bluuebunny/crossref_metadata_2025_split huggingface dataset:
Environmental cues during infection determine effector CD8 T cell differentiation (LYM4P.748)
Authors: Plumlee Courtney , Lefrançois Leo , Khanna Kamal 
Published: January 2023

Abstract
Naive antigen-specific CD8 T cells expand in response to infection and can be phenotypically separated into distinct effector populations, which include memory precursor effector cells and short-lived effector cells. In the days before the peak of the T cell response, a third population called early effector cells (EECs) predominate the antigen-specific response. However, the contribution of the EEC population to the CD8 T cell differentiation program during an immune response is not well understood. Here we show that EECs were pre-programmed to differentiate based on early signals received from distinct infectious environments, but surprisingly, when these same EECs were transferred into infected hosts, the EECs could be diverted from their original fate by a disparate inflammatory environment. After observing plasticity at the population level, we also examined the antiviral and antibacterial responses of single CD8 T cells from the polyclonal repertoire. The progeny of naive clonal CD8 T cells displayed unique profiles of differentiation based on extrinsic pathogen-induced environmental cues, with some clones demonstrating extreme bias toward a single developmental pathway. These results, taken together, demonstrate that stochastic and instructive events differentially contribute to shaping the CD8 T cell response and provide insight into the underlying forces that drive effector differentiation and protective memory formation.
Resistance of rhesus monkeys to homotypic challenge after Chlamydia trachoma tis genital tract infection (MPF2P.816)
Authors: Qu Yanyan , O'Connell Catherine , F. Tarantal Alice , Andrews Charles , O'Connor Shelby , Vallejo Abbe , Darville Toni 
Published: January 2023

Abstract
Chlamydia trachomatis is the most prevalent sexually transmitted bacterial infection for which a vaccine is needed. Here, 2 groups of 5 female rhesus monkeys were infected with either plasmid-cured attenuated CTD153 or wildtype virulent C. trachomatis D/UW-3/Cx, and then challenged with D/UW-3/Cx. Cervical infection was monitored, blood and cervical secretions were collected at intervals for T cell and antibody assays and genital tissues were harvested at wk 12 for histopathology. Results showed 3/5 monkeys infected with D/UW-3/Cx or CTD153 were resistant to challenge. However, 2/5 in each group were susceptible to challenge and 2 monkeys primarily infected with CTD153 exhibited prolonged challenge infections. No chronic genital tract pathology was observed. Serum anti-elementary body (EB) IgG was detected in all; mucosal anti-EB IgG/IgA were detected only in the 2 with prolonged infection. Bioassays showed enhanced proliferation in CD4 and CD8 central (TCM) and effector (TEM) memory compartments, and IFNγ production in response to EB at wk 4, that declined to baseline by wk 12. In vivo expansion of CD4 TCM, CD8 TEM, and NKG2A cells at wk 4 coincided with infection resolution in 8/10. T cell proliferative responses to EB were highest in monkeys with prolonged infection. Three reinfection-resistant monkeys shared Mamu-DRB1*03 and DRB1*10. Resistance to chlamydial reinfection is likely linked to MHC II and driven by chlamydial antigens minimally expressed by inactivated EBs.
Biomarkers of pulmonary tuberculosis identified in multiplexed proteomic assay (SOMAscan) of human serum (MPF2P.808)
Authors: Ochsner Urs , Green Louis , De Groote Mary , Sterling David , Ostroff Rachel , Janjic Nebojsa 
Published: January 2023

Abstract
Background: A rapid, accurate, and inexpensive tuberculosis (TB) diagnostic test in the developing world would allow earlier treatment and reduce transmission. Methods: We used slow off-rate modified aptamers (SOMAmers) in a highly multiplexed proteomic assay (SOMAscan) to measure 1129 human proteins and 16 M. tuberculosis proteins in serum samples collected in S. Africa, Vietnam and Peru, provided by the Foundation for Innovative New Diagnostics. Results: Among the top host serum biomarkers distinguishing TB from non-TB were factors involved in cell-matrix interactions such tissue remodeling and fibrosis (Kallistatin, TSP4, gelsolin, CDON) which were lower in TB compared to non-TB, and factors of innate immunity, acute phase reactants, and inflammation (LBP, ITI heavy chain H4, NPS-PLA2, IP-10), which were higher in TB, regardless of the HIV status. A 9-marker model performed well in a training set of 173 TB vs. 160 non-TB samples (sensitivity 90% / specificity 85%, AUC=0.94), which was confirmed in a blinded verification set of 132 TB vs. 118 non-TB samples (sensitivity 80% / specificity 84%, AUC=0.88). Conclusions: The discovery of robust, quantitative, non-culture based diagnostic biomarkers of active pulmonary TB has great potential to facilitate the rapid and accurate diagnosis of TB disease.
Epigallocatechin-3-gallate Reduces Mast Cells Activity TNF-α and NFKB in Colitis by Interrupting an Inflammatory Cascade (MUC2P.827)
Authors: Hajj Hussein Inaya , Zgheib Zeina , Zeenny Marie Noel , Chams Sana , Bou Assi Tarek , Chams Nour , Jurjus Abdo 
Published: January 2023

