bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of swine immune reagents for swine immunity and biomedical research
Authors: Lunney Joan K , Loving Crystal , Kenney Scott , LaBresh Joanna , Suresh Raksha , Byrne Kristen , Walker Kristen , Aquino Jovan , Renukaradhya Gourapura 
Published: August 2023

Abstract
The USDA-NIFA Swine Immune Toolkit Initiative has been involved in generating priority immune reagents for swine immunity and biomedical research and pipelining them for marketing. With the help of commercial partners we expressed each target immune molecule as a soluble protein using yeast expression systems and then produced panels of monoclonal antibodies (mAbs) against each target. Most recently we generated panels of mAbs reactive with porcine IL-5, IL-6, IL-13, IL-21, IL-28B, CXCL10, and BAFF and screened their reactivity in multiple immune assays. We determined mAb reactivity with orthologous proteins for most panels of mAbs. A sensitive sandwich ELISA is now available for IL-13 and CXCL10; other targets are being screened for best mAb pairs. Reactivity tests of labeled α-CXCL10, α-IL-6 and α-IL-13 mAbs for intracellular staining of porcine immune cells using flow cytometry assays are underway using different cell stimulation conditions. Immunohistochemistry analyses for binding of α-CXCL10 and other mAbs on formalin fixed pig lymph nodes and spleen tissues are in progress. Further, analysis of a large panel of commercial α-human CD antigen mAbs for cross-reactivity with porcine cells is ongoing. For each target, our goal is to provide the veterinary community with new commercial reagents and standardized assay techniques for their research efforts. Finally, efforts are proceeding to develop SLA tetramers for antigen presentation studies. Tools and reagents generated by this project will undoubtedly advance our understanding of swine immune responses to disease and vaccine and use for biomedical research efforts.
Supported by USDA-NIFA AFRI grant # 2019-67015-29815 and USDA ARS project 8042-32000-117.
Evaluating immunological outcomes associated with histotripsy treatment in canine osteosarcoma: a comparative human oncology model
Authors: Wagner Alayna , Ruger Lauren , Vickers Elliana , Coutermarsh-Ott Sheryl , Patwardhan Manali Nitin , Gannon Jessica , Allen Irving C , Vlaisavljevich Eli , Tuohy Joanne 
Published: August 2023

Abstract
Osteosarcoma (OS), the most commonly occurring primary bone tumor in dogs and humans, has deficient treatment options and dismal survival outcomes. Spontaneously occurring OS in dogs shares numerous similarities with humans and has greater prevalence, allowing the dog to serve as an informative comparative oncology model. The objective of this study was to investigate the non-thermal focused ultrasound technique histotripsy as an innovative treatment option for OS. Histotripsy has the potential to both ablate the primary tumor and stimulate an antitumor immune response to mitigate metastatic disease, the leading cause of death in OS. To investigate the immunological outcomes associated with histotripsy and OS, we delivered histotripsy to client-owned canine OS patients (n=29) and utilized an in-vitro model for cross species comparisons between human, dog, and mouse. Histotripsy ablation led to a significantly greater expression of activation markers CD80 and CD62L on circulating monocytes, at 1- and 5-days post histotripsy (DPH), respectively. We observed a significantly greater production of TGFβ in circulating monocytes at 3 DPH compared to baseline. On an independent patient basis, we observed a greater population of CD5+ CD4+ and CD5+ CD8+ cells within the treated region of the tumor compared to untreated region and changes in genetic signature also indicated immune activation. Across species in-vitro we observed macrophage activation (CD80, CD62L, and CD86), and upregulation of genes associated with immune cell chemotaxis and pro-inflammatory response. Our results indicate that histotripsy ablation leads to immune activation and has strong potential to target both primary OS and metastatic disease.
NIH (1R21 EB030182-01), Focused Ultrasound Foundation (PLL7GBVK and FUSF-RAP-823R1), and American Kennel Club (Canine Health Foundation No. 02773)
Development of a ferret model of maternal antibody passive transfer to suckling neonates
Authors: Baker Pari , Beverly Christopher , Byrne Jake , Sun Francis , Langel Stephanie 
Published: August 2023

