bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibiting a mutagen: mitigating Activation-Induced Deaminase activity
Authors: Alvarez-Gonzalez Juan Antonio , Crawford Daniel J , Maul Robert W , Kohli Rahul M , Gearhart Patricia J 
Published: January 2023

Abstract
Activation-Induced Deaminase (AID) is a cytosine deaminase which converts cytosine into uracil within stretches of ssDNA. The presence of uracil in DNA initiates a cascade of mutagenic DNA repair to introduce point mutations and DNA double strand breaks. Specific targeting of AID to the immunoglobulin heavy chain loci (Igh) promotes mutagenesis in the antibody variable region for somatic hypermutation (SHM) or switch regions leading to class switch recombination (CSR). However, AID also plays a role in accumulating unnecessary mutations across the genome. Mis-targeting of AID results in the initiation of tumor development and leads to greater drug resistance among cancer cells when continually expressed. To identify an AID inhibitor, we screened ~400,000 compounds in conjunction with the NCATS core facility at NIH. Using FRET based analysis of cytosine deamination, we have identified approximately 150 potential inhibitors of AID catalytic activity. To confirm biological function, we have examined the compounds’ effects on CSR in an in vitro murine B-cell activation assay. Of 30 confirmed inhibitor candidates, we have selected the top 7 to proceed with further characterization to determine if they prevent uracil accumulation in primary mouse cells after ex vivo stimulation. Using these approaches, we hope to identify a small molecule with great efficacy and low toxicity for use as a molecular probe to further characterize AID’s intricacies, or even as a therapeutic.
Harnessing tissue resident memory T cells to combat solid tumors
Authors: Rosato Pamela , Manlove Luke , Nelson Christine , Pennell Christopher , Vezys Vaiva , Masopust David 
Published: January 2023

Abstract
The immunosuppressive tumor microenvironment is barrier to successful cancer therapies. Newly discovered tissue resident memory CD8+ T cells (TRM) function to create a potent immunostimulatory environment to protect against local reinfection. As TRM are present in abundance in nearly every tissue and can be triggered by cognate peptide alone, without adjuvant, we tested whether we could hijack infection-specific TRM in tumors to reverse the immunosuppressive tumor microenvironment and enhance existing immunotherapies. Mice with established vesicular stomatitis virus (VSV)-specific skin TRM were challenged with the transplantable B16 melanoma cell line. We find VSV-specific CD8+ T cells within established tumors and by 12 hours of cognate peptide delivery, these cells upregulate IFNγ, CD25, and granzyme B. This led to 1) a 10-fold increase in intratumoral NK cells, 2) bystander memory CD8+ T cell recruitment to the tumor, 3) tumor NK cell and CD8+ T cell activation through granzyme B upregulation and 4) DC activation in tumor draining lymph nodes through CCR7 and CD86 upregulation. Ongoing studies using transplantable and autochthonous melanoma models are testing whether TRM immunotherapy synergizes with CAR T cell and checkpoint blockade immunotherapies for eradication of recalcitrant tumors. These results demonstrate proof of principal efficacy for exploiting infection-specific TRM as a tumor immunotherapy.
Eomesodermin together with KIR/NKG2A membrane receptors and CD49d as an useful strategy to identify NK-like CD8(+) T cells with innate features in humans: focusing on cancer diseases
Authors: Herbelin Andre , Cayssials Emilie , Jacomet Florence , Nunez Nicolas Gonzalo , Basbous Sara , Lefevre Lucie , Abdallah Myriam , Piccirilli Nathalie , Lavoue Vincent , Catros Véronique , Piaggio Eliane , Gombert Jean-Marc , Barbarin Alice 
Published: January 2023

