bluuebunny/crossref_metadata_2025_split huggingface dataset:
Choline acetyltransferase+ lymphocytes regulate endothelial nitric oxide synthase (MUC2P.939)
Authors: Olofsson Peder , Hanes William , Steinberg Benjamin , Oswald Michaela , Ahmed Mohamed , Hudson LaQueta , Valdes-Ferrer Sergio , Liu Shu , Pavlov Valentin , Chavan Sangeeta , Andersson Ulf , Diamond Betty , Miller Edmund , Gregersen Peter , Tracey Kevin 
Published: January 2023

Abstract
Mammalian blood vessels are innervated by adrenergic neurons that increase blood pressure. The neurotransmitter mechanism is dependent upon norepinephrine that mediates contraction of vascular smooth cells to increase arterial resistance. Vasodilation is mediated by acetylcholine, which lowers blood pressure through a mechanism dependent upon relaxation of vascular smooth muscle cells in response to endothelial release of nitric oxide. Endothelial cells are not innervated by cholinergic neurons, and it was unknown whether acetylcholine release by lymphocytes might modulate blood pressure. Here, we observed lymphocytes expressing choline acetyltransferase (ChAT), the enzyme that catalyzes biosynthesis of acetylcholine, in the blood. Co-culture of ChAThi lymphocytes with endothelial cells significantly increased calcium concentration and phosphorylation of eNOS in endothelial cells (p&lt;0.05 vs. ChATlow lymphocytes). Passive transfer of engineered ChAThi Jurkat T cells resulted in an immediate significant decrease in blood pressure in recipient mice to 83±5 % of baseline, while blood pressure after transfer of ChATlow Jurkat T was 104±4 % of baseline (p=0.02). Pre-treatment with the NO synthesis inhibitor L-NG-monomethyl arginine citrate prevented the blood pressure reduction by ChAThi Jurkat T cells, and blood pressure remained at 98%±5% of baseline in these animals (p=0.21). These findings suggest a novel mechanism of lymphocyte-mediated blood pressure regulation.
Antimicrobial peptide gene expression and functionality in P-glycoprotein deficient models of colitis (MUC3P.943)
Authors: Lorenz Robin , Tanner Scott , Daft Joseph , Little Marguerite 
Published: January 2023

Abstract
Inflammatory bowel diseases (IBD), such as Crohn’s disease and ulcerative colitis, are chronic inflammatory conditions of the digestive tract. Antimicrobial peptides (AMPs) are produced in the intestines and function to control bacterial growth. AMP deficiency can result in unfavorable microbiota, a proposed IDB risk factor. FVB.mdr1a-/- mice, lacking the multidrug resistance gene (MDR) and its functional product, P-glycoprotein (P-gp), develop spontaneous colitis. We hypothesized that P-gp deficiency alters AMP expression, resulting in changes in the microbiota. To investigate this, intestinal AMP mRNA expression from control and FVB.mdr1a-/- were compared. This revealed altered expression of small intestinal and colonic AMPs. Next, FVB.mdr1a-/- mice were treated with antibiotics to determine if these changes are due to P-gp deficiency or secondary to microbial colonization. Intestinal AMP mRNA expression was analyzed and no changes were observed, suggesting that the absence of P-gp alters AMP expression, regardless of microbiota. Functionality of these AMPs were investigated, showing higher killing capabilities in the FVB.mdr1a-/- mice. For insight into human IBD, AMP mRNA expression were measured in the human colon adenocarcinoma cell line (Caco2-C) and a P-gp knockdown (Caco2-MR) after treatment with bacterial products. Here, we observed changes in AMP expression. These studies provide insight into the role of the MDR gene in the development of colitis.
A vital control of intracellular Chlamydia infection requires interleukin-17 receptor-mediated signaling (MUC3P.946)
Authors: Tram Cynthia , Kampen Rachel , Halperin Scott , Wang Jun 
Published: January 2023

