bluuebunny/crossref_metadata_2025_split huggingface dataset:
Clinical and Immunologic Biomarkers for Regression of High Grade Cervical Dysplasia and Clearance of HPV16 and HPV18 Infection after Immunotherapy with VGX-3100 in a Phase IIb Clinical Trial
Authors: Morrow Matthew P , Kraynyak Kimberly A , Sylvester Albert J , Dallas Michael , Knoblock Dawson , Boyer Jean D , Yan Jian , Vang Russell , Khan Amir , Humeau Laurent , Sardesai Niranjan , Kim J Joseph , Plotkin Stanley , Weiner David , Trimble Cornelia Liu , Bagarazzi Mark 
Published: January 2023

Abstract
We have previously demonstrated proof-of-principle of VGX-3100, an immunotherapy which targets HPV16/18 E6 and E7, in a Phase IIb study in women with premalignant cervical lesions caused by HPV16 or HPV18. After completion of the treatment regimen, cervical lesions underwent complete regression (CR) concomitant with elimination of underlying HPV16/18 infection in 40.0% of VGX-3100-treated patients and in only 14.3% of patients receiving placebo (p=0.002). Here we detail characteristics present that either correlated with or predicted therapeutic benefit with VGX-3100. Combining HPV typing and Pap smear data collected following the third dose (Week 14) was predictive for both CR and the elimination of HPV16/18 at the efficacy assessment twenty two weeks later (predictive value 94%, sensitivity 96%). Patients who had a significant elevation of peripheral blood CD137+ Perforin+ CD8+ T cells specific for the HPV type causing disease (p=0.001) were more likely to have lesion regression and HPV elimination than women without a detectable peripheral blood response. In comparisons of the intensity of cervical mucosal CD103+ and Perforin+ cell infiltrates pre- and post-vaccination, significant increases were observed in patients displaying exhibiting CR with elimination of HPV infection (p=0.027 and p=0.043, respectively). In sum, we found that quantitative measures associated with an effector response to VGX-3100 antigens were associated with subsequent lesion regression. These analyses indicate that certain immunologic responses are associated with successful resolution of HPV 16/18-induced pre-malignancy, with particular emphasis on the upregulation of Perforin in the immunotherapy induced immune response.
Controlled release of mTOR inhibitors to enhance CD8+ T cell memory
Authors: Zeng Qin , Jewell Christopher M 
Published: January 2023

Abstract
Recent studies demonstrate, they frequent, low-dose administration of rapamycin (Rapa) during vaccination can help maintain the plasticity of antigen-specific CD8+ T cells, polarizing the phenotypes of these cells toward central memory T cells (TCM). However, these effects required multiple, systemic doses of Rapa. Here we hypothesized that local, intra-LN controlled release of Rapa from these depots during vaccination would polarize T cells toward highly proliferative TCM during T cell expansion. to enhance T cell immunity. To test this idea, vaccine were formulated by mixing free ovalbumin (OVA) and PLGA microparticles loaded with a TLR9 agonist (CpG). Mice were first immunized with the vaccine, and then received daily i.p injection of Rapa at low doses (75 μg/kg) for 28 days. This regimen confirmed i.p injection of Rapa increased the percentage of OVA-specific CD8+ T cells in PBMCs by 2.4 times on day 14 and specific TCM levels by 4.3 times on day 49 compared to vaccine without Rapa. We next assess the effects of controlled release by using the vaccine above but treating mice with a single dose of controlled release depots loaded with defined doses of Rapa. In mice, this approach showed functional as similar as daily i.p injection of Rapa. While high dose of Rapa (20.9 μg to 36.4 μg per dose) suppressed the immunity, Rapa at a low dose (0.07 μg to 4.17 μg per dose) in MP significantly promote the CD8+ T cell immunity over 7 weeks, where specific CD8+ T cells was promoted by 2.9 times on day 14 and specific TCM on day 49 by 2.4 times of vaccine without Rapa. These result suggested that sustained release of immunomodulator in biomaterials depots directly in the LN microenvironment can significantly enhance the antigen specific CD8+ T cells immunity.
The QuickSwitch Quant HLA-A*02:01 Tetramer Kit can be used for determining the biological activity of a cancer vaccine.
Authors: Delcommenne Marc C , Hrytsenko Olga , Tram Cynthia , Weir Genevieve , Stanford Marianne M. 
Published: January 2023

