bluuebunny/crossref_metadata_2025_split huggingface dataset:
Blocking CD13 in combination with a toll like receptor agonist increases the efficacy of cancer vaccine adjuvants
Authors: Qendro Veneta , Ghosh Mallika , Shapiro Linda 
Published: January 2023

Abstract
Vaccines have been an extraordinary tool in the fight against human disease. Recent cancer research has revealed promising results leading to approved cervical and prostate cancer vaccines. Despite the success, a major downfall of these vaccines remains poor immunogenicity.
MPLA (monophosphorylated lipid A) is a synthetic analog of LPS that binds to TLR4 leading to activation of the adaptive immune response without triggering deleterious inflammatory consequences. As such, MPLA has recently been FDA approved as an adjuvant of the prophylactic vaccine against HPV-driven (Human Papilloma Virus) cervical cancer. However, this vaccine is limited to non-preexisting HPV cases while protecting against only two out of the dozen HPV strains that can potentially lead to cervical cancer.
Pertinent to this subject, we have found that the absence of CD13 in antigen presenting cells (APCs), leads to increased TLR4 endocytosis and bias toward the same endocytic-signaling pathway triggered by MPLA. CD13 is a large, multifunctional cell surface peptidase constitutively expressed on all lineages of myeloid cells. We have previously shown that lack of CD13 increases tumor antigen uptake and presentation, resulting in enhanced activation of tumor-specific cytotoxic T cells.
The combination of these facts has led to the intriguing possibility that blocking CD13 may amplify the efficacy of MPL as an adjuvant and ameliorate the efficacy of prophylactic and therapeutic cancer vaccines. Therefore, we hypothesize that CD13-blocking agents will act as dual-action adjuvants by enhancing MPL-TLR4 endocytosis, and as a delivery system through increased endocytic antigen uptake and subsequent cytotoxic T cell activation.
Combination of Peptide-based Vaccine and Chinese Herbal Medicine as a Strategy to Enhance Immune responses in Virus-associated Cancer
Authors: Song Ying-Chyi , Yen Hung-Rong 
Published: January 2023

Abstract
Virus infections may account for the development of several cancers, such as HPV16/18 are known to cause around 70% of cervical cancer cases. Until now, several immunotherapeutic approaches for virus-induced cancer are under development. Peptide-based vaccines have several advantages over conventional whole-protein vaccines in terms of purity, lot-to-lot consistency, production costs, and the high antigenic specificity. However, the use of peptide antigens in vaccine development has been hampered by problems, such as weak immunogenicity coupled with a paucity of potent adjuvants. Specific T cell immune response is crucial for anti-tumor immunity. Manipulating specific T cell immunity by Chinese herbal medicine (CHM) is a promising field to explore. Therefore, we aimed to investigate whether CHM can modulate specific T cell responses to apply for the development of cancer vaccine. We have identified a CHM extract that could increase expression of maturation cytokines and activation markers of murine bone marrow-derived dendritic cells. Furthermore, in a murine TC-1 tumor-bearing model, we found that the CHM extract could act as an adjuvant to induce cellular immune responses and anti-tumor effect in peptide vaccine strategy. We suggested that immune-stimulator CHM combined with cancer vaccine endows them with increased immunologic activity, which may be used to bypass the requirement for the conditional adjuvant. Further delineation of the mechanism may provide new clues for vaccination strategy.
Dendritic cell CD40 targeting vaccines elicit CD8+ T cell-mediated therapeutic immunity against cancers
Authors: Yin Wenjie , Duluc Dorothée , Joo HyeMee , Xue Yaming , Gu Chao , Wang Zhiqing , Wang Lei , Oxford Lance , Clark Amelia , Parikh Falguni , Thompson-Snipes LuAnn , Woo Jung-Hee , Zurawski Sandra , Sikora Andrew G , Zurawski Gerard , Oh SangKon 
Published: January 2023