Abstract
Among the burgeoning family of unconventional T cells, innate-like CD8(+) T cells bear a memory phenotype without having encountered a foreign antigen and can respond to innate-like IL-12+IL-18 stimulation. Although the concept of innate memory (IM) CD8(+) T cells is now well-established in mice, whether an equivalent memory NK-like T-cell population exists in humans remains under debate. We recently reported that CD8(+) T cells responding to innate-like IL-12+IL-18 stimulation and co-expressing the transcription factor Eomesodermin (Eomes) and KIR/NKG2A membrane receptors with a memory/EMRA phenotype may represent a new, functionally distinct innate T cell subset in humans.
Here, we proposed Eomes together with KIR/NKG2A and CD49d as a signature to standardize the identification of this innate CD8(+) T-cell subset in humans. Focusing on cancer diseases, we have provided new insights into the potential role of these IM CD8(+) T cells in a physiopathological context in humans. Based on empirical data obtained in cases of chronic myeloid leukemia, a myeloproliferative syndrome controlled by the immune system, and in solid tumors, we have observed both the possible contribution of IM CD8(+) T cells to cancer disease control and their susceptibility to tumor immune subversion. Finally, we have investigated the involvement of IL-4 and IL-15 in the generation of the IM CD8(+) T cell pool and its dependency on promyelocytic leukemia zinc finger (PLZF)-expressing invariant Natural Killer T (iNKT) cells.
All in all, this study significantly contributes to understanding of the physiopathological role of NK-like CD8(+) T cells in humans and raises the question of the possible involvement of an iNKT/IM CD8(+) T cell axis in cancer.
Calnexin induces impairment of proliferation and effector function of CD4+ and CD8+ T cells and promotes tumor growth
Authors: Ma Da , Chen Yi Chen , Wang Zhi 
Published: January 2023

Abstract
Elucidation of molecular mechanism underlying the inhibited infiltration effector function of T cells induced is essential for improvement of anti-tumor immunotherapy. Here, we found that Calnexin, a ER chaperon protein, is significantly up-regulated in OSCC tumor cells and multiple other tumors. Up-regulation of Clanexin in tumor cell membrane links with limited infiltration of T cells in OSCC tumor core and poorer survival of OSCC patients. Importantly, we found that Calnexin mediates inhibition of proliferation of CD4+ and CD8+T cells and effector functions of expression of IFN-γ, TNF-α, IL-2 by T cells. While knock-down of Calnexin enhanced the infiltration and effector functions of T cells in tumor cores and conferred better control of tumor growth, treatment of recombinant Calnexin induced impairment of infiltration and effector functions of T cells in tumor cores and promotes tumor growth in mice. Thus, this work uncovers a previously unknown mechanism in which infiltration of T cells and effector functions of T cells are inhibited by Calnexin expressed in tumor cells, and implicates that Calnexin may serve as an important target for improvement anti-tumor immunotherapy.
NKG2D signaling certifies CD8 T cells to become memory cells
Authors: Perez Cynthia , Prajapati Kushal , Burke Brianna , Plaza-Rojas Lourdes , Guevara-Patiño Jose Alejandro 
Published: January 2023

Abstract
The ability to form memory is a major characteristic of the adaptive immune system. NKG2D, as a costimulatory receptor for CD8+ T cells, was shown to rescue memory formation in absence of CD4+ T cell help. However, the exact function of NKG2D on CD8+ T cells and the importance of NKG2D signaling for memory formation are not known. To recapitulate a full course of CD8+ T cell activation, from priming to memory recall, we immunized C57BL/6 mice with peptide-pulsed dendritic cells and followed the CD8+ T cell responses at priming, effector and memory phases. Using NKG2D deficient (NKG2D-KO) CD8+ T cells, we found that NKG2D signaling was not necessary to achieve CD8+ T cell priming. However, NKG2D-KO CD8+ T cells showed lower effector functions (reduced in vivo kill and reduced cytokine secretion) compared to wildtype CD8+ T cells. Blocking NKG2D signaling at the effector phase with a single injection of a non-depleting anti-NKG2D antibody recapitulated the results observed with NKG2D-KO CD8+ T cells. Importantly, the CD8+ T cells that lacked NKG2D signaling at effector phase failed to differentiate into protective memory cells. These results indicate that NKG2D signaling certifies CD8+ T cells to become memory cells and that this certification occurs during the effector phase. This study was supported by the PO1 CA154778, the RO1 CA188395 and the Vogol Foundation.
Administration of Sparstolonin B- a TLR4 antagonist, attenuates hepatic fibrosis by inhibiting TLR4 mediated hepatic fibroblast proliferation.
Authors: Dattaroy Diptadip , Chandrashekaran Varun Chandrashekaran , Seth Ratanesh Kumar , Alhasson Firas , Fan Daping , Nagarkatti Mitzi , Nagarkatti Prakash , Diehl Anna Mae , Chatterjee Saurabh 
Published: January 2023

