bluuebunny/crossref_metadata_2025_split huggingface dataset:
Transcriptomic profiles of NK cell function indicate impaired responsiveness in chronic HIV-1 infection and enhanced functional activity post-vaccination
Authors: Costanzo Margaret C , Kim Dohoon , Creegan Matthew , Lal Kerri G , Ake Julie , Currier Jeffrey R , Streeck Hendrik , Robb Merlin L , Michael Nelson L , Bolton Diane L , Steers Nicholas James Robert , Eller Michael A. 
Published: January 2023

Abstract
Natural Killer (NK) cells limit viral replication by direct recognition and killing of infected cells, performing Fc mediated ADCC and releasing cytokines to modulate other effector activity. Despite growing evidence that supports the importance of NK cells in HIV-1 disease progression, understanding how these cells engage in vivo remains unclear. Here, we combined flow cytometry and gene expression analysis of NK cells responding to three in vitro models of NK stimulation to reveal distinct transcriptional profiles. While 76 gene transcripts were commonly upregulated following stimulation by direct cellular recognition, ADCC, and cytokine activation, 276, 36, and 293 genes were uniquely upregulated, respectively. Subsequently, a targeted gene expression panel was used to assess NK cell transcriptional content, directly ex vivo, in HIV-1 chronic infection. Transcriptional profiles in HIV-1 chronically infected individuals showed higher expression of genes associated with global activation and lower expression of genes associated with direct cell recognition and cytokine activation, compared to healthy individuals. We next looked at the transcriptional content of NK cells in response to MVA-CMDR vaccination, a live-attenuated viral vectored HIV-1 vaccine. NK cell transcriptional profiles of vaccinated individuals revealed upregulation of genes associated with direct cell recognition. These findings demonstrate that targeted transcriptional profiling provides a sensitive assessment of NK cell activity directly ex vivo that will improve our understanding of how NK cells respond to viral infections and after vaccination.
A New Melanoma Vaccine Based on Conditioned tumor Cell Lysates and Adjuvants Inhibits Tumor Growth
Authors: Onfray Flavio A Salazar 
Published: January 2023

Abstract
Clinical strategies using immune-checkpoint blockers, such as anti-CTLA4, (Ipilimumab) or anti-PD1 antibodies (Nivolumab), have demonstrated durable survival benefits in patients with melanoma and other solid tumors. Nevertheless, an important percentage of patients treated with anti-checkpoint proteins remain refractory. In this context, cancer vaccines become again a complementary alternative as cancer treatment. The optimal delivery of antigens and the use of adequate adjuvants are crucial for vaccine success. Here, a prototype of a generic therapeutic vaccine for the treatment of malignant melanoma named TRIMELVax™ was tested in an experimental model. This vaccine is based on conditioned melanoma tumor lysates combined with a specific adjuvant already tested in patients in the context of a DC vaccine, without adverse effects. The vaccine is intended to directly activate in vivo response against tumors inhibiting their growth. TRIMELVax was evaluated, in terms of safety and efficacy at pre-clinical level, in C57Bl/6 murine model of melanoma (B16). In short, immunocompetent mice were vaccinated with three doses of TRIMELVax, and then challenged with B16 melanoma cells. Alternative, immunization was also tested therapeutically in tumor bearing mice. Our results showed that only TRIMELVax was capable to reduce drastically the occurrence of tumor, inhibiting tumor growth in immunized mice. In contrast the separated use of tumor lysate, or adjuvant did not impact tumor growth. This response was associated to CD8+ T cells mediated immune response and antibody production. Immune-checkpoint blockers therapy may be strongly potentiated by the combined use of TRIMELVax, which need to be proved in future clinical trials.
Epstein-Barr Virus Latent Membrane Protein 2A (LMP2A) increases IL-10 via Bruton’s tyrosine kinase and STAT3: Effect on T cell function
Authors: Swanson-Mungerson Michelle Anne , Nelson Theodore , Stone Amanda , Incrocci Ryan 
Published: January 2023