Abstract
Dendritic cells (DCs) are major antigen-presenting cells that can efficiently cross-prime antigen-specific cytotoxic T lymphocytes (CTLs). Antigen targeting to DCs through surface receptors is therefore a promising strategy to mount CD8+ CTL-mediated immunity against viral infections and cancers. We have previously reported that viral (Influenza HA and NP) and tumor antigen (MART-1) targeting to human DCs via CD40 resulted in greater antigen-specific functional CD8+ CTL responses in vitro. In this study, we have tested CD40 targeting prototype vaccines for HPV16-associated malignancies (anti-CD40-HPV16.E6/7) and prostate cancer (anti-CD40-PSA). Our data show that both anti-CD40-HPV16.E6/7 and anti-CD40-PSA prototype vaccines are able to activate antigen-specific CD8+ T cells in the blood of head-and-neck and prostate cancer patients, respectively. We further demonstrate that the in vivo administration of anti-CD40-HPV16.E6/7 or anti-CD40-PSA plus poly(I:C) can mount tumor-specific CTL-mediated therapeutic and preventive immunity against the respective tumors in human CD40 transgenic mice. Data from this study support the clinical development of CD40 targeting vaccines against cancers and viral infections.
MHC class II-peptide complexes displayed on activated T cells guide Treg suppression
Authors: Germana Sharon K , Fryer Madeline L. , Junius Steffie , Turka Laurence A , LeGuern Christian A 
Published: January 2023

Abstract
Regulatory T cell (Treg) therapy is a promising strategy for extending allograft survival. Treg suppression requires initial T cell receptor (TCR)/peptide-MHC-II (pMHC-II) contacts, the nature of which remains elusive. As inflammation facilitates transfer of pMHC-II from antigen presenting cells (APCs) to activated T cells, we hypothesize that during transplantation associated inflammation, pMHC-II relocate on alloreactive T effector cells and serve as Treg activation signals. To test this, suppression assays were performed using murine CD4+ CD25− (Teff), CD90 depleted APCs, and CD4+ CD25+ Treg cells of different MHC haplotypes. We also used a transplantation model of skin allografts in RAG1 deficient mice injected with Teff and Treg cells from various MHC backgrounds. Data from more than 150 co-culture experiments show that Treg suppression occurs only when Treg and Teff share the same MHC-II, but independently of Treg matching with APC MHC-II. Further experiments demonstrate that pMHC-II complexes, displayed on activated Teff cells, are crucial to the induction of Treg suppression. In vivo results recapitulate the in vitro data: Treg suppression, leading to graft survival, occurs only when Teff and Treg cells are MHC-II matched. Collectively, these findings suggest that suppression of alloreactivity is directed by Treg TCR recognition of pMHC-II complexes exposed on Teff cells. The nature, origin (host vs. donor), and significance of MHC-II loaded peptides have yet to be established. Future studies to improve targeted Treg therapies for transplantation tolerance will focus on mechanisms coupling Treg function with Teff cell activation.
Imaging flow cytometry enhances a new model of dynamic FOXO1 nuclear localization in human CD4 T cells
Authors: Golding Amit , Hritzo Molly 
Published: January 2023