Abstract
We have devised a fast and user-friendly assay (QuickSwitch™ Quant) that can both help determine binding of novel peptides to MHC class I molecules and generate new specificity MHC class I tetramers for peptide specific T cell detection. This study aimed to determine whether the QuickSwitch™ Quant HLA-A*02:01 Tetramer Kit can also be used for evaluating the biological activity of a vaccine. The tested vaccine was DPX-Survivac, an ovarian cancer vaccine candidate which consists of several survivin peptide antigens that are each restricted to a different human class I allele. We sought to evaluate the specificity and sensitivity of the QuickSwitch™ Quant HLA-A* 02:01 Tetramer Kit-PE for assessing the biological activity of SurA2.M, an HLA-A2-restricted peptide, in DPX-Survivac. The complete vaccine also contains non-HLA-A2 restricted peptides, lipids and a polynucleotide adjuvant. We tested the detection of the peptide prepared individually in a buffered solution or in the DPX-Survivac vaccine prepared in an aqueous formulation. Results indicate that peptide exchange rate of SurA2.M is similar whether it is dissolved individually in a buffered solution or mixed with other components of the vaccine. Results also may be dependent on the affinity of the peptide for HLA-A2. Thus, by optimizing a concentration curve using individual peptides, the QuickSwitch™ Quant HLA-A*02:01 Tetramer Kit can be used to quantify the concentration of HLA-A2 restricted peptides in simple solutions or more complex formulations.
Genetic linkage analysis of cancer vaccine response in Diversity Outbred mice
Authors: Gibson Heather , Reyes Joyce , Jones Richard , Dyson Greg , Wei Wei-Zen 
Published: January 2023

Abstract
Recent advances in cancer immunotherapy demonstrate the power of anti-tumor immunity, however only a fraction of patients maintain durable responses. To test the hypothesis that genetic variation determines cancer immunotherapy response, we conducted genetic linkage analysis of HER2 vaccine response in Diversity Outbred (DO) mice. Our HER2 DNA vaccine has shown varying anti-tumor activity in mice, cats and human patients. Differential vaccine response was found in HER2 transgenic (Tg) mice of BALB/c versus C57BL/6 backgrounds. DO mice, produced by non-sibling crossing of 8 mouse stains (A/J, C57BL/6, 129S1/Svlm, NOD, NZO, CAST/EiJ, PWK/PhJ and WSB/EiJ) incorporate &gt;90% of all genetic variation in mice. The fully sequenced founder strain genomes enable genetic linkage analysis using Mouse Universal Genotyping Array (GigaMUGA), which consists of &gt;150,000 genetic markers, mostly single nucleotide variants. DO mice were bred with C57BL/6 HER2 Tg mice, generating HER2 DO mice that express human HER2 as a self-antigen. A wide range of IgG (1.8 – 50 μg/mL) and IFNg producing T cells (0 – 348/106 splenocytes) were detected after 2× vaccination with an adenoviral vaccine encoding human HER2. Genetic loci associated with positive response (p&lt;10−5) were sought by linear regression analysis. Immune-related genes within identified loci were examined. Preliminary analysis links IgG response to NFkB, IL6, and IFNg-related gene clusters, and IFNg producing T cell response to NFkB, IL6, IL10, Akt and p38 MAPK-related gene clusters. Further analysis will define and rank genetic variants of candidate immune regulators to help identify vaccine responsive individuals.
Foxp3+ regulatory T cell expression of IL-10 is required for IL-33-mediated expansion of regulatory B cells.
Authors: Roessing Anna , Cherukuri Aravind , Rothstein David , Turnquist Heth Roderick 
Published: January 2023

Abstract

BACKGROUND
FoxP3+ regulatory T cells (Tregs) are crucial to self- and antigen-specific tolerance. IL-33 drives expansion of Tregs expressing the IL-33 receptor, ST2, and secreting high levels of IL-10. Regulatory B cells (Bregs) are recently identified negative regulators of the immune system and depend on IL-10 to suppress effector T cell responses. In new data, we find that IL-33 also expands Bregs, however, the relationship between ST2+ Treg and Bregs cells remains unknown. Herein we elucidated if a directional relationship exists between IL-33-expanded Bregs and Tregs.

METHODS
B6 eGFP-FoxP3-Diptheria Toxin (DT) receptor mice were used to establish whether IL-33-expanded Treg were needed for IL-33-aided increases of TIM-1+ IL-10+ Bregs. B6 Foxp3-Cre X IL-10fl/fl mice were used to determine the need for IL-10 expression by Tregs for IL-33-mediated expansion of Bregs.