Abstract
IL-17A is a pleiotropic proinflammatory cytokine that has a vital role in host resistance against extracellular bacterial and fungal infections. However, its role in controlling intracellular bacterial infection remains elusive. Here, we examined the IL-17/IL-17 receptor-mediated signaling in controlling intracellular Chlamydia muridarum infection in non-hematopoietic cells in vitro and in vivo. Remarkably, Chlamydia replication in mouse embryonic fibroblasts (MEF) was significantly increased in IL-17RC-knockout (IL-17RCKO) MEF and, to a lesser extent, in IL-17RAKO MEF compared to WT MEF at 6 and 12 hrs post-infection. To verify this finding in vivo, a respiratory Chlamydia infection model was established in bone marrow (BM) chimeric mice using WT BM donor cells transferred into γ-irradiated WT, IL-17RAKO and IL-17RCKO mice. Both IL-17RAKO and IL-17RCKO BM chimeric mice had significantly elevated bacterial burden in the lung compared to WT mice at days 5 and 11 post-infection. However, IL-17RCKO, but not IL-17RAKO, BM chimeric mice exhibited significant body weight loss compared to WT mice during early infection. While IL-17RAKO BM chimeric mice displayed an increased IL-17A response, IL-17RCKO mice showed an increased IFN-γ response in the lung. Our data demonstrates an indispensable role of the IL-17RC-mediated signaling in controlling Chlamydia infection and suggests differential roles of IL-17RA- and IL-17RC-mediated signaling in shaping Chlamydia-induced host responses.
Substance P and IL-33 synergistically stimulate mast cells to release IL-1β and TNF-α implicated in psoriasis; inhibition by the flavonoid methoxyluteolin (THER2P.960)
Authors: Taracanova Alexandra , Theoharides Theoharis 
Published: January 2023

Abstract
Psoriasis is an autoimmune skin disease characterized by keratinocyte hyperproliferation and inflammation. The pathogenesis of psoriasis appears to involve interactions among keratinocytes, mast cells (MC) and cytokines. MC are immune cells that secrete various cytokines and chemokines including interleukin (IL)-1 family of cytokines (IL-1β, 18, 33), which can stimulate keratinocyte activation. IL-33 also augments activation of MC with further release of several mediators such as TNF-α, IL-6, and prostaglandin D2. Also, IL-33 has synergistic effect with a neuropeptide substance P (SP) on VEGF release in MC. Both IL-33 and SP are upregulated in psoriatic skin. Moreover, SP stimulates keratinocytes to secrete IL-1β. Our findings show that SP and IL-33 synergistically stimulate MC to produce and release IL-1β and TNF-α, pro-inflammatory mediators involved in psoriasis pathogenesis. The combination of SP and IL-33 also significantly up-regulates IL-1β and TNF-α gene expression. The natural flavonoid methoxyluteolin (Metlut) is a potent inhibitor of both keratinocyte and MC activation. Here we report that Metlut significantly inhibits IL-1β and TNF-α gene expression, production and release triggered by SP and IL-33 combination. Therefore, Metlut-based formulations could be a novel treatment for psoriasis.
High-throughput functional assay to screen anti-BDCA2 candidates for systemic lupus erythematosus therapeutics (THER3P.973)
Authors: Li Jenny , Ladi Ena , Pappu Rajita , Chan Pamela 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a clinically heterogeneous disease characterized by a large number of aberrations in the immune system. Studies with SLE patients have shown that heightened levels of the cytokine interferon-α (IFNα) can interfere with immune regulation and effect lupus pathogenesis. The plasmacytoid dendritic cells (pDCs) are the main producer of type I IFNs and have the capacity to synthesize up to 109 IFNα molecules per cell after activation. Crosslinking BDCA2, a typeII C-type lectin specific to pDCs, results in the inhibition of IFNα production in pDCs stimulated with TLR7/9. To develop an agonistic therapeutic antibody to BDCA2, a large number of antibody candidates have to be screened in a functional assay that detects blocking of IFNα production in pDCs isolated from peripheral blood mononuclear cells (PBMC). pDCs constitute less than 0.4% of PBMCs making pDC purification inefficient and challenging. Here we report an automated high-throughput BDCA2-agonistic functional assay using fraction 5 (F5) cells from an elutriated leukopak as substitute for purified pDCs coupled with a robust INFα detection method. The Integrated automation program in 384well format has provided a powerful tool for antibody screening and can be applied to other projects requiring a similar screening strategy.
Molecular profiling of CD8 T cells from autochthonous melanoma identifies Maf as driver of T cell exhaustion (TUM2P.1004)
Authors: Verdeil Gregory , Giordano Marilyn , Henin Coralie , Maurizio Julien , Bourdely Pierre , Buferne Michel , Auphan-Anezin Nathalie , Speiser Daniel , Schmitt-Verhulst A 
Published: January 2023

