bluuebunny/crossref_metadata_2025_split huggingface dataset:
Developmental plasticity allows outside-in immune responses by resident memory T cells
Authors: Quarnstrom Clare F , Fonseca Raissa , Beura Lalit K , Ghoneim Hazem E. , Fan Yiping , Zebley Caitlin C , Scott Milcah C , Fares-Frederickson Nancy J , Wijeyesinghe Sathi P , Thompson Emily A , da Silva Henrique Borges , Vezys Vaiva , Youngblood Benjamin A , Masopust David 
Published: January 2023

Abstract
Central memory T (TCM) cells patrol lymph nodes and perform conventional memory responses upon re-stimulation: proliferation, migration, and differentiation into diverse T cell subsets while also self-renewing. Resident memory T (TRM) cells are parked within single organs, share properties with terminal effectors, and contribute to rapid host protection. We observed that reactivated TRM cells rejoined the circulating pool. Epigenetic analyses revealed that TRM cells align closely with conventional memory T cell populations, bearing little resemblance to recently activated effectors. Fully differentiated TRM cells isolated from small intestine epithelium exhibited the potential to differentiate into TCM, TEM, and TRM cells upon recall. Ex-TRM cells, former intestinal TRM that rejoined the circulating pool, heritably maintained a predilection for homing back to their tissue of origin upon subsequent reactivation and a heightened capacity to re-differentiate into TRM cells. Thus, TRM cells can rejoin the circulation but are advantaged to re-form local TRM when called upon.
Outcome of Chlamydia muridarum genital infection and immune response in a beta2-adrenergic receptor knockout stress mouse model
Authors: Belay Tesfaye , Martin Elisha 
Published: January 2023

Abstract
The beta-2-adrenergic receptor (β2-AR) is one of the β-AR subsets found in most immune cells and is stimulated by norepinephrine (NE) binding. We showed previously that exposure of a mouse model to cold water stress leads to high level production of NE that may result in increased intensity of Chlamydia muridarum genital infection but the mechanism is not well defined. The purpose of this study was to develop and characterize a β2-AR knockout (KO) stress mouse model to investigate its susceptibility to C. muridarum genital infection. We hypothesized that lacking the β2-AR leads to a decreased C. muridarum shedding from the genital tract of stressed mice compared to that of stressed wild type (WT) mice. Stressed and non-stressed C57BL/6J WT and β2-AR KO mice were infected intravaginally with C. muridarum. Swabbing at 3-day intervals during primary infection and C. muridarum isolation was performed by culturing in McCoy tissue culture following standard methods. β2-AR KO stressed mice had a mean of 8540 inclusion forming units/ml (IFU/ml) compared to a mean of 18000 IFU/ml of stressed wild type at day 9 post infection. The overall course of primary infection in the β2-AR KO mouse model showed slightly reduced Chlamydia shedding. In addition, transfer of CD4+ T cells from black J6 WT to β2-AR KO resulted in a significant increase of shedding compared to that of non-stressed WT transfer (6000 versus 1000 IFU/ml). Secretion of interferon gamma was restored in β2-AR KO mice compared to WT. Overall, these results imply that deficiency in β2-AR leads to decreased C. muridarum shedding compared to the WT. However, other members of the alpha- and beta-adrenergic receptor family may compensate for the absence in β2-AR KO mice thus more investigation is needed.
Expediting antibody discovery using Bioelectronica’s HypercellTM platform
Authors: Yu Tian , Hull Jonathan , Ruiz Andrea , Bhat Ashwini , Basu Amar 
Published: January 2023

Abstract
Antibody-based drugs have been successful in a range of therapeutic categories. However, generating monoclonal antibodies is time-consuming and expensive. A common approach is Hybridoma technology, which overcomes the short life-span of IgG-secreting plasma B cells in vitro. However, many plasma B cells are lost due to the low efficiency of hybridoma cell fusion (typically &lt;10%).
Direct single B cell screening strategies have emerged to bypass hybridoma fusion and recombinatorial display, coupled with the generation of recombinant monoclonal antibodies through mammalian expression systems. Obtaining expression systems with the required productivity, specificity and stability for clinical or commercial use requires screening millions of cells and thousands of clones.
Bioelectronica’s HypercellTM platform is an emerging technology used throughout antibody discovery and cell-line development to identify and isolate single, high-antibody secreting cells from large pools (~10,000,000 cells) in short periods (ca. 48 hrs). This scalable “electrofluidic” sorting system reduces time and cost by integrating antigen-detection reagents and real-time computer vision analysis to expedite single cell sorting.
In this paper antigen-specific IgG-secreting hybridoma cells are identified and sorted by their secretion rate. The cells and reagents are encapsulated in a Polydisperse Oblate Dispersion system (PODs), incubated for 1–4 hours for signal gain, and loaded into the HypercellTM device for cell sorting. Alternatively, the mixture can be analyzed without sorting to produce single cell secretion “finger print” signatures that can help identify unique expression patterns and monitor cell line stability over culturing time.
Development of a Chinese Herbal Immunogen Assay for the Measurement of T cell Anti-Tumor Immunity
Authors: Yen Hung-Rong , Song Ying-Chyi 
Published: January 2023

