bluuebunny/crossref_metadata_2025_split huggingface dataset:
New cross-strain protective influenza vaccine using self-attenuated influenza virus
Authors: Wu Lihong , Li Junwei , Arévalo Maria T , Chen Yanping , Yan Yongyong , Wang Haiyan , Zeng Mingtao 
Published: January 2023

Abstract
Live attenuated vaccines (LAVs) are derived from disease-causing pathogens and are effective way for prevention of infectious disease. Classical LAVs are produced experimentally by repeated passaging of a virus in cultured cells, but this method is not always reliable and safe because in some cases, there is a reversion to wild-type virulence. Several methods have been employed to improve them, but the efficacy and safety are still a challenge. In this study, we have developed a self-attenuated RNA virus expressing a functional and mammalian species-specific artificial microRNA. Using influenza virus as a model, we designed an artificial miR-93 cassette for insertion into NS gene segment of influenza viral genome, which produces a specific amiRNA for inhibiting NP gene expression that would result in a virus which is attenuated in mammalian cells. The resulting engineered self-attenuated influenza virus, PR8-amiR-93NP, produced mature and functional artificial microRNA against NP in mammalian cells, but could be propagated at a high yield with the traditional influenza virus production method in embryonated chicken eggs. Furthermore, PR8-amiR-93NP was attenuated by 100 fold in mice compared with its wild-type counterpart. In animal experiments, intranasal vaccination with PR8-amiR-93NP induces antigen-specific antibody responses and protective immunity against heterologous influenza virus strains. In conclusion, our results demonstrated that this self-attenuation approach provides a safe and effective platform for generation of live attenuated influenza vaccine, as a next generation nasal spray influenza vaccine.
A practical approach to high complexity multiplexed chromogenic immunohistochemistry.
Authors: Blenman Kim RM 
Published: January 2023

Abstract
Histology is a pathologist’s first step to diagnosing and monitoring diseases but we still rely heavily on methods created in the mid-1800s. A major gap in histology is the ability to assess multiple cellular targets within the tumor microenvironment simultaneously and quantitatively. Immunohistochemistry (IHC) is an essential tool for clinical diagnostic and translational research laboratories. It is typically used to qualitatively assess morphology and cell types present within tissue. IHC involves staining a tissue section with a purified primary antibody followed by a peroxidase- or phosphatase-conjugated secondary antibody and developing the stain with a chromogenic substrate such as 3,3′-diaminobenzidine tetrahydrochloride (DAB). Currently, for clinical diagnostic purposes the majority of IHC staining is a two-color stain, which includes DAB chromogen and hematoxylin dye. Many diseases such as autoimmunity and cancer require assessment of multiple cell types simultaneously in order to fully unravel the complexity surrounding the disease, metastasis, treatment, and management. We describe a high complexity 7-color IHC chromogen-based staining method to assess in situ tumor microenvironments and show how the staining method can be utilized for quantitative analysis. In comparison with current standard clinical laboratory techniques, this method could reduce the amount of biospecimens used, reduce time for processing, and increase the amount of clinically meaningful data extracted from a single biospecimen. The method can be used broadly to assess any disease and any tissue type with an intact tissue microenvironment.
Optogenetic stimulation of brain cholinergic networks suppresses inflammation
Authors: Pavlov Valentin A , Lehner Kurt R. , Silverman Harold A. , Tsaava Tea , Chavan Sangeeta S. , Tracey Kevin J. 
Published: January 2023

Abstract
Brain cholinergic muscarinic acetylcholine receptor (mAChR) signaling regulates peripheral inflammation via the inflammatory reflex (Nat Rev Endocrinol, 2012, 8:743). Here we provide further insight into this regulation by utilizing optogenetic stimulation, a method allowing precise spatiotemporal neuronal activation. Laser light (473 nm) and transgenic mice expressing light activated channelrhodopsin-2 coupled to a fluorescent protein (ChR2-YFP) under the control of the choline acetyl transferase (ChAT) promoter were used. Optogenetic stimulation of the medial septum, a major source of brain cholinergic output to areas with abundant M1 mAChR localization significantly suppressed serum TNF levels, as compared to sham stimulation (P&lt;0.0398) in ChAT-ChR2-YFP mice during endotoxemia. In contrast to ChAT-ChR2-YFP mice, using the same approach in control (non-carrier) endotoxemic mice did not alter serum TNF. In addition, intracerebroventricular administration of the selective M1 mAChR agonist benzyl quinolone carboxylic acid (5 μg/kg) significantly decreased serum TNF, as compared to vehicle administration (P&lt;0.0180) in C57Bl/6 mice during endotoxemia. These results from ongoing studies reveal a role for forebrain cholinergic M1 mAChR-mediated signaling in controlling peripheral inflammation. These findings may be of interest for development of new approaches for selective brain modulation for the treatment of inflammatory conditions.
Sequence-specific detection of different strains LCMV in a single sample using tentacle probes
Authors: Franco Lina S , Holechek Susan A , Caplan Michael , Blattman Joseph N 
Published: January 2023

