bluuebunny/crossref_metadata_2025_split huggingface dataset:
Bortezomib impacts Notch—miR-155 mediated augmentation of CD8+T Cell antitumor immunity
Authors: Renrick Ariana N. , Thounaojam Menaka C. , Thomas Portia L. , Shanker Anil 
Published: January 2023

Abstract
The immunosuppressive tumor microenvironment dampens host antitumor immunity by multiple mechanisms including interference with the Notch system, which is important for various cell fate decisions and hematopoietic cell differentiation and function. We observed that treatment with bortezomib, a proteasome inhibitor, in mice bearing subcutaneous tumors resulted in an upregulated expression of various Notch signaling components in lymphoid tissues and increased CD8+T lymphocyte IFNγ secretion and expression of effector molecules, perforin and granzyme-B, as well as the T-box transcription factor eomesodermin. Of note, bortezomib reversed tumor-induced downregulation of Notch receptors, Notch1 and Notch2, as well as increased the levels of cleaved Notch intracellular domain (NICD) and downstream targets Hes1 and Hey1 in tumor-draining CD8+T cells. These data suggest that bortezomib can reverse tumor-induced dysfunction of CD8+T cells by its intrinsic stimulatory effects. Our preliminary data also suggest that bortezomib can positively regulate miR-155 expression in CD8+ T cells from mice bearing tumor. Further, miR-155 suppression was found to downregulate bortezomib-induced increase in Notch target genes in T cells. We are currently elucidating how bortezomib affects the expression of miR-155 and its target genes, such as suppressor of cytokine signaling 1 (SOCS1) and inositol polyphosphate-5-phosphatase (SHIP1) that are associated with T cell function. These data provide novel insights on using bortezomib not only as an agent to sensitize tumors to cell death, but also to provide lymphocyte-stimulatory effects, thereby overcoming immunosuppressive actions of tumor on antitumor T cell functions.
The role of thymocyte selection-associated HMG box protein (TOX) in CD8 T cell differentiation and dysfunction
Authors: Scott Andrew Crosbie , Philip Mary , Camara Steven , Shakiba Mojdeh , Levy Olivier , Glickman Michael , Chang Kenneth , Schietinger Andrea 
Published: January 2023

Abstract
CD8 T cells are powerful adaptive immune cells that specifically recognize infected cells and eliminate them. However, in the context of tumors, antigen-specific T cells enter a state of dysfunction allowing tumors to progress. By employing genome-wide transcriptome analysis of dysfunctional tumor-specific T cells (TST) we found that a nuclear factor, thymocyte selection-associated HMG box protein (TOX), was uniquely upregulated during T cell differentiation/dysfunction in progressing tumors, but not during normal CD8 T cell differentiation (e.g. acute infections). TOX is a nuclear DNA-binding factor and member of the high-motility group box superfamily. TOX has been shown to be required for CD4 T cell and NK cell development but its role in peripheral CD8 T cells is not known. We set out to investigate the role of TOX in tumor-specific CD8 T cell differentiation. Using various murine tumor models we found that TOX was uniquely overexpressed in TST within the tumor (TIL) and its overexpression was driven by chronic TCR/antigen stimulation and not by the tumor microenvironment. To understand the role of TOX in CD8 T cell differentiation and dysfunction we are currently conducting TOX genetic knock-out and overexpression studies in vitro and in vivo. The identification and characterization of novel transcriptional regulators such as TOX will provide important insights into the molecular underpinnings defining CD8 T cell dysfunction in tumors.
Circadian Clock disruption promotes TH17 Inflammation and Colon Cancer
Authors: Osman Abu , Saadalla Abdulrahman , Gounari Fotini , Turek Fred , Keshavarzian Ali , Rakshit Kuntol , Matveyenko Aleksey , Khazaie Khashayarsha 
Published: January 2023