Abstract
T cells are the master regulators of adaptive immune responses. Proper T cell responses are essential for the hosts to orchestrate sufficient adaptive immunity. T cell immunity is crucial for anti-tumor immunity. Manipulating T cell immunity by Chinese herbs is a promising field to explore. Many herbs were reported to regulate the T cell activation. Modulation of T cell immunity by Chinese herbs may be an applicable intervention to modulate immunity, for example, to promote anti-tumor immunity against cancer. In this study, we developed a Chinese Herbal Immunogen Assay to measure the T cell immunity by screening the commonly used Chinese herbal formulas and single herbs from our Chinese herbal medicine extracts database. In the preliminary test, we collected human peripheral blood mononuclear cells (PBMC) were from fresh whole heparinized blood of healthy donors or cancer patients to proof the concept. The isolated PBMCs were stimulated with DMSO or a panel of various herbal extracts, respectively. Cytokines secretion from the activated PBMCs in culture supernatants were quantified at various time points by using standard ELISA for TNF-a, IL-6 and IFN-γ. At baseline, PBMCs from cancer patients had a lower average level of TNF-a, IL-6 and IFN-γ, compared to the healthy controls. After stimulation with a panel of herbal extracts, we identified the herbs that can consistently enhance cytokine productions. These herbs could be used for further development of new drug for boosting anti-tumor immunity.
Immunosenescent characteristics of T cells in young patients following haploidentical stem cell transplantation from parental donors
Authors: Lee Ga Hye , Hong Kyung Taek , Kim Bonah , Kang Yeon Jun , Lee Su Jeong , Choi Jung Yoon , Shin Hee Young , Lee Won-Woo , Kang Hyoung Jin 
Published: January 2023

Abstract
Pediatric and adolescent patients in need of allogeneic hematopoietic stem cell transplantation (HSCT) generally receive stem cells from older, unrelated or parental donors when a sibling donor is not available. Despite encouraging clinical outcomes, it has been suggested that immune reconstitution accompanied by increased replicative stress and a large difference between donor and recipient age may worsen immunosenescence in pediatric recipients. Therefore, in this study paired samples were collected at the same time from donors and recipients of haploidentical hematopoietic stem cell transplantation (HaploSCT). We then conducted flow cytometry-based phenotypic and functional analyses and telomere length (TL) measurements of 21 paired T-cell sets from parental donors and children who received T cell-replete HaploSCT with post-transplant cyclophosphamide (PTCy). Senescent T cells, CD28− or CD57+ cells, were significantly expanded in patients. Further, not only CD4+CD28− T cells, but also CD4+CD28+ T cells showed reduced cytokine production capacity and impaired polyfunctionality compared with parental donors, whereas their TCR-mediated proliferation capacity was comparable. Of note, the TL in patient T cells was preserved, or even slightly longer, in senescent T cells compared with donor cells. Regression analysis showed that senescent features of CD4+ and CD8+ T cells in patients were influenced by donor age and the frequency of CD28− cells, respectively. Our data suggest that in pediatric HaploSCT, premature immunosenescent changes occur in T cells from parental donors and therefore, long-term immune monitoring should be conducted.
Vesicular stomatitis virus expressing destabilized tumor associated antigens with enhanced MHC class I presentation to improve anti-tumor T cell responses
Authors: Huff Amanda , Evgin Laura , Thompson Jill , Kottke Tim , Wongthida Phonphimon , Driscoll Christopher , Shim Kevin G. , Scheulke Matthew , Vile Richard G. 
Published: January 2023