Abstract
Viral infections often result in quasi-species of virus strains that can have dramatic impacts on disease outcomes or experimental results. However, sequencing of viruses to determine strain composition is time consuming and often cost-prohibitive. Rapid, cost-effective methods are needed for accurate measurement of virus diversity in order to design appropriate treatment, and can be useful for many experimental systems. We have developed a novel molecular method for simple and accurate detection of RNA virus genetic variants using Tentacle probes coupled with quantitative PCR. Tentacle probes are modified molecular beacons that are less expensive and dramatically improve false positive rates in routine qPCR. To validate this approach we have designed two tentacle probes for two strains of Lymphocytic Choriomeningitis Virus (LCMV): the parental Armstrong strain and the more virulent clone-13 strain, which differ by only 3 nucleotide substitutions. Using this approach we were able to distinguish these two strains of LCMV. Furthermore, in virus mixing experiments we were able to detect down to 5% of clone-13 diluted in Armstrong, and vice versa. Thus, we have developed a fast, cost-effective approach for differentiating between closely related viruses.
Diferuloylmethane prevents type 1 diabetes development via inhibition of IFNγ-producing T cells.
Authors: Alard Pascale , Harder James W , Fausnaught Thomas , Kosiewicz Michele M 
Published: January 2023

Abstract
Diferuloylmethane (cucurmin), a natural phenol chemical isolated from the plant Curcuma longa (turmeric, is thought to be anti-inflammatory, and has been shown to ameliorate autoimmune diseases, i.e., multiple sclerosis, rheumatoid arthritis, psoriasis, and inflammatory bowel disease. We examined the effects of diferuloylmethane on type I diabetes development and the immune response in NOD mice. Treatment of NOD dendritic cells (DC) with diferuloylmethane decreased pro-inflammatory cytokines (e.g., IL-12) in vitro, but had no effect on the anti-inflammatory cytokine, IL-10, or DC viability. Interestingly, the ability of diferuloylmethane-treated NOD DC to induce IFNγ-producing T cells in vitro was decreased by comparison to controls. In in vivo studies, NOD mice treated with diferuloylmethane three times weekly exhibited significantly delayed disease (type 1 diabetes) onset, and dramatically decreased disease incidence and increased survival. This protection was associated with a significant decrease in IFNγ production by T cells in the spleen and pancreatic LN. Moreover, the frequencies of CD4+Foxp3+ and CD4+Foxp3+Neuropilin− cells in the pancreatic LN were not increased by this treatment. Overall, our in vitro and in vivo data show that diferuloylmethane suppresses the differentiation of the IFNγ-producing cells that are involved in type 1 diabetes development. Diferuloylmethane may, therefore, have therapeutic potential for the prevention of the development of type 1 diabetes. Experiments are ongoing to test a more effective delivery system that could enhance bioavailability, i.e., diferuloylmethane-encapsulated nanoparticles.
Prolongation of the remission period and reversal of new onset type 1 diabetes with the selective elimination of autoreactive T cells
Authors: Carroll Kaitlin , McNally Jonathan P , Elfers Eileen , Millen Scott , Hildeman David A , Jordan Michael B , Katz Jonathan D 
Published: January 2023