Abstract
Previous studies have shown that TLR4 activation accentuates hepatic inflammation and fibrosis in nonalcoholic steatohepatitis (NASH). Here we report an anti-fibrotic activity of Sparstolonin B (SsnB)-a TLR4 antagonist, derived from Chinese herb Sparganium stoloniferum. We hypothesize that, SsnB decreases TLR4 induced inflammation and fibrosis in liver by inhibiting TLR4 signaling, TGFβ signaling and by inducing senescence in fibrogenic hepatic stellate cells. Here we used Bromodichloromethane (BDCM) induced liver fibrosis model where diet induced obese (DIO) mice were administered with BDCM (a CYP2E1 activator) for 4 weeks. A similar group were administered SsnB for 4 weeks along with BDCM. Picrosirius stain and IHC of fibronectin images showed a robust decrease of extracellular matrix deposition upon SsnB administration. Mechanistically, we found that SsnB decreased fibroblast proliferation by inhibiting TLR4-PI3K akt mediated fibroblast proliferation-as was shown by increased PTEN, p53, p21 and decreased cyclin D1, MDM2 protein expression in SsnB treated mice. Interestingly, we found a parallel mechanism in which SsnB increased BAMBI (a TGFβ pseudo-receptor) protein expression. Inhibition was possibly mediated by targeting the TLR4 dependent MYD88-NfkB pathway and TGFβ signaling was decreased following attenuated SMAD2/3-SMAD4 co-localization in the nucleus. SsnB increased gene expression of cell senescence markers (p53, p21), decreased alpha SMA gene expression and increased anti-fibrotic matrix metallo protease 2 (MMP2) gene expression in rat stellate cells in vitro. BAMBI gene and protein expression- a negative regulator of pro-fibrogenic TGFβ signaling, were also increased by SsnB treatment in vitro.
The transcriptional response of Kupffer Cells to fat and cholesterol is non-canonical and distinct from bone-marrow derived macrophages
Authors: McGettigan Brett M , McMahan Rachel H , Rosen Hugo R 
Published: January 2023

Abstract
Diets high in fat and cholesterol can cause hepatic lipid accumulation and inflammation known as non-alcoholic steatohepatitis (NASH). Liver macrophages putatively drive disease by responding to PAMPS/DAMPS and producing pro-inflammatory cytokines. However, recent studies show that there are two principle subsets of macrophages in a NASH liver: Tissue-resident macrophages that are embryonically derived (Kupffer Cells; KCs) and infiltrating macrophages that are adult-derived from bone marrow (IMs). Furthermore, the role that each subset plays in disease pathogenesis is unclear. We FACS sorted the KC (F4/80hiCD11blow) and IM subsets (F4/80lowCD11bhi) from mice fed a high fat/cholesterol (HFHC) diet as well as those from healthy controls. RNA was extracted and gene expression was analyzed using RNA sequencing techniques. HFHC mice had a prominent IM population that was absent in controls. IMs were the primary producers of cytokines and chemokines. KCs did not upregulate pro-inflammatory cytokines in response to dietary fat and cholesterol, yet upstream analysis revealed that lipid metabolism and bile acid signaling pathways were more active than IMs. 3,617 genes were differentially expressed KCs, while 3,421 genes were differentially expressed in IMs. Genes known to be expressed by restorative macrophages were enriched in KCs (CD163, MARCO, TIMD4, MMP12, MMP13), while pro-inflammatory and M1 genes predominated in the IM population (TNF, SPP1, CXCL2, CCL2, S100A8). In conclusion, bone marrow-derived macrophages have a transcriptional profile consistent with the previously reported role of liver macrophages in NASH pathogenesis. However, Kupffer Cells respond to dietary stress in a non-canonical manner.
Adipokine Leptin mediated NOX-2 promotes kidney inflammation in nonalcoholic fatty liver disease via miR21-dependent mesangial cell immune activation
Authors: ALHASSON FIRAS A , DAS SUVARTHI , Chandrashekaran Varun , Dattaroy Diptadip , Seth Ratanesh Kumar , Nagarkatti Mitzi , Nagarkatti Prakash , Chatterjee Saurabh 
Published: January 2023