Abstract
We have previously shown that vesicular stomatitis virus (VSV) vectors encoding truncated tumor antigens, NRAS, Cytochrome C1, and TYRP1, prime superior systemic anti-tumor T cell responses and provide enhanced therapeutic benefit compared to vectors expressing full-length forms of the protein. Mapping of the roughly 20 amino C-terminal truncated regions onto structural domains of each protein suggested the truncations disrupted the stability and localization of each protein. Indeed, immunofluorescent staining and radiolabeled pulse chase found that truncated TYRP1 (ΔTYRP1) had an altered localization pattern and a significantly reduced half-life compared to full-length TYRP1. Thus, we hypothesized that expression of destabilized antigens from VSV would provide superior therapy compared to expression of wild-type antigens. To test this, we introduced point-mutations into structurally important domains of the model antigen ovalbumin (OVA). We identified one mutant which was significantly destabilized (OVAmut), which resulted in enhanced MHC Class I presentation of the immunogenic SIINFEKL epitope on transfected cell in vitro. Expression of OVAmut resulted in better recognition by cognate CD8 OTI T cells, enhanced T cell activation, proliferation, and effector cytokine production. Finally, a VSV vector expressing OVmut was better able to prime OTI T cells in vitro as well as in vivo. Ongoing studies aim to identify the changes in the quantity and repertoire of immunogenic epitopes presented from cells infected with VSV vectors expressing destabilized antigens. These studies provide insight into the capacity of the VSV platform to successfully encode unstable antigens with the goal improve immunogenic responses.
Tumor membrane vesicle-based vaccine immunotherapy inhibits tumor growth and metastasis of squamous cell carcinoma of head and neck in mice
Authors: Bommireddy Ramireddy , Munoz Luis E , Pack Chrstopher , Ramachandiran Sampath , Reddy Shaker , Chen Georgia , Saba Nabil , Shin Dong M , Selvaraj Periasamy 
Published: January 2023

Abstract
Head and neck cancer is a leading cause of cancer related deaths accounting for approximately 3% of all cancer related mortalities in the US. Currently, there is no cure for the advanced squamous cell carcinoma of the head and neck (SCCHN) thus development of efficacious therapies is urgently needed. To test whether vaccine-induced immunity inhibits tumor growth, we investigated efficacy of a tumor membrane-based vaccine (TMV) immunotherapy in murine SCCHN models. The MOC1, MOC2 and SCC VII tumors grown subcutaneously in syngeneic mice were harvested to generate TMVs. TMVs were then protein transferred with glycolipid-anchored immunostimulatory molecules GPI-B7.1 and GPI-IL-12 to generate the TMV vaccine. Mice were vaccinated with TMV vaccine after tumor cell challenge (therapeutic) and tumor growth was monitored every 3 days. Survival was then assessed using a Kaplan-Meier survival curve and significance determined using a Log-rank test for comparison analysis. The TMV vaccine inhibited tumor growth and improved the survival of mice challenged with MOC1 or SCC VII tumor cells. Interestingly, combination of anti-PD1 antibody with TMV vaccine decreased MOC1 tumor growth. However, MOC2 (poorly immunogenic tumor) did not respond to anti-PD1 or anti-CTLA-4 antibody therapy but significantly inhibited by TMV vaccine. MOC2 cells metastasize to the lungs in control mice but it is significantly inhibited in mice administered with TMV vaccine. TMV vaccine increased the infiltration of CD4+ and CD8+ T cells into the tumors suggesting that TMV vaccine converts cold tumors into hot tumors. These observations suggest that TMV vaccines can be harnessed to develop an effective immunotherapy for squamous cell carcinoma of the head and neck.
Characterization of innate and adaptive immune impacts of chemically dissimilar STING adjuvants
Authors: Boehm Dylan T , Mizuno Nobuyo , DeFilippis Victor 
Published: January 2023

