter cells in allergic and inflammatory responses. When the mast cells are activated they undergo regulated secretion of intracellular granules by exocytosis. This includes inflammatory cytokines such as IL-6, TNF-α and other chemokines. In this study we examined mast cell activation using a microscope slide based pico-well array at a single cell resolution.
The human umbilical cord blood derived mast cells were cultured in the mast cell medium. The cells were counted and 2uL of cell suspension were added to the pico-wells of the array (Livecell array, NUNC). The cells in the array were covered with a sliding cover slip allowing each cell to settle into an individual well. The cover slip also allowed for repeated treatment and washing of reagents via capillary flow. The cells were treated with substance P (SP) to induce activation directly on the array. Live cell imaging showed cells were distributed evenly on the array as one cell per well. After activation, the cells secreted granules which are clearly visible in the pico-wells surrounding the cells. Additionally, LAD2 cells were activated with compound 48/80 (0.1ug/ml). Use of anti-CD63 PE permitted single cell investigation of degranulation. The ability to culture, treat and image non-adherent mast cells in the array allowed us to study single cell responses in a mast cell population, and permits fast screening of potential inhibitors.
The neurotrans mechanism of electronic-acupuncture in DNP-KLH immunized mice. (B21)
Authors: Lee Youngseop , Lee Heekyung , Min Ji-young , Oh Banggul , Lee Mi-Kyung , Hong Moo-Chang , Bae Hyunsu , Kim Yangseok , Shin Min-Kyu 
Published: January 2023

Abstract
Our previous study demonstrated that successive electroacupuncture (EA) at the ST36 acupoint reduces IgE production in BALB/c mice immunized with 2,4-dinitrophenylated keyhole limpet protein (DNP-KLH) by suppression of the Th2 cell lineage development. Here we report that pretreatment of phentolamine (¥á-adrenergic receptor antagonist, 10 mg/kg, i.p.) completely blocks the EA-induced suppression of antigen-specific and total IgE levels in serum, and IL-4 production in anti-CD3 mAb-activated splenocytes in DNP-KLH immunized mice. And pretreatment of Y-25130 (5HT3 receptor antagonist, 3 mg/kg, i.p.) partly blocks EA-induced suppression effect shown above. These results suggest that ¥á-adrenergic receptor play an important role in mediating the suppressive effects of EA on IgE production and Th2 cell response in DNP-KLH immunized mice, and 5HT3 receptor play as a minor factor.
This work was supported by the Brain Korea 21 project in 2007
Simultaneous Determination of Phenotypic Expression and Electronic Volume in Mouse Cells by the Cell Lab Quanta® SC-MPL Analyzer. (B210)
Authors: Cabana Elisa R , Daly Maria , Cheetham Mark , Shankey Vincent T , Thomas Richard , Krishan Awtar 
Published: January 2023

Abstract
The Cell Lab Quanta® SC analyzer, a unique platform with two excitation light sources; a 488 nm diode laser and a HBO arc lamp, can simultaneously measure Electronic Volume (EV), side scatter (SS) and three color fluorescence. This system has an innovative multiplate loader (MPL), which can undertake automatic sample preparation and high throughput analysis.
We have used this instrument to analyze cells from various murine tissues with the objective to describe and identify changes in cellular volume and morphological complexity during development, activation and differentiation. Immunophenotypic analysis of cells isolated from bone marrow, peripheral blood, spleen, thymus and lymph nodes of C57 BL/6 mice was done with conventional combinations of MABs conjugated with FITC, PE and PE cy-5. The MPL was used for sample preparation in 96 well plates.
EV, which is not used at present as a relevant parameter in flow cytometry, is a more precise measure of cell size than the conventional Forward Scatter (FS). EV can be calibrated with NIST traceable beads or appropriate biological calibrators with known Mean Corpuscular Volume.
We conclude that this new automated platform expands the possibilities for researchers by providing means for identifying and differentiating between different cell types on the basis of EV/SS and also by improving the throughput with rapid sample analysis.