Abstract
High insulin and leptin resistance followed by underlying inflammation is often ascribed to the ectopic manifestations in nonalcoholic fatty liver disease (NAFLD) but the exact molecular pathways remain unclear. We have shown previously that CYP2E1-mediated oxidative stress and circulating leptin in NAFLD is associated with renal disease severity. Extending the studies, we hypothesized that high circulatory leptin in NAFLD causes renal mesangial cell activation and tubular inflammation via a NOX2 dependent pathway that upregulates proinflammatorymiR21. High fat diet (60%kcal) was used to induce fatty liver phenotype with parallel insulin and leptin resistance in WT, Leptin knockout (ob/ob) and p47 phox knockout mice. Kidneys were collected after pathology showed NASH-like phenotype and accelerated oxidative stress in the liver. The kidneys were probed for mesangial cell activation and tubular inflammation. Results showed that NAFLD kidneys had significant increases in α-SMA, a marker of mesangial cell activation, miR21 levels, tyrosine nitration and renal inflammation while they were significantly decreased in leptin and p47 phox knockout mice. Micro RNA21 knockout mice showed decreased tubular immunotoxicity and proinflammatory mediator release. Mechanistically, use of apocynin or phenyl boronic acid (FBA) or DMPO or miR21 antagomir inhibited leptin primed-miR21-mediated mesangial cell activation in vitro suggesting a direct role of leptin-mediated NOX-2 in miR21-mediated mesangial cell activation. Finally JAK-STAT inhibitor completely abrogated the mesangial cell activation in leptin-primed cells suggesting that leptin signaling in the mesangial cells depended on the JAK-STAT pathway.
Characterisation of gene-modified CD3-enhanced CD4+ T cells for cancer immunotherapy
Authors: Piapi Alice , Nicholson Emma , Morris Emma C , Stauss Hans J 
Published: January 2023

Abstract
TCR gene transfer is used to redirect the antigen specificity of T lymphocytes towards known tumour antigens. TCR gene therapies in murine studies have shown promising results. However, in the clinic they often generate sub-optimal responses, when compared to treatments with tumour infiltrating lymphocytes. Previous work to improve TCR gene therapy has demonstrated that transferring additional CD3 genes increases TCR expression of both endogenous and introduced TCR in CD4+ and CD8+ T cells. In vivo experiments demonstrated that CD8+ T cells, transduced with TCR and additional CD3 were more effective in tumour protection than T cells transduced with the TCR alone. Whilst, CD4+ T cells transduced with TCR and additional CD3 initially showed improved tumour protection, lethal toxicity, unrelated to tumour burden, was later observed. To investigate the effects of CD3 overexpression, CD3 genes only (no TCR genes) were transferred into purified CD4+ and CD8+ T cells. Following adoptive transfer, CD3-enhanced CD4+ T cells survived for longer and were recovered in higher percentages in spleen, lymph nodes, bone marrow and liver, compared to CD3-enhanced CD8+ T cells and mock transduced CD4+ T cells. The same trend was also seen in competition experiments where mice received a 1:1 ratio of CD3-enhanced CD4+ T cells and mock-transduced CD4+ T cells. Interestingly, this was observed despite a twofold downregulation of TCR levels in the CD3-enhanced CD4+ T cells, compared to their pre-transfer TCR levels. Current experiments are aimed at dissecting the mechanisms responsible for, and the physiological implications of the observed TCR downregulation.
Ovarian cancer ascites induces a T cell suppressive phenotype in mature neutrophils: a potential barrier to anti-tumor immunity
Authors: Singel Kelly L , Khan ANM Nazmul H , Moysich Kirsten B , Odunsi Kunle , Segal Brahm H 
Published: January 2023