Abstract
The forkhead box O (FOXO) family contains transcription factors critical to differentiation and homeostasis in T cells. FOXO1, acts as a key integrator of proliferation and differentiation signals in both effector T cells (Teff) and regulatory T cells (Treg). In Teff, when the phosphoinositide 3-kinase (PI3K)/Akt pathway is activated downstream of T cell receptor (TCR) signaling and co-stimulation, Akt phosphorylates FOXO1 and excludes it from the nucleus. This promotes T cell proliferation and effector functions. In contrast, nuclear FOXO1 is essential for Treg thymic development and peripheral stability. Multiple studies have examined the effects of FOXO1 deficiency or overexpression in T cells, but to date, few have been able to directly examine the dynamics of the native FOXO1 signaling in human T cells. The accepted method of examining FOXO1 signaling dynamics is nuclear/cytoplasmic protein extraction and subsequent western blotting (WB). In order to assess native FOXO1 signaling dynamics, we obtained the human CD4+ T cell line, HuT102. Using the classic WB approach, we demonstrate that HuT102 cells have more nuclear FOXO1 compared to a similar CD4+ T cell line, HuT78. However, WB does not allow us to examine FOXO1 signaling dynamics in subpopulations. To answer this question, we developed a staining protocol to assess FOXO1 nuclear translocation in HuT102 cells using imaging flow cytometry on an Amnis Imagestream MKII. We show that addition of PI3K and Akt inhibitors increases nuclear FOXO1, while TCR signaling mimics such as PMA/ionomycin and anti-CD3/CD28 decrease nuclear FOXO1 kinetically. Using this protocol, we are now able to directly examine native FOXO1 signaling dynamics in human CD4+ T cells.
Myeloid derived suppressor cells are controlled by regulatory T cells via TGF-b during murine colitis
Authors: Park Sung-Gyoo , Lee Cho Rong 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of cells composed of immature myeloid cells including precursors of macrophages, granulocytes, dendritic cells, and myeloid cells at early stages of differentiation. In mice, MDSCs are defined as CD11b+Gr1+ cells and are normally present in low numbers in the bone marrow and spleen of healthy mice. However, in response to inflammatory diseases, MDSCs accumulate in greater numbers in the spleen, blood, and lymph nodes where they suppress immune responses. Moreover, MDSCs are well-known regulators of regulatory T-cells (Treg cells); however, the direct regulation of MDSCs by Treg cells, has not been well characterized. Interestingly, we found that colitis caused by functional deficiency of Treg cells leads to altered expansion and reduced function of MDSCs. In addition, during differentiation of MDSCs in vitro from bone marrow cells, Treg cells enhanced MDSC function and controlled their differentiation in a mechanism involving TGF-β. Moreover, TGF-β-deficient Treg cells were not able to regulate MDSC function in an experimentally-induced model of colitis. Finally, we evaluated the therapeutic effect of TGFb-mediated in vitro differentiated MDSCs on colitis. Adoptive transfer of MDSCs that differentiated with TGF-β was more capable of preventing colitis than the transfer of those MDSCs that differentiated without TGF-β. Our results demonstrate an interaction between Treg cells and MDSCs that contributes to regulation of MDSC proliferation and acquisition of immunosuppressive functions.
TULIPES: Tn5-mediated ultra low input parallel epigenetic sequencing
Authors: Patterson Dillon , Scharer Christopher , Boss Jeremy M 
Published: January 2023

Abstract
Accumulating evidence reveals that changes in the epigenome regulate gene expression profiles of developing, differentiating, and effector immune cells. Importantly, disturbances in this developmental road map may contribute to aberrant immune responses. The recent development of the Assay for Transposase Accessible Chromatin sequencing (ATAC-seq) allows for the epigenetic programming of rare immune cells to be determined. This method utilizes the Tn5 transposase to simultaneously fragment and tag genomic DNA by inserting Illumina sequencing adapters. Unique sequencing indexes are then added to the adapter-modified genomic fragments during the PCR enrichment step of sample preparation. This tagmentation-based approach has emerged as a popular approach to generating sequence-ready libraries. Here, we describe a novel strategy in which we have customized the tagmentation process by using in-house purified Tn5 transposase pre-loaded with indexed adapters. This method allows for a more efficient analysis of samples which increases experimental throughput while significantly reducing sequencing cost.
Tools to simplify panel design for multi-color flow cytometry experiments.
Authors: Bashratyan Roman , Luan Feng-Jun , Widmann Stephanie , Corselli Mirko , Semyonova Maria , Rabenstein Jacob , Stall Alan M 
Published: January 2023

Abstract
While flow cytometry continues to be a critical tool for immunologists, one of the more difficult aspects of this practice is panel design. Fully resolving dim populations of interest can be challenging. Low antigen density, fluorescence spillover, available fluorochromes and limited catalog offerings are just a few of the factors that make panel design difficult often leading to sub-optimal resolution. Previously, we have shown that following a basic set of principles can reduce the number of iterations needed to maximize the resolution for populations of interest. Here we show how recent developments, including the BD Horizon™ Guided Panel Solution (GPS) and BD OptiBuild™ reagents, help simplify panel design even further. The BD GPS was designed with the best practices of panel design at its core. By guiding the user through critical steps in this process and flagging potential problem areas in a proposed panel, the GPS minimizes the need for early panel iteration. BD OptiBuild reagents vastly improves the catalog offering of fluorochrome/specificity combinations, thus reducing the need for panel redesign due to limited reagent availability. To demonstrate the utility of these new tools, we walk through panel design for an 8 color assay on both 3 laser (8 parameter) and 5 laser (18 parameter) systems. The analysis of these panels shows that by using these tools, the chances of obtaining a successful panel increase. Additionally, panel design can be made easier and population resolution improved when utilizing a 5 laser system where impacts of spillover and spread can be minimized. Together, using these tools and approaches the goal is to minimize the number of iterations required to develop panels that meet the needs of the investigator.
Umbilical cord derived monocytes and platelet rich plasma for diabetic wound healing
Authors: Greene-Roos Jennifer A , Laughlin Mary 
Published: January 2023

