bluuebunny/crossref_metadata_2025_split huggingface dataset:
An effective peptide vaccine strategy circumventing heterogeneity of murine myeloid leukemia
Authors: Cho Hyun-Il , Shin A-Ri , Lee Sang-Eun , Choi Haeyoun , Sohn Hyun-Jung , Kim Tai-Gyu 
Published: January 2023

Abstract
Therapeutic cancer vaccines are an attractive approach for treating malignant tumors, and successful tumor eradication depends primarily on controlling tumor immunosuppression status as well as heterogeneity of tumor cells driven by epigenetic alterations. Here we describe an effective peptide vaccination approach capable of circumventing antigenic heterogeneity by targeting multiple shared tumor antigens simultaneously in murine myeloid leukemia. Peptide-loaded dendritic cell (DC) prime and noninfectious peptide booster heterologous immunizations were assessed for the immunogenicity of polo-like kinase-1 (PLK1)-derived peptides, and targeting multiple shared tumor antigens simultaneously with PD-L1 blockade was evaluated for antitumor effects against murine myeloid leukemia. A synthetic PLK1122 (DSDFVFVVL)-based heterologous vaccination generated large numbers of long-lasting antigen-specific CD8 T-cells eliciting therapeutic effects against various established tumors. The therapeutic efficacy of single antigen-targeting PLK1122-based vaccine with sufficient endurance of PD-L1 blockade toward C1498 leukemia relied on the heterogeneous clonal levels of MHC-I and PD-L1 expression. A novel multi-peptide-based vaccination targeting PLK1 and survivin simultaneously along with PD1 blockade led to complete tumor eradication and long-term survival in mice with clonally heterologous C1498 myeloid leukemia. Our findings suggest that PLK1 could be an attractive immunotherapeutic target antigen for cancer immunotherapy, and that similar strategies would be applicable for the optimization of cancer vaccines for the treatment of numerous viral diseases and malignant tumors.
Exploiting metabolism of vaccine-elicited T cells to enhance tumor immunotherapy
Authors: Willett Benjamin A. S. , Klarquist Jared , D’Alessandro Angelo , Kedl Ross M. 
Published: January 2023

Abstract
Given the highly glycolytic nature of most cancers, glycolysis inhibition remains a strong potential treatment strategy. However, effective glycolysis blockade may also inhibit the function and survival of infiltrating T cells. Unlike T cells activated by infections and cancer, T cells activated by subunit vaccination do not rely on aerobic glycolysis, either during their initial proliferative burst or thereafter. Instead, these vaccine-elicited T cells primarily rely on mitochondrial metabolism of fatty acids and OXPHOS. Thus, we hypothesize that vaccine-elicited T cells will maintain anti-cancer therapeutic efficacy even under conditions of glycolytic inhibition. Additionally, vaccine-elicited T cells are characterized by robust expression of the transcription factor Tcf1, which has been shown to identify a stem-like pool of T cells capable of sustaining an anti-cancer response. We show that a therapeutic regimen combining subunit vaccination with glycolysis inhibition significantly impairs tumor growth in mice. These effects are further enhanced by the addition of clinically-relevant αPD-1 inhibitors. Collectively, these data show the potential of controlling tumor progression by exploiting the distinct metabolic and transcriptional program of vaccine-elicited T cells through a novel combination immuno/metabolomic therapeutic regimen.
Development of AAV6-based vaccine with prime-boost capacity by using exosome-protected viral particles
Authors: Aslanidi George , Krotova Karina 
Published: January 2023