Abstract
Ovarian cancer (OC) is often diagnosed at advanced stages and presents with ascites. Necrosis is a hallmark of advanced cancer and releases DAMPs that activate innate immune responses (e.g., neutrophils). While neutrophils are critical in host defense, their role in the context of tumor is less understood. CTLs correlate with better outcomes in advanced OC, and immunotherapy for cancer is based on augmentation of CTL responses. Our prior studies showed that granulocytes from ascites of patients with newly diagnosed OC suppressed donor T cell proliferation ex vivo. The current work probes mechanisms for this suppression.
In ascites from patients with newly diagnosed OC, ~90% of cells were inflammatory (CD45+). Neutrophils comprised ~15% of CD45+ cells and the neutrophil:CD8+ T cell ratio was 1.5:1. In ex vivo studies, cell-free ascites (ASC; 500g) attracted normal donor neutrophils (NDN). While neither ASC nor NDN alone impaired CD3/CD28-stimulated normal donor T cell proliferation, ASC (10/15 patients) when combined with NDN (1:1) completely suppressed T cell proliferation. Suppression by ASC + NDN required cell contact and was lost if NDN were added after overnight CD3/CD28 stimulation of T cells. Finally, heat-inactivating ASC (56C, 1h) abrogated the ability of NDN to suppress T cell proliferation.
Here we provide evidence that ascites recruits neutrophils to the tumor microenvironment and induces a suppressive phenotype of neutrophils that requires cell contact and heat-labile cell-free constituents. Ongoing studies aim to identify the putative protein(s) in ascites and the signaling effects on neutrophils. Our long-term goal is to develop novel approaches to overcome this suppression and improve durable anti-tumor immunity.
Rosiglitazone, a peroxisome proliferator-activated receptor-gamma agonist, reduces but does not prevent, the pulmonary CD4+ effector T-cell response in a murine model of asthma
Authors: Maslanka Tomasz , Otrocka-Domagała Iwona , Zuśka-Prot Monika , Gesek Michał 
Published: January 2023

Abstract
Nuclear factor (NF)-κB is regarded as a promising target for the development of a novel therapeutic strategy in asthma. Peroxisome proliferator-activated receptor gamma (PPAR-γ) agonists inhibit the activity of NF-κB. Therefore, these agents are potent therapeutic agents for the treatment of asthma. It was found that rosiglitazone (ROS), a PPAR-γ agonist, attenuated airway inflammation in a murine model of asthma. The present study attempts to shed light on the involvement of CD4+ effector (Teff) and regulatory (Treg) T cells in the anti-asthmatic action of ROS in ovalbumin (OVA)-induced airway allergic inflammation (AAI) in BALB/c mice. We studied the effect of a 10-day treatment with ROS on the development of AAI and the absolute counts of Treg (Foxp3+CD25+CD4+) cells and activated Teff (Foxp3−CD25+CD4+) cells in the mediastinal lymph nodes (MLNs) and lungs of OVA-sensitized/challenged mice. The severity of AAI was only reduced, but not abolished, by the treatment with ROS. The absolute number of CD4+ Treg and Teff cells was increased in the MLNs and lungs of OVA-sensitized/challenged mice treated with the vehicle as compared to healthy mice. No differences were found in the count of CD4+ Treg cells between vehicle- and ROS-treated OVA-sensitized/challenged mice. The treatment with ROS reduced, but did not fully prevent, the OVA-induced increase in the count of activated CD4+ Teff cells in the MLNs and lungs. The results suggest that the impairment of the activation and clonal expansion of CD4+ Teff cells in the MLNs is involved in the anti-asthmatic action of PPAR-γ agonists. This effect does not seem to be mediated through increased recruitment of CD4+ Treg cells into the MLNs and lungs and/or local generation of Treg cells.
T cell mediated mechanisms underlying HSC dysfunction and bone marrow failure
Authors: Hoyer Katrina K , Gravano David M 
Published: January 2023

