bluuebunny/crossref_metadata_2025_split huggingface dataset:
Overexpression of βarrestin-2 in prostate cancer cells from African American males may contribute to aggressive tumor growth (P2075)
Authors: Thomas Ariel , Rivers E. , Smith Nikia , Richardson Ricardo 
Published: January 2023

Abstract
Prostate cancer is the second leading cause of death in African American males. Its incidence is ~60% higher in African American males relative to Caucasian American males. More aggressive prostate cancer is attributed to cancer that is androgen independent resulting in lack of response to androgen replacement therapy. βarrestins are scaffold proteins that promote G protein coupled receptor desensitization and internalization. βarrestin-2 was recently shown to reduce Androgen Receptor expression in prostate cancer cells. The goal of this study was to determine the levels of expression of βarrestin-1/2 in prostate cancer cells from both racial groups and assess their roles in tumor aggressiveness. To that end, prostate cancer cell lines from African Americans (MDAPCa2B, E006AA), Caucasian Americans (LNCaP, PC3), and control (RWPE-1) were assayed for βarrestin-1/2 expression and cell proliferation. βarrestin-1 expression was elevated in all prostate cancer cell lines whereas βarrestin-2 overexpression was observed only in MDAPCa2B. Interestingly, MDAPCa2B showed greater cell proliferation relative to LNCaP and PC3. Suppression of βarrestin-2 in MDAPCa2B cells reduced cell proliferation and tumor growth in nude mice. Altogether the data indicate that 1) βarrestins modulate prostate cancer growth, and 2) overexpression of βarrestin-2 in prostate cancer cells from African Americans relative to Caucasian Americans may contribute to the prostate cancer disparity.
Collagen matrix deposition by hepatic stellate cells protects hepatocellular carcinoma from NK-mediated cytotoxicity (P2013)
Authors: Mailloux Adam , Epling-Burnette Pearlie 
Published: January 2023

Abstract
Liver fibrosis (LF), a leading risk factor for hepatocellular carcinoma (HCC), is caused by collagen-producing hepatic stellate cells (HSC) activated during chronic inflammation secondary to hepatitis-C infection, or alcohol liver disease (ALD). NK cells promote disease resolution via RAE1-dependant HSC clearance. A defect in NK function by an ill-defined mechanism contributes to a pro-fibrotic response. Using an engineered bioartifical collagen matrix, the expression of inhibitory leukocyte associated IgG-like receptor-1 (LAIR-1) was increased on dividing NK cells and IL-2-induced NK proliferation was inhibited. Thus, we hypothesized that the accumulation of HSC-derived collagen directly inhibits NK activity and protects both HSC and HCC from innate clearance. We found that deposition of native collagen matrix for 72h protected HSCs from NK lysis in Cr51 cytotoxicity assays. A HCC cell line (HEPG2), which produced no collagen when cultured in the absence of HSCs, was susceptible to activated NK killing and was unaffected by collagenase. To test if HSC-derived collagen protects HEPG2 cells, the populations were differentially labeled with ipophilic dyes and co-cultured for 72h for use in flow-based assays. Collagen matrix protected both targets, while collagenase pretreatment restored NK sensitivity. These results suggest that HSC-derived collagen may contribute to NK dysfunction in LF and HCC, and identifies LAIR-1 as a potential mediator of tumor-induced immune suppression.
Impact of PPV-23 vaccine in HIV patients under long term HAART (P4262)
Authors: Iyer Anita , Khaskhely Noor , Leggat David , Westerink M A Julie 
Published: January 2023

Abstract
HIV infection is characterized by profound changes in the B cell compartment, which includes, impaired antigen specific response and a significant loss of IgM memory B cells, amongst other defects. Despite HAART’s success in reducing the incidence of opportunistic infections, invasive pneumococcal disease in the HIV infected population continues to be high. PPV-23 vaccine is recommended as a prophylactic measure in the HAART treated population; however, its serological benefit is currently unclear. Although, HAART has been highly successful in reconstituting T cell function, the effect of HAART on antigen specific B cell reconstitution remains to be elucidated. We developed a novel method to identify pneumococcal polysaccharide responding B cells in circulation by conjugating vaccine serotype 14 and 23 to fluorescent molecules. Using this method, we investigated the polysaccharide responding B cells in HIV cohorts on long term HAART. Preliminary data suggests severe depletion of polysaccharide responding IgM memory B cells in HAART treated adults comparable to HAART naïve group; a striking contrast to HIV uninfected adults. This is in alignment with the diminished anti pneumococcal antibody response observed in the HAART treated group. HAART is thus unable to fully reconstitute the functional B cell subset required for protection against S.pneumoniae.
Comparative analysis of CD8 T cell response to cytomegalovirus or influenza virus in healthy human adults (P3002)
Authors: Chen Guobing , Solokina Arina , Oelke Mathias , Madara Karen , Wersto Robert , Schneck Jonathan , Weng Nan-ping 
Published: January 2023