Abstract
STING is a PRR for cyclic dinucleotides (CDN) synthesized by the cytosolic enzyme cGAS following exposure to microbial or cellular DNA. This stimulates innate responses characterized by localized type I IFN and proinflammatory cytokines. It is thus key to establishing adaptive immunity by facilitating APC recruitment and function. As such, STING is an attractive target to direct adjuvants and anti-tumor drugs. However, STING also drives processes such as apoptosis and autophagy that are certain to shape immune responsiveness. The degrees to which the protein elicits phenotypes can also vary substantially between STING-inducing stimuli including CDN species and small molecule agonists. While pharmacologic STING activation enhances immunogenicity of vaccine antigens, the impact of differences in STING-associated intracellular responses on overall immunity has not been explored. To address this, we utilize a mouse model of vaccination against Chikungunya virus. This is done in combination with chemically diverse STING agonists including CDNs and small molecules. Despite the direct engagement of STING by each, the characteristics of the immune reactivity to the antigen varied significantly. In addition, our exploration of STING-mediated innate correlates of these differences led to unexpected observations regarding agonist-specific cytokine induction, APC maturation, antigen trafficking and presentation. This has important implications in at least two areas: 1) It is crucial to filling the numerous gaps that exist with regard to understanding mechanisms that drive STING-associated immune enhancement; 2) It will reveal properties desirable for STING-based adjuvants to help direct future optimization.
Evaluation of Cancer-testis Antigens in Osteosarcoma and Dedifferentiated Liposarcoma as Targets for Immunotherapy
Authors: Jirovec Anna , Flaman Ashley , Purgina Bibianna , Werier Joel , Diallo Jean-Simon 
Published: January 2023

Abstract
T cell based immunotherapies targeting tumor antigens are a promising alternative to traditional cancer treatments due to their ability to eliminate malignant cells, but require highly immunogenic tumor-associated proteins as targets for CD8+ T cell recognition. Additional parameters to consider in a T cell based immunotherapy include tumor T cell infiltration and expression of HLA-peptide complexes on the surface of cancer cells. The goal of this study is to identify targetable immunogenic tumor antigens and to characterize the immune profile of human dedifferentiated liposarcoma (DDLPS) and osteosarcoma (OS) sarcoma subtypes by screening for cancer-testis antigen (CTA) expression, HLA expression, and tumor T cell infiltration by immunohistochemistry (IHC).
Human tissue micro-arrays composed of 80 cores of OS and 49 cores of DDLPS were obtained and stained by IHC for selected CTAs (MAGE-A3, NY-ESO-1 and SSX2) and various immune markers. In DDLPS, 11% of samples express MAGE-A3 and 2% express SSX2 and NY-ESO-1 in low percentages of tumor cells. In OS, 100% of samples express MAGE-A3 and 89% express SSX2, both with &gt;80% of positive cases showing moderate to high expression. NY-ESO-1 is expressed in 78% of OS samples, predominantly at low levels. Brisk infiltration of CD8+ T cells is observed in over 70% of both sarcoma types. Furthermore, all sarcoma samples tested are positive for HLA expression.
These results show promising expression of CTAs MAGE-A3 and SSX2 in OS, which may be used as targets for immunotherapy for OS. The data generated throughout this study will provide insight into the immune profile of these sarcomas - information that is critical for immunotherapy design.
Dietary β-glucan reduced Salmonella shedding, shifted intestinal microbiome, and altered intestinal integrity
Authors: Byrne Kristen A , Trachsel Julian M , Bond Zahra F , Slate Jamison R , Kerr Brian J , Bearson Bradley L , Bearson Shawn M , Loving Crystal L 
Published: January 2023

Abstract
β-glucan is a prebiotic dietary fiber with immune and microbiota modulating properties. Dietary β-glucan can shift intestinal microbial populations and concentrations of short chain fatty acids (notably, butyrate and propionate), which may alter local populations of regulatory T-cells. Additionally, epigenetic reprograming of monocytes by β-glucan impacts subsequent monocyte responses to heterologous agonists, a form of innate training. Here, β-glucan from Saccharomyces cerevisiae was evaluated as a non-antibiotic dietary additive to limit shedding of the foodborne pathogen, Salmonella enterica server Typhimurium, via changes to the intestinal microbiota, intestinal integrity, and/or the local and peripheral immune system in pigs. Overall Salmonella shedding and cecal mucosa burden was significantly reduced in pigs fed dietary β-glucan compared to Salmonella challenged pigs on control diet. Shifts in microbial communities were associated with reduced Salmonella burden, as pigs fed β-glucan exhibited enhanced cecal epithelial barrier function, altered fecal bacterial communities, and increased short chain fatty acid concentrations. However, no significant differences in regulatory T cell population nor ex vivo monocyte responses were found between groups. Collectively, the non-antibiotic dietary additive β-glucan altered microbial communities and host epithelial cell function in conjunction with reduced Salmonella shedding in pigs.
Intraepithelial T cell phenotype and function diverge by intestinal location with age
Authors: Wiarda Jayne E , Trachsel Julian M , Bond Zahra F , Byrne Kristen A , Gabler Nicholas K , Loving Crystal L 
Published: January 2023

