bluuebunny/crossref_metadata_2025_split huggingface dataset:
Function of HCMV derived chemokines on NK cells (IRC4P.614)
Authors: Yamin Rachel , Mandelboim Ofer 
Published: January 2023

Abstract
Natural killer (NK) cells belong to the innate immune system and are able to control various microbial infections as well as tumor cell burden. NK cells can be divided into two different subsets based on the expression of the two surface markers: CD16 and CD56. Two major NK cell populations are present in the blood; one that express high levels of CD56 and low levels of CD16; named CD56Bright cells and a bigger population (90%) that express CD56 in inermediate levels and express high levels of CD16, named CD56Dim. The latter poultion is more mature and more cytotoxic, while the CD56Bright cells produce high levels of cytokines. The human Cytomegalovirus (HCMV) is a β herpesvirus with a double-stranded DNA genome which encodes a broad range of genes that involved in immune evasion and regulation. Among these, are 3 viral chemokines named vCXCL1, vCXCL2 and UL128. The function of these 3 viral chemokines is not completely understood. Using fusion and recombinant viral proteins we showed that all three chemokines are capable of binding to the CD56Dim CD16Pos NK cell population. We demonstrate that only one of them, the vCXCL1, can surprisingly cause NK cells migration and we identified the chemokine receptors involved in these migrations.
Expression of CD40L on CD8 T cells and its role in influenza A infection (IRC5P.618)
Authors: Tay Neil , Prabhu Padubidhri Nayana , Wong Hok Sum , Chua Yen , Kemeny David 
Published: January 2023

Abstract
CD8 T cells play an important role in protective immunity against a wide range of pathogens and the factors that control their activation are clearly important. CD40L-mediated CD40 licensing of dendritic cells by CD4 T cells is known to be necessary for the generation of a robust CD8 T cell response. Less clear is the contribution of CD40L on CD8 T cells to their activation. We have previously shown that CD8 T cells are able to induce the production of IL-12 p70 by dendritic cells in a CD40L-dependent manner. To better understand the role of CD40L on CD8 T cells responses, we generated and characterized CD40L-expressing CD8 T cells both in vitro and in vivo. We found that CD40L is expressed on 30-50% of activated effector CD8 T cells and the differentiation of CD8 T cells into CD40L-expressing cells is strongly induced by IL-12, suggesting the presence of a positive feedback mechanism mediated by CD40L and IL-12. Using an influenza A mouse model, we examined whether this mechanism is involved in the primary expansion of CD8 T cells during an infection and whether CD40L-expressing CD8 T cells are able to license dendritic cells for optimal memory CD8 T cell programming. In the absence of CD40L signaling, the ability to upregulate CD25 and downregulate CD62L is diminished in CD8 T cells, causing an impairment in their primary expansion and also their ability to accumulate at the site of infection.
Pulmonary vascular heterogeneity: microvascular vs. arterial generation of angiopoietin-2 (IRC5P.633)
Authors: Lomas-Neira Joanne , Heffernan Daithi , Ayala Alfred , Chung Chun-Shiang 
Published: January 2023