Abstract
We recently showed that the vaccine based on optimized AAV6 vector generates high level of antigen (Ag)-specific immune response. However, certain limitations in use of viral vectors as vaccine carriers need to be overcome to ensure its wide use for clinical application. Particularly, the primary injection of AAV lead to generation of neutralizing antibodies (NAB) against the virus capsid, hampering AAV re-administration. To address this limitation, we exploited the ability of AAV to accumulate in exosomes. AAV vectors associated with exosomes (exoAAV) were purified from conditioned media of HEK 293 cells, along with the standard AAV production (stAAV) from the same lysed cells. Here we report several significant observations that highlight the advantages of using exoAAV for development in a prime-boost protocol. First, we confirmed comparable infectivity between stAAV and exoAAV using YFP and luciferase reporter genes. Second, we showed that exoAAV is highly resistant to NAB compared to stAAV. Third, we developed a prime-boost protocol with injections of stAAV followed by exoAAV expressing OVA Ag as a model system. This protocol led to significantly higher levels of OVA-specific CD8+ T cells compared to the single injection method. The therapeutic potential of prime-boost was evaluated in a melanoma B16F10 tumor prevention model in comparison to single injection vaccination. While a single injection was able to significantly delay tumor growth, the prime-boost protocol completely prevented tumor development. In summary, we use exosome-protected AAV to allow for repeated vaccination, and this method produced a superior protective immune response compared to single vaccination.
A novel TLR adjuvant increases the efficacy of vaccines against fentanyl-induced toxicity and overdose
Authors: Miller Shannon M , Robinson Christine , Whitacre Margaret , Buhl Cassandra , Crouse Bethany , Pravetoni Marco , Evans Jay T 
Published: January 2023

Abstract
The widespread illicit use of fentanyl and its analogs is associated with the increased incidence of fatal drug overdoses in the United States in recent years. Pre-clinical studies with vaccines targeting opioids show selectivity and efficacy but clinical data suggests vaccine efficacy against drugs of abuse may be limited by individual variability and a requirement for high and sustained antibody levels. New strategies are needed to enhance vaccine efficacy and increase the quantity and quality of the anti-drug antibody response. We investigated the ability of novel synthetic TLR adjuvants to increase anti-fentanyl antibody titers and protect from fentanyl challenge in mice. Fentanyl-based haptens were conjugated to CRM and combined with alum, novel synthetic TLR adjuvants, or combinations thereof. The most promising TLR adjuvants and combination adjuvants increased anti-fentanyl hapten total IgG, IgG1, and IgG2a antibody concentrations. Splenocyte restimulation with CRM protein showed adjuvant combinations significantly increased secretion of Th1-type cytokines compared to antigen only or single adjuvants alone. Likewise, in a fentanyl challenge model, combination adjuvanted vaccines increased protection from respiratory depression compared to mice vaccinated with antigen alone or single adjuvant controls. These data demonstrate that TLR adjuvants improve the anti-fentanyl adaptive immune response and increase protection against fentanyl challenge in mice, and may enhance vaccine-induced protection against fentanyl overdose in humans.
LAG-3: a potentially important checkpoint of the humoral immune response
Authors: Haun Brien K , To Albert , Wong Teri Ann , Nakano Eileen , Ndhlovu Lishomwa , Lehrer Axel T 
Published: January 2023

Abstract
Emerging infectious viruses such as Zika virus (ZIKV) pose serious threats to human health. Currently, there are no FDA-approved therapeutics or vaccines available for ZIKV. Our laboratory has developed a vaccine candidate using recombinant envelope protein (E) of the ZIKV. Our previous work has shown that neutralizing antibody responses are the protective factor upon viral challenge in BALB/c mice and non-human primates. Since lymphocyte-activation gene 3 (LAG-3) is an immune checkpoint receptor that directly interacts with MHC-II during antigen presentation, we decided to use the mouse model to investigate the effects of administering a LAG-3 blockade prior to immunization. We hypothesized that this pre-treatment may increase the number of CD4+ T helper (Th) cells available to interact with B cells in germinal centers, thus increasing the antibody response. We observed an increase in CD4+ T cells among the treatment group, changes in overall Th phenotypes, and a reduction of phenotypically exhausted CD8+ T cells. This alteration in cell populations was accompanied by an increase in vaccine potency based on ZIKV neutralizing titers. Based on preliminary observations, we believe that pre-treatment with LAG-3 blockade prior to immunization may modulate the magnitude of humoral responses by increasing Th cell activity in germinal centers. Overall, we think that LAG-3 may be an important controller of the quality of adaptive immune responses and work to further investigate these phenomena is ongoing.
The synergy of TLR and STING in cancer immunity
Authors: Taylor David R , Young Kim , Korrer Michael J 
Published: January 2023