Abstract
Chronic non-healing ulcers are a significant complication present in approximately 15% of diabetic patients. Despite standard clinical treatment including wound dressing, debridement of necrotic tissue, and offloading, 15–27% of patients with diabetic ulcer will require amputation. Deep infection is a risk factor for amputation. Platelet rich plasma (PRP) and stem cells have been tested for potential application to treat diabetic ulcers and have been shown to improve wound healing. Umbilical cord blood provides robust cellular and secreted products with therapeutic potential, but remains yet to be fully exploited as a resource for wound healing strategies. Our initial proof of concept studies indicate that umbilical cord blood derived monocytes and platelet rich plasma enhance wound healing processes measured by in vitro assays. When compared to monocytes or platelet rich plasma alone, monocytes and platelet rich plasma significantly enhanced neovascularization as measured by the matrigel angiogenesis assay. Monocytes and platelet rich plasma enhanced fibroblast migration in a wound scratch assay. Additionally, monocytes and platelet rich plasma enhance proliferation of endothelial cells and fibroblasts. Furthermore, these monocytes have potent antibacterial effects as measured by their ability to phagocytose P. aeruginosa in vitro. Collectively, this in vitro data indicates that this product could heal wounds by a mechanism involving enhancing neovascularization, proliferation, migration, and antibacterial activities, all processes which are dysregulated in diabetic wound healing.
Combining gene expression data from inflamed tissue and machine learning for blood biomarker discovery
Authors: Milanez-Almeida Pedro , Martins Andrew J , Narayanan Manikandan , Torabi-Parizi Parizad , Franco Luis , Tsang John S , Germain Ronald N 
Published: January 2023

Abstract
Inflammation is common in virtually all diseases. Since immune cells travel through the blood to inflamed tissues, peripheral blood reflects inflammation at distant sites. However, blood biomarkers are hard to detect in global gene expression data since blood is not the site of perturbation and statistical corrections for multiple testing reduce sensitivity. We hypothesized that combining transcriptional signatures from inflamed tissue and machine learning would be useful to derive blood biomarkers. We first tested this in the mouse model of influenza infection. We identified a set of early genes in the lungs associated with lethality after infection. We then tested their differential abundance early in the blood of mice infected with a range of influenza doses. We applied elastic net to build a biomarker able to predict disease outcome (4 classes: early lethal, late lethal, non-lethal vs. non-infected; accuracy on training and validation sets: 91% and 79%). We next confirmed our hypothesis in lung cancer and breast cancer in humans. We identified sets of genes associated with tumors but not with tumor-free tissues and tested their differential abundance in the blood of patients compared to healthy controls. Using elastic net, we built two blood biomarkers to classify subjects according to disease status (case vs. control; accuracy on training and validation sets in breast cancer: 92% and 67%; in lung cancer: 89% and 78%). In conclusion, combining inflamed tissue signatures and machine learning, we were able to derive blood biomarkers for three different diseases in two different species. We expect that this novel strategy can be applied in other diseases as well.
This work was supported by the Intramural Research Program of NIAID, NIH.
IFN-γ-deficient transplant induces acute gut GVHD via dysregulation of RagIIIγ and intestinal microbiota
Authors: Song Qingxiao , zeng Veronica , Wu Xiwei , Riggs Arthur D. , Zeng Defu 
Published: January 2023