Abstract
Circadian clocks regulate numerous physiological processes that vary across the day-night (diurnal) cycle, but whether the circadian clock regulates the adaptive immune response to cancer is mostly unclear. In mouse models of polyposis, and human colorectal cancer patients, we showed that tumor infiltrating Tregs overexpressed b-catenin and RORgt, and secreted IL-17. These Tregs suppressed tumor reactive cytotoxic T cells, and promoted tumor growth. Therefore, we examined how the expression of a dominant negative ClockΔ19 gene affects inflammation and susceptibility to cancer in the APCΔ468mouse model of hereditary polyposis. ClockΔ19 mutant mice had high expression of RORgt+ and b-catenin+ Tregs. These mice showed a notable expansion of Th17 inflammation in vivo upon injection of aCD3. Functionally these Tregs showed pro-inflammatory properties. In addition to these effects, APCΔ468ClockΔ19 mice had increased numbers of CD11b+ cell, Ly6C+CXCR1− monocytes, and Ly6C+Ly6G+ myeloid derived suppressor cells (MDSC) as compared with APCΔ468 mice. APCΔ468ClockΔ19 mice had higher tumor burden in the small intestine and colon than the APCΔ468 mice. Both T-cells and tumors had intense stabilization of nuclear b-catenin and were more aggressive as indicated by abundance of stromal mMCP6+ mast cells (MC), which typically appear at the invasive borders of tumors. Our finding indicating increase in tumor load and worsening inflammation caused by direct ClockΔ19 effects in the epithelium and enhanced TH17 bias in the immune system.
Generation of a therapeutic monoclonal antibody for acute myeloid leukemia by employing the U937 cell line as a surrogate antigen
Authors: Chen Bethany , Kandov Evelyn , Ghebrehiwet Berhane 
Published: January 2023

Abstract
The prognosis of acute myeloid leukemia (AML) has improved for younger patients, but it still remains grim for patients over 65 years old with a likelihood of death within a year of diagnosis. Here, we evaluated the effects of novel anti-gC1qR monoclonal antibodies (mAbs) that were generated against intact U937 cells, which are used here as a model for AML. The U937 cell line is known to overexpress gC1qR, which in turn serves not only as a proliferative molecular signal but also provides a platform for the generation of angiogenic and metastatic factors. Cell viability results reveal that all of the mAbs significantly decrease (p&lt;0.05) the percentage of viable AML cells although the mixture of those mAbs was not significant when compared to the individual mAbs themselves. The anti-U937 antibodies, monoclonal and polyclonal, showed a significant difference (p&lt;0.05) against the anti-purified recombinant gC1qR antibodies. The anti-U937 mAb methodology of immunizing with the mammalian cell instead of the purified protein allowed the mAbs to better target the native endogenous protein present on human cells. Our study not only provided novel and potent mAbs against AML, but also implicated a potentially more efficient technique of creating therapeutic mAbs against cancer cells.
Cryptotanshinone has curative dual anti-proliferative and immunotherapeutic effects on Lewis lung carcinoma
Authors: Yang De , Liu Shuo , Han Zhen , Trivett Anna , Lin Hongsheng , Oppenheim Joost J. 
Published: January 2023

Abstract
Lung cancer is the leading cause of cancer-related mortality, with very limited effective therapeutics. Screening of a variety of Traditional Chinese Medicines (TCMs) for capacity to inhibit the proliferation of human lung cancer A549 cells and to induce the maturation of human dendritic cells (DCs) led to the identification of cryptotanshione (CT), a compound purified from the TCM Salvia miltiorrhiza Bunge. CT can be chemically synthesized. Here CT was shown to inhibit the proliferation of mouse Lewis lung carcinoma (LLC) cells by upregulating p53 and downregulating cyclin B1 and Cdc2 and consequently inducing G2/M cell-cycle arrest of LLC cells. In addition, CT promoted phenotypic maturation of mouse and human DCs with upregulation of costimulatory and MHC molecules, and stimulated DC to produce TNFa, IL-1b and IL-12p70, but not IL-10. CT-induced maturation of DCs depended on MyD88 and also involved the activation of NF-kB, p38, and JNK. CT was effective in the treatment of established LLC tumors and, when used in combination with low-dose anti-PD-L1, can cure LLC-bearing mice with the induction of subsequent anti-LLC specific immunity. Our data indicate that CT has novel unique antitumor and immunotherapeutic activities that may provide a new promising therapeutic for the treatment of human lung cancers.
Altering the cytokine profile in the pancreatic cancer microenvironment with heat shock protein-90 inhibitors to enhance immunotherapy.
Authors: Lesinski Gregory B. , Zhang Yuchen , Farren Matthew R. , Komar Hannah , Ware Brandon , Olson Brian , Zaidi Mohammad , Nagaraju Ganji P. , Akce Mehmet , Alese Olatunji B. , Shaib Walid Labib , Wu Christina Sing-Ying , El-Rayes Bassel 
Published: January 2023