Abstract
Endothelial cell (EC) growth factors, Angiopoietin (Ang)-1 and Ang-2, are important mediators of EC function. Bound to a shared receptor, Tie2, expressed on ECs, they have opposing effects on EC activation. Ang-1/Tie2 promotes a quiescent while Ang-2/Tie2 promotes an activated phenotype and increased vascular permeability. Ang-2, stored preformed in EC Weibel Palade bodies (WPbs), is rapidly released from activated ECs. We have reported increased Ang-2 in the blood of trauma patients with ARDS, representing a potential therapeutic target. To assess lung EC activation and release of Ang-2, we used human pulmonary arterial ECs (hPAECs) and microvascular ECs (hPMECs) grown on electric cell-substrate impedance sensing arrays. These arrays measure changes in EC monolayer integrity in response to inflammatory stimulus. We show that, in response to TNF-a stimulation, hPMECs exhibited increased loss of barrier function compared to hPAECs. Also, Ang-2 in culture supernatant increased in both hPAECs and hPMECs compared to untreated controls. While ECs in larger vessels (arteries), contain WPbs, they are not observed in microvascular ECs (&lt;10um diam). To further investigate, we inhibited WPb vesicle exocytosis. Our findings show that inhibition of WPb release decreased Ang-2 in supernatant from hPAECs, but not significantly from hPMECs, suggesting that the microvascular is a region with distinct responses potentially associated with their location and anatomic structure.
Role of tumor-infiltrating regulatory T cells according to the presence of tertiary lymphoid structures in human lung cancer (IRC5P.619)
Authors: Dieu-Nosjean Marie-Caroline , Devi Priyanka , Goc Jeremy , Germain Claire , Knockaert Samantha , Validire Pierre , Damotte Diane , Fridman Wolf Herve , Sautes-Fridman Catherine 
Published: January 2023

Abstract
Regulatory T cells (Tregs) exert various suppressive mechanisms to dampen the host immune response which can help tumor cells to escape immune surveillance. However, the prognostic value of tumor-infiltrating Tregs (Ti-Tregs) is controversial based on the expression of FoxP3 marker in cancer patients. Here, we decipher the phenotype of Tregs infiltrating human lung cancer to determine their role in shaping the immune response against tumor cells. Our aim was to study the differentiation, activation, and immunosuppression status of Ti-Tregs in different areas of human lung tumors. Immunohistochemistry was performed on paraffin-embedded tissue sections for CD3 and FoxP3 stainings, and phenotypic analysis was carried out on fresh human lung cancer specimens (n=50) by flow cytometry. In lung cancer patients, Ti-Tregs localize in the different part of the tumor i.e. tumor nests, stroma, and tertiary lymphoid structures (TLS). Ti-Tregs are CD4+ and show central-memory and effector-memory phenotype. Interestingly, Ti-Tregs exhibit different phenotypes based on the differential expression of activation and immunosuppression molecules. In conclusion, the presence of several Ti-Treg subsets in lung tumor suggests that they may play different roles in distinct tumor areas. Thus, the next step will be to decipher the mechanisms and functions used by these Treg populations in order to manipulate them for therapeutic intervention.
PHLDA1 is a negative regulator for TLR2-mediated inflammatory response (IRC5P.623)
Authors: Baek Suk-Hwan , Park Dae-Weon 
Published: January 2023

Abstract
Atherosclerosis is a slow and complex disease that causes cardiovascular complications and stroke. toll like receptor 2 (TLR2)-mediated signaling pathways and gene regulation are mainly involved in the early development and progression of this complex disease. We found that pleckstrin homology-like domain family A member 1 (PHLDA1) is a unique gene upregulated by TLR2 stimulation and determined the detail mechanisms the signaling pathway for expression in macrophages. Treatment with Pam3CSK4, a TLR2 agonist, increased PHLDA1 mRNA and protein expression levels. Gain- and loss-of-function analyses revealed that PHLDA1 modulates foam cell formation by TLR2 stimulation and foam cell formation-related molecule such as MCP-1. In sequence analysis revealed the presence of typical g-activated sequence (STAT3) element in the promoters of mice PHLDA1 and confirmed the STAT3 binding sites. Subsequently, we also confirmed the detailed mechanism of PHLDA1 expression through the JAK2-ERK1/2-STAT3 pathway. These findings suggest that PHLDA1 is a novel negative regulator aimed at TLR2 and may play a vital role eliciting TLR2-mediated responses in atherosclerosis.
Loss of NADPH oxidase-derived superoxide induces CD4 T cell antigen-specific hyporesponsiveness by modulating IL-12 signaling (IRM5P.637)
Authors: Padgett Lindsey , Tse Hubert 
Published: January 2023