Abstract
Gut GVHD is critical for determining the outcome of allogenic hematopoietic cell transplantation (HCT). Damage of Paneth cells, loss of defensins, and increase of anti-bacterial lectin regenerating islet-derived IIIγ (RagIIIγ) can lead to dysregulation of intestinal microbiota and gut GVHD pathogenesis. RagIIIγ can be produced by Paneth cells and intestinal epithelial cells and its secretion is augmented by IL-22 and MyD88-mediated signaling. However, the circumstances leading to its dysregulation remains largely unknown. Here we show that, in a MHC-mismatched model of C57BL/6 donor to BALB/c recipient, one injection of depleting anti-CD4 effectively prevents acute gut GVHD in recipients given wild-type (WT) transplants but has no effect in recipients given IFN-γ−/− transplants. Most of latter recipients died with diarrhea and severe colitis by two weeks after HCT. As compared to recipients given WT transplants, recipients given IFN-γ−/− transplants showed expansion of gut tissue infiltrating CD8+ Tc17 cells that produced both IL-17 and IL-22, although no damage of Paneth cells. Recipients given IFN-γ−/− transplants had markedly increased mRNA levels of RagIIIγ in the intestinal tissue and markedly increased protein levels in the serum as well as significant reduction of anaerobic bacteria Barnesiella in feces. Co-housing recipients given IFN-γ−/− transplants with recipients given WT transplants significantly increased the survival of the former recipients. These results indicate that IL-22-producing Tc17 cells can augment intestinal tissue secretion of RagIIIγ, leading to imbalance of microbiota and gut GVHD pathogenesis, even in the absence of Paneth cell damage.
Mucosal T-Bet+CD8+T cells are implicated in the pathogenesis of human chronic Graftversus- Host Disease
Authors: da Silva Ana C Costa , Dodge Joshua T , Rose Jeremy J , Dhamala Susan , Hakim Frances T , Pavletic Steven Z , Mays Jacqueline 
Published: January 2023

Abstract
Allogeneic hematopoietic stem cell transplantation is a curative cancer immunotherapy whose use is limited by the high incidence of post-transplant chronic graft-versus-host disease (cGVHD), an alloimmune disease that manifests in multiple organ systems including the oral cavity where buccal mucosa (BM) and minor salivary glands (MSG) are affected. As little is known about the immune network that drives pathogenesis of cGVHD, therapeutic agents are limited, and this study used a multiparameter approach to query the relationship between salivary gland gene expression, salivary proteins and the immune cell network within oral tissues with regard to oral cGVHD severity. Microarray analysis of labial MSG from oral cGVHD patients revealed significant up-regulation in gene expression pathways involving immune/inflammatory responses (IL-1 and IL-6 signaling, IL-32), response to oxidative stress (superoxide dismutase 2) and wounding/fibrosis (TIMP-1, TGF-β) in patients versus non-sicca controls. Immunohistochemistry revealed that MSG of these patients had increased numbers of activated T-bet+ CD8+ T cells which were directly associated with acinar structure alteration and glandular infiltrates. Flow cytometric analysis also demonstrated high frequency of CXCR3+ cells in cGVHD-affected MSG, consistent with increased levels of CXCL9 and CXCL10 detectable in the whole saliva of oral cGVHD patients. Parallel flow cytometric analyses in a mouse model of cGVHD in the submandibular salivary gland demonstrated an increased percentage of CD103+ DCs, which can cross-present antigens to activate CD8+ T cells. These studies are ongoing, but implicate activation of T cytotoxic (Tc1) cells in the pathogenesis of oral cGVHD.
Alloreactive T cell proliferation is suppressed by 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR) independent of AMP-activated protein kinase
Authors: Beezhold Kevin J , Byersdorfer Craig A 
Published: January 2023

Abstract
The benefits of hematopoietic stem cell transplantation, a therapy used to treat a variety of hematologic and immunologic disorders, are often outweighed by the risks of acute graft-versus-host disease (GVHD), where donor T cells attack and destroy host tissues. Our lab has previously shown that targeting lymphocyte metabolism is a promising and novel strategy for the treatment of GVHD. More recently, we have found that AMP-activated protein kinase (AMPK), a major cellular energy sensor, plays a significant role in T-cell proliferation during disease pathogenesis. To explore this relationship further, we treated alloreactive T cells generated during a mixed leukocyte reaction (MLR) with 5-aminoimidazole-4-carboxamide ribonucleoside (AICAR), an AMP mimetic that strongly activates AMPK. AICAR treatment of MLR cultures strongly repressed T cell proliferation, with a 70% decrease in total cells by 96 hours. Interestingly this decrease in T cell proliferation was independent of AMPK, as similar results were obtained using AMPK−/− cells. Mechanistically AICAR repressed AKT signaling in stimulated T cells, with a 50% decrease in AKT phosphorylation when cells were concurrently treated with 100 μM AICAR. Cytokine driven STAT5 activation was also significantly blunted by simultaneous AICAR exposure. In summary, despite its wide use as an AMPK agonist, we found that AICAR exerts significant inhibitory effects on T cell allo-activation independent of AMPK. This inhibition occurs, at least in part, through decreased phosphorylation of AKT and down-regulation of STAT signaling, making AICAR a potential therapy to combat GVHD and similar inflammatory disorders.