Abstract
Neonates and infants are at increased risk of severe disease from influenza. While placentally-transferred IgG reduces the risk of influenza disease in neonates, maternal antibodies (matAbs) can also impact the ability of infants to mount an immune response post immunization, referred to as matAb interference. It is not fully understood how matAb interference impacts protection against influenza once matAbs have waned and if these effects are influenza lineage-dependent. Understanding preexisting influenza immunity may allow for the development of strategies to overcome preexisting antibody interference in neonatal and adult vaccination. To study this, we developed a ferret model of maternal anti-influenza antibody passive transfer to suckling kits. Pregnant and lactating ferrets were orally and intranasally immunized 2 weeks prepartum and 2 weeks postpartum with an H1N1 hemagglutinin (HA) adenovirus type 5 (Ad5) vaccine. Serum and breast milk from dams and serum from kits were assessed via ELISA. We performed a regression analysis and identified a positive relationship between peak levels of HA-specific IgG antibodies in dam milk and HA-specific IgG antibodies in kit serum at 5–6 weeks of age (P = 0.0001, R2 = 0.47). Additionally, peak concentrations of HA-specific IgG antibodies in dam milk correlated with the levels of HA-specific IgG antibodies in dam serum at the same time point (P = 0.0012, R2 = 0.89). Our data suggest matAbs traffic from dam blood to milk, are ingested by kits and translocate through the neonatal intestine into serum. We will use this model to study the role of influenza lineage-specific matAb passive transfer to offspring and their role in de novo antibody responses in suckling kits.
Supported by the Bill & Melinda Gates Foundation Grant (INV-22595D).
Regulation of fetal tolerance by KIR+ regulatory CD8+ T cells in human pregnancy
Authors: Li Jing , Li Lin , Prunicki Mary , Narasimhan Purnima , Mallajosyula Venkata , Kirosingh Adam , Jagannathan Prasanna , Winn Virginia , Nadeau Kari Christine , Gaw Stephanie , Davis Mark M 
Published: August 2023

Abstract
Immune responses during pregnancy need to be precisely regulated to protect the fetus from microbial infections and to maintain tolerance for the semi-allogeneic fetus. However, the mechanisms regulating fetal tolerance during pregnancy are not well understood. Recently we have identified KIR+CD8+ T cells as a previously unappreciated subset with regulatory functions in humans to suppress harmful self-reactivity. Therefore, we asked whether KIR+CD8+ T cells also play a role in inducing immune tolerance during pregnancy. We first observed an increased frequency of KIR+CD8+ T cells, but not CD4+ regulatory T cells (Tregs), in the peripheral blood of pregnant women at second trimester compared to age-matched nonpregnant females. Interestingly, those with a male fetus had an even higher level of KIR+CD8+ T cells than the ones with a female fetus. In vitro, we found that KIR+CD8+ T cells can suppress CD8+ T cells specific to H-Y antigens (encoded by the Y chromosome) only in mothers with a male fetus. Therefore, the higher induction of KIR+CD8+ T cells in mothers carrying a male fetus may suppress the additional allogenic immune responses triggered by the Y chromosome from the male fetus. Moreover, KIR+CD8+ T cells expand, lose memory features and gain cytotoxic capacity along gestation and their levels are maintained postpartum. In addition, we found the numbers of these suppressive CD8+ T cells correlated with pregnancy disorders, such as spontaneous abortion or pre-eclampsia. Taken together, our findings suggest an important role of KIR+CD8+ T cells in the maintenance of fetal tolerance by suppressing alloreactive fetus-specific T cells, therefore they may be useful as predictive biomarkers or drug targets for human pregnancy disorders.
Supported by Howard Hughes Medical Institute.
ILC2-dependent induction of small intestinal memory and systemic mucosal resilience
Authors: Cortez Victor S , Locksley Richard M 
Published: August 2023