Abstract
Adjuvants are essential components of cancer vaccine formulations to promote effective immune responses. We initially screened multiple immune-activating targets, including TLR and STING adjuvants using in-vitro assays to test their ability to activate bone marrow DC (BMDC) and showed STING had the most potent activation (i.e., MHC and co-stimulatory molecules) on DCs. Since STING and TLR signaling is non-redundant, TLR and STING adjuvants were combined to enhance DC activation. We utilized the multidimensional synergy of combinations (MuSyc), a novel synergy algorithm, to assess molecular synergy in combining STING and TLR adjuvants, and we noted that the combination of R848 (TLR7-8) plus STING agonists provided a synergistic efficacious and potent response on BMDCs. From this data-set, we generated a MuSYC synergy strategy where we will utilize max signal dose for STING adjuvant and ten times less of max signal dose for R848 (TLR7-8) to determine synergy for other assays. We tested this same MuSYC strategy for activation of human monocytic cell line THP-1 and determined that the synergy strategy induced similar or better activation effects compared to the max signal dose for both adjuvants. Finally, we tested the in-vivo anti-tumor immune response for the B16 melanoma and MOC2 head/neck tumor-bearing vaccination models that resulted in an improved anti-tumor response for the combinatorial STING-TLR adjuvanted vaccines compared to the single agents.
Porcine messenger RNA and microRNA pathway analysis of Porcine Reproductive and Respiratory Syndrome Virus infection
Authors: Miller Laura C , Fleming Damarius 
Published: January 2023

Abstract

OBJECTIVE
Studies of the host-pathogen interaction between pigs and porcine respiratory and reproductive syndrome virus (PRRSV) have tried to uncover the process that allows the virus to evade the immune response to its benefit. This study examined the effects messenger RNA (mRNA) and microRNA (miRNA) expression has on host biological networks.

METHODS
Analysis was carried using whole blood from both mock infected and PRRSV infected pigs. Transcriptomic techniques were then applied to examine the differential expression of miRNAs using Hisat2 for alignment to the Sus scrofa 10.2 genome and changes in expression analyzed using DeSeq2. Statistical significance for pathway analysis was set at FDR ≤ 0.5. The resulting list of miRNAs were examined for their effects on porcine biological networks using the mirPATH software to generate KEGG pathways affected by changes in miRNA differential expression.

RESULTS
The results of the statistically significant (FDR ≤ 0.15) miRNAs were defined by days post inoculation (dpi) to examine biological networks inhibited or activated by the identified miRNAs. Post-transcriptional control of host gene expression appeared to take place over several host biological pathway groupings which effect homeostasis that included immune, metabolic, and structural pathways. This included pathways such as the proteoglycan in cancer pathways that showed multiple genes targeted for inhibition that would cause host dysregulation and impair the ability to properly maintain homeostasis during infection.

CONCLUSION
The analyses suggests inhibition and activation of networks involved in viral entry, proliferation, and pro-inflammatory signaling may underlie the ability of PRRSV to hinder homeostasis.
Characterization of systemic, mucosal and lymphoid granulocyte subsets in rhesus macaques
Authors: Manickam Cordelia , Jones Rhianna , Ram Daniel , Kroll Kyle , Hueber Brady , Varner Valerie , Reeves R. Keith 
Published: January 2023

Abstract
Granulocytes mediate immune protection by phagocytosis, NETs, degranulation, antibody mediated effector functions and recruitment of innate and adaptive cells against pathogens. However, limited research describing granulocytes in HIV infection and in mucosal tissues exists. In particular, the role of granulocytes in the rhesus macaque (RM) model of HIV infection and vaccine/treatment studies have been understudied. Our goal was to characterize granulocyte subsets in systemic, mucosal and lymphoid tissues of naïve and infected RM. Mononuclear cells from jejunum, colon, cervix, vagina, lymph nodes, spleen, liver, and whole blood of naïve, SIV+, and SHIV+ RM were characterized using flow cytometry and Imagestream. Phenotype of granulocyte subsets were identified as- CD45+CD66+CD49d+ eosinophils, CD45+CD66+CD14+ neutrophils, and CD45+CD123+FcRE+ basophils. Polynuclear and mononuclear visualization with DAPI staining and surface marker images by Imagestream analysis further confirmed granulocytic phenotypes. Flow cytometry data showed that differential expression of FcRgs (CD64, CD32 and CD16) can be helpful in distinguishing granulocyte subsets while FcRE was only expressed by basophils. Interestingly, frequencies of granulocyte populations were generally higher in mucosal compared to lymphoid tissues in both naïve and infected animals. Eosinophils were particularly enriched within jejunal tissue, with a median of 45% of CD45+ cells. High frequencies of granulocytes in mucosal tissues indicate their importance in gut and reproductive tract immunity. Overall, further investigation is warranted to understand the significance of granulocytes in tissue homeostasis and infection.
Oclacitinib induces Foxp3 expression in CD4+ T cells under activation conditions
Authors: Jasiecka-Mikolajczyk Agnieszka , Socha Piotr , Jaroszewski Jerzy Jan , Maslanka Tomasz 
Published: January 2023