Abstract
Efficient T cell adaptive immune activation requires innate immune-derived reactive oxygen species (ROS) and pro-inflammatory cytokines. Our laboratory previously demonstrated blunted CD4 T cell autoreactivity in the absence of NADPH oxidase (NOX)-derived superoxide, as NOX-deficient Non-obese diabetic (NOD.Ncf1m1J) mice exhibited spontaneous Type 1 diabetes-resistance, partly due to decreased Th1 cytokine responses. To examine the role of ROS deficiency on antigen-specific T cells in a non-autoimmune setting and further test the hypothesis that ROS exacerbates Th1 T cell responses, we generated the OT-II.Ncf1m1J mouse containing superoxide-deficient OVA323-339-specific CD4 T cells. Stimulated OT-II.Ncf1m1J splenocytes displayed decreased Th1 T cell cytokines IFN-γ and TNF-α (p&lt;0.05) and diminished expression of the Th1 transcription factor T-bet and Th1 signal transducer IL-12Rβ2, compared to OT-II CD4 T cells. IFN-γ synthesis and IL-12 signaling by OT-II.Ncf1m1J T cells were rescued by addition of exogenous superoxide and co-culture with superoxide-sufficient antigen-presenting cells; conversely, Th1 responses were blunted in wild type OT-II T cells via free radical dissipation with a superoxide dismutase mimetic. This data highlights the importance of NOX-derived ROS in providing a third signal for adaptive immune maturation by modulating IL-12 signaling and implicates that targeting ROS may represent a promising therapeutic strategy in curbing CD4 T cell autoreactivity.
Ethanol induced expression of forkhead box O3 and associated tolerance is amplified in female derived dendritic cells. (IRM5P.642)
Authors: Thompson Matthew , Vidrine Amy , Watkins Stephanie 
Published: January 2023

Abstract
Dendritic cells (DCs) are often referred to as the bridge between innate and adaptive immunity and are a critical player in inducing a robust immune response to antigen. However they also can act to dampen the immune response by a process called tolerance. Tolerance is critical in preventing autoimmune disorders but can be manipulated by tumors to prevent anti-tumor immunity. Ethanol has previously been shown to reduce the ability of DCs to present antigen and to induce a pro-inflammatory response but the mediator of this reduced DC function is not yet known. One of the key players in DC tolerance is the transcription factor Forkhead Box O3 (FOXO3). FOXO3 is a transcriptional regulator of a variety of processes associated with cell cycle progression as well as regulation of nuclear receptors, including the androgen and estrogen receptors. In these studies we investigated the effects of ethanol on FOXO3 expression in dendritic cells. We also examined the ability of dendritic cells to uptake antigen and mature into functional APCs in the presence of ethanol. and on the ability of ethanol treated dendritic cells to become mature antigen presenting cells. Additionally we examined if the effects of ethanol on DCs were increased due to gender specific differences. Our results indicate that exposure of dendritic cells to ethanol increases the expression of FOXO3 and decreases the expression of co-stimulatory and pro-inflammatory molecules.
T helper subset distribution changes with age and is influenced by IL-2 (LYM2P.724)
Authors: Hopkins Jacob , Masters April , Haynes Laura 
Published: January 2023