Abstract
Data from our laboratory demonstrate that Epstein-Barr Virus Latent Membrane Protein 2A (LMP2A) promotes B cell lymphoma survival by enhancing IL-10 production, which has important implications in the treatment of EBV-associated B cell tumors. To determine the pathway used by LMP2A to increase IL-10, pharmacological inhibitors were added to LMP2A-expressing B cells and IL-10 was assessed. Our findings indicate for the first time, that LMP2A activates BTK to phosphorylate STAT3 in B cell tumors, which mediates the LMP2A-dependent increase in IL-10. Additionally, we assessed whether the LMP2A-mediated increase in IL-10 is immunosuppresssive to cytotoxic T cells that are critical for controlling the growth of EBV-associated tumors. Purified human CD8+ T cells activated with anti-CD3/CD28 mAbs were exposed to supernatants from either LMP2A-negative or –positive B cell lines for 48 hours. Our findings indicate that there is no difference in proliferation and IFN-γ secretion by activated CD8+ T cells exposed to supernatants from LMP2A-positive B cell lymphomas when compared to CD8+ T cells exposed to supernatants from LMP2A-negative B cell lymphomas. Although the LMP2A-mediated increase in IL-10 does not significantly influence the CD8+ T cell response, these findings highlight the possibility of using BTK and STAT3 inhibitors in clinical trials to treat EBV-associated lymphomas that express LMP2A. This work was supported by NIH grant 1R15CA149690-01.
Immune and metabolic regulation mechanism of Dangguiliuhuang decoction against insulin resistance
Authors: Cao Hui , Xiang Ming 
Published: January 2023

Abstract
Insulin resistance (IR) is recognized as a chronic inflammatory and metabolic disease, playing a pivotal role in the development of diabetes. Obesity and abnormal lipids metabolism increase infiltration of inflammatory cells, affect immune homeostasis, and reduce insulin sensitivity, consequently leading to the occurrence of insulin resistance. In this study, we found that Dangguiliuhuang decoction (DGLHD), a traditional Chinese medicine formula used for the treatment of diabetes, presented a beneficial effect on the progression of IR in ob/ob mice characterized by leptin gene deficiency. DGLHD exerted anti-inflammatory and immunosuppressive actions on ob/ob mice through altering function and metabolism of DCs and T cells with activated PPAR-γ molecule or inhibited PI3K/Akt signaling pathway in cells. Additionally, DGLHD also ameliorated glucolipid metabolism in liver and adipose tissue via regulating the expression of metabolic genes and activating PI3K/Akt signaling pathway. Importantly, metabolomics analysis revealed that DGLHD could decrease glutamate, phenlalanine, branched-chain fatty acids and glyoxylate as well as cholesterol and triglyceride in adipose and hepatic tissues of ob/ob mice, thereby improving the insulin sensitivity of target organs. Therefore, we first elucidated that DGLHD has anti-insulin resistant effects by maintaining immune and metabolic homeostasis.
Negative regulation of Th17 cell development and function by the REV-ERBs
Authors: Solt Laura A , Amir Mohammed , Chaudhari Sweena M. , Wang Ran , Campbell Sean 
Published: January 2023

Abstract
A significant amount of work has identified key factors that drive Th17 cell development, including the nuclear receptors (NRs) RORα and RORγ. However, cell-intrinsic mechanisms that negatively regulate Th17 cell development and inflammation have received less attention. Two other members of the NR superfamily, the REV-ERBs (REV-ERBα and REV-ERBβ), are often co-expressed in the same tissues as the RORs, bind the same DNA response elements, and co-regulate their shared target genes. The REV-ERBs have well described roles in the regulation of the circadian rhythm and metabolism, but their roles in immune cell function are poorly understood. Therefore, we hypothesize that the REV-ERBs are key negative regulators of Th17 cell development and function.
We we aim to understand the role of the REV-ERBs in Th17 cell development as well as determine whether targeting these NRs is a viable therapeutic option for the treatment of Th17-mediated autoimmune diseases. Our results indicate that the REV-ERBs are differentially expressed during Th17 cell development. Overexpression and pharmacological modulation of REV-ERB activity suppresses Th17 cell development whereas genetic deletion of the REV-ERBs perturbs Th17 cell development in vitro and induces autoimmunity in vivo. Using REV-ERB-specific small molecules that we have developed, we have found that pharmacological modulation of REV-ERB activity suppresses Th17 cell development in vitro and development of autoimmunity in vivo. Collectively, our data suggest that the REV-ERBs are key negative regulators of Th17 cell development and function and represent a novel therapeutic target for the treatment of Th17-mediated autoimmune diseases.
Funding support – NIH (R01AI116885-01A1)
The resolutive factors issued from apoptotic cell clearance allow the termination of ongoing inflammation
Authors: Perruche Sylvain , Bonnefoy Francis , Couturier Mélanie , Martin-Rodriguez Omayra , Gauthier Thierry , Saas Philippe 
Published: January 2023