Abstract
There is increasing evidence that (-)-epigallocatechin-3-gallate (EGCG) inhibits carcinogenesis and inflammation among other properties. However, its mechanism is not fully elucidated.To investigate the mechanism of action of EGCG in improving experimental colitis. 35 male Sprague-Dawley rats were randomly divided into 4 groups: Normal control group (n=5), EGCG group (n=9), TNBS group (n=9), and TNBS+EGCG group (n=12). For both, TNBS treated group and EGCG group, 1 mg/Kg EGCG was administered daily by intraperitoneal injection, starting one week before the induction. At days 3, 10 and 17, rats were sacrificed and the descending colon was collected. The score of histological alterations of the colonic mucosa was evaluated. The mast cells were assessed by toluidine blue staining. The mRNA expression of Tumor Necrosis Factor-α (TNF-α), and nuclear factor (NF-KB) was measured and ROS was tested by immunofluorescence. EGCG decreased significantly the erosions and the inflammatory cellular infiltration of the mucosa and submucosa. In addition, the mast cells were markedly reduced in number and degranulation. The increased levels NF-KB and TNF-α after TNBS administration were significantly and concurrently reduced in colonic scrapings after EGCG treatment at all the time points and did ROS. EGCG probably prevented the transcription of inflammatory genes by stabilizing mast cells and reducing its secretory pro-inflammatory factors such as TNF-α.
Role of non-hematopoietic β-arrestin1 in intestinal inflammation (MUC2P.828)
Authors: Lee Taehyung , Lee Eunhee , Obey Kimberly , Michelin Erika , Chung Misoo , Vlachos Elena , Parameswaran Narayanan 
Published: January 2023

Abstract
β-arrestin1 (β-arr1), an intracellular scaffolding protein has been shown to regulate inflammation by modulating Toll-like receptor (TLR) signaling in immune cells. We recently demonstrated that β-arr1 deficiency attenuates experimental colitis in mice. However the cell-type-specific role of β-arr1 in colitis is not known. To test this we generated bone marrow chimeras with β-arr1 deficiency only in non-hematopoietic compartment (non-hβ-arr1KO), and subjected the WT and non-hβ-arr1KO mice to DSS-induced colitis. Interestingly, our results demonstrate that β-arr1 expression in the non-hematopoietic compartment protects mice from early weight loss during colitis. To understand the signaling mechanisms, we knocked down β-arr1 in human intestinal epithelial cells (SW480) and tested the effect of TNF-α on MAPK and NFκB signaling pathways. Even though β-arr1 knockdown did not affect TNF-α-induced NFκB activation, p38 and JNK pathways were enhanced in the knockdown cells. Intriguingly, ERK1/2 activation by TNF-α was significantly inhibited in β-arr1-knockdown cells. Differential modulation of the MAPK signaling by β-arr1 could lead to higher apoptosis of intestinal epithelial cells in non-hβ-arr1 deficient mice. Together, our results demonstrate that β-arr1-dependent signaling in non-hematopoietic cells likely plays a critical role in the pathogenesis of colitis.
Loss of regulatory intestinal dendritic cell subsets during gut inflammation in humans and mice (MUC2P.830)
Authors: Mann Elizabeth , Shi Jie , Wang Dezhi , Zhai Huihong , Jin Shi , Jiang Weiwei , Li Xuhang 
Published: January 2023