Abstract
To better understand how aging impacts the immune response to influenza infection, we have examined CD4+ T helper (Th) subsets and how they are impacted by aging. Th cells are important for helping Ab production by B cells and are required for the generation of cytotoxic and memory CD8+ T cells. Influenza nucleoprotein (NP)-specific Th subsets can be distinguished by PSGL1 and Ly6C expression: PSGL1hiLy6Chi cells are terminally differentiated Th1 effectors; PSGL1hiLy6Clo cells are more memory-like; PSGL1loLy6Clo cells are mostly T follicular helper (Tfh)-like. With age, the proportion of PSGL1hiLy6Chi Th1 effectors is reduced and Tfh-like PSGL1loLy6Clo cells are increased both in naïve mice and during infection. Additionally, Tfh cells (defined as CXCR5+PD-1+) are also increased with age. This change in subset distribution with age is also associated with a decrease in ex vivo IL-2 levels in aged animals. In order to further examine how IL-2 can impact subset distribution, an in vitro system was used in which CD4 T cells from young, middle aged and aged mice were cultured with anti-CD3/anti-CD28. Addition of exogenous IL-2 to Th cells from all age groups resulted in reduction of the PSGL1loLy6Clo Tfh population. Furthermore, blocking IL-2 with anti-CD25 Ab increased the PSGL1loLy6Clo Tfh population, regardless of age. Thus, the availability of IL-2, independent of aging, influences Th subset distribution and can account for the increase in Tfh with aging.
Examining pathogen specific T cell responses during antibiotic intervention (MPF2P.743)
Authors: Benoun Joseph , Fogassy Zachary 
Published: January 2023

Abstract
Salmonella enterica serovar Typhi can cause recurrent and relapsing infection in antibiotic-treated individuals, suggesting that bacterial clearance may hinder the development of adaptive immunity. We have developed an antibiotic-treatment model in mice to examine this issue using antigen-specific reagents. Infection with Salmonella Typhimurium strain BRD509-2W1S caused expansion of 2W1S-specific CD4 T cells that were easily detected in peripheral blood and lymphoid tissues. In contrast, antibiotic-treated mice displayed lower 2W1S-specific CD4 clonal expansion in blood and lymphoid tissues that persisted for months after infection. This reduced clonal frequency correlated with diminished protective immunity to re-challenge infection. We are currently examining the phenotype and functional differences in CD4 T cell memory populations to determine if central, effector, and tissue-resident memory cells are equally impacted by antibiotic-intervention. Greater understanding of how antibiotics affect CD4 memory development may allow for therapeutics to boost protective immunity to secondary or relapsing Salmonella infections.
A novel role for Irgm1 in immune homeostasis (MUC1P.906)
Authors: Azzam Kathleen , Madenspacher Jennifer , Lai Lihua , Gowdy Kimberly , Cain Derek , Clayton Natasha , Cunningham Willie , Janardhan Kyathanahalli , Taylor Gregory , Fessler Michael 
Published: January 2023

Abstract
Irgm1 is an IFN-inducible GTPase that mediates host resistance to intracellular pathogens. We sought to define the role of Irgm1 in pulmonary immune cell homeostasis by profiling steady state and pathogen-induced immunity in the lungs of Irgm1-/- mice. The lungs of naïve Irgm1-/- mice displayed abnormal bronchiolocentric cellular infiltration composed predominantly of B cells with focal collections of T cells, associated with increased lung and airway cytokines. Compared to WT controls, naïve Irgm1-/- mice had elevated serum and airway levels of immunoglobulins, suggesting dysregulated B cell function. B cells were found to be reduced in the blood of Irgm1-/- mice, and B cell precursors were also decreased in the bone marrow. Whereas B2 and B1a cells were elevated in the Irgm1-/- lung, B1a cells were reduced in the peritoneal cavity, suggesting abnormal trafficking of B cell subsets. Irgm1-/- mice had increased influx of neutrophils, chemokines, cytokines, and IgM into the airspace after exposure to inhaled LPS and/or S. pneumoniae. Pathogen clearance from the lung and survival were increased in Irgm1-/- mice upon challenge with S. pneumoniae. Future studies are planned to better define the mechanism of B cell dys-homeostasis, determine whether Irgm1-/- mice have autoimmunity, and define whether the enhanced host defense response and survival of Irgm1-/- mice against S. pneumoniae derives from increased number and function of innate B cells and their products.
Increasing intestinal reactive oxygen species reverses diabetes-induced inflammation (MUC2P.928)
Authors: Chen Lee-Wei , Wu Ying-Ying , Chen Pei-Hsuan , Hsu Ching-Mei 
Published: January 2023