Abstract
One feature of pancreatic ductal adenocarcinoma (PDAC) is the fibrotic stroma, which is a result of interactions between tumor cells, immune suppressive cells and pancreatic stellate cells (PSC). Stroma-derived cytokines including IL-6 represent a significant barrier restraining immunity against PDAC. Targeting pathways in the tumor microenvironment (TME) that regulate cytokine secretion can be an innovative means to improve access of effector immune cells to PDAC. We hypothesize that targeting Hsp90 can modulate the TME, through its ability to target inflammatory signaling and cytokine production by PSC and enhance the efficacy of immunotherapy. The Hsp90 inhibitor XL888 led to a concentration-dependent inhibition of IL-6 and MCP-1 production by PSC in vitro. Concurrently, PSC displayed growth inhibition as determined by MTT assay, and reduced alpha-SMA expression, a marker of cell activation. Inhibition of Jak/STAT and MAPK signaling was confirmed via immunoblot. These data compliment our published results indicating that Hsp90 inhibitors modulate survival pathways and PDAC growth in vitro and in vivo. This has led to an investigator-initiated Phase Ib/II clinical trial of XL888 (Hsp90i) and pembrolizumab (anti-PD-1) now accruing at our institution. Extensive laboratory correlative studies are ongoing to examine the impact of Hsp90 inhibition on anti-PD-1 mediated T cell proliferation, cytokine production, and impact on PSC-derived cytokine signatures. This work is being performed in paired biopsy samples (pre- and on-treatment), and peripheral blood from expansion cohorts of patients with metastatic pancreatic cancer (n=16) or colorectal cancer (n=16).
Transcriptome Responses to Respiratory Virus Infection of Pigs within the Tracheobronchial Lymphnode Following Infection with PRRSV, PCV2 or IAV
Authors: Miller Laura C. , Fleming Damarius S. , Harhay Gregory P. , Kehrli Marcus E. , Lager Kelly M. 
Published: January 2023

Abstract
Porcine reproductive and respiratory syndromevirus (PRRSV) is a major respiratory pathogen of swine that has become extremely costly to the swine industry worldwide, often causing losses in production and animal life due to their ease of spread. However, the intracellular changes that occur in pigs following viral respiratory infections are still scantily understood for PRRSV, as well as, other viral respiratory infections. The aim of this study was to acquire a better understanding of PRRS disease by comparing gene expression changes that occur in tracheobronchial lymph nodes (TBLN) of pigs infected with either porcine reproductive and respiratory syndrome virus (PRRSV), porcine circovirus type 2 (PCV2), or swine influenza virus (IAV) infections. The study identified and compared gene expression changes in the TBLN of 16 pigs following infection by PRRSV, PCV2, IAV, or sham inoculation. Total RNA was pooled for each group and time-point (1, 3, 6, and14 DPI) to make 16 libraries, for analysis by Digital Gene Expression Tag Profiling (DGETP). The data underwent standard filtering to generate a list of sequence tag raw counts that were then analyzed using multidimensional and differential expression statistical tests. The results showed that PRRSV, IAV and PCV-2 infections followed a clinical course typical of experimental infection of young pigs with these viruses. Gene expression results echoed this course, as well as, uncovered genes related to shared and unique host immune responses to the 3 viruses. By testing and observing the host response to other respiratory viruses, our study has elucidated similarities and differences that can assist in development of vaccine and therapeutics that shorten or prevent a chronic PRRSV infection.
The fish adaptive immune response and its suppression by helminths
Authors: Steinel Natalie C. , Bolnick Daniel 
Published: January 2023

Abstract
Helminth infections can immunosuppress hosts, leaving them vulnerable to co-infection. This talk will present a model of helminth immunosuppression: the teleost, stickleback (Gasterosteus aculeatus), and its helminth, Schistocephalus solidus. The teleost adaptive immune response is thought to occur in the melano-macrophage center (MMC). This structure is the proposed site in which B cells proliferate and generate high affinity antibody, homologous to germinal centers in mammals. Stickleback MMCs transiently increase in size following immunization, suggesting that the MMC likely participates in the adaptive immune response. Infection of stickleback with S.solidus, however, suppresses MMC size, suggesting helminth-mediated immunosuppression. To investigate the mechanisms underlying parasite-mediated immunosuppression in teleosts, wild populations of Vancouver Island stickleback were identified that vary in their magnitude of helminth-mediated immunoregulation. Helminth-infected stickleback from Gosling lake show suppressed MMC responses, while Roberts lake fish are refractory to helminth-induced immunosuppression. The resistance of Roberts lake fish to immunosuppression, however, is helminth genotype specific, with some helminth genotypes able to suppress MMC size. F2 hybrid fish were generated from Gosling and Roberts populations and QTL analysis was performed to identify loci associated with differential immunoregulation of the MMC response by helminths. As MMCs are found in most teleosts (and reptiles and amphibians), these findings highlight a new and broadly applicable assay of immunity in both laboratory and wild animals, which does not require the development of species-specific reagents.
Galectin-3 enhances avian H5N1 influenza A virus-induced pulmonary inflammation by promoting NLRP3 inflammasome activation
Authors: Chen Huan-Yuan , Chen Yu-Jung , Wang Sheng-Fan , Lo Tzu-Han , Liu Fu-Tong 
Published: January 2023