Abstract
Background: Dendritic cells (DC) and macrophages (Mφ) mediate intestinal immune tolerance. Recent evidence suggests intestinal CD103+CD11b- DC are involved in generation of regulatory T cells (T-reg), with CD103+CD11b+ DC stimulating Th17 cell differentiation. Information regarding gut DC function during inflammation is scarce. Methods: We compared murine and human gut DC and Mφ in the steady state and during intestinal inflammation, by flow cytometry. Results: Proportions of CD103-CD64+ Mφ were enhanced in the inflamed IL-10 knockout (KO) murine colon compared to WT, alongside a loss of CD103+CD11b- DC. Proportions of CD4+FoxP3+ T-reg were reduced in the inflamed IL-10 KO colon, with enhanced production of IL-17A and TNFα by T cells but a loss of regulatory TGFβ production by CD103+ DC. Proportions of CD103-CD64+ Mφ were also enhanced in the human Crohn’s disease colon compared to non-inflamed control tissue (diverticulitis), alongside a loss of CD103+ DC. CD103 was co-expressed with either CD141 or CD1c on human intestinal DC, corresponding to CD103+CD11b- and CD103+CD11b+ murine DC respectively. Proportions of all CD103+ gut DC subsets were reduced in human Crohn’s disease. Conclusions: Lack of IL-10 may skew DC and Mφ differentiation in the gut, leading to loss of DC able to generate T-reg. Further experiments will determine the ability of intestinal DC and Mφ subsets to stimulate specific T cell responses in vitro and the roles of IL-10 and TGFβ during this process.
The microbiota conditions CX3CR1+ intestinal mononuclear phagocytes to limit intestinal inflammation. (MUC2P.822)
Authors: Diehl Gretchen , Galan Carolina , Longman Randy , Egawa Takeshi , Yang Yi , Littman Dan 
Published: January 2023

Abstract
Regulation of intestinal immune responses requires distinguishing normal beneficial bacteria (the microbiota) from pathogens capable of inducing disease. While it is clear that the intestine and intestinal immune system recognize and respond to the microbiota, it is unclear how this recognition influences subsequent immune responses. One intestinal antigen presenting cell population, characterized by expression of the chemokine receptor CX3CR1, has been described as tissue resident macrophages. In contrast to previous reports, we recently demonstrated that CX3CR1+ cells migrate from the intestine to the mesenteric lymph node, suggesting that they can behave as dendritic cells (DCs). By utilizing both genetically modified mice and antibiotics to manipulate the intestinal microbiota, we demonstrated that the microbiota influences intestinal immunity by modulating CX3CR1+ cells. In the presence of an intact microbiota, CX3CR1+ cells limit the induction of inflammatory immune responses. In contrast, in conditions where the microbiota has been perturbed, CX3CR1+ cells promote the induction of inflammatory immune responses. We further demonstrate that antigen presentation by CX3CR1+ cells is critical for priming T cell responses to intestinal antigens in vivo, confirming CX3CR1+ cells can act as DCs. Together, our findings indicate that CX3CR1+ cells play a central role in integrating signals from the microbiota to maintain intestinal immune homeostasis.
Antimicrobial peptides are induced in the lower bowel after acute Helicobacter hepaticas infection (MUC2P.824)
Authors: Wang Chuanwu , Fox James , Horwitz Bruce 
Published: January 2023

Abstract
Antimicrobial peptides (AMPs) play an essential role in innate immunity to enteric bacterial infection. Previous studies have demonstrated higher antimicrobial peptides RegIIIβ and RegIIIγ expression in the colon of AOM treated 129 RAG-deficient mice 12 weeks post Helicobacter hepaticus (Hh) infection. Blocking of IL-22 interferes with AMP induction. However, whether Hh infection alone acutely induces AMP expression, and whether the ability of Hh to induce AMP expression varies in the different regions of the large bowel has not been evaluated. To examine these issues, several anatomical regions of the lower bowel from 129 Rag2-/- mice were examined at 2 and 6 weeks post Hh infection. Surprisingly, we found that while AMP expression at baseline was markedly lower in the cecum than the colon, AMP expression in the cecum was strongly induced within 2 weeks of Hh infection. This was associated with higher expression of IL-22, and cecal tissue infiltration by neutrophils and inflammatory monocytes. In contrast, Hh infection markedly inhibited IL-25 mRNA expression in both the cecum and colon. Interestingly, AMP expression is not regulated by production of IL-10 within the innate immune system. We hypothesize that Hh mediated induction of AMP may lead to alterations in the cecal microbiome. Further, we propose that this rapid model of Hh-induced AMP expression will be useful for delineating key cellular and molecular pathways that regulate intestinal innate inflammatory pathways.
A strategy to improve current MSC-based therapies (TECH1P.833)
Authors: Li Yan , Lin Feng 
Published: January 2023