RESULTS
Assessment of splenocytes for Tregs and Bregs by flow cytometry showed that IL-33 administration led to a 3–5-fold increase in ST2+ Tregs. IL-33 also led to a 3-fold increase in the frequency of IL-10+ Breg, most of which expressed ST2. A lack of Tregs hindered the ability of IL-33 to increase Bregs. While Treg expression IL-10 was not required for ST2+ Treg expansion by IL-33, IL-10 expression by Tregs is crucial for Breg expansion by IL-33.

CONCLUSIONS
We have uncovered a relationship between Tregs and Bregs in response to IL-33. Specifically, while we show that Bregs express ST2, our data suggest that IL-33 expansion of Bregs is indirectly mediated by Treg IL-10 production.
A new model to investigate cell death-independent mechanisms of direct Treg suppression of B cells
Authors: Golding Amit , Weingartner Elizabeth 
Published: January 2023

Abstract
Tregs are well-known for their role in maintaining peripheral self-tolerance and immune homeostasis. One such role is regulation of the humoral response, which is most clearly demonstrated by abundant auto-antibody production in Scurfin mice and IPEX patients, both of which lack functional Tregs. Indirect Treg regulation of the humoral response via their influence on helper T cells, in particular in germinal centers, is well established. Prior demonstration of direct Treg inhibition of B cells has also been demonstrated, but is mostly attributable to Treg killing of B cells. We have re-addressed the role of Tregs in directly suppressing Ig secretion specifically, and we have begun to elucidate the mechanism behind this suppression. Because it is likely that regulation is dependent on the developmental stage of the B cell, we used a homogeneous, immature, antibody secreting cell line, Ramos B cells. We demonstrate that human Tregs can directly suppress basal and stimulated IgM secretion by Ramos through a death- and contact-independent mechanism. This suppression is enacted by a soluble, anti-inflammatory factor and does not appear to alter the rate of IgM transcription but instead specifically affects the amount of secreted IgM mRNA by altering RNA processing. The finding that Tregs suppress IgM secretion by immature B cells via a soluble factor is novel. We plan to further characterize this suppression and determine if it is altered in disease states that present with high immunoglobulin titers.
Restoration of Immune Tolerance in Type 1 Diabetes by Modulating Interleukin-2 Receptor Signaling
Authors: Yuan Xiaomei , Dong Yi , Tso J. Yun , Fu Wenxian 
Published: January 2023

Abstract
Type 1 diabetes (T1D) is an organ-specific autoimmune disease, characterized by the destruction of insulin-producing beta cells in pancreatic islets. A variety of lymphocytes, including T cells and natural killer (NK) cells infiltrate into the islets. Their roles in T1D are rather different--effector T cells and NK cells accelerate, while regulatory T cells, dampen the development of T1D. Signaling through interleukin-2 (IL-2) receptor beta (IL-2Rβ, also known as CD122) is critical for the differentiation and function of both T cells and NK cells. However, it remains poorly understood how these cells respond to IL-2Rβ signaling during the development of T1D. Here we show that in non-obese diabetic (NOD) mice, the most commonly used animal model for human T1D, administration of a non-immunogenic monoclonal antibody (mAb) selectively ablates pathogenic cells, including memory CD4+ T cells, CD8+ T cells and NK cells. In contrast, CD4+ Foxp3+ regulatory T (Treg) cells are only mildly affected. This leads to significantly elevated ratios between Treg cells and pathogenic cells. This reset balance preferentially occurs in pancreatic islets, without systemic immunosuppression. Importantly, IL-2Rβ blockade suppresses both insulitis and diabetes in NOD mice. Moreover, IL-2Rβ blockade significantly suppresses interferon gamma (IFN-γ) production in T cells and NK cells, but promotes interleukin-17 (IL-17) production in CD4+ T cells. Therefore, the modulation of IL-2Rβ signaling represents a novel and efficient approach to restore immune tolerance in T1D.
Modulation of T cell function by CD271
Authors: Lebert-Ghali Charles-Etienne , Bonkoungou Carole , Fournier Marilaine , Sood Aditi , Melichar Heather J 
Published: January 2023