Abstract
Oclacitinib (OCL) is a novel immunosuppressive agent approved for dogs that controls inflammation in allergic disease via the inhibition of the JAK/STAT pathway. The present study attempts to shed light on the involvement of Foxp3-expressing regulatory CD4+ T (Treg) cells and inducible type 1 regulatory T (Tr1) cells in the anti-inflammatory and anti-allergic actions of OCL. Our previous study demonstrated that exposure of unstimulated canine lymphocytes to OCL did not affect Foxp3 expression in T cells. This study investigates the effect of OCL on Foxp3 expression by canine CD4+ T cells in the course of in vitro activation. Effect of OCL on generation of Tr1 (i.e. CD223+CD49b+CD4+) cells was studied using murine lymphocytes due to the lack of appropriate antibodies for the induction and detection of these cells in dogs. The exposure of canine lymphocytes to both concentration of OCL, with a concomitant stimulation, led to a significant increase in the percentage of Foxp3-expressing cells within the CD4+ T cell subset. In the light of our previous results, it can be suggested that the effect of OCL on Foxp3 expression in CD4+ T cells may depend on T cell activation status. The mean percentage of CD3/CD28-induced CD223+CD49b+CD4+ T cells was approximately 6- and 2-fold lower in the cultures exposed to OCL 10−6 and 10−7M, respectively, compared to untreated cells. Thus, the results strongly suggest that the generation of inducible Foxp3-expressing CD4+ T cells may constitute one of mechanisms responsible for the production of anti-inflammatory and anti-allergic effects of OCL. These effects are not mediated by generation of Tr1 cells. What is more, the present results indicate that OCL may counteract the induction of these cells.
Code Blue: Nurse Cells Can Resuscitate Lymphocyte Receptors in Nurse Shark Thymus by Secondary Rearrangement
Authors: Mitchell Christian Danielle , Criscitiello Michael F 
Published: January 2023

Abstract
The thymus is the most conserved primary lymphoid organ present, in some form, in mammals to all the way back in lamprey. It is composed of a densely-packed outer cortex and an exiguous inner medulla. The thymus functions to provide a microenvironment conducive to T lymphocyte maturation and receptor repertoire formation. Sharks are situated at a pivotal location in evolutionary immunology. They are the oldest vertebrates to have similar adaptive immune characteristics to mammals. Shark lymphocyte antigen receptors are diversified by V(D)J recombination via RAG and heavy chain class switch recombination catalyzed by AID. Somatic hypermutation occurs in both B and T cell antigen receptors of shark, mediated by AID as well. Shark T cell receptors also employ antibody-like NARTCR and IgHV components. Looking at the oldest lymphoid tissue in the oldest vertebrate ancestor with adaptive characteristics similar to mammals will yield important information on the evolution of the adaptive immune response as it relates to T lymphocytes. Nurse cells are cortical thymic epithelial cells that play an important role in selection of functional T cells during the maturation process. Nurse cells have been shown, in mice, to surround non-functional T cells in order to allow them to reconfigure their receptors and attempt selection another time via RAG and/or AID. Through the use of immunohistochemistry and in situ hybridization, we will investigate whether nurse cells provide a microenvironment for shark thymocyte receptor revision in sharks as in mammals.
Transcriptomic profiling of Strongylocentrotus purpuratus coelomocyte populations
Authors: Fugmann Sebastian , Cheng Jun Fang , Huang Hsiao Han , Buckley Katherine M 
Published: January 2023