Abstract
The competency of antigen specific (Ag+) lymphocytes plays a critical role in the adaptive immune response. However, little is known about the differences in the competency of Ag+ T cells against different viruses. Here, we compared the CD8 T cell responses of 23 healthy adults to CMV (pp65) or Flu (M1) peptides using an in vitro artificial antigen presenting system. We found that CD8 T cell responses to CMV or Flu varied in both proliferation and Granzyme B production and were independent of the frequency of initial CMV+ or Flu+ CD8 T cells. In the comparison of the responses of Naïve (Tn) and central memory (Tcm) CD8 T cells, Tcm exhibited a greater expansion than did Tn for both CMV and Flu antigen. Intriguingly, the variations of the expansion between CMV+ and Flu+ CD8 T cells were observed only in Tcm but not Tn cells, indicating the importance of antigen experience. Furthermore, the expansion ability was correlated with the telomere length in the initial CD8 T cells positively and expanded Ag+ CD8 T cells negatively, suggesting a role for telomere length in cell proliferation. Taken together, this study showed that incompetency of Ag+ CD8 T cells is associated with antigen experience and short telomere length in healthy adults. Currently, we are analyzing the usage of the TCR variable genes (alpha and beta) of CMV+ or Flu+ CD8 T cells by a single cell RT-PCR-Sequencing method to gain additional information about the structure of these TCR against CMV and Flu antigens.
Rate of AIDS progression is associated with gastrointestinal dysfunction in SIV-infected pigtail macaques (P3045)
Authors: Klatt Nichole , Canary Lauren , Vinton Carol , Morcock David , Estes Jacob , Brenchley Jason 
Published: January 2023

Abstract
During HIV/SIV infection, mucosal immune system dysfunction and systemic immune activation are associated with progression to AIDS, however it is unclear to what extent pre-existing gastrointestinal damage relates to disease progression after infection. Pigtail macaques (PTM) are an excellent model in which to assess mucosal dysfunction in relation to HIV/SIV pathogenesis, as the majority of these animals have high levels of gastrointestinal damage, immune activation, and microbial translocation prior to infection, and rapidly progress to AIDS upon SIV infection. Here we characterized the mucosal immune environment prior to and throughout SIV infection in 13 uninfected PTM and 9 SIV-infected PTM, of which 3 were slow progressors. This small subset of slow progressors had limited innate immune activation in mucosal tissues in the periphery, which was associated with a more intact colonic epithelial barrier. Furthermore, we found that pre-infection levels of microbial translocation, as measured by lipopolysaccharide binding protein (LBP), in PTM correlated with the rate of progression to AIDS. These data suggest that pre-existing levels of microbial translocation and gastrointestinal tract dysfunction may influence the rate of HIV disease progression.
Comparison of the early innate immune responses of Vibrio parahaemolyticus and Vibrio cholerae in human intestinal epithelial cells (P1237)
Authors: Winstead Candace , Buck Evan , El Adaimi Jad , Moores Thomas , Slocum Tyler , Moyle Misty 
Published: January 2023

Abstract
Diarrheal disease is one of the leading causes of death in low-income countries. Two pathogens responsible for these diseases are Vibrio parahaemolyticus and Vibrio cholerae. Although from the same genus, they have distinct virulence factors and induce different host responses. This study evaluates the early immune response of human intestinal epithelial cells (HT-29) to Vibrio pathogens by comparing changes in gene expression induced by the bacteria using quantitative PCR (qPCR) arrays. 84 different genes in an anti-bacterial inflammatory response were compared in cells stimulated in triplicate for two hours with V. parahaemolyticus and V. cholerae. Overall, V. cholerae upregulated many more genes (24 total) than V. parahaemolyticus (11 total). Both species induced expression of the pattern recognition receptor TLR1 and the chemokine CCL5 as well as unexpectedly increasing expression of several putative antiviral mediators including DDX58, IFNGR1 and MHC I. Genes upregulated by only V. parahaemolyticus include IL-8, IRAK-1, MAPK-8, and TLR-5, whereas V. cholerae upregulated NFKB1A and ICAM-1. Interestingly, many more genes were downregulated by V.parahaemolyticus (31 total) compared to V. cholerae (15 total), and several genes downregulated by V. parahaemolyticus (CCR4, TLR4, TLR6 and IL13) were upregulated by V. cholerae. This comparative expression approach with related pathogens may allow patterns to emerge that will guide future experiments.
LC3-associated phagocytosis is a critical regulator of innate immunity. (P1261)
Authors: Martinez Jennifer , Oberst Andrew , Devi-Kanneganti Thirumala , Green Douglas 
Published: January 2023