Abstract
T-cells infiltrating neoplasms express surface molecules typical of chronically virus-stimulated T-cells, often termed “exhausted” T-cells. We analyzed the transcriptome of “exhausted” CD8 T-cells infiltrating autochthonous melanomas compared to those of naïve and acutely stimulated CD8 T-cells. In spite of strong similarities between transcriptional signatures of tumor- and virally-induced exhausted CD8 T-cells, notable differences appeared. Among transcriptional regulators, Nr4a2 was expressed in both exhaustions, whereas Maf was highly over-expressed only in tumor-exhausted T-cells. We confirmed Maf and nr4a2 up-regulation in Melan-A/MART-1 specific CD8 T cells from tumor infiltrated LN from melanoma patients. Anti-tumor CD8 T-cells transduced to express Maf showed dampened anti-tumor activity upon adoptive transfer, whereas Nr2a4 over-expression was without effect. Maf-expressing CD8 T-cells showed unaltered homeostasis but failed to accumulate in tumor-bearing hosts and developed defective anti-tumor secondary responses. We also found that Maf expression in CD8 T cells induced part of the transcriptional program associated with tumor-induced exhaustion. We further identified TGFβ as a main contributor to Maf expression in CD8 T-cells. Therefore the melanoma microenvironment contributes to skewing of CD8 T-cell differentiation programs, in part by TGFβ-mediated induction of Maf.
Longitudinal analyses of CD3ζ-chain expression in the correlation of the disease status in a cohort of head and neck cancer patients (TUM2P.1033)
Authors: Upreti Deepak , Zhang Man-li , Bykova Elena , Pathak Alok , Kung Sam 
Published: January 2023

Abstract
Purpose: Despite advances in multimodality treatment, the 5-year survival rate of Head and neck squamous cancer (HNSCC) patients has unimproved over the last 4 decades. CD3ζ has emerged as a clinically important immunological molecule in HNSCC. Its relevance as a prognostic biomarker of HNSCC, however, has not been formally addressed in a longitudinal study. Methods: Peripheral blood mononuclear cells (PBMC) were collected from 46 patients and 53 controls at the time of diagnosis and post treatments (upto 2 year period). Expression of CD3ζ in the T cells of the PBMC samples were analyzed in flow cytometry. Results: We standardized a method for longitudinal analyses of intracellular CD3z expressions in the PBMC samples. We considered a &lt;10% baseline increase in the normalized MFI of CD3ζ expression as a predictor of disease status in evaluating the follow-up samples of the HNSCC patients. Correlation analysis showed that 27/29 HNSCC patients who showed an increase in the CD3ζ expression relative to their baselines were disease free (negative predictive value. 93.1%). 10/17 HNSCC patients who showed a reduced/no change in the CD3ζ expression died or had recurrent disease (positive predictive value, 58.8 %). Overall accuracy of the assay was 80.43%. The sensitivity and specificity were respectively 83.3% and 79.41%. Conclusion: Our longitudinal analyses supported that a &gt;10% increase in the CD3ζ expression against baseline could be a good prognostic biomarker in HNSCC patients.
Effector CD8+ T cells abrogate fibroblasts-mediated chemoresistance in ovarian cancer (TUM2P.1007)
Authors: Wang Weimin , Kryczek Ilona , Tan Lijun , Zhao Lili , He Gong , Vatan Linda , Szeliga Wojciech , Zou Weiping , Liu Rebecca 
Published: January 2023