Abstract
Organ-specific autoimmune diseases such as type 1 diabetes (T1D) are principally mediated by autoreactive T lymphocytes targeting self-antigens. The rapid proliferation and concomitant genomic stress that these T cells undergo during their active immune response to self-antigens affords a unique opportunity for their elimination, since these autoreactive T cells have distinct apoptotic sensitivity as a result of intrinsic DNA damage response upon activation. This sensitivity can be harnessed with a combination of p53 potentiation and cell-cycle checkpoint abrogation (PPCA), selectively eliminating these pathogenic cells while leaving naïve and regulatory T cell populations largely untouched. Using this therapy at the onset of T1D, we are able to significantly reverse new onset T1D (≥200 mg/dL blood glucose) and prolong the remission period in the Nonobese Diabetes (NOD) mouse, representing a novel therapeutic approach with significant translational potential for not only type 1 diabetes, but also other T cell-mediated autoimmune diseases.
Gut bacteria modulate dendritic cell priming of diabetogenic T cell effector functions in a mouse model of autoimmune diabetes
Authors: Mohamed Zaynab Sidi , Lau Kenneth , Ardissone Alexandria , Wasserfall Clive , Atkinson Mark , Triplett Eric , Larkin Joseph 
Published: January 2023

Abstract
Increasing evidence implicates a contribution of the microbiome, within the gastrointestinal tract, in the initiation and progression of type 1 diabetes (T1D). Notably, our understanding of mechanisms whereby gut flora interaction with the host immune system can influence T1D pathogenesis remain poorly understood. Comparison of stool samples from the non-obese diabetic (NOD) mouse model of spontaneous T1D and non-obese resistant (NOR) mice revealed distinct bacterial flora communities present within the two strains correlated to a Th17 bias. Given that dendritic cells (DC) are critical in sampling luminal antigens and priming naïve T lymphocyte effector functions, we next interrogated the populations of CD11c+ DC within the lamina propria (LP) of NOD and NOR mice. Phenotypic characterization of LP DC revealed that NOR mice possessed more mature DC denoted by a higher frequency of CD11c+ MHCII+ cells and CD103+ CD86+ cells. We have previously shown that the oral administration of a gut bacterium Lactobaccillus johnsonii (LjN6.2), present in diabetes resistant rodents, was capable of conferring diabetes resistance to diabetes prone rodents while another strain Lactobacillus reuterii (LrTD1) had no effect. Notably, T1D resistance was correlated to a Th17 bias. Hence, we sought to investigate the influence of LjN6.2 on NOD and NOR DC in comparison to LrTD1 in vitro. We found that LjN6.2, but not LrTD1 mediated an increase in IL6 and IL10 cytokine production by DC. Together, these data suggest that gut bacteria have the important ability to shape the maturation and priming abilities of DC. These results could have critical implications regarding the ability of bacterial flora to regulate the generation of a diabetogenic immune repertoire.
Targeting antigen-specific B cells using BAR-transduced cytotoxic and regulatory T cells
Authors: Zhang Ai-Hong , Parvathaneni Kalpana , Yoon Jeongheon , Kim Yongchan , Scott David W 
Published: January 2023

Abstract
Eliminating adverse humoral immune responses in an antigen-specific manner is highly desirable. However, effective means for achieving this goal is still lacking. Analogous to the chimeric antigen receptor (CAR) strategy successfully used in cancer immunotherapy, we have created a chimeric receptor comprising a protein antigen or its domain, linked with the signal transduction domains, CD28-CD3ζ. We termed this receptor as BAR for B-cell receptor (BCR)/antibody receptor. Human CD4 T cells transduced with a BAR containing OVA (OVA-BAR) proliferated in the presence of anti-OVA antibodies. The functional utility of BAR was confirmed in that human CD8 T cells transduced with a C2-BAR that contained the C2 domain of human clotting factor VIII (FVIII) demonstrated killing activity towards co-cultured FVIII C2-specific B cell hybridomas. To test the hypothesis that BAR transduced regulatory T cells (Tregs) could directly and effectively suppress the activity of specific B cells, C2-BAR and OVA-BAR transduced human CD4+ Tregs have been successfully expanded in vitro, and shown to maintain Treg phenotypic markers in terms of co-expression of Foxp3 and Helios. Functional suppression of specific B-cell activity will be tested using a murine FVIII-specific B cells in a xenogeneic model in vitro and in vivo.
Tetrandrine (TET), an immunosuppressive component of Chinese herb, induces tolerogenic dendritic cells and consequently expands regulatory T cells
Authors: Chen Xin , Oppenheim Joost J 
Published: January 2023