Abstract
The IL-25-dependent ILC2-tuft cell circuit drives altered intestinal physiology in response to chronic helminth and protist infections. We optimized an induction regimen with recombinant IL-25 that resulted in long-lasting alterations of the small intestine, which we termed small intestinal memory. This state was characterized by expanded ILC2s and epithelial tuft cells, small intestinal lengthening, and enhanced resistance to helminth infection. ILC2s were necessary for the establishment of small intestinal memory, as this state was induced in Rag-deficient, but not Rag x gc-deficient mice or mice with deletion of IL-17RB on ILCs. ILC2s also maintained small intestinal memory, as this state was rapidly lost upon ILC2 ablation. Transcriptomic analysis revealed dynamic changes in ILC2 identities from IL-25-treated mice, demonstrative of innate memory. Strikingly, although these alterations were induced by IL-25, the ILC2 memory state was sustained in the absence of IL-25, tuft cells, or multiple alarmins like IL-33, TSLP, or IL-18. Finally, we found that small intestinal memory conferred resistance to bacterial and viral infections at distant mucosal sites and notably impacted the metabolic fitness of mice. The induction of enhanced mucosal resilience accompanied by increased metabolic fitness by IL-25 mimics the positive effects of chronic helminth infestation, revealing a mechanistic pathway that might underpin the widespread prevalence of these infections among vertebrates and that could be therapeutically adapted to improve mucosal health.
You are what you vape: Implications of THC chronic self-administration in changes in the gut microbiome of rats
Authors: Rosado-Franco Jose J , Ellison Alysha , Moore Catherine , Weerts Elise , Williams Dionna 
Published: August 2023

Abstract
Vaping of cannabis and its constituents, such as Δ9-tetrahydrocanabinol (THC), has been promoted as a safer alternative to smoked cannabis. Nevertheless, chronic effect of THC vaping in gut microbiome modulation and its effect of health and or disease is still largely unknown. Hence, this project intends to study the effect of chronic THC vapor self-administration on changes in the gut microbiome of Sprague-Dawley rats. Animals were randomly assigned to either vehicle vapor (100% propylene glycol [PG]) or THC vapor groups and were allowed to self-administer PG or THC vapor puffs in operant chambers for one hour on intermittent days for a total of nine months. At the end of this period, a subset of rats was tested for anxiety-like behavior using the open field test. Afterwards, rats were euthanized with isoflurane overdose and rapid decapitation. Feces, metabolic and immune organs, and serum were collected at the time of necropsy. Fecal DNA was extracted, and relative bacterial gut abundance was determined sequencing 16S hypervariable region V4. Rats exposed to THC vape show anxiety-like behavior in the open field test, when compared with the PG exposed group. This change in behavior is also accompanied by an imbalance in their Firmicutes/Bacteroides ratio, having less Firmicutes and more Bacteroides, specifically members of the Muribaculaceae family. Currently, the effect of this dysbiosis on immune markers expression, such as: CCL2, IL-6, TNF-a, IFNa, IFNb, MX1 and TRAIL, are being studied to determine if there is any disruption on immune homeostasis caused by altered microbiome abundance.
R01DA052859-03S1
RORγt+ Tregs are not the sole mediators of tolerance to systemic vaccination following oral antigen pre-exposure
Authors: Potchen Nicole B , Johnson Andrew M F , Hager Kevin W , Graham Jessica B , Lund Jennifer M , Kublin James G 
Published: August 2023

Abstract
Many promising vaccine candidates fail to elicit protective immune responses against critical human pathogens. Exposure to antigens in the gut leads to a reduction in systemic antibody responses upon subsequent vaccination due to immune tolerance, which could contribute to low efficacy of some vaccines clinically. Tolerogenic mechanisms to avoid inappropriate inflammation are partly achieved by the high abundance of regulatory T cells (Tregs) in the gut. RORγt+ FoxP3+ Tregs are a largely peripherally derived subset that depend on stimulation from microbial antigens for development and could control type-2 intestinal immune responses in some disease models. Currently, it is unclear which Treg subsets control systemic vaccine responses following oral antigen pre-exposure. By utilizing a variety of adjuvants in our tolerance model, we have determined that vaccine-specific IgG1 is suppressed following oral antigen exposure, but not IgG2c, regardless of adjuvant strength or mechanism of action. We have used a RORγt+ Treg conditional knock-out (cKO) mouse model to determine that lack of RORγt+ expression within Tregs leads to similar suppression of vaccine-specific IgG1, but not IgG2c, in groups orally exposed to vaccine antigen compared to WT controls. cKO mice exhibit higher frequencies of IL-33R+ and Gata3+ Tregs at baseline in the gut, which suggests other subsets could compensate for the loss of function of RORγt+ Tregs. Analysis of local and systemic sites by flow cytometry also suggest the T follicular-helper compartment could be altered in RORγt+ Treg cKO mice. Together, we hypothesize that RORγt+ Tregs are not solely responsible for controlling tolerance to oral antigens in a model of tolerance to a systemic vaccine.
Supported by grants from NIH (R01AI127100-01, 5TL1TR002318-05)
Investigating persistent factor VIII-specific IgM in the humoral immune response to factor VIII
Authors: York Elizabeth S , Ito Jasmine , Batsuli Glaivy 
Published: August 2023