Abstract
Tumor infiltrating effector CD8+ T cells have been described to play a major role in antitumoral activity and to be a favorable prognostic factor in ovarian cancer. The strategies of immunotherapy to improve effector T cells response have also shown potential clinical benefit in subset of ovarian cancer patients. Currently platinum-based chemotherapy is still the most active regimen for ovarian cancer treatment. However, the therapeutic resistance is a major challenge to limit the therapeutic efficiency. Here we found primary fibroblasts isolated from ovarian tumor speciman could induce resistance against cisplatin in cancer cells when the two cell types were co-cultured in vitro or co-inoculated in vivo. Activated CD8+ T cells abolished fibroblasts-mediated cisplatin resistance through IFNγ secretion. Next, we found fibroblasts induced cisplatin resistance through increasing intracellular Glutathione (GSH) level and reducing the cisplatin accumulation in cancer cells. We also identified both intact GSH and Cysteine released by fibroblasts contributed to cisplatin resistance in cancer cells. IFNγ producted by activated CD8+ T cells promoted fibroblasts extracellular GSH degradation through up-regulation of Gamma-glutamyl transpeptidase, and also diminished Cysteine generation by fibroblasts through down-regulation of System xc⁻ cystine/glutamate antiporter.These date implicate effector T cells immunotherapy as a powerful regimen to overcome the resistance to chemotherapy.
Optimizing the efficacy of dual-costimulation cancer therapy by engaging tumor-unrelated CD4 T cell help (TUM2P.1009)
Authors: Mittal Payal , Wang Xi , Uthaya Dinesh , St. Rose Marie-Clare , Vella Anthony , Adler Adam 
Published: January 2023

Abstract
Tumor escape following immune therapy can be caused by loss of specifically targeted MHC class I-restricted epitopes. It would thus be beneficial to develop therapies that engage multipronged anti-tumor responses. Agonist mAbs to the TNF/TNFR family costimulatory receptors CD134 plus CD137 (dual costimulation) not only elicits CD8+ CTL, but also cytotoxic CD4 Th1 cells that can directly target MHC-II+ tumors. Further, dual costimulated cytotoxic CD4 Th1 cells not only provide linked help to CD8 T cells responding to cross-presented antigen, but can also provide non-linked help that programs Th1/Tc1 functions in antigen-non-responding (bystander) T cells. Here we tested whether engagement of tumor-unrelated CD4 help augments dual costimulation-mediated control of established B16 melanoma that can be induced to express both MHC class I and II. Notably, engagement of tumor-unrelated CD4 T cells dramatically enhanced the ability of dual costimulation to control B16 tumor growth, this effect depended upon both CD8 and CD4 cells, and therapeutic efficacy correlated with increased tumor infiltration of GzmB-expressing effector CD8 and CD4 T cells with a concomitant decrease in Foxp3+CD4+ cells. Interestingly, the dual costimulated tumor-unrelated CD4 helper T cells themselves appear to require help from a CD134-responsive but presumably antigen-non-responding cell to attain their maximal function.
Role of natural killer cells in ovarian cancer chemoresistance (TUM3P.1050)
Authors: Scholler Nathalie , Repellin Claire 
Published: January 2023