Abstract
Mesenchymal stem cell (MSC) hold promise in the therapy of many inflammatory and degenerative disease. However, result from recent clinical trials using these cells are disappointing, suggesting that current MSC-based therapies need to be improved. We previously demonstrated that, immediately after their contact with serum following infusion, MSCs active complement of the innate immunity and are injured by the activated complement despite the presence of native cell surface complement regulators. In this work, we found that systemic administration of heparin, a widely used drug which is also a potent complement inhibitor, protects MSCs from serum-mediated damage after i.v. infusion. To minimize the side effects of systemic heparin administration, we also developed a method to paint heparin onto the surface of MSCs, We found that chemically modified heparin can be painted onto MSCs, and that these heparin-painted MSCs are better protected from serum (complement)-medicated attack and retain better immunosuppressive activity than regular MSCs. These data suggest that the cell surface engineering of MSC with heparin would be a simple and effective approach to improve the outcomes of current MSC-based clinical trials.
An efficient way to analyze function of immune cells (TECH1P.843)
Authors: Wang Zhong 
Published: January 2023

Abstract
Cellular assays for suspension cells, especially immune cells, provide investigators an opportunity to interrogate the biology of cellular function of immune cells or immunotherapeutic drugs. Despite significant technical advances made during the last few decades, working with suspension cells, such as primary blood cells, PBMCs (peripheral blood mononuclear cells), leukemia, myeloma, or lymphoma cells, current cellular assay approaches still suffer from limitations. For example, significant cell loss can occur during assay process. This loss can be further exacerbated when multi-step staining procedures are required. We applied DropArray plates, which employ surface tension to retain cells on the plate surface, for our suspension cellular assays. For a variety of suspension cells, including PBMCs, AML cells, or B cells, efficient retention (50% to 80%) is achieved during multiple washes. Reproducibility and quality of the experimental results are improved with DropArray plates. Furthermore, specific cytotoxic action of therapeutic antibodies on tumor cells can be also observed in real time. In summary, this system presented an efficient approach for phenotypic study of suspension cells, especially rare population of cells.
Profiling secreted cytokines from human Th2 and Th17 cells using antibody arrays (TECH1P.845)
Authors: Finkel David , James Amy , Brumbaugh Kathy , Dhawan Sanjeev , Goetz Christine , Bonnevier Jody , Wegner Greta 
Published: January 2023

Abstract
Naive T Helper cells can differentiate into Th1, Th2, and Th17 lineages which each secrete a characteristic signature of molecules. In this study, we demonstrate the utility of antibody arrays to monitor T cell differentiation processes. Naive T cells were isolated from PBMC preparations obtained from different donors. Following activation, the generation of each T cell subtype was confirmed by flow cytometry. Secreted proteins present in the cell culture supernatants were analyzed with a Human XL Cytokine Array containing 102 analytes. More than 30 proteins were identified, including cytokines specific to each Th subtype. ELISA data were used to quantitatively confirm the increases and decreases of multiple Th-specific cytokine levels observed using the array. These data demonstrate that antibody arrays provide an efficient tool to simultaneously study multiple parameters in complex biological processes where alternate differentiation protocols are being tested along with evaluating the impact of donor-to-donor variability.
Cohort-wide automated identification of morbid alleles with the human gene connectome (TECH1P.835)
Authors: Itan Yuval , Zhang Shen-Ying , Abel Laurent , Casanova Jean-Laurent 
Published: January 2023

Abstract
To determine the disease-causing allele(s) underlying primary human inborn errors, high-throughput genomic methods are applied and provide thousands of gene variants per patient. We recently reported a novel approach, the “human gene connectome” (HGC) - the set of all in silico-predicted biologically plausible routes and distances between all pairs of human genes. We demonstrated that the HGC is the most powerful approach for prioritizing high-throughput genetic variants in Mendelian disease studies. However, there is currently no available method for automating the selection of candidate disease-causing mutant alleles in the absence of a known morbid gene in at least one patient with the disease of interest, posing a major bottleneck in the field in high-throughput clinical genomics. We hypothesized that within a cohort of patients with the same Mendelian (or nearly Mendelian) disease, the cluster that contains the key disease-causing gene for each patient is the HGC-predicted biologically smallest cluster. We then developed and applied a Mendelian clustering algorithm, which estimates the biologically smallest HGC-predicted cluster that contains one allele per patient. By that we estimated and statistically validated a set of disease-causing alleles in a whole exome sequencing cohort herpes simplex encephalitis patients. This new unbiased approach should facilitate the discovery of morbid alleles in patients with primary inborn errors that lack a genetic etiology.
Th22 cells shape colorectal cancer stemness via epigenetic regulation (TUM2P.877)
Authors: Kryczek Ilona , Lin Yanwei , Nagarsheth Nisha , Peng Dongjun , Zhao Lili , Zhao Ende , Vatan Linda , Szeliga Wojciech , Dou Yali , Zgodzinski Witold , Majewski Marek , Wallner Grzegorz , Fang Jingyuan , Huang Emina , Zou Weiping 
Published: January 2023