Abstract
Immune-mediated inflammatory diseases have in common a dysregulation of the immune response. Resolution of inflammation is a powerful process involving a numerous of factors including specialized pro-resolutive lipid mediators and proteins such as TGF-β, resulting in inflammation termination and the initiation of tissue healing. A critical step of inflammation resolution is efferocytosis of apoptotic effector cells by phagocytes which produce factors allowing the termination of inflammation and initiating tissue healing. TGF-β has been shown has mandatory in such process since it is produced by apoptotic cell and phagocytes clearing them. We hypothesize whether the reintroduction of pro-resolutive factors issued from apoptotic cell efferocytosis would allow a termination of ongoing inflammation. Injection of such resolutome in ongoing collagen-induced experimental arthritis allowed the long term reduction of clinical symptoms and was accompanied by the emergence of highly collagen-specific Tregs and antigen presenting cells (APC) reprograming through a tolerogenic state. When injected in ongoing T cell-transfer-induced Inflammatory Bowel Disease experimental model, again, clinical symptoms and colon mucosa lesions were severely reduced, accompanied by a higher proliferation, migration and wound healing capacities of both epithelial cells and fibroblasts. TGF-β demonstrated a prominent role in the reprogramming of APC and regulatory T cell emergence. So far the pro-resolutive factors issued from apoptotic cell efferocytosis demonstrated the potential to terminate ongoing inflammation. This opens clinical drug development opportunities in the treatment of refractory chronic inflammatory diseases.
Temporally controlled B cell depletion with universal chimeric antigen receptor (CAR) T cells for pemphigus vulgaris (PV) therapy
Authors: Ellebrecht Christoph T , Mao X , Melenhorst J J , Lacey S F , Zhao Y , Milone M C , Payne A S 
Published: January 2023

Abstract
Therapy of PV relies on chronic immunosuppression, which results in significant morbidity. Complete, transient B cell depletion should cure PV, since autoreactive clones do not recur upon regeneration of the B cell repertoire. Engineered CAR T cells (CARTs) are the most potent means for total B cell depletion, but for therapy of autoimmunity, temporal control of CART activity is necessary to prevent lasting immunosuppression. Here we validate 3 novel strategies to control CART survival and function. We combined a B cell targeting CAR with an inducible caspase 9 suicide gene (sCAR), a reverse (constitutively active) suicide gene (revCAR), or a molecular on-switch that permits CAR surface expression (onCAR). sCAR, revCAR and onCARTs showed potent in vitro killing equivalent to conventional CARTs. Activation of the respective regulatory system resulted in in vitro depletion of &gt;95% of sCART and revCARTs and loss of CAR expression in onCARTs. In an in vivo leukemia model, &gt;90% of sCARTs were eliminated (p&lt;0.01) as compared to vehicle treated mice, while preserving their efficacy before suicide gene activation (p&lt;0.001). RevCARTs showed complete loss of leukemia control in vivo in the absence of a suicide-preventing compound (p&lt;0.01), indicating their functional depletion in vivo. Finally, to allow universal (allogeneic) CART therapy, we used CRISPR genome editing to disrupt endogenous T cell receptor and MHCI expression. Universal sCARTs caused total B cell depletion in a humanized allogeneic BLT mouse model (p&lt;0.05) with complete in vivo depletion of sCARTs. In summary, we present 3 novel strategies to regulate CARTs, which provide a platform for curative, universal, and large-scale applications in PV and other autoimmune diseases.
A pharmacological protein kinase C/D inhibitor Gö6976 ameliorates collagen-induced arthritis
Authors: Yoon Taewon , Cho Hongsik , Kim Young-In , Brand David , Hasty Karen , Stuart John , Yi Ae-Kyung 
Published: January 2023