Abstract
Systemic inflammation appears to persist in diabetic patient. To examine the role of reactive oxygen species (ROS) of intestine in DM-induced inflammation, streptozotocin (STZ)-induced diabetes and Ins2Akita (Ins2Akita mutation mutant) mice were used. STZ mice demonstrated a significant decrease of Lacobacilli as well as ROS production and a marked increase of p-JNK as well as iNOS expression in the intestinal mucosa. Oral supplementation of fructooligosaccharides (FOS) or dead Lacobacilli increased the production of ROS of intestine and decreased IL-1β expression of Kupffer cells in STZ and Ins2Akita mice. Feeding of L-NAME, an iNOS inhibitor, but not D-NAME increased the production of ROS of intestinal mucosa and decreased endotoxin in portal vein and IL-1β expression of Kupffer cells in STZ mice. Oral feeding of N-Acetylcysteine, a ROS scavenger, decreased ROS production and inhibited the effects of FOS as well as L-NAME on reversing DM-induced ICAM expression of liver. Using STZ JNK1-/- and Ins2Akita JNK1-/- mice to reverse DM-induced pJNK expression of intestinal mucosa induced a significant decrease of IL-1β expression of Kupffer cells and ICAM expression of liver. Collectively, diabetes induces systemic inflammation through decreasing ROS production and increasing pJNK expression of intestine. Increasing intestinal ROS production could reverse diabetes-induced systemic inflammation.
Impact of a high energy diet on obesity-associated immune biomarkers in Sprague Dawley rats (MUC2P.929)
Authors: Clarke Sandra , Kalmokoff Martin , Brooks Stephen , Green Judy , Caldwell Don , Inglis G. , Green-Johnson Julia 
Published: January 2023

Abstract
A trial was designed to examine effects of a high energy diet on obesity-associated biomarkers in Sprague-Dawley (SD) rats. Male rats (n = 176) were fed an AIN93G high-energy diet containing 16% dietary fat for 2 weeks, and segregated into obese-prone and obese-resistant phenotypes. Necropsies of animals displaying the highest (n = 10) and those displaying the lowest weight gain (n = 10) were conducted to examine potential differences in the concentration of immune biomarkers. Adipose, mucosal (ileum, ileal Peyer’s patches, cecum, proximal and distal colon), mesenteric lymph nodes (mln) and systemic (serum, liver, spleen) tissues were comparatively analyzed for cytokine or adipokine profiles. Obese rats had higher levels of leptin (P = 0.0001) and IL-22 (P = 0.059) in the distal colon and higher levels of IL-10 (P = 0.032) and TNF-α (P = 0.016) in the proximal colon compared to their obese-resistant counterparts. In obese-resistant rats, higher levels of CINC-2αβ (P = 0.048) occurred in mesenteric adipose tissue along with higher levels of soluble ICAM-1 (P = 0.018) in the mln. These findings suggest a high energy diet imparted differential effects on mucosal cytokine and adipokine production in colonic and mesenteric tissue between obese-prone and obese-resistant SD rats. Elevated levels of pro-inflammatory and regulatory cytokines are evident as little as 2 weeks after the initiation of a high-energy diet.
Disruption of the endocrine system leads to local mucosal responses favoring allergic sensitization (MUC2P.919)
Authors: Berdnikovs Sergejs , Loffredo Lucas , Abdala-Valencia Hiam 
Published: January 2023