Abstract
Novel immunotherapeutic approaches revolutionize cancer prognosis but the role of innate immunity in chemoresistance remains poorly understood. For example, toll-like receptor (TLR) ligands in combination with anticancer treatments yielded unpredictable effects, either beneficial or deleterious. We hypothesized that the function of tumor-infiltrating immune cells depends on constitutive TLR variants. We characterized healthy donor T cells, B cells, monocytes and natural killer (NK) for functional polymorphism after transwell co-culture with chemosensitive (OVCAR5) vs. chemoresistant (SKOV3TR) ovarian tumor cells +/- (1) medium only; (2) LPS; (3) PamCys3; (4) Paclitaxel (PTX); (5) OVCAR5; (6) OVCAR5+PTX; (7) SKOV3TR; (8) SKOV3TR+PTX. CD8+ T cell activation was measured by perforin staining, monocyte and B cell maturations by cytokine expression array, and NK-mediated cytotoxicity by PI staining of GFP-transfected K562 after 4 hour-incubation. We observed minor functional variations between donor T cells, B cells and monocytes. However, while all NK cells killed GFP-K562 cells as expected, in some donors NK cytotoxicity was specifically inhibited by SKOV3RT co-culture. NK genotyping and phenotyping are ongoing including tumor-dependent NKG2D expression, and PTX-dependent MICA release by tumor cells. These results support the hypothesis that chemoresistant tumors release soluble factors that modulate NK function though specific interactions with innate immune receptor variants.
A novel class of small molecule agonists with preference for human over mouse TLR4 activation (VAC3P.1056)
Authors: Maynard Sean , Marshall Jason , Heeke Darren , Rao Eileen , Hornigold David , McCrae Chris , Fraser Neil , Williams Nicola , King Sarah , Cooper Martin , Frantz Chris , Hajjar Adeline , Woo Jennifer 
Published: January 2023

Abstract
The best-characterized Toll-like receptor 4 (TLR4) ligands are lipopolysaccharide (LPS) and its chemically modified and detoxified variant, monophosphoryl lipid A (MPL). After a high throughput screening process of small molecule libraries, we have discovered a new class of TLR4 agonist with a species preference profile differing from MPL. Products of the 4-component Ugi synthesis reaction were demonstrated to potently trigger human TLR4-transfected HEK cells but not mouse TLR4, although inclusion of the human MD2 with mTLR4 was able to partially recover activity. The species preference profile for the panel of Ugi compounds was found to be strongly active for human and cynomolgus monkey primary cells, with reduced but still substantial activity for most Ugi compounds on guinea pig cells. Mouse, rat, rabbit, ferret, and cotton rat cells displayed little or no activity when exposed to Ugi compounds. However, engineering the human versions of TLR4 and MD2 to be expressed in mTLR4/MD2 deficient mice allowed for robust activity by Ugi compounds both in vitro and in vivo. These findings extend the range of compounds available for development as agonists of TLR4 and identify novel molecules which reverse the TLR4 triggering preference of MPL for mouse TLR4 over human TLR4. Such compounds may be amenable to formulation as more potent human-specific TLR4L-based adjuvants than typical MPL-based adjuvants.
Therapeutic immunization with a mixture of herpes simplex virus type 1 Glycoprotein D derived “asymptomatic” human CD8+ T cell epitopes decreases spontaneous ocular shedding in latently infected HLA transgenic rabbits: association with low frequency of local PD-1+TIM-3+CD8+ exhausted T cells (VAC4P.1114)
Authors: Benmohamed Lbachir , Srivastava Ruchi , Khan Arif , Nesburn Anthony 
Published: January 2023

Abstract
Most blinding ocular herpetic disease is due to reactivation of herpes simplex virus type 1 (HSV-1) from latency rather than to primary acute infection. No herpes simplex vaccine is currently available for use in humans. In this study, we used the HLA-A*02:01 transgenic (HLA Tg) rabbit model of ocular herpes to assess the therapeutic efficacy of a therapeutic vaccine based on HSV-1 gD epitopes that are mainly recognized by CD8+ T cells from “naturally” protected HLA-A*02:01 positive, HSV-1 seropositive healthy asymptomatic (ASYMP) individuals (who have never had clinical herpes disease). Three ASYMP CD8+ T cell epitopes (gD53-61, gD70-78 and gD278-286) were linked with a promiscuous CD4+ T-cell epitope (gD287-317) to create 3 separate pairs of CD4-CD8 peptides, which were then each covalently coupled to an Ne-palmitoyl-lysine moiety, a toll like receptor 2 (TLR-2) ligand. This resulted in the construction of 3 CD4-CD8 lipopeptide vaccines. Latently infected HLA-Tg rabbits were immunized with a mixture of these 3 ASYMP lipopeptide vaccines, delivered as eye drops in sterile PBS. The ASYMP therapeutic vaccination: (i) significantly reduced HSV-1 shedding detected in tears; (ii) boosted the number and function of local HSV-1-specific CD8+ T cells; and (iii) was associated with fewer exhausted HSV-1 gD-specific PD-1+TIM-3+CD8+ T-cells. The results underscore the potential of an ASYMP CD8+ T cell epitope-based therapeutic vaccine strategy against recurrent ocular herpes.
Reversal of peanut anaphylaxis by retinoic acid-differentiated dendritic cell immunotherapy. IL-27-dependent induction of Foxp3- non-Tr1 regulatory T cells (VAC4P.1100)
Authors: Gordon John , Li Chunyan , Churchman Laura , Dawicki Wojciech 
Published: January 2023

