bluuebunny/crossref_metadata_2025_split huggingface dataset:
Development of therapeutics to maintain vaccine immunity following exposure to ionizing radiation.
Authors: McFarland Hugh I , Surujdin Ryan T , Aryankalayil Joseph , Berkson Julia D. , Lee Jay P. , Rosenberg Amy S. 
Published: January 2023

Abstract
Exposure to γ radiation from nuclear power plant disasters, terrorist acts, and medical procedures devastate the peripheral immune system, killing both naïve and memory T cells. Although, naïve T cells recover, memory T cells mediating vaccine immunity are permanently lost rendering a victim highly susceptible to infection. There is no treatment to globally preserve vaccine immunity following ionizing radiation exposure. We have recently published that CD8+ memory T cell responses to Listeria can be preserved in the aftermath of radiation exposure by revaccination within 3–4 days after irradiation. A correlation between survival and T cell activation prior to irradiation has been demonstrated in vitro. Comprehensive revaccination in a radiation mass casualty situation would be problematic from many perspectives. We are working to develop a therapeutic regimen to replicate the pro-survival effects of revaccination. Among the signaling pathways stimulated by T cell activation is the Akt “survival pathway” which inhibits apoptosis, and enhances the rate and fidelity of repair of damaged DNA. Agonists or inhibitors targeting the various receptors and signaling molecules that interact within this pathway may result in survival of memory T cells and are potential therapeutic candidates. In addition, revaccination rescue upregulates p21 mRNA. p21 is strongly anti-apoptotic, and enhances DNA strand break repair. In preliminary studies, a number of candidate therapeutics have been identified including nicotine, which virtually eliminates splenic Listeria in immunized and irradiated mice following challenge with wild-type bacteria. Testing is underway with nicotine patches which would provide an ideal method of administration.
Radiation Therapy and Talimogene Laherparepvec (TVEC) Synergize in the Treatment of Melanoma
Authors: Jhawar Sachin R , Goyal Sharad , Thandoni Aditya , Bommareddy Praveen , Hassan Suemair , Schiff Devora , Haffty Bruce G , Kaufman Howard , Zloza Andrew 
Published: January 2023

Abstract
Talimogene laherparepvec (TVEC), an oncolytic virus, is an FDA-approved therapy that increases the overall survival of locally advanced and metastatic melanoma patients. Radiation therapy (RT) in combination with immunotherapies improves response rates (compared to either modality alone). Therefore, we hypothesized that combination RT and TVEC results in a synergistic therapeutic effect on melanoma. We treated 8 human melanoma cell lines with radiation (0, 4, or 8 Gy) and/or T-VEC (0, 0.01, 0.1, or 1 MOI) and determined its therapeutic effects using the AlamarBlue cell viability assay. Treatment of UACC257 melanoma cells with combination RT (8 Gy) and TVEC (1 MOI) resulted in a significant decrease in tumor cell viability (a decrease of 44.9%) compared to no treatment, RT alone (33.2%, P=0.0004), or T-VEC alone (24.2%, P=0.0088), with similar findings in other cell lines. We then injected a B16 mouse melanoma cell line, permissive to T-VEC infection, into B6 mice to assess in vivo synergy. Mice were treated when tumors were ~7mm × 7mm. RT resulted in an early 20% decrease in tumor size within 4 days, followed by an increase thereafter. Intra-tumoral TVEC resulted in no clear benefit until 9 days post-treatment, when it showed a slight decrease in size sustained for ~10 days. Combination treatment, however, led to an early response that was sustained until 18 days after the initial treatment. When tumors were assessed 18 days after initial treatment, a 3-fold or more increase in tumor size was lowest in the combination T-VEC + RT group (17%) compared to the control (100%), RT (60%), or TVEC (50%) groups. These data propose that combination T-VEC + RT may be beneficial for cancer patients, and studies establishing the mechanism of synergy are ongoing.
RNAdjuvant®, a novel, highly-potent RNA-based adjuvant, combines strong immunostimulatory capacities with a favorable safety profile
Authors: Lutz Johannes , Heidenreich Regina , Mehr Keyvan Tadjalli , Noth Janine , Koch Sven D , Döner Fatma , Hong Henoch S , Melber Karl , Dähling Angelika , Roos Tilmann , Kowalczyk Aleksandra , Baumhof Patrick , Scheel Birgit , Voss Söhnke , Kallen Karl-Josef , Fotin-Mleczek Mariola , Gnad-Vogt Ulrike 
Published: January 2023

