bluuebunny/crossref_metadata_2025_split huggingface dataset:
Small molecule inhibitor screen of MHCII processing pathways using influenza virus A/PR/8/34 (130.11)
Authors: Miller Michael , Eisenlohr Laurence 
Published: January 2023

Abstract
During a viral infection, antigen presenting cells (APCs) process foreign intracellular material into short peptides (epitopes) and present them on the cell surface for recognition by CD4+ and CD8+ T cells. We have previously identified three immunodominant epitopes derived from the surface proteins of the influenza virus strain A/PR/8/34. S1 and S3 are derived from endocytosed hemagglutinin protein whereas NA79 is derived from endogenously-produced neuraminidase. Although these three epitopes are restricted to the same MHCII molecule I-Ed, their processing pathways are reported to be dependent on different cellular components such as H2-DM, TAP, and endosomal proteases. To discover additional components of these distinct antigen presentation pathways, we screened a small molecule library for compounds that would selectively inhibit presentation of S1, S3, and NA79. We screened 8,000 compounds from the NCI/DTP Open Chemical Repository and identified 239 primary leads for specific inhibition of epitope presentation ranging from 50% - 100 %. To further validate the primary leads, we performed dose responses to identify 27 potent inhibitors of our known epitopes. Future work will involve purification methods to identify cellular targets that directly interact with our small molecule hits.
Human invariant chain Iip35 isoform expressed in Ii-deficient mice fully restores thymic selection (130.16)
Authors: Genève Laetitia , Labrecque Nathalie , Thibodeau Jacques 
Published: January 2023

Abstract
During biosynthesis, invariant chain (Ii) associates with MHC II molecules, prevents peptide binding in the endoplasmic reticulum (ER) and targets the complexes to the endocytic pathway. Iip35, a human-specific isoform, displays a cytoplasmic ER retention motif that is masked upon MHC II binding. Little is known on the role of this ER retention motif and we asked if Iip35 alone was capable of supporting the functions of Ii. We generated a Tg mouse expressing the human Iip35 isoform (Tgp35). Breeding with murine Ii deficient mice produced mice expressing only Iip35 (Tgp35/mIiKO). In these mice, Iip35 is highly expressed by about 30 % of B cells, 50% of splenic DC and 74% of IFNγ-induced macrophages. Interestingly, B cells expressing Iip35 were lost during the maturation process and not found in the T2 stage. We have demonstrated that Iip35 interacts with both murine Ii and MHC II. Also, we observed MHC II compacts forms typical of tight peptide binding. Iip35 partially rescued MHC II cell surface expression in mIiKO mice and was sufficient to fully restore thymic selection as judged by normal CD4+ T cell numbers in thymus and periphery. In contrast, as observed in mIiKO mice, no antigen-specific antibody production was observed in Tgp35/mIiKO following injection of Ova. Altogether, our results suggest that the Iip35 isoform is able to restore thymic selection but may not be sufficient to perform all Ii functions.
B cell receptors micro-cluster by antigen cross-linking and macro-cluster in a cap by biased transport (130.2)
Authors: Alla Srinivas , Raychaudhuri Subhadip 
Published: January 2023

Abstract
It is known from experiments that, in the presence of soluble antigens B-cell receptors (BCRs) assemble into small micro clusters and then structure into a cap macro cluster. However the underlying mechanisms of the antigen interaction with the BCRs and their cluster formation remain unclear. Such mutual attraction between two adjacent BCR molecules could arise, among other possibilities, due to cross-linking by bivalent soluble antigens. Recently, we have developed and studied a Monte Carlo model of B cell receptor clustering caused by binding and cross-linking of soluble antigens. The results of our study demonstrate the formation of small micro-clusters of BCR molecules (typically of size 2-10 molecules). But antigen cross-linking only is not adequate enough for the formation of large macro-clusters. A biased diffusion may drive the BCRs to form large macro-cluster. A simple model of biased diffusion where BCR molecules experience a biased directed motion towards the (a) largest cluster or (b) towards a random BCR micro-cluster is then applied, which results in a single macro cluster (cap) of receptor molecules. The mechanisms for both types of biased transport are compared using suitable network based metrics. We also develop and use appropriate network measures to analyze the effect of BCR and antigen concentrations on the receptor clustering, the stability of the formed clusters over the time, and size of BCR-antigen cross-linked chains.
Drug Controlled Antigen Degradation Defines Sources of MHC Class I Peptides and Enables Pathway Discovery (130.29)
Authors: Dolan Brian , Bennink Jack , Yewdell Jonathan 
Published: January 2023