Abstract
Tetrandrine (TET) is a chemical component derived from the Stephania Tetrandra, a traditional Chinese medicine with anti-inflammatory and immunosuppressive properties. The underlying mechanism of its immunosuppressive effect remains incompletely understood. There is now compelling evidence that CD4+Foxp3+ regulatory T cells (Tregs) play an indispensable role in the maintenance of immune homeostasis by actively suppressing the activation of effector immune cells. It is also well established that the dendritic cells (DCs) with immunosuppressive property, termed tolerogenic DCs (Tol-DC), are instrumental for the generation and expansion of Tregs. Therefore, we examined the effect of TET on the differentiation of Tol-DCs and its effect on Tregs. We found that, after pre-treatment with non-toxic concentrations of TET, LPS-activated human monocyte-derived DCs failed to mature. TET-treated DCs (TET-DC) expressed high levels of IL-10 and low levels of IL-12, and expressed high levels of ILT3 mRNA, indicative of a typical Tol-DC phenotype. The allostimulatory effect of TET-DC was markedly reduced. Importantly, co-culture with TET-DC, Foxp3 expression by CD4 cells was increased ~2-fold. Intra-peritoneal (i,p.) injection of TET into normal mice, TET treatment also increased the number of functional suppressive Tregs which produced ~4-fold higher IL-10 than Tregs isolated from control mice. Taken together, our study revealed that TET had the capacity to promote the generation of Tol-DC and consequently to expand Tregs. Therefore, induction of immunosuppressive network of Tol-DC and Tregs represent a novel mechanism underlying the anti-inflammatory effect of this Chinese herb-derived naturally occurring compound.
CTLA-4-targeted proteins in differentiating CD8+ T lymphocytes
Authors: Brunner-Weinzierl Monika C , Wissing Josef , Arra Aditya , Pierau Mandy , Klawonn Frank , Jänsch Lothar , Lingel Holger 
Published: January 2023

Abstract
The blockade of inhibitory receptors like Cytotoxic T Lymphocyte-associated molecule-4 (CTLA-4, CD152) on T cells is a potent approach to regain functionality of ineffective immune responses against tumor cells. However, the underlying mechanisms of CTLA-4 mediated attenuation in CD8+ T cells are still incompletely explored. To discover proteins targeted by CTLA-4, we used a CD8+ T cell in vitro activation system combined with iTRAQ proteomic analysis. The mass spectrometry data revealed that CTLA-4-controls central changes in phosphorylation of proteins involved in T cell signaling, RNA processing, DNA replication and microtubule polymerization. Beside other molecules, we identified and verified CTLA-4-mediated post-translational modifications of the AP-1 family transcription factor Fos-related antigen 2 (Fra-2) as well as the regulation of an inhibitor of protein translation and set up a network of proteins altered by CTLA-4. Consequently, CD8+ T cells deficient for CTLA-4-induced targets showed enhanced production of otherwise repressed effector molecules like IFN-gamma and Granzyme B and ultimately proofed to be superior in control of tumor growth. These findings delineate how CTLA-4 is able to modulate CD8+ T cell differentiation and provide new strategies to improve anti-tumor immune therapy.
Nrf2 induces IL-17D to mediate tumor and virus surveillance
Authors: Saddawi-Konefka Robert , Seelige Ruth , Gross Emilie T.E. , O’Sullivan Timothy Edward , Harismendy Olivier , Bui Jack D 
Published: January 2023

Abstract
Cells undergoing xenobiotic or oxidative stress activate the transcription factor Nrf2, which initiates an intrinsic “stress surveillance” pathway involving detoxifying enzymes. We recently found that the cytokine IL-17D effects a form of extrinsic stress surveillance by inducing antitumor immunity, but how IL-17D is regulated remains unknown. Here, we show that Nrf2 induced IL-17D directly in primary tumors as well as during viral infection in vivo. Expression of IL-17D in tumors and virally infected cells is essential for optimal protection of the host as il17d−/− mice experienced a higher incidence of tumors and exacerbated viral infections compared to WT animals. Moreover, activating Nrf2 to induce IL-17D in established tumors led to NK cell-dependent tumor regression. These data demonstrate that Nrf2 can initiate both intrinsic and extrinsic stress surveillance pathways and highlight the use of Nrf2 agonists as immune therapies for cancer and infection.
AK4 influences mTOR activation and the modulation of platelet-derived growth factor receptors
Authors: Larson Candice , Bautista Jonah , Camille Nelianie , Lanning Nathan 
Published: January 2023