Abstract
Estrogen receptors play fundamental roles in health and disease by integrating systemic hormonal signals and local factors for maintenance of proper tissue homeostasis. Using a bioinformatics approach, we identified a genetic network reflecting aberrant regulation of mucosal homeostasis by estrogen receptors in allergic disease. To determine whether disruption of the estrogen system directly affects mucosal immune responses, we exposed mice to environmentally relevant doses of estrogen mimetic bisphenol A (BPA) in drinking water. Remarkably, systemic exposure to BPA facilitated allergic sensitization to inhaled ovalbumin and subsequent development of airway allergic inflammation without the use of adjuvants. Consistent with this effect, we identified a tissue stress response signature to endocrine disruption in the absence of tissue damage or noticeable inflammation, which persists for several weeks after cessation of BPA treatment. This includes increased tissue levels of chemokines CCL2, CXCL9, CXCL10 and CXCL11 at multiple barrier sites and elevated systemic levels of alarmins. Moreover, systemic exposure to BPA during postnatal development reduced differentiation potential of primary lung mesenchymal cells in adulthood, further supported by decreased expression of epithelial differentiation factors Fgfr2 and Sox2 in vivo. These findings demonstrate that environmental estrogens can induce tissue stress favoring allergic response in the absence of tissue damage.
High-throughput direct cell counting-based natural killer cell-mediated cytotoxicity assay using Celigo Imaging Cytometry (TUM2P.1027)
Authors: Chan Leo , Somanchi Srinivas , Rosbach Kelsey , Lee Dean 
Published: January 2023

Abstract
Cytotoxicity assays play a central role in studying the function of immune effector cells such as cytolytic T lymphocytes (CTL) and natural killer (NK) cells. Traditionally, cytotoxicity assays have been performed using 51Chromium (51Cr) and Calcein release assays. The assays involve labeling tumor cells (target) with radioisotope or fluorescent dyes, when the target cells are subjected to cytolysis by CTLs or NK cells, they releases the entrapped labels into the media upon lysis. These traditional methods may generate inconsistent results due to low sensitivity caused by poor loading efficiency and high spontaneous release of the reagents. In this work, we demonstrate a novel high-throughput direct cell counting method for cytotoxicity assay using the Celigo imaging cytometry method. Human NK cells from one healthy donor were used as effectors, and K562 and IMR32 were used as the target cells. Both target cells were first stained with Calcein AM, and seeded at in a 96-well microplate. NK cells were then added to each well at various Effector-to-Target (E:T) ratios. The 96 well plate was then scanned and analyzed using the Celigo to measure the % lysis of target cells at different time points. The results showed increasing % lysis as incubation time and E:T ratio increased. The propose Celigo imaging cytometry is an accurate and simple method for direct quantification of cytotoxicity, which can be an attractive method for both academic and clinical research.
Immunotherapeutic targeting of HSP90 client proteins overexpressed in chemo-/radio-resistant melanoma (TUM2P.1012)
Authors: Raveendran Subhara , Rao Aparna , Storkus Walter 
Published: January 2023

Abstract
Amongst the numerous genetic alterations that occur in cancer cells, is overexpression of DNA repair proteins (DNA-RP) in a disease-stage associated manner which has also been correlated to chemo/radiotherapy (CRT) resistance and poor overall prognosis. DNA-RP overexpression has been reported to involve post-translational protein stabilization mediated by the molecular chaperone Heat shock protein 90(HSP90), thereby preventing proteasome-dependent degradation of these HSP90 “client” proteins. Hence, HSP90 inhibitors (HSP90i) have been heralded as co-therapy agents for cancer patients that have developed resistance to first-line treatments such as temozolomide (TMZ). Under conditions in which its function is blocked by HSP90i, HSP90 client proteins rapidly become (poly)ubiquinated and undergo degradation resulting in improved sensitization of cancer cells to CRT. We observed that HSP90i promote the proteasome-dependent (i.e. blocked by inhibitors PS-341 and MG-132) degradation of a range of DNA-repair client proteins. We have also identified H-2Kb/Db-presented peptide epitopes derived from these DNA-RPs that may be recognized by Type-1 CD8+ T cells after specific vaccination in C57BL/6 mice. We are currently investigating the capacity of HSP90i to conditionally sensitize melanoma cells to the tumoricidal action of CRT as well as CD8+ T cells reactive against overexpressed DNA-RP that constitute HSP90 client proteins.