Abstract
Toll-like receptor (TLR) signaling has been implicated with the initiation and progression of rheumatoid arthritis (RA). Disruption of TLR signaling at early stages of RA might thereby provide an opportunity to halt the RA progression and ameliorate outcomes. We previously found that protein kinase D1 (PKD1) is essential for proinflammatory responses mediated by MyD88-dependent TLRs, and TLR-dependent PKD1 activation is inhibited by Gö6976. However, it is currently unknown whether PKD1 plays a role in pathogenesis of RA and whether Gö6976 has ameliorating effects on arthritis. Here, we investigated whether PKD1 contributes to the TLR-mediated proinflammatory responses in human synovial cells, and to development and progression of type II collagen (CII)-induced arthritis (CIA; an experimental animal model for human RA). We found that PKD1 is constitutively hyperactivated in synoviocytes of RA patients, and is rapidly activated in response to TLR/IL-1R ligands. Silencing PKD1 expression in RA synoviocytes resulted in substantial inhibition of both spontaneous and TLR/IL-1R-mediated expression of cytokines/chemokines. Although Gö6976 had no effect on the TCR-induced cell proliferation and cytokine production, synergistic effects of TCR and TLR2, as well as TLR2 alone, on the spleen cell cytokine production and proliferation were significantly inhibited in the presence of Gö6976. In addition, daily treatment with Gö6976 significantly reduced serum levels of IFNg and anti-CII IgG, and the incidence and severity of arthritis in humanized HLA-DR1 transgenic mice immunized with CII. Our data suggest that PKD1 might play an important role in proinflammatory responses in RA and CIA, and could be a therapeutic target for arthritis.
Glucocorticoid induced leucine zipper in Alzheimer’s disease.
Authors: Srinivasan Mythily , Lahiri Debomoy K 
Published: January 2023

Abstract
Aging and stress are significant risk factors for Alzheimer’s disease (AD). Stress induced glucocorticoids accelerate beta amyloid (Aβ) plaque formation and tau phosphorylation, aggregates of which upregulate reactive oxygen species in vulnerable neurons. The oxidative stress negatively regulates glucocorticoid receptor, preventing nuclear translocation and suppression of the transcriptional factor, nuclear factor-κB (NF-κB) by glucocorticoids. The ensuing inflammatory distress enhance further Aβ accumulation initiating a vicious cycle. Glucocorticoid induced leucine zipper (GILZ) is a recently identified protein that mediates many of the cellular effects of glucocorticoids. Exogenous GILZ ameliorated disease in models of chronic inflammation. Mechanistically GILZ inhibits NF-κB by binding the p65 subunit of NF-κB in the cytoplasm and thereby preventing transactivation of inflammatory and apoptosis mediators. Microglial GILZ expression has been shown to correlate negatively with inflammatory cytokines. Increased p65, upregulated inflammatory cytokines and apoptotic markers are frequently observed in postmortem AD tissues. Hence selective targeting of activated p65 is an attractive strategy to modulate neuro-inflammation and suppress neurodegeneration. Recently we reported the design and functional potential of structural analogs of GILZ that selectively blocked activated p65 and suppressed Aβ induced cytotoxicity in mixed brain cell cultures. Here we evaluated the efficacy of GILZ mimics in lipopolysaccharide induced mouse model of AD. Our data suggest that mice treated with select mimics of GILZ exhibited reduced inflammation and Aβ deposits suggesting a therapeutic potential.
Teaching students how to think like an immunologist: flipping a classroom without flipping out.
Authors: Parkin Kirstin 
Published: January 2023

Abstract

Study
A traditional lecture-based 400 level immunology course was converted into a team-based learning environment. Activities were used to teach students how to analyze published data from research labs at MSU and thus, learn how to “think” like MSU scientists.

Objectives
For the main concepts covered in the course, the goal was to improve students’ ability to discuss the concept orally and in written format, diagram the concept, use drawings to portray the concept, graph the concept, and identify techniques and measurements used in current research related to the concept.

Methods
Learning objectives, guided textbook readings, weekly on-line formative assessments with feedback, in-class team based activities, Top Hat in-class response questions, and in-class summative assessments were used to meet the course objectives.

Assessments
35 on-line formative assessments, 35 Top Hat in-class response questions, and 4 in-class summative assessments were provided during the semester. Online student feedback regarding exam questions and course satisfaction were also used.

Results
Student performance on short-answer questions on formative assessments improved throughout the semester. Students identified multiple choice questions as the least likely to assess course objectives. Students with low attendance earned the lowest scores.