Abstract
Defective Ribosomal Products (DRiPs) are an important source of MHC class I peptide ligands. How and where DRiPs are generated is an unanswered question with important implications for rationale design of T cell vaccines. To better understand DRiP antigen processing, we developed a novel antigenic protein, based upon the ProteoTuner technology, whose metabolic stability is controlled by a small molecule termed Shield-1. We show robust MHC class I antigen presentation derived from this shield controlled recombinant antigenic protein (SCRAP) regardless of the overall stability conveyed by shield. We use Shield-1 to generate a large pool of SCRAP, that is rapidly degraded upon Shield-1 removal, and show that these “retirees” are a much less efficient source of peptides than DRiPs. These findings demonstrate that true DRiPs are the source for a large proportion of peptide-MHC class I complexes. We also exploit the unique properties of this system to screen for chemicals and siRNAs that specifically inhibit DRiP antigen presentation, and have generated results demonstrating a role for deubiquitinating enzymes in DRiP antigen presentation.
Mechanisms of antigen presentation in recombinant adeno-associated virus (rAAV) vector mediated vaccination and gene therapy (130.36)
Authors: Xu Dan , Velazquez Victoria , Schaffer David , Kaspar Brian , Walker Christopher 
Published: January 2023

Abstract
Recombinant adeno-associated virus (rAAV) vectors are being considered for vaccination against infectious diseases and gene replacement therapy. Direct transduction of dendritic cells (DC) is thought to be required for initiation of CD8+ T cell immunity, but a role for antigen cross-presentation has not been excluded. Here we evaluated the requirement for DC transduction in T cell priming important to the outcome of vaccination and gene therapy strategies. Persistent eGFP expression was established in murine skeletal muscle after rAAV vector transduction. CD8+ T cells were primed but failed to clear transduced myocytes. Remarkably eGFP-positive DC were detected in draining lymph nodes (DLN) after vector transduction. They expressed co-simulatory and inhibitory molecules that regulate T cell priming. To determine if the DC were transduced by rAAV, F1 (H2bXd) mice were lethally irradiated several weeks after vector transduction of muscle and reconstituted with parental (H2b or H2d) bone marrow. eGFP-positive DC expressing only parental (H-2b or H-2d) class I molecules were visualized in the DLN after bm reconstitution. This result indicates that DC were not transduced by the vector, but instead captured antigen in the muscle before migrating to the DLN. Moreover T cells primed in these bm-reconstituted animals were restricted only by parental class I molecules, indicating a critical role for parental DC but not transduced myocytes in antigen presentation.
Humoral responses to haptenated and native proteins: The role of MyD88 (130.39)
Authors: Snowden Pilar 
Published: January 2023

Abstract
Recently, Palm and Medzhitov claimed that antibody responses to native proteins require MyD88 whereas responses to haptenated protein do not. Haptens are small antigenic molecules that are capable of inducing humoral responses only when covalently coupled to larger, “carrier” molecules; most carrier molecules are proteins. Antibodies elicited by carrier-hapten can be hapten- or carrier specific, or can recognize the hapten-carrier junction. Often, the majority of antibody induced by hapten-carrier is hapten-specific, but haptenation was not thought to alter fundamentally the nature of the carrier antigen. To compare the capacity of native and haptenated protein to elicit antibody responses in MyD88 deficient and congenic control mice, we immunized cohorts of animals with chicken γ-globulin (CGG) or CGG haptenated with (4-hydroxy-3-nitrophenyl)acetyl (NP9-CGG) in alum. Subsequently, we compared the humoral responses of control and MyD88-/- mice by quantifying serum antibody and germinal center (GC) responses. Under these experimental conditions, control and MyD88-/- mice mounted comparable IgG and GC responses to CGG and NP9-CGG. In all cases, NP9-CGG elicited higher amounts of NP-specific antibody compared to that against CGG; native CGG elicited higher levels of carrier antibody but less than the anti-NP response. By FACS, GC responses were comparable in both strains also. Our findings suggest that immune responses to native CGG and NP9-CGG are MyD88 independent.
Carbohydrate Tumor Antigen Vaccines using Unique Strategies (131.27)
Authors: Rittenhouse-Olson Kate , Abdulla Julia , Heimburg-Molinaro Jamie , Sahoo Padmini , Almogren Adel , Roy Rene , Chang Richard , Morey Susan , Barchi Joseph 
Published: January 2023

Abstract
The Thomsen-Friedenreich antigen (TF-Ag), a carbohydrate tumor associated antigen, is differentially expressed on carcinomas including those of the breast, colon, and prostate. It is involved in adhesion and metastasis and even low levels of naturally formed antibody to TF-Ag is related to better prognosis. We hypothesize that vaccination to create antibody to TF-Ag may create a survival advantage for patients with TF-Ag+ tumors. The innovation of this work is that both peptide mimics and other novel constructs have been utilized as immunogens. Peptide mimics of TF-Ag, with and without C3d N- terminal peptide constructs and synthetic TF-Ag-Muc 4 