Abstract
Bone marrow failure occurs when the bone marrow cannot produce sufficient numbers of red blood cells, white blood cells or platelets. Bone marrow failure disorders are rare, yet serious conditions, and are often thought to be the result of immune destruction within the bone marrow. IL-2-deficient mice, which have a deficit in functional regulatory T cells, develop spontaneous bone marrow failure. We show that disease occurs in these mice with a specific accumulation of T cells in the bone marrow, a shift in the CD4/CD8 ratio, IFNγ-mediated hematopoietic stem cell dysfunction, and antibodies directed against bone marrow and bone tissue, recapitulating human disease, and suggesting that this disease may be caused by self-reactive immune responses. Disease onset is rapid at 12 days of age with death by 27 days, and T cell responses are essential for disease progression, providing a novel model for defining the immune cell dysregulation in this autoimmune disease. We use this model to define the mechanisms underlying the deficit in myelopoiesis observed during bone marrow failure, including the contribution of activated lymphocyte subsets to disease induction. Our study describes the influence of IFNγ, CD4 and CD8 T cell subsets, and local bone marrow proliferation and migration on the kinetics and severity of disease.
S100a9-deficiency accelerates lupus disease and renal failure in male (NZB x NZW)F1 mice
Authors: Dand Hardik A , Davison Laura M , Alberto Andres , Vogl Thomas , Jorgensen Trine N 
Published: January 2023

Abstract
All (NZB x NZW)F1 (BWF1) female mice develop lupus-like disease, compared to ~30% of male BWF1 mice. Our previous research described an immunosuppressive effect of Gr1hiCD11bhi cells in male BWF1 mice. Specifically, as BWF1 mice age, female Gr1hi cells lost their ability to suppress altogether, while antibody-mediated depletion of Gr1hi cells accelerated disease development in male mice only. Reverse transcriptase-PCR studies revealed upregulated S100a9 and S100a8 mRNA levels in male flow-sorted Gr1hi cells as compared to cells from female BWF1 mice and non-autoimmune stains, leading us to hypothesize that S100a8/a9 produced by male Gr1hi cells is immunosuppressive. To study this, we created S100a9-deficient BWF1 mice. Our results showed spleen weight and splenocyte count, indicators of lupus advancement, were significantly higher in S100a9−/−male mice. Additionally, increased IgG-IC deposition in kidney glomeruli of S100a9−/−male mice was observed along with a significant increase in serum anti-chromatin IgG levels. Deficiency of S100a9 also resulted in increased frequencies of memory B cells and plasma cells in male mice, but not in female mice. Interestingly, S100a9-deficiency caused an accumulation of Gr1hiCD11b+ splenocytes in male mice. We speculated that this is due to a lack of trafficking of Gr1hi cells into the glomerulus of the kidney due to S100a9-deficiency. Subsequent IHC confirmed the lack of S100a9 expression in kidneys from S100a9-deficient mice. Finally, H/E and Trichrome stainings indicated accelerated renal failure in S100a9−/−mice. In conclusion, S100a9 may function as a chemoattractant of Gr1hi cells exerting immunosuppression and protection against lupus-like renal disease in male BWF1 mice.