Abstract
Virus-induced excessive inflammatory response contributes to severe disease and high mortality rates. Galectin-3, a β-galactoside-binding protein widely distributed in immune and epithelial cells, regulates various immune functions and modulates microbial infections. Here we describe galectin-3 upregulation in mouse lung tissue following challenges with the H5N1 influenza virus. We investigated the effects of endogenous galectin-3 on H5N1 infection and found that survival of galectin-3 knockout (Gal-3KO) mice was comparable to Wild-type (WT) mice following infections. Compared to infected WT mice, infected Gal-3KO mice exhibited less inflammation in the lungs and reduced interleukin-1 beta (IL-1β) levels in bronchoalveolar lavage fluid. We also found that bone marrow-derived macrophages (BMMs) from Gal-3KO mice exhibited reduced oligomerization of apoptosis-associated speck-like proteins containing caspase-associated recruitment domains (ASC), and secreted less IL-1β compared to BMMs from WT mice. However, we observed similar levels of the inflammasome component of nucleotide oligomerization domain-like receptor protein 3 (NLRP3) in two genotypes of BMMs. Co-immunoprecipitation data indicate galectin-3 and NLRP3 interaction in BMMs infected with H5N1. We also observed an association between galectin-3 and NLRP3-ASC complex. Our results indicate that galectin-3 promotes host inflammatory responses and regulates IL-1β production by macrophages via interaction with NLRP3. It suggests that endogenous galectin-3 enhances the effects of H5N1 infection.
Interferon signal transduction in airway barrier epithelial cells is key to controlling human picornavirus infection in the lung
Authors: Zhang Yong , Mao Dailing , Keeler Shamus P. , Wu Kangyun , Agapov Eugene , Holtzman Michael J. 
Published: January 2023

Abstract
Interferon (IFN) signal transduction is required for defense against viral infection, and this signal in airway barrier epithelial cells is assumed to be responsible for controlling common respiratory viruses. However, these conventional concepts have not been rigorously tested in an experimental model given the relative lack of native human respiratory viruses that replicate efficiently in non-human models and the absence of tools to analyze IFN signal transduction specific to airway epithelial cells. To address these issues, we generated a new line of Scgb1a1-Foxj1-Cre-Stat1f/f mice to selectively target IFN signaling in airway epithelial cells and tested these mice for control of infection with the human picornavirus pathogen EV-D68 that might cause more severe infection than human rhinoviruses. Our results show significant replication of EV-D68 with a 1×105-fold increase in viral RNA in lungs of Scgb1a1-Foxj1-Cre-Stat1f/f mice compared to wild-type mice and was similar to viral titer and accompanying lung inflammation and mucous cell formation in conventional Stat1−/− mice. We also found increases in viral titer and inflammation in Scgb1a1-Foxj1-Cre-Stat1f/f mice compared to wild-type mice after the natural mouse pathogen murine parainfluenza virus aka Sendai virus (SeV) infection, but in this case the defect was significantly more pronounced in Stat1−/− mice, consistent with evidence of extra-airway spread of SeV infection. Together, these results indicate a critical requirement for airway epithelial cell capacity for IFN signaling in protection against respiratory picornavirus infection, and to our knowledge, the best evidence to date for the proposed role of airway barrier epithelial cells in host defense.
Early transcriptional responses to influenza virus infections in vivo
Authors: Langlois Ryan A. , Sjaastad Louisa , Fay Elizabeth , Maccietto Marissa , Shen Steve 
Published: January 2023