Abstract
Coelomocytes are important cellular mediators of immunity in echinoderms. In the purple sea urchin (Strongylocentrotus purpuratus) these cells can readily be divided into phagocytes, vibratile cells, red spherule cells, and colorless spherule cells based on their morphology. While some information has been reported about genes that are expressed in specific types of coelomocytes, a comprehensive analysis of the gene expression pattern in each cell type was not available. Using a density gradient coelomocytes from individual adult urchins were separated into three populations (phagocytes, a mixture of vibratile and colorless spherule cells, and red spherule cells), RNA was isolated, and next generation sequencing was employed to determine the transcriptome of each cell population. All three cell populations showed clear differences in their gene expression patterns, and the differences in transcript levels for selected genes that were found to be uniquely expressed in one cell type were confirmed by qRT-PCR using cDNA obtained from additional sea urchins. We now complemented this bulk population data with single-cell RNAseq to define coelomocyte identity and function based on their gene expression profiles. The new findings from this study will be discussed.
This work was supported by the Chang Gung Memorial Hospital (CMRPD1C0242, CMRPD1F0421, and CORPD1F0051) and the Ministry of Science and Technology Taiwan (MOST 107-2320-B-182-002 and 108-2320-B-182 -018 -MY3)
Development of a multiplex assay for the bovine inflammatory cytokines IL-1β, IL-6, and TNF-α
Authors: Chitko-McKown Carol G , Bierman Stacy L. 
Published: January 2023

Abstract
Commercially available bovine-specific assays are limited in number, and multiplex assays for this species are rare. Our objective was to develop a multiplex assay for the bovine inflammatory cytokines IL-1β, IL-6, and TNF-α using the Meso Scale Discovery U-PLEX platform.
"Do-It-Yourself" ELISA kits that contained polyclonal antibodies, both unlabeled and biotinylated, and the specific recombinant bovine cytokine, were purchased for each of these three cytokines. The biotinylated antibodies that would normally be used as reporters in a traditional ELISA were coupled to linkers that bind to specific locations within each well of the U-PLEX plate. Unique linkers were used for each of the cytokines. The unlabeled antibodies were conjugated with electrochemiluminescent labels to serve as detection antibodies. Each cytokine assay was optimized individually prior to performing an optimization on the multiplex assay containing reagents for all three cytokines. To calculate cytokine concentrations, standard curves were developed using the recombinant cytokines and were run concurrently on each plate.
Standard curves for IL-1β and TNF-α were run at concentrations ranging from 0 – 50,000pg/ml, and for IL-6 from 0 – 10,000pg/ml. The minimum average concentrations measured by the standard curves were 5.3pg/ml, 0.92pg/ml, and 22.34pg/ml for IL-1β, IL-6, and TNF-α respectively as determined by data from seven plates. Only 37 out of 280 unique samples assayed had IL-β or TNF-α concentrations above the upper limits of these standard curves.
In our hands the U-PLEX platform was a viable means to develop analyte- and species-specific multiplex assays using privately developed or purchased sets of commercially available reagents.
Mitigation of Equine Recurrent Uveitis through topical Suppressor of Cytokine signaling-1 (SOCS1) mimetic peptide: Open Label Safety and Efficacy Pilot study
Authors: Larkin Joseph , Polk Timothy , Sharma Jatin , Bae Sarah , Barr Olivia , Jones Amari , Kitchen Holly , Wihelmy Michelle , Devine Katherine , Plummer Caryn 
Published: January 2023

Abstract
Recurrent Uveitis (RU), is a relapsing/remitting disease that accounts for more than 10% of severe visual handicaps in US patients per year, despite treatments that often include steroids. Even when effective, prolonged steroid use is accompanied by significant negative side effects. As such, there is a critical need for the development of novel therapeutic strategies to treat RU. Notably, equine recurrent uveitis (ERU) is the only spontaneous model of human recurrent uveitis, and is also a leading cause of equine blindness. RU is driven by aberrant inflammation from the immune system. Notably, the body naturally generates a family of intracellular proteins, known as suppressors of cytokine signaling (SOCS), which prevent excessive inflammatory responses. We have previously shown that topical application of an eye drop, containing a cell-penetrating peptide that mimics SOCS1 (SOCS1-KIR), could mitigate experimental uveitis in mice. In order to evaluate the ability to translate our preclinical findings we conducted a first in equine, open label study in which we evaluated the safety and efficacy of topical SOCS1 peptide administration in ERU patients. We found that topical SOCS1-KIR administration was well tolerated in equine patients. Moreover, topical administration of SOCS1-KIR peptide twice a day was associated with reductions in ocular discomfort, hyperemia, and aqueous flare. Additionally, biochemical analyses of SOCS1-KIR revealed an enrichment strategy that could potentially be utilized to optimize ocular peptide dosing. Together, these results prove justification for conducting larger additional treatment studies in horses, and exploring SOCS1-KIR use in humans.