Abstract
Hemophilia A is an X-linked bleeding disorder resulting in bleeding events due to insufficient levels of factor VIII (FVIII). The most significant complication in the management of hemophilia A is the formation of polyclonal neutralizing IgG (i.e., inhibitors) which impede FVIII procoagulant activity. In a hemophilia A (FVIII KO) murine model injected with multiple doses of FVIII, FVIII-specific IgM were found to persist despite an established IgG response. In attempt to characterize the role of IgM in FVIII immunity, 18 IgMs were purified from immortal FVIII-specific hybridomas and FVIII binding was confirmed via ELISA. IgMs demonstrated weak binding to FVIII in a static ELISA model with mean optical densities (OD) at 405 nm of 0.33 ± 0.43 compared to anti-FVIII IgG with a mean OD of 1.85 ± 0.02. A novel fluid-phase ELISA, developed in our lab, utilizing N-hydroxysuccinimide (NHS)-activated magnetic beads indirectly conjugated to FVIII increased IgM-FVIII binding with mean OD of 1.15 ± 0.60. IgM binding also demonstrated porcine FVIII cross-reactivity. To evaluate the effect of IgM on antibody titers in vivo, FVIII KO mice received 5 weekly injections of either FVIII or 4 separate immune complexes (IC) of IgM-FVIII. Mice injected with IgM-FVIII IC showed slightly higher median IgG ELISA titers than FVIII alone. These findings suggest a potential role for persistent anti-FVIII IgM in propagating FVIII inhibitor development. Further studies determining the effect of IgM-FVIII IC on complement activation, as well as anti-FVIII IgM epitope specificity and clonality utilizing computational methods of RNA-sequencing, may provide additional mechanistic insight into the role of persistent IgM in the humoral immune response to FVIII.
Hemostasis and Thrombosis Research Society- Student Research Award (EY), NIH K99HL150595 (GB), and Hemophilia of Georgia Clinical Scientist Development Grant (GB).
Differential sensitivity to replication stress of human NK cells
Authors: Guilz Nicole , Ahn Yong-Oon , Mace Emily 
Published: August 2023

Abstract
Natural killer cells are critical components of the innate immune system that kill virally infected or malignant cells. Though rare, natural killer cell deficiency (NKD) occurs when NK cell development and function are impaired and individuals with NKD are susceptible to severe and recurrent viral infections. Several gene deficiencies have been described to result in NKD, including variants in MCM4, GINS1, MCM10 and GINS4, which are components of the CMG helicase. The helicase unwinds DNA during replication and is found in any actively proliferating cell. NK cells are more strongly impacted by mutational deficiencies in helicase proteins than other lymphocytes, though the mechanisms underlying this are not completely understood. NK cells from individuals with NKD as a result of helicase deficiency have increased DNA damage, cell cycle arrest, and replication stress. Here, we induced replication stress during NK or T cell specific activation by chemical methods, using aphidicolin, and genetic methods like siRNA targeting helicase proteins. We found that the CD56bright subset of NK cells accumulates more DNA damage and replication stress during activation than CD56dim NK cells or activated T cells. The drug aphidicolin increases apoptosis of CD56bright NK cells through caspase activation while simultaneously decreasing intracellular perforin expression, a protein important for killing of target cells. This effect is modeled by siRNA mediated knockdown of CMG proteins, thus linking decreased protein expression to replication stress and impaired NK cell function. These findings lend important insight into the mechanisms by which helicase deficiencies selectively affect human NK cell function and cause NK cell deficiency.
NIH R01A137275
CD8+Helios+ T cells: A unique functional T cell subpopulation?
Authors: Jones Madalyn , Shevach Ethan 
Published: August 2023