Abstract
Influenza A virus (IAV) infects a broad range of cell types within the respiratory tract. The cells initially targeted by the virus in a naïve host are the site of primary replication and virus spread. These cells are difficult to detect using replication competent IAV as the virus rapidly spreads to secondary cells. To overcome this obstacle we utilize a fluorescence expressing single cycle IAV (scIAV) to identify the cells initially infected in the mouse lung. Using this tool, we observed two distinct populations of epithelial cells during the early stages of infection: cells with high virus replication and cells with low virus replication. This suggests that some cells within the lung are innately permissive to virus replication while others are able to blunt replication. We have determined that this phenotype is not due to coinfection of a cell with multiple virions, and it appears to be independent of type I IFN. The level of replication is also not dependent on epithelial cell type. There are distinct cellular pathways that are significantly impacted in each population of infected cells compared to uninfected cells. Additionally, there are subsets of interferon-stimulated genes that are specifically upregulated in each population. These data indicate that different levels of virus replication may activate distinct cellular responses to infection. Fluorescent reporter scIAVs could be used to further elucidate the mechanism of cellular permissibility and the early innate immune response to IAV infection.
ZIKV-specific CD8 T cell immunity in humans is affected by DENV pre-exposure
Authors: Grifoni Alba , Pham John , Peters Bjorn , de-Oliveira-Pinto Maria Luzia , de Silva Aravinda , Durbin Anna , Diehl Sean A. , Harris Eva , Crowe James , Busch Micheal , Vivanco-Cid Hector , Graham Barney S. , Turtle Lance , Kallas Esper , Watkins David , Weiskopf Daniela , Sette Alessandro 
Published: January 2023

Abstract
Several studies have characterized humoral immunity to Zika virus (ZIKV) in humans but little is known regarding the corresponding T cell responses to ZIKV. In this study we sought to investigate the ZIKV-specific T cell response in terms of kinetics and viral epitopes targeted and whether pre-existing dengue virus (DENV) T cell immunity can affect these responses. Our results shows that memory T cell responses elicited by prior infection with DENV infection or vaccination recognize ZIKV-derived peptides. The cross-reactive response is explained by the sequence similarity of the two viruses, as the ZIKV peptides recognized are either identical or highly conserved between DENV and ZIKV. In addition, DENV exposure also influences the timing and magnitude of the ZIKV-specific T cell response. In the acute phase of infection, ZIKV-reactive T cells are detected earlier and in greater magnitude than those in DENV pre-exposed patients. Conversely, while the frequency of ZIKV-reactive T cells continues to rise in the convalescent phase in DENV-naive donors, it declines in DENV pre-exposed donors. In addition, ZIKV-specific CD8 T cells form DENV pre-exposed donors selectively up-regulated granzyme B and PD1, as compared to DENV-naïve donors. These results overall suggest a more efficient control of ZIKV replication and/or clearance of ZIKV antigen in patients with previous DENV exposure. Finally, when we focused on the viral epitopes targeted by DENV naïve donors, ZIKV structural proteins (E, prM and C) are major targets of CD8 T cell responses, whereas DENV pre-exposed individuals skew the T cell epitopes primarily in nonstructural proteins.
What have we learned about the dynamics of peptide loading from structures of TAP binding protein, related (TAPBPR)?
Authors: Margulies David H. , Jiang Jiansheng , Natarajan Kannan 
Published: January 2023

Abstract
Understanding the molecular details of peptide loading in the MHC-I pathway is crucial to approaches to manipulate recognition of peptide/MHC-I complexes in treating autoimmunity, tumors, and for vaccine design. Recent insights have been provided by two X-ray structures of TAPBPR, in complex with MHC-I molecules. Although structures determined in the two laboratories are remarkably similar (pdb ID: 5WER and 5OPI), the structural models based on X-ray data obtained at similar resolution, 3.4 and 3.3 Å respectively, reveal a discrepancy in the disposition of the TAPBPR 22–36 loop. This loop was not modeled in 5WER due to a lack of electron density, whereas it was modeled as a 3,10 helix in 5OPI and ascribed a significant function in peptide loading. In an attempt to resolve this structural discrepancy, we reanalyzed 5OPI based on the authors’ deposited structure factors. We calculated the refined average B-factor, an indication of positional uncertainty, as 247 Å2 and 153 Å2 for the 22–36 loop and the entire TAPBPR/MHC-I complex, respectively. Prompted by the unusually high B-factors we reexamined the 22–36 loop by calculating composite omit-simulated annealing maps from deposited structure factors of 5OPI as well as of 5WER. When model bias was eliminated in this way, continuous electron density was not visible in this loop region. Thus, we conclude that although the 22–36 loop of TAPBPR may play a role in peptide exchange, there presently is no reliable crystallographic evidence that this loop is a 3,10 helix and is positioned as modeled in 5OPI. Further structural studies of TAPBPR and of tapasin should help to resolve in detail the mechanism by which these molecules facilitate MHC-I stabilization and peptide exchange.