Abstract
Little is known about the direct impact of immune elements on human colorectal cancer invasiveness and stemness. We investigated the interaction between IL-22+ cells and cancer (stem) cells in colorectal cancer patients. We have demonstrated that Th22-derived IL-22 targets disruptor of telomeric silencing 1-like (DOT1L). DOT1L methylates histone 3 lysine 79 (H3K79) and, in turn, leads to core stem cell gene activation, resulting in increased colon cancer stemness and tumorigenic potential. In addition, increased levels of DOT1L and H3K79me2 in colorectal cancer predict poor patient survival. Collectively, this work identifies that Th22 cells control colon cancer stemness via epigenetic regulation that affects patient outcome. Efforts to target this network might be a strategy in treating patients with colorectal cancer.
Very low avidity endogenous memory CD8+ T cells control tumor growth (TUM2P.893)
Authors: Stagliano Katherine , Zhu Ziqiang , Hurwitz Arthur 
Published: January 2023

Abstract
In this study, we describe mice bearing a very low avidity TCR transgene (TCRvlo) specific for a melanoma antigen. Remarkably, a majority of peripheral TCRvlo CD8+ T cells display a phenotype consistent with the expression pattern for endogenous memory (EM) T cells: CD44hi, CXCR3hi, Gr1hi, GITRhi, CD69lo, CD25lo, CD49dlo. This phenotype was not observed among thymic CD8+CD4- TCRvlo T cells. EM-T cells are proposed to arise from naïve T cells in the absence of activation by foreign antigen and are a very rare population of cells which may contribute to primary immune responses, but have not been studied in cancer models. We hypothesize that EM-T cells may provide a unique reservoir of cells that can control tumor growth. Our previous observations indicate that T cells bearing a high avidity TCR with identical antigenic specificity initially control tumor growth but are prone to tumor-induced tolerance. In our preliminary studies, compared to wildtype animals, TCRvlo transgenic mice displayed delayed subcutaneous B16 melanoma growth. In an adoptive transfer setting, TCRvlo T cells also delayed B16 tumor growth compared to untreated animals. Ongoing studies are focused on characterizing the functional responses of the TCRvlo CD8+ T cells to antigen and cytokines as well as determining whether TCRvlo T cells resist tolerization in the tumor microenvironment. Results from these studies may enable us to use EM-T cells as a resource for generating more durable tumor immunity.
Cancer immunotherapy: treatment of established breast cancer or prevention of tumor relapse and metastasis? (TUM2P.903)
Authors: Payne Kyle , Graham Laura , Bear Harry , Manjili Masoud 
Published: January 2023

Abstract
Cancer vaccines and adoptive immunotherapies (AITs) are being tested both in preventive and therapeutic settings. In order to determine the most effective immunotherapeutic strategy, we investigated the efficacy of AIT against established advanced carcinoma and minimal residual disease. We used Decitabine to improve immunogenicity of the tumor cells by inducing the expression of cancer testis antigens (CTA), in vivo. Tumor-sensitized immune cells were activated and reprogrammed, ex vivo, using bryostatin 1 and ionomycin, along with IL-2, IL-7, and IL-15; this produced a central memory phenotype in 73% of T cells, and an activated phenotype in 27% of NK cells and 94% of NKT cells. FVBN202 transgenic mice were protected from developing lung metastases following AIT, but failed to cure established lung metastases. The reprogrammed immune cells also demonstrated immunological memory that protected animals from recall tumor challenge. In vitro studies showed that reprogrammed immune cells were able to kill 82% of viable Adriamycin-treated quiescent tumor cells. Our results suggest that AIT could be efficacious against chemotherapy-resistant dormant tumor cells in a setting of low tumor burden and prevent distant metastasis of breast cancer. These findings are significant because breast cancer mortality results from metastatic disease that develops from residual tumor cells remaining dormant, sometimes for prolonged periods following conventional therapies.