Implications
By providing students with examples of how MSU scientists design experiments, display data, and analyze data, students learn how to “think” like a scientist rather than learning a large number of scientific facts. The active learning environment also provides an opportunity for students to improve teamwork skills that are sought by corporations hiring college graduates.
Aggregation and innate immune response modulating impurities
Authors: Polumuri Swamy kumar , Haile Lydia , Rao Roshni , Verthelyi Daniela 
Published: January 2023

Abstract
Therapeutic proteins can induce immune responses that affect the safety and efficacy even if derived from human sequences. The factors that contribute to the immunogenicity of this therapeutics are poorly understood, but protein aggregation is thought to facilitate it. Previously, our lab showed that trace levels of product or process derived innate immune response modulating impurities (IIRMIs) can also increase product immunogenicity. In this study we hypothesized that protein aggregates and IIRMIs may synergize to facilitate product immunogenicity and explored it using clinical grade Human Serum Albumin (HSA) and Intravenous Immunoglobulin (IVIG) proteins as model. Protein aggregations formed by mechanical stress (shaking and stirring) were characterized by SDS-PAGE, turbidity, MFI and FlowCAM. Cellular responses to aggregated proteins with trace levels of IIRMs were tested in human PBMC. We show that stirring and shaking resulted in the formation of stable aggregates with size ranges of 2 – 50 μm for stirred HSA; and 2–100 μm for IVIG. Despite the similarity in size, the functional assays showed that aggregates of IVIG result in increased mRNA expression of IL-8, IL-6, IL-1β and CCL-2 whereas HSA aggregates did not. Further, we analyzed how the aggregated product changed the transcriptome using Nanostring and showed that aggregated IVIG (shaken or stirred) significantly increased the mRNA of CCL2, CCL7, CCL3, CCL24, CSF1, CXCL2, IRAK1, EGR2, CEBPβ, PPARg, TNFSF15 and whereas HSA aggregates did not. Similarly MAPKs (pp38, pERK and pJNK) and pAKT were activated when PBMC were stimulated with IVIG aggregates but not HSA aggregates. Lastly, we examined whether the effect was mediated by Toll like receptors.
Human astrocytes and microglia-like cells express cGAS-STING viral sensing components
Authors: Jeffries Austin , Marriott Ian 
Published: January 2023

Abstract
NeuroAIDS is a major health problem in AIDS patients and long-term HIV survivors, and evidence is accumulating that the innate immune responses of glial cells to this virus play a critical role in disease pathology within the central nervous system (CNS). While astrocytes and microglia are uniquely positioned to respond to CNS pathogens and these cells have been shown to produce key inflammatory and antiviral mediators following exposure to retroviruses including HIV, the molecular mechanisms by which glia perceive such viruses remain incompletely understood. It has become increasingly apparent that innate immune sensing of nucleic acids underpins many physiological and pathological responses to viral pathogens, and the recent identification of a key role for cyclic GMP-AMP synthase (cGAS) as a cytosolic DNA sensor has been one of the most exciting developments in viral immunology in recent years. Importantly, cGAS has been demonstrated to contribute to the sensing of replicative HIV-1 DNA intermediates and eliciting cytokine responses in a human monocytic cell line. Here, we demonstrate that human glial cell lines and primary human astrocytes constitutively express robust levels of cGAS, and we show that such expression is maintained in these cells following challenge with either viral ligands or a laboratory strain of HIV-1. Furthermore, we show that human glial cells express the critical cGAS downstream adaptor molecule STING, and confirm that the robust levels of expression of this molecule are similarly maintained following viral challenge. The data therefore support a potential role for the cGAS-STING cytosolic viral sensing pathway during acute and/or chronic HIV infection of the CNS.
An Essential Role for TAGLN2 in Phagocytosis of Lipopolysaccharide-activated Macrophages
Authors: Jun Chang-Duk , Kim Hye-Ran 
Published: January 2023

Abstract
Activated macrophages have a greater ability of phagocytosis against pathogens that is mediated by large-scale actin rearrangement. However, molecular machineries that conduct this task have not been fully identified. Here, we demonstrate an unanticipated role of TAGLN2, a 22-kDa actin-binding protein, in Toll-like receptor (TLR)-stimulated phagocytosis. TAGLN2 was greatly induced in macrophages in response to lipopolysaccharide (LPS), a ligand for TLR4, partly via the NF-κB pathway. TAGLN2-deficient macrophages (TAGLN2−/−) showed defective phagocytic functions of IgM- and IgG-coated sheep red blood cells as well as bacteria. Cell signaling pathways involved in actin rearrangement—PI3 kinase/AKT and Ras-ERK—were also down-regulated in LPS-stimulated TAGLN2-deficient macrophages. Moreover, TAGLN2−/− mice showed higher mortality after bacterial infection than wild-type littermates. Thus, current results provide a novel function of TAGLN2 as a molecular armament required for host defense.