Abstract
Co-signaling molecules are essential modulators of the immune system. These molecules belong to the immunoglobulin and tumor necrosis factor receptor superfamilies (IgSF and TNFRSF, respectively). Despite intensive study of these protein families, the functions and binding partners for many IgSF and TNFRSF members remain largely unknown. In particular, CD271 (NGFR and TNFRSF16) is expressed on neural crest-derived cells, several immunomodulatory cell types, as well as on diverse cancers cells, and our data attribute new immunoregulatory properties to CD271 that may play a role in both cancer evasion and autoimmune disease. Indeed, in vitro stimulation of mouse and human T cells in the presence of recombinant CD271-Fc protein inhibits T cell activation, proliferation and cytokine production. Additionally, we co-cultured antigen-specific mouse T cells with artificial antigen presenting cells overexpressing CD271 and observed a partial inhibition of T cell proliferation in the presence of CD271. Further, although the proportion and activation state of immune cells are similar in adult CD271 knockout (KO) and control mice at steady-state, our preliminary data suggest that CD271 KO mice have a more severe phenotype in autoimmune models as compared to control animals. Together, these results demonstrate that CD271 modulates T cell function and may inform therapeutic approaches in autoimmunity and cancer.
Human gingiva-derived mesenchymal stem cells inhibit xeno-graft-versus-host disease through CD39 and IDO signals
Authors: Wang Julie , Huang Feng , Olsen Nancy J , Zheng Song Guo 
Published: January 2023

Abstract
Mesenchymal stem cells have the capacity to maintain immune homeostasis and prevent autoimmunity. We recently reported that human-derived gingival mesenchymal stem cells (GMSCs) have strong capacity to suppress immune responses and T cell-mediated collagen-induced arthritis in animals. However, it is unclear whether these cells can suppress human T cell-mediated diseases. Here, we used a xeno-GVHD model in the NOD/SCID mouse, which is a useful preclinical construct for evaluating the therapeutic and translational potential of this approach for applications in human disease. We found that GMSCs potently suppressed the proliferation of PBMC and T cells in vitro. Co-transfer of GMSC with human PBMC significantly suppressed human cell engraftment and markedly prolonged the mouse survival. Moreover, we demonstrated that GMSCs inhibited human PBMC-initiated xenogenic responses via CD39 and IDO signals. These findings suggest the potential for GMSCs to suppress human immune responses in immune system-mediated diseases, offering a potential clinical option to be used for modulating GVHD and autoimmune diseases.
Exosomes from B lymphocytes are a major source of Fas ligand
Authors: Lundy Steven K , Taitano Sophina H , van der Vlugt Lucien EPM 
Published: January 2023

Abstract
Exosomes are nanovesicles released by a variety of cell types, including antigen-presenting cells. The ability of exosomes to induce immunity has prompted great interest in their clinical use. Murine studies have also indicated that circulating FasL+MHCII+ exosomes can be tolerogenic in allergic and delayed-type hypersensitivity responses. B cells are capable of expressing FasL and are a major in vivo source of exosomes, but their role in producing tolerogenic exosomes is incompletely explored. In the current study, we evaluated B cells as a source of FasL+ exosomes in vivo and signals that induced B cells to release FasL+ exosomes in vitro.
In B-cell deficient mice, a major loss in total splenic exosomes and exosome-associated FasL was observed, suggesting B cells are the major source of FasL+ exosomes in vivo. After sorting B cells into CD107a+, CD5+ or CD9+ cells, all subsets showed some intracellular storage of FasL, yet FasL was predominately found in the CD9+ subset. Searching for signals that induced FasL+ exosomes in vitro, we stimulated B cell signaling through Toll-like receptors (TLRs), the B-cell receptor (BCR), and/or CD40. In response to BCR ligation, B cells produced 20% more FasL compared to medium controls, which increased to 50% in the presence of any TLR ligand. Interestingly, B cells increased their intracellular FasL storage, but failed to increase their FasL+ exosome release when stimulated through CD40 alone. TLR-4, TLR-7(/8) or TLR-9 ligation with CD40 activation induced the FasL+ exosome release, which was further enhanced by BCR ligation. These data suggest that signals known to induce the differentiation of IL-10-producing B cells, also enhance FasL+ exosome production as another potential suppressive mechanism.
Fast and easy immunomagnetic positive selection of PE- or biotin-conjugated antibody-labelled cells with Releasable Rapidspheres results in highly purified, functional and particle-free cells
Authors: Ewen Catherine , Kokaji Andy I , Clarke Samuel J , Lambertz Ulrike , De Jong Susan , Woodside Steven M 
Published: January 2023