Abstract
Cell growth is important for organisms; however, uncontrolled growth results in their demise. The mTORC1 pathway, when turned on, results in the following functions protein and lipid synthesis, lysosome biogenesis, autophagy, and energy metabolism. The inability to adequately turn off mTORC1 leads to cancer. Adenylate kinase 4 has shown to be both a modulator and regulator for maintaining cellular homeostasis in cancer cells. It interacts with the mitochondrial ADP/ATP exchange and promotes stress response proteins for cell proliferation. Platelet-derived growth factor receptors are also ligands that are involved in stimulating growth. They are another extracellular cue shown to drive the cell cycle. I hypothesize that when AK4 is knocked down the mTORC1 pathway will be turned off and inhibit cell growth while down regulating platelet-derived growth factor receptors on the surface of the cell. To test this hypothesis AK4 will be knocked down and the effects of cell growth as well as platelet-derived growth factor receptors will be observed quantitatively. The knockdown of AK4 expression will be modulated by siRNA and the expressions will be visualized via western blot and luminescent-based assay. I expect to find evidence to suggest AK4 in combination of platelet-derived growth factor receptors modulate a role in cancer biology.
The role of STAT3 (signal transducer and activator of transcription 3) in non-small cell lung cancer (NSCLC)
Authors: Heim Lisanne , Andreev Katerina , Balabko Ljubov , Trufa Denis Iulian , Sirbu Horia , Hartmann Arndt , Finotto Susetta 
Published: January 2023

Abstract
Non-small cell lung cancer (NSCLC) is associated with high mortality worldwide underlining the need for novel therapeutic approaches. Signal transducer and activator of transcription 3 (STAT3) is activated via pro-inflammatory cytokines and growth factors (e.g. IL-6, EGF, VEGF) and may promote tumorigenesis by regulating cellular senescence, proliferation and apoptosis. We recently found an upregulation of the IL-6R expression in the tumoral region of patients with lung adenocarcinoma (ADC) as compared to the tumoral region of patients with squamous cell lung carcinoma (SCC). Consistently, increased levels of pSTAT3 were observed in the tumoral region of ADC as compared to SCC patients. Moreover, stimulation of A549 lung adenocarcinoma cells with IL-6 activates pSTAT3 and promotes cell proliferation.
STAT3 is also important for the differentiation of both Foxp3+ T regulatory cells which suppress the anti-tumor immune response and RORγt+ T helper 17 cells, a T cell subtype expanded in lung adenocarcinoma. Indeed, ablation of STAT3 in hematopoietic cells triggers an intrinsic immune-surveillance system that inhibits tumor growth and metastasis.
However, the role of STAT3 in tumor-infiltrating lymphocytes (TIL) in NSCLC is largely unknown. We therefore will investigate the functional effects of STAT3 inactivation in T cells in a murine model of lung adenocarcinoma by using mice with conditional inactivation of STAT3 (STAT3fl/flCD4Cre). To detect STAT3-regulated genes we will isolate TIL of tumor-bearing STAT3 conditional knockout mice and control littermates.
These studies will provide new insights into the functional role of STAT3 in NSCLC and may lead to novel immunotherapeutic approaches for this disease.
Exogenous IL-33 overcomes T cell tolerance in acute myeloid leukemia
Authors: Qin Lei , Dominguez Donye , Chen Siqi , Fan Jie , Long Alan , Zhang Minghui , Fang Deyu , Kuzel Timothy , Zhang Bin 
Published: January 2023

Abstract
Leukemic cells can be recognized and attacked by the immune system. However, emerging studies suggest that dominant peripheral tolerance is a major mechanism of immune escape in disseminated leukemia. Therefore, novel strategies are needed to overcome the tolerance and restore the anti-leukemia T cell function for both leukemia clearance and relapse prevention. Using an established murine acute myeloid leukemia (AML) model, we have demonstrated that systemic administration of recombinant IL-33 dramatically inhibits leukemia relapse and prolongs the survival of leukemia-bearing mice in a CD8+ T cell dependent manner. Exogenous IL-33 treatment enhanced anti-leukemia activity by increasing the expansion and IFN- g production of leukemia-reactive CD8+ T cells. Moreover, IL-33 induced dendritic cell (DC) maturation and activation in favor of its cross presentation ability to evoke a vigorous anti-leukemia immune response. Finally we found that the combination of PD-1 blockade with IL-33 further prolonged the survival with half of the mice achieving complete regression. Our data establish a role of exogenous IL-33 in reversing T cell tolerance and suggest its potential clinical implication into leukemia immunotherapy.