Abstract
The magnitude and quality of the innate immune response is shaped by many factors, both cell-intrinsic and -extrinsic. The process of autophagy is a cell-intrinsic pathway that has been implicated in the suppression of innate pro-inflammatory cytokines, and polymorphisms in autophagy genes (ATG16L) have been linked to inflammatory disease. A similar but distinct pathway, LC3-associated phagocytosis (LAP), is also important in shaping innate immune responses, largely by linking the sensing of PAMPS to the autophagic machinery, a process critical for the clearance of both pathogens and dead host cells. As autoimmunity may arise from a failure to engulf or degrade dying cells, we sought to determine if LAP might function to dampen inflammatory responses by halting otherwise constitutive signals from within the phagosome. Indeed, we show that mice with phagocytes deficient for LAP machinery (such as ATG7) exhibit increased pro-inflammatory cytokines levels in the serum when challenged with exogenous dead cells. Moreover, successful clearance of the pathogen Listeria monocytogenes requires LAP, rather than conventional autophagy. In order to fully understand this distinct and vital mechanism for effective host defense, it will be necessary to identify molecular events that are unique to LAP, which will allow us to further distinguish the physiological and pathological roles for LAP.
LPS-induced autophagy is mediated by nitric oxide in macrophages (P1285)
Authors: Park Hye-Jin , Pyo Suhkneung 
Published: January 2023

Abstract
Activation of macrophages is essential for the host defense system and accumulating evidence suggests that autophagy is an important component of macrophage activity. Lipopolysaccharide activates Toll-like receptor 4 leading to the release of pro-inflammatory cytokines and inflammatory mediators, which are necessary to activate innate immune response. Another downstream response to LPS/TLR4 signaling is the induction of autophagy, but the signal transduction pathway leading to autophagy is unclear. Here we demonstrate that nitric oxide, which is induced by LPS, induces autophagy via a number of mechanisms in macrophages. To verify the role of NO in autophagy, we compared autophagy-related gene expression in LPS-stimulated and SNP-stimulated macrophages by western blotting. Both LPS and SNP stimulate autophagy in macrophages. In addition, inhibition of NO production by NOS inhibitors, L-NMMA and SMT, significantly reduced LPS-induced autophagy. These finding imply that LPS-induced autophagy might be mediated via NO production. The inhibition of MAPK or NF-κB by pharmacological inhibitors also impaired activation of autophagy by LPS in macrophages. Overall, these finding indicate that LPS induces autophagy in macrophages via activating MAPK/NF-κB/iNOS signaling pathway.
Role of STAT1 in Chlamydia-induced type-I interferon production in murine oviduct epithelial cells (P1245)
Authors: Derbigny Wilbert , Hosey Kristen , Hu Sishun 
Published: January 2023

Abstract
We previously reported that C. muridarum-infected murine oviduct epithelial cells (OE cells) secrete IFN-β in a mostly TLR3-dependent manner. However, C. muridarum-infected TLR3-deficient OE cells were still able to secrete detectable levels of IFN-β into the supernatants, suggesting that other signaling pathways contribute to Chlamydia-induced IFN-β synthesis in these cells. We investigated the role of STAT1 as a possible contributor in the Chlamydia-induced type-1 IFN production in wild-type and TLR3-deficient OE cells. Our data show that C. muridarum infection significantly increased STAT1 gene expression and protein activation in wild-type OE cells; however, TLR3-deficient OE cells showed diminished STAT1 protein activation and gene expression. There was significantly less IFN-β detected in the supernatants of C. muridarum-infected OE cells derived from mice deficient in STAT1 when compared to wild-type OE cells; which suggest that STAT1 is required for the optimal synthesis of IFN-β during infection. Quantitative real-time PCR analyses of signaling components of the type-1 IFN signaling pathway demonstrated equal upregulation in the expression of STAT2 and IRF7 in the wild-type and TLR3-deficient OE cells, but no upregulation in these genes in the STAT1-deficient OE cells. Finally, experiments in which INFAR was blocked with antibody revealed that IFNAR-mediated signaling was critical to the Chlamydia-induced syntheses of IFNα-2 and IFNα-4, but not IFN-β.
The essential, non-redundant roles of RIG-I and MDA5 in detecting and controlling West Nile virus infection (P1385)
Authors: Errett John , Suthar Mehul , McMillan Aimee , Gale, Jr. Michael 
Published: January 2023