Abstract
Intestinal intraepithelial T cells (IETs) provide a first line of defense against intestinal pathogens and play an important role in immune tolerance versus activation. In pigs, intestinal T cell populations stabilize at ~8 weeks of age, but IET phenotypes and function are poorly understood during the early postnatal period. Better understanding age- and location-dependent characteristics of porcine IETs may be useful in developing strategies to maintain intestinal integrity and improve health. To assess impacts of age and intestinal location on abundance and composition of IETs, sections of jejunum, ileum, cecum, and colon from 4-, 6-, and 8-week-old pigs were analyzed by histologic staining and flow cytometric analyses. IET abundance increased with age, and CD2+CD8α+γδ and CD4−CD8α+αβ T cells were the primary IET populationsin all samples (&gt;78%). Compositions of IETs were similar at all locations in 4-week-old pigs. As pigs aged, cecal and colonic IET compositions remained similar to each other but diverged from small intestinal IETs, while jejunal and ileal IET compositions diverged from both large intestinal IETs and each other. These findings indicate both intestinal location and age are drivers of IET heterogeneity. Small intestinal IETs had lower percentages of γδ IETs in 6- and 8-week-old pigs, and CD4−CD8α+αβ IETs increased in abundance at all intestinal locations as pigs aged. Percentages of CD27+IETs in ileum and large intestine decreased over time, but decreases were more drastic in large intestine, suggesting IETs in the distal but not proximal intestinal tract were activated as pigs aged. Small intestinal IET compositions closely resembled human IET populations, highlighting pigs as a biomedical model for IET research.
Immune fitness as a measure of animal health, welfare and productivity in the feedlot
Authors: Purdie Auriol C , Plain Karren , de Silva Kumudika 
Published: January 2023

Abstract

Background
Animals have long been bred for production traits that enable high-yield, but this does not consider the overall immunological ‘fitness’. What is the physiological basis that enables some animals to thrive or fail when exposed to production stresses, are there any measures of the immune component to identify these animals? Further, are there commonly related genes associated with immune fitness?

Methods
50 mixed breed beef cattle were randomly selected at feedlot intake. Blood and saliva samples were collected at arrival (high stress), post- acclimatization and pre-slaughter (60 days post intake or ≥400kg). Body condition and other measures (weight, height, disposition, coat colour) were collected at each sampling and at slaughter. Samples were processed for cell counts, RNA (transcriptomics analysis) and serum (protein analysis). Transcriptomic and proteomic analysis was by 24k Bovine Affymetrix ST1 GeneChip and RayBiotech Bovine Cytokine Array Q30.

Results and conclusion
Analysis correlated to outcome measures identified genes and proteins associated with immune fitness including those correlated to animals with susceptibility to disease and/or low weight gains. These results suggest that immune measures may be utilized to identify animals at risk of poor production performance facilitating improved animal management to optimize animal health and welfare.
TARGETING METABOLIC REPROGRAMMING OF RESPIRATORY SYNCYTIAL VIRUS INFECTION
Authors: Bahadoran Azadeh , Bezavada Lavanya , Rezinciuc Svetlana , Devincenzo John , Cormier Stephania , Smallwood Heather 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) causes severe respiratory disease and infects virtually most children by 2 years of age. It is the leading cause of hospitalization of infants worldwide and remerges later in life to be a serious lower respiratory tract illness in the elderly. Recently, significant emphasis focused on changes in host cellular metabolism in response to viral infection. We performed a retrospective study of pediatric patients infected with RSV and found this infection correlated with increased glucose uptake in the lungs. We characterized cells freshly isolated from naturally RSV infected pediatric patients’ nasopharyngeal aspirates (NPA) and quantified their bioenergetics. Next, we will determine RSV infection induced changes in metabolic responses of different cell types including primary human epithelial cells (NHBE), dendritic cells (DCs), macrophages (MOs) and A549 cell line. Additionally, we will characterize the infect co-cultures of NHBE and DCs/MOs with RSV to determine if their crosstalk can help regulate aberrant NHBE metabolism. We found a dramatic increase in glycolysis and mitochondria respiration in cells isolated from patients’ NPA. RSV infection increases basal respiration, ATP production, and proton leak while reducing mitochondrial oxygen consumption and respiratory capacity in pediatric patients’ NPA. Significant increases in metabolism in the infected patient’s upper respiratory cells ex vivo support our in situ findings in RSV infected patients lungs. Understanding and defining the metabolic changes in the host during RSV infection may lead to novel therapeutic approaches through targeted inhibition of specific cellular metabolic pathways.