Abstract
The zinc finger protein, Helios, is a member of the Ikaros transcription factor family and is expressed in CD4+T cells, CD8+T cells and in some NK cells in both mice and humans. While it has been found to have a role in the homeostasis and suppressive function of CD4+Foxp3+T regulatory cells (Treg), its function in CD8+ T cells and NK cells is not well characterized. We have demonstrated that ~10–40% of CD8+ T cells from normal donors express Helios. Expression of Helios in CD8+ appears to be downregulated with TCR stimulation alone but is maintained in the presence of excess IL-2 signaling. Thus far, extensive flow cytometry studies have failed to show a correlation between Helios expression and any other surface or intracellular markers. Mouse CD8+Helios+Ly-49+ T cells have been reported to inhibit the activation of B cells in germinal centers and have been claimed to be the CD8+ counterpart to CD4+Treg. While these studies have been performed in mice, there is very little data to suggest that human CD8+Helios+ T cells exhibit T suppressor function. In humans, the killer-cell immunoglobulin-like receptors (KIRs), a family of transmembrane glycoproteins, are said to be the functional equivalent of the Ly49 proteins. KIR+CD8+T cells have previously been said to highly express Helios and have a function in autoimmune conditions. Data we have obtained from a CITE-seq analysis has shown that Heliosexpression in CD8s is highly correlated with many NK-cell associated markers. This, along with data showing that these cells produce cytotoxic effector molecules, leads us to speculate that these cells have a role in immune suppression via cytotoxic killing of self-activated immune cells. This work was supported by the Intramural Research Program of NIAID, NIH.
GIMAP5 deficiency reveals a human cellular longevity assurance pathway
Authors: Park Ann Y , Gabrielski Justin Q , Lenardo Michael J 
Published: August 2023

Abstract
Studies in model organisms have revealed that lifespan and senescence are genetically controlled, but how cellular longevity assurance pathways are regulated is unknown. Here, we report new insights into cell longevity and senescence through the investigation of a novel disease caused by mutations in GTPase of the immunity-associated protein 5 (GIMAP5). We identified 17 affected individuals in 9 kindreds with lymphopenia, liver disease, splenomegaly, thrombocytopenia, lymphadenopathy, lung infections, and lymphoma. GIMAP5 is highly expressed in T and NK lymphocytes as well as endothelial cells (EC) and is famous as the gene mutated in the Biobreeding diabetic rat and Sphinx lymphopenic mice. Given the strong disease association, the mechanism of GIMAP5 is an important biological and medical question. We found that loss of GIMAP5 leads to the uncontrolled accumulation of long chain ceramides, which is associated with premature senescence and death of lymphocytes, resulting in severe and recurrent infections in patients. We further showed that Gimap5 deficiency in mice increases senescence markers in T cells and ECs of the lung and liver, suggesting that GIMAP5 is involved in the senescence mechanisms affecting multiple tissues. Therefore, our results illustrate the role of the GIMAP5 regulatory pathway in cell longevity and senescence and reveal new molecular targets for increasing the healthy functional lifespan of human cells. This research was supported by the Intramural Research Program of NIAID, NIH.
Thioacetamide-induced liver fibrosis is reduced in humanized immune system mice
Authors: Chen Wen-Yu 
Published: August 2023

Abstract
Fibrosis is defined as a consequence of the abnormal wound-healing response lading to excessive deposition of extracellular matrix components. Chronic liver diseases accompanied with liver fibrosis have caused increased morbidity and mortality worldwide. Carbon tetrachloride (CCl4) and thioacetamide (TAA) are the well-known toxins used to induce liver fibrosis in murine. Compared to CCl4, TAA displays a longer latency between exposures to the drug and less safety concerns with operators. In addition, the translation of murine experimental data to human events often fails due to differences in the immune system of both species, indicating that humanized mice may be a better choice to model human diseases. The effect of TAA in humanized mice is still unknown. In our study, liver fibrosis in wild type mice was induced by escalating doses of TAA. Results showed that liver fibrosis evaluated by Sirius red staining was significantly induced (Metavir score, F2). In addition, serum levels of AST, ALT and ALP were significantly increased after TAA injection. Humanized mice were generated by injection of human hematopoietic stem cells (hHSC) into NOD.Cg-PrkdcscidIl2rgtm1Wjl/YckNarl (Advanced Severe ImmunoDeficiency (ASID)) mice, termed as hHSC-ASID. The escalating doses of TAA were introduced into hHSC-ASID and ASID mice. Although the serum levels of AST, ALT and ALP were increased in both hHSC-ASID and ASID mice following TAA injection, the degree of liver fibrosis was reduced in hHSC-ASID mice than in ASID mice (Metavir score, F1 vs. F2). These results indicated that human immune cells reduced TAA-induced liver fibrosis. It is worthwhile to assess which immune cell(s) participates in this process.