Abstract
Virus recognition by the innate immune system represents a critical component of host defense against infection. Proper activation of innate immunity and full immune protection to the emerging infectious disease of West Nile virus has previously been shown to be dependent on viral recognition by the host RIG-I-like receptor (RLR) protein family. RIG-I and MDA5 are the two members of the RLR family that have been shown to recognize viral RNA and to activate innate immune programs that suppress infection. In the current study we have expanded our understanding of the role of the RLRs in protection against West Nile virus infection by assessing the individual roles of RIG-I and MDA5 both in vitro and in vivo. The loss of either gene alone results in a deficit of innate immune signaling and virus control in vitro and increased mortality in vivo. Loss of both RIG-I and MDA5 results in a near complete absence of innate immune gene induction in key target cells of infection and a more severe pathogenesis very similar to that of cells and animals lacking the RLR central signaling adaptor MAVS. Additionally, we have shown that cells infected with West Nile virus contain two distinct pathogen associated molecular patterns that are sensed by RIG-I and MDA5 respectively. Our results highlight the necessity of multiple pattern recognition receptors in the same pathway acting in concert to affect a successful immunological outcome.
Differential effects of TGF-β1 on IL-5 and eotaxin-induced chloride currents in human blood eosinophils (P5130)
Authors: Agrawal Devendra , Kim Min-Jung , Gaurav Rohit 
Published: January 2023

Abstract
TGF-β1 is critical in the pathogenesis of chronic asthma and acts on eosinophils and other cells in the airways. Recently, we found TGF-β1 increases CLC3 expression in human blood eosinophils and induces migration. Here, we examined the interaction of TGF-β1 with IL-5 and eotaxin in the activation of chloride currents in human blood eosinophils. Eosinophils were purified (&lt;99% pure, &gt;98% viable) from the blood of healthy volunteers. Chloride current was measured at pH 7.4 by whole cell patch clamping in freshly isolated eosinophils using Axopatch 200B amplifier and analyzed by pClamp 10 software. Both eotaxin-1 and eotaxin-3 at low doses (0.075-10 ng/ml) increased outwardly rectified Cl- current in a time-dependent manner with maximum response at 500msec plus 100 mV. The eotaxin response at low doses (0.075-0.1 ng/ml) was slower in onset than at high dose. SB328437, a CCR3 antagonist (100 nM), inhibited eotaxin-induced Cl- current. Effect of eotaxin-3 was about 7-times larger than eotaxin-1. TGF-β1 (10ng/ml) decreased eotaxin-induced Cl- current. Incubation of eosinophils with IL-5 (10ng/ml) for 3hr decreased basal level of Cl- current. However, the addition of TGF-β1 with IL-5 significantly enhanced Cl- current. These data suggest differential effect of TGF-β1 in the activation and migration of human blood eosinophils in the presence of eotaxin or IL-5 in allergic airway inflammation and chronic asthma.
The role of CCR2 and CCR7 on dendritic cell migration to and from mycobacterial granulomas (P5096)
Authors: Harding Jeffrey , Schreiber Heidi , Clarkson Ben , Sandor Matyas 
Published: January 2023

Abstract
Dendritic Cells have a complex and critical role in mycobacteria-containing granulomas. Understanding the mechanisms that regulate DC responses during mycobacterial infection will aid the generation of novel vaccines and therapies. Using a novel murine model of granuloma transplantation, combined with the use of CD11c-YFP donors or recipients, we have reported on the incoming and outgoing migration of fluorescent inflammatory dendritic cells from acute and chronic BCG-containing granulomas. Here, we extend our studies to dissect the molecular components of DC involvement during murine mycobacterial infection. We examined CCR2, a chemokine that regulates migration into inflamed tissue, and CCR7, which regulates migration of dendritic cells from the granuloma to the lymph nodes. First, we crossbred CCR2 and CCR7 KO mice with CD11c-YFP mice and analyzed the resulting granulomatous response to BCG infection. We measured granuloma size and frequency, recruitment of different cell types, bacterial load of infected tissues, and spatial organization of CD11c-YFP cells in the granuloma. Then, using transplantation of CD11c-YFP CCR2 and CCR7 KO mice, we measured the chemokine-specific regulation on rates of DC migration in and out of tissue, as well as their role in bacterial dissemination.
The dietary component, curcumin, inhibits the proliferation, survival and function of bone-marrow derived mast cells (P6032)
Authors: Mathias Clinton , Shah Sagar , Statham Rebecca , Kinney Shannon 
Published: January 2023