Abstract
Immunomagnetic cell isolation is a cost-effective and gentle method of obtaining purified cell populations. However, isolating novel or rare cell populations often relies on relatively time consuming and expensive cell sorting techniques due to the lack of off-the-shelf immunomagnetic cell isolation kits. To bridge this gap, we have developed the EasySep™ Release isolation kits for PE- or biotin-conjugated antibodies. EasySep™ Release uses magnetic particles with low non-specific binding characteristics that can be rapidly removed from isolated cells after positive selection.
The system is highly customizable, targeting cells through the use of any biotin or PE-conjugated primary antibody or ligand. Starting with a variety of sample materials, including single cell suspensions from mouse spleen, lymph nodes, lungs, rat spleen, or human PBMCs, the EasySep™ Release kits have demonstrated purities consistently above 90%.
The EasySep™ Release technology can also be used to isolate subsets of cells when used in combination with our other cell separation products. For example, functional and particle-free mouse CD25+ or CD304+ CD4+ regulatory T cells were isolated by using our EasySep™ PE Positive Releasable Rapidspheres™ kit and our EasySep™ Mouse CD4+ T Cell Isolation kit. We have used EasySep™ Release with EasySep™ Dextran Rapidspheres™ to isolate CD4/CD8 double positive mouse thymocytes (double positive selection), and to isolate mouse lung epithelial cells by EpCAM positive selection followed by the removal of contaminating leukocytes via CD45 depletion.
These kits offer a customizable, and cost-effective magnetic isolation approach for the isolation of virtually any cell population.
A novel real-time imaging technique to quantify mechanisms of cell death in neutrophils
Authors: Chan Diana Wen , Zaal Kristien J , Ralston Evelyn , Gupta Sarthak , Kaplan Mariana J 
Published: January 2023

Abstract
Neutrophils have distinct mechanisms of cell death, including the formation of neutrophil extracellular traps (NETs- a process called NETosis). NETosis is characterized by reactive oxygen species generation, chromatin decondensation, membrane rupture and extrusion of a meshwork of chromatin bound to granule peptides. Objective, efficient techniques to quantify NETosis are required. We developed a new method to automatically quantify the percentage of neutrophils undergoing NETosis using the IncuCyte ZOOM™ (Essen BioScience, Inc.) platform to image neutrophils at 20× in a 96-well plate and a dual-dye system dependent on membrane integrity to stain DNA, collecting both phase and fluorescence images. DNA in healthy control neutrophils were stained with a cell membrane permeable NUCLEAR-ID Red DNA dye. Neutrophils were then incubated with Sytox, a membrane-impermeable DNA dye, used to assess cell death. Conditions were run in triplicate and 3 images per well were captured at 10-minute intervals. A processing definition was set and optimized to count all neutrophils (NUCLEAR-ID stained) and neutrophils undergoing cell death (Sytox stained) using fluorescence intensity and stained area size to identify cells undergoing NETosis. Changes in nuclear morphology, quantified by the processing definition, distinguished PMA, Ionomycin and SLE sera induced NETosis from staurosporine induced apoptosis. Findings were confirmed and correlated with the gold-standard of immunofluorescence microscopy. This novel real time assay distinguishes types of neutrophil cell death and quantifies NETosis in a rapid, accurate and reproducible way, significantly improving the study of neutrophils.
Cyanine TruStain™, an effective blocking buffer to eliminate non-specific cyanine-like dye-mediated monocyte binding
Authors: Ransom John T , Elia Jeanne , Godfrey William , Wang Jing , Jiang Nan , Yang Xifeng , Monell Craig 
Published: January 2023

Abstract
Cyanine dyes and their analogs are commonly used as acceptors in commercially available antibody-tandem fluorophore conjugates. However, it has been confirmed that the dyes have a tendency to bind monocytes or macrophages nonspecifically, which limits the ability to do multi-color flow cytometric analysis for lower density antigens. We are introducing Cyanine TruStain™ Buffer which eliminates this nonspecific binding. This reagent blocks nonspecific binding of monocytes and macrophages to Cyanine dyes, (PE/Cy7, PE/Cy5, PercpCy5.5, APC/Cy7, APC/Fire™ 750, PE/Dazzle™ 594). It does not affect binding of, e.g., CD64, CD14 and other tested antibodies. Multiple Cyanine dyes can be blocked at once and stability data shows no decrease in signal intensity compared to the conjugate alone. In addition the reagent has no impact on cell viability.
Mock sorting experiments were performed with PBMC to determine if the Cyanine TruStain™ Buffer stimulates cells to produce cytokines and chemokines. Our data indicates minimal to no stimulation above conjugated antibody alone for IL-6, IL-8, IL-10 and MCP-1 production. We also tested Cyanine TruStain™ Buffer for its ability to negatively affect PBMC proliferation in response to anti-CD3/anti-CD28 stimulation. We observed minimal to no effect on proliferation by measuring BrdU incorporation. In summary, we are introducing a blocking reagent to eliminate nonspecific binding to Cyanine dyes which shows minimal perturbation of cell functions.