Abstract
Purified recombinant proteins and peptides, which are currently under development in various anti-cancer vaccination approaches, lack sufficient immunogenicity. Therefore, potent adjuvants are needed to induce strong and persistent anti-tumor immunity. However, currently only few adjuvants are licensed, most of which primarily enhance antibody, but not T cell responses.
Here, we demonstrate that a novel, well defined, and thoroughly characterized RNA-based adjuvant mediates balanced and long-lasting humoral and cellular immune responses. Our adjuvant significantly enhances anti-tumor immunity, and even complete tumor rejection can be achieved as shown for the syngeneic TC-1 tumor model, a murine model of human HPV-induced cervical cancer.
Our adjuvant acts locally, promoting strong but transient up-regulation of anti-viral and pro-inflammatory cytokines, CXCR3-ligands and cytoplasmic RNA sensors at the injection site, avoiding any systemic cytokine release. A phase I first in man clinical trial testing different doses of RNAdjuvant® alone and in combination with reduced doses of the licensed rabies vaccine Rabipur® was successfully conducted in 43 subjects. Healthy volunteers received 2 intramuscular injections of RNAdjuvant® on days 0 and 21, either alone or in combination with 1/20 or 1/10 of the licensed Rabipur® dose. Virus neutralizing antibody titers (VNTs) measured on days 14 and 28 revealed a significant increase in median VNTs in subjects with RNAdjuvant® compared to their respective control group with 1/10 dose Rabipur® alone.
In summary, our data suggest that RNAdjuvant® represents a novel, highly efficacious adjuvant candidate that can enhance cellular and humoral immune responses.
Revealing the mechanism of regulatory T-cell generation in vivo by novel Fluorescent Timer reporter
Authors: Ono Masahiro , Bending David , Martin Paz Prieto 
Published: January 2023

Abstract
Foxp3+ regulatory T-cells (Treg) are considered to mainly differentiate in the thymus and protect against autoimmunity. However, the physiological significance of peripheral Treg differentiation is still unclear, due to a lack of methods to identify newly generated Treg in vivo. In addition, the mechanism of Treg differentiation in the thymus is still obscure, because of difficulties in defining temporal sequences of developmental events in vivo.
Here we develop a ground-breaking technology to reveal the time-dependent mechanisms of Treg generation and maturation in vivo. Using Fluorescent Timer protein that spontaneously and irreversibly changes its emission spectrum, we establish a novel Foxp3Timer reporter, and and visualize the dynamics of Foxp3 transcription and thereby identify de novo Foxp3 expression in vivo. Here we show that antigenic recognition immediately induces de novo Foxp3 expression in activated effector T-cells in vivo. Surprisingly, most of Foxp3-expressors in inflamed tissue are not derived from the pre-existing Treg pool, but are newly generated upon antigen recognition. RNA-seq analysis reveals novel genomic mechanisms for peripheral Treg generation and identifies new markers for antigen-reactive Treg, by which we successfully manipulate Treg differentiation dynamics.
Collectively, we show that Foxp3 constitutes a negative feedback mechanism for activated effector T-cells, which induces the resolution of inflammation. Our novel Timer technology is applicable to many outstanding problems in immunology.
Controlling T cell development and function via innate Toll-like receptor signaling for autoimmune therapy
Authors: Liu Xia , Ye Jian , Wang Yadan , Zhong Xiaoli , Li Lingyun , Opejin Adeleye , Hsueh Eddy C. , Luo Huanle , Wang Tian , Hawiger Daniel , Peng Guangyong 
Published: January 2023

Abstract
Innate regulation through Toll-like receptor (TLR) signaling has been shown to be important for promoting T cell subset development and function. However, limited information is known about whether differential TLR signaling can selectively inhibit Th17 and/or Th1 cells, important for controlling excessive inflammation and autoimmune responses. Here we demonstrate that activation of TLR7 signaling in T cells can inhibit both Th17 cell differentiation from naïve T cells and IL-17 production in established Th17 cells. We further report that down-regulation of STAT3 signaling is responsible for TLR7-mediated inhibition of Th17 cells due to induction of SOCS3 and SOCS5. TLR7-mediated suppression of Th17 cells does not require DC involvement. In addition, we show that TLR7 signaling can suppress Th1 cell development and function due to a mechanism different from Th17 cell suppression. Importantly, our complementary in vivo studies demonstrate that treatment with the TLR7 ligand imiquimod can inhibit both Th1 and Th17 cells, resulting in prevention of and immunotherapeutic reduction in experimental autoimmune encephalomyelitis (EAE). These studies identify a new strategy to manipulate Th17/Th1 cells through TLR7 signaling, with important implications for successful immunotherapy against autoimmune and inflammatory diseases.
Unique interaction dynamics and peptide-MHC class II (pMHC II) transendocytosis lead to antigen-specific T regulatory cell (Treg)-mediated suppression
Authors: Akkaya Billur , Akkaya Munir , Kamenyeva Olena , Kabat Juraj , Dorward David W , Holstein Amanda H , Maz Mitra P , Shevach Ethan M 
Published: January 2023

Abstract
Tregs can employ numerous mechanisms for inhibiting immune responses. Antigen-specific Tregs exhibit enhanced capacity to suppress compared to polyclonal Tregs, but unique suppressor mechanisms used by antigen-specific Tregs have not been elucidated. We first compared the interaction of antigen-specific induced Tregs (iTregs) and antigen-specific T effector cells with dendritic cells (DC) using scanning electron microscopy and intravital two-photon microscopy. Antigen-specific iTregs uniquely and rapidly formed dense clusters around DC suggesting that they may prevent the access of antigen-specific CD4+ T cells to the DC surface. In an adoptive transfer model, this rapid interaction was accompanied by a decreased ability of co-transferred naïve T cells to interact with the DCs as indicated by their failure to decrease their motility. More importantly, antigen-specific iTregs were found to specifically transendocytose cognate peptide-MHCII complexes from the DC surface to a greater extent than T effector cells resulting in diminished levels of antigen on the DC surface. Transmission electron microscopy of DC-Treg co-cultures showed that Tregs engulfed parts of DC processes as membrane invaginations within two hours and then internalized the p-MHCII complexes into endosomal vesicles. Taken together, these results indicate a two-step process by which antigen-specific Tregs inhibit antigen presentation. They rapidly and efficiently adhere to the DC surface and then deplete the pMHC II complexes from the DC resulting in potent suppression of the capacity of the DC to activate antigen-specific T cells.
Mer tyrosine kinase (MerTK) deficiency drives the loss of marginal zone macrophages with subsequent accumulation of apoptotic cells and dysregulation of germinal centers
Authors: Soni Chetna , Schell Stephanie L. , Fasnacht Melinda , Rahman Ziaur S.M. 
Published: January 2023

Abstract
MerTK is a scavenger receptor involved in anti-inflammatory efferocytosis. We previously reported that MerTK deficiency on a mixed B6 and 129 background (B6-129.MerKO) led to dysregulated germinal center (GC) responses and accumulation of apoptotic cells (ACs), primarily in the GCs. Here we report that the dysregulation of GCs in B6-129.MerKO mice strongly correlates with the loss of marginal zone macrophages (MZMs) starting at two months of age. In addition to MerTK, MZMs also express other AC-scavenger receptors like SIGNR1, MARCO and Tim-4, which are also lost from the splenic MZ of B6-129.MerKO mice. We do not observe a loss or translocation of marginal zone B cells (MZBs) or marginal zone metallophillic macrophages (MMMs) from the marginal zone, indicating the exclusive role of MZMs in deregulated GC responses observed in this model. Preliminary results using splenic and bone marrow derived macrophages show reduced expression of a pro-survival heat shock protein Hsp27 and its phosphorylation through p38 and MAPKAPK-2 in the absence of MerTK. We also observe increased cell death in MerTK deficient macrophages vs. WT macrophages, when challenged with ACs. Our data indicate that reduced Hsp27 expression due to MerTK deficiency may lead to cell death of MZMs.
We are further investigating if the increased availability of self-antigens through AC accumulation, due to the loss of MZMs can lead to positive selection of self-reactive B cells in the GCs, causing autoimmunity. Our data indicate that MerTK deficiency and ACs enhance antigen presentation by macrophages, DCs and B cells. In accordance, we observe an increase in activated T cells in MerTK deficient mice, which may promote dysregulation of GC responses observed in this model.
Bright blue fluorescent nanoparticles for live cell imaging by fluorescence microscopy
Authors: Richey Kristen H , Zhuang Meng , Fraser Cassandra L. , Pompano Rebecca R. 
Published: January 2023

Abstract
Fluorescence imaging of labeled cells is an important method to monitor long-term cell behaviors. Long-lasting green stains such as CFSE (carboxyfluorescein succinimidyl ester) and its derivatives are routinely utilized for cell migration studies, but currently there are no commercially available long-lasting (days), non-cytotoxic blue cell stains compatible with a standard DAPI filter that is brighter than the autofluorescence of tissue. Here, we fabricated two bright blue emitting boron dye-polymer nanoparticles (BNPs) (e = 32100 M−1cm −1) from difluoroboron dibenzoylmethane conjugated to poly(lactic acid) (BF2dbmPLA) or poly(e-caprolactone) (BF2dbmPCL). Particles were &lt; 100 nm diameter, and their excitation and emission wavelengths (lex = 381 nm; lem = 439 nm) corresponded to the DAPI filter. When incubated with mouse splenocytes, the blue emitting nanoparticles were taken up by the cells and readily detected by fluorescence microscopy (EVOS microscope) or a plate reader. Viability of the stained cells was assessed by calcein/propidium iodide staining. These data showed that BNPs have the capability to stain cells brightly without significantly diminishing viability and therefore show promise as a new tool for live cell tracking in the blue fluorescence channel. We are currently measuring cell fluorescence and viability over time, determining which types of cells take up the particles, and comparing the staining capabilities of the nanoparticles to commercial blue dyes.
CD47 antibody-induced engulfment of human T cell leukemia cells by bone marrowderived macrophages
Authors: APPLEDORN DANIEL , Lovell Gillian , Szybut Clare , Patel Kalpana , Campwala Hinnah , Bevan Nicola , Dale Tim , Trezise Derek 
Published: January 2023

Abstract
CD47 is a trans-membrane “don’t-eat-me” signaling protein that enables tumor cells to evade clearance by neighboring phagocytes. Blocking CD47 allows phagocytes to clear tumor cells and is a promising new approach for cancer immunotherapy. In this study, we characterized anti-CD47 antibody-mediated engulfment of living tumor cells (CCRF-CEM) by mouse bone-marrow derived macrophages (BMDMs) or immortalized mouse macrophages (J774A.1). Phagocytosis was quantified using a pH-sensitive cell-labeling fluorescent probe, pHrodo, and automated kinetic live-cell analysis (IncuCyte). CCRF-CEM cells were labeled, washed and then treated with antibody. Target cells were then added to BMDMs or J774A.1 cells in 96-well plates. Phase- and fluorescence images were captured and quantified over time. Anti-CD47 antibody, but not IgG-control, produced time- and concentration-dependent engulfment of CCRF-CEMs by BMDMs (30’-4h), as evidenced by a significant increase in intracellular fluorescence. Close inspection of the time-lapse images revealed clear cellular engulfment coincident with the appearance of the fluorescent signal. Similar observations were made with J774A.1 macrophages. Interestingly, the rate and degree of engulfment was effector cell-dependent. The mechanism of engulfment was not via induction of target cell apoptosis since anti-CD47 did not induce PS externalization (Annexin V) or activate caspase 3/7. Our experimental findings substantiate the known pro-phagocytic effects of anti-CD47 antibodies, and provide a model system and method for quantitative functional analysis and mechanistic insight of CD47 modulators as cancer therapeutics.
Analysis of Mixed Microbial Populations and Deep Immunotyping of Peripheral Blood on the ZE5 Cell Analyzer
Authors: Green Angela M , Scott Michelle , Kroeger Kelly 
Published: January 2023

Abstract
With up to 5 lasers and 30 analysis parameters, the ZE5 flow cytometer is equipped for complex multiplex analysis of cells. Excellent fluorescence sensitivity (&lt;100MESF values in FITC, PE and APC channels) enables detection of antigens expressed at low levels. The three scatter options; forward scatter (FSC), side scatter (SSC) and forward scatter small particle detector (FSCspd) provide optimal resolution of particles &gt;0.3um to 50um in size. Furthermore, the option to set a dual trigger using any scatter or fluorescence channels allows for optimal background reduction. For this study, the ZE5 was used to evaluate size beads and mixed microbial populations, and to perform immunophenotyping of multiple cellular subsets in human blood using multi-color panels. Beads sized 0.3um and larger were discriminated from noise when using scatter or fluorescence as a trigger. Analysis of mixed microbial populations clearly identified the unique populations within the mixture. For immunophenotyping experiments, PBMCs from normal healthy donors were stained using a 14 color panel to identify T cell, B cell, NK cells, dendritic cells and monocyte sub populations. The panel data was compensated accurately on the ZE5 and each sub-cellular population was well identified.
Development and testing of a novel multiplex serodiagnostic assay for Zika and other arboviruses
Authors: Venkateswaran Neeraja , Sarwar Jawad , Parameswaran Nishnath , Fecteau Tracy , O’Connor David , Nelson William M , Venkateswaran Kodumudi 
Published: January 2023

Abstract
Human immunoglobulin (IgM) antibody capture enzyme linked immunosorbent assay (MAC-ELISA) is recommended by US Centers for Disease Control (CDC) to detect the Zika Virus (ZIKV) infection in samples collected after few days of the onset of disease. However, the cross-reactivity of IgM and immunoglobulin G (IgG) antibodies to ZIKV with dengue virus (DENV), West Nile virus (WNV), yellow fever virus (YFV) and some other related flaviviruses poses a significant challenge for an accurate detection of ZIKV infection in a singleplex serological assay. To overcome these challenges we developed a novel multiplex serological assay that will simultaneously detect antibody response to ZIKV, other related flaviviruses and Chikungunya virus (CHIKV). Various recombinant arboviral antigens were coupled to optically-coded microspheres from Luminex Corporation. This fluorescent labeled microsphere assay was used to differentiate between recent, past arbovirus infections, or co-infections that may occur in endemic regions. We used plasma samples from non-human primates (NHPs) prior to and after ZIKV infection for longitudinal assessment of IgM and IgG antibody responses for evaluation of this method. 37 blood samples from 5 different animals challenged with either African strain or Asian lineage strain of ZIKV were analyzed for antibody responses to multiple antigens included in this test. We developed a detection algorithm for automation of IgM result analysis to differentiate ZIKV IgM response from other arboviruses. Thus this multiplex arbovirus assay has great promise for testing human samples for detecting ZIKV infection.
Control of alloreactivity using transient expression of a monoclonal antibody directed chimeric antigen receptor in conventional and regulatory T cells
Authors: Pierini Antonio , Iliopoulou Bettina , Peiris Heshan , Perez-Cruz Magdiel , Baker Jeanette , Hsu Katie , Strober William , Zheng Ping Ping , Erkers Tom , Alvarez Maite , Ring Aaron , Velardi Andrea , Negrin Robert S. , Kim Seung , Meyer Everett 
Published: January 2023

Abstract
The genetic modification of T cells offers many promising cellular therapies, but little known about how the transient targeting of T cell or T cell subsets such as CD4+CD25+FoxP3+ regulatory cells (Treg) could be used to control alloreactivity. We used the expression of a chimeric antigen receptor that recognizes labeled therapeutic monoclonal antibodies, to control T cell activation in vivo. By specifying the location of activation of donor T cells during hematopoietic cell, we modulated graft-versus-host disease and anti-leukemia effects. We also controlled the activation of Treg cells which remain immunoregulatory. When recipient Treg are targeted to activate in response to MHC Class I mismatch, we observed significantly prolonged allogeneic islet graft survival in an antigen-specific manner, as secondary skin grafts were specifically protected against rejection. The transient genetic modification of T and Treg can have lasting effects on the immune system and could be used therapeutically to regulate alloreactivity.
Bone marrow-derived mesenchymal stem cells (MSCs) to enhance nerve regeneration and functional recovery in vascular composite allotransplantation (VCA)
Authors: Salgar Shashikumar K , Bingham Jason , Kniery Kevin , Jorstad Nikolas , Horkayne-Szakaly Iren , Hoffer Zachary 
Published: January 2023

Abstract

Introduction
Optimizing nerve regeneration and mitigating denervation atrophy of target muscles are the keys to achieving successful VCA such as limb and face transplants.

Methods
Lewis rat was anesthetized, sciatic or individual sciatic nerve branches (tibial, peroneal and sural) of the right hind-limb were transected and surgically repaired. MSC (5 million; passage ≤6) or vehicle were administered locally and intravenously.

Results
Rat MSCs expanded ex vivo were CD29+, CD90+, CD34−, CD31−, CD45low, MHC Class I+, Class II−, and were pluripotent. Total sensory function recovery at 2 weeks post sciatic or individual nerve repair (SNR or INR) with or without MSC was ~ 1.2 on a scale of Grade 0–3 (0=No function; 3=Normal); by 12 weeks it was 2.6–2.8 (n≥6/group). Peroneal sensory function recovery was as early as 1 week but not tibial or sural. MSC treatment accelerated sensory function onset. At 8 weeks post-INR, the sciatic nerve function index (SFI), a measure of motor function (0=Normal; −100=Nonfunctional) was −34 and −77 in MSC and vehicle groups, respectively (n≥9); with SNR it was −72 and −92 in MSC and vehicle groups, respectively. Long-term motor function (24 weeks) was apparent in MSC treated INR (SFI −63) but not in SNR (SFI −100) model. Gastrocnemius muscle atrophy was lower (P ≤0.05) in INR compared to SNR model. Significant reduction in axonal area (P ≤0.002), g ratio/axonal myelination (P ≤0.05), reduced nerve fiber density, and increased degenerating axons in the distal segment of the transected/repaired tibial nerve was observed compared to the contralateral naive tibial nerve. Histology and molecular studies are ongoing.

Conclusion
MSC therapy appears to promote peripheral nerve regeneration and functional recovery.