Abstract
IgE antibodies and mast cells play critical roles in the establishment of allergic responses. Curcumin, the active ingredient of the naturally occurring curry spice turmeric, has anti-allergic properties, both in vitro and in vivo, and thus may have the capacity to modulate mucosal mast cell activation and function during allergic responses. Here, we show that curcumin inhibits the proliferation, survival and function of bone-marrow derived mast cells (BMMCs). BMMCs were grown in the presence of IL-3+SCF or monomeric IgE for 6 days. Co-culture with curcumin completely inhibited the proliferation and long-term survival of BMMCs, suggesting that curcumin has suppressive effects on mast cell expansion. Similarly, curcumin also suppressed the proliferation of peritoneal mast cells. Flow cytometric analysis of the BMMCs demonstrated that curcumin-treated BMMCs were undergoing apoptosis, as indicated by expression of Annexin V and propidium iodide. Intracellular cytokine staining further suggested that these BMMCs produced lower levels of TNF-α. Finally, curcumin-treated BMMCs, activated in the presence of antigen, exhibited reduced β-hexosaminidase activity in comparison with vehicle-treated controls. Our data demonstrate that dietary components such as curcumin can regulate the expansion, activation and function of mast cells, and thus may affect the outcome of mast-cell-dependent allergic responses in vivo.
Quantifying the unanticipated diversity of the human NK cell repertoire (P1456)
Authors: Strauss-Albee Dara , Horowitz Amir , Dogan Ozge , Mackey Sally , Swan Gary , Dekker Cornelia , Davis Mark , Parham Peter , Blish Catherine 
Published: January 2023

Abstract
Like their T and B lymphocyte counterparts, natural killer (NK) cells exist in small and specialized subpopulations. However, while T and B cell repertoire diversity is generated primarily through DNA rearrangement to produce a single antigen-specific receptor, the diversity of the NK cell repertoire is determined by the expression of a spectrum of activating and inhibitory receptors. To evaluate the human NK cell repertoire on a single-cell basis, we simultaneously measured more than 25 NK cell receptors in the peripheral blood of 22 healthy individuals via mass cytometry. We used Boolean gating to classify cellular phenotypes. Based on the Simpson index, an ecological measure designed to quantify population diversity, we found that total NK cell repertoire diversity was approximately normally distributed across individuals. Using rarefaction curves and non-parametric species estimators, we calculate the expansiveness of the NK cell repertoire at a minimum of 100,000 unique phenotypes. Furthermore, within an individual, the ex vivo addition of the homeostatic cytokine IL-15 biased the repertoire toward highly common and uncommon phenotypes, increasing the total population richness but decreasing the overall diversity. These data show, for the first time, that the receptor-based NK cell repertoire is exquisitely diverse. They further suggest an optimal baseline diversity that is highly sensitive to optimization by exogenous factors.
Transgenic mice expressing functional CD8αβ without an active CD8αα implies the contribution of CD8αα in the biological function of CD8αα+TCRαβ+ iIELs (P1466)
Authors: Kang Yoon-Joong , Chang Hsiu-Ching 
Published: January 2023

Abstract
To understand the functional significance of CD8αα complexes in vivo, we generated Tg mice carrying a variant CD8αβ (CD8αm3β) capable of forming inactive CD8αα/pMHCI complexes. These mice show a sub-optimal thymic differentiation with reduced populations of CD8 single-positive thymocytes. Tg CD8 T cells exhibit a compromised developmental capacity when competing with CD8 T cells from B6 mice in the adoptive bone marrow transferred host. However, once emigrated to the peripheral lymphoid organs, these CD8 T cells exhibit normal effector function against viral infection. Furthermore, TCRαβ+ T cell populations expressing CD8αβ+ or CD8αα+ reduced to about 50% in our Tg mice compared to normal B6 mice, and none of these CD8αα+TCRαβ+ T cells were stained by TL tetramer. Furthermore, this mice shows reduced infmmatory responses in DSS-induced colitis model. The findings obtained from our Tg animal models are going to be important to dissect the distinct functions of CD8αα molecule.