Abstract
Background. Semi-mature IL-10-induced DC (DC10) induce human or mouse effector Th2 cells (Teff) to differentiate into Foxp3+ Treg and thereby induce asthma tolerance. Retinoic acid (RA) and TGFβ-induced gut DC similarly induce tolerance to gut contents via Foxp3+ Treg, while immature hDC-10 induce IL-10-dependent Foxp3- Tr1 cells. Methods. We assessed the abilities of mature RA-induced DC to reverse anaphylaxis sensitivity, characterizing LPS-matured DC-RA and their tolerogenic activities in mouse models of OVA and peanut systemic anaphylaxis. Results. Mature DC-RA strongly expressed CD103 (αE integrin), MHCII, co-stimulatory markers, PDL1 & -L2, ICOSL, TGFβ, IL-27 and the RA-metabolizing enzyme Aldh1A2. They induced Th2 cells to convert into CD25+LAG3+CD49b-IL-10-Foxp3- Treg in an IL-10-independent, IL-27-dependent fashion, although IL-27 was dispensable for Th2 suppression by DC-RA. Wild-type, but not IL-27-/- DC-RA therapy reversed systemic anaphylaxis, including clinical scores, diarrhea, mast cell activation and Th2 responses, and lowered serum allergen-specific IgE and IgG1 levels. Conclusions. Allergen-presenting DC-RA induce food allergen tolerance by activating CD25+Foxp3-, non-Tr1 responses in an IL-27-dependent fashion. This data indicates that, like DC10, DC-RA can also be used for allergic disease immunotherapy, but that these two populations of DCreg induce distinct Treg responses, and this potentially provides us with important options DCreg immunotherapy
Forskolin increases propagation of influenza virus in Madin-Darby canine kidney cells (VAC5P.1118)
Authors: Lee Sang-Yeon , Sohn Sung-Hwa , Park Hye-Lim , Nam Jae-Hwan 
Published: January 2023

Abstract
Influenza virus is a pathogenic negative-sense RNA virus belonging to the Orthomyxoviridae, which causes severe respiratory illness worldwide. A vaccine against the influenza virus is the best way to control the spread of influenza. Recently, a cell-based influenza vaccine was developed to overcome the disadvantages of egg-based vaccines. Here, we investigated whether forskolin, an adenylyl cyclase activator, could increase the output of the cell-based influenza vaccine. We found that forskolin increased the propagation of three influenza virus subtypes, A/H1N1/California/4/09, A/H3N2/Mississippi/1/85, and B/Shandong/7/97, in MDCK cells. We performed a microarray analysis to identify the mechanism whereby forskolin enhances influenza virus replication. We compared a forskolin-treated group with untreated group after infection with influenza virus. The expression of importin α5 and Erk1/2, which can regulate the import and export of viral ribonucleoprotein complex (vRNP), is upregulated in the forskolin-treated group. By contrast, the expression of innate immunity factors and antiviral factors such as TLR3, RIG-I/MDA5, IRF3, MxA and viperin, is downregulated. Taken together, we suggest the reduction of the antiviral response and the increase of translocation of vRNP by forskolin enhance the propagation of influenza virus. Therefore, the addition of forskolin to cultures may be helpful to increase the production efficiency of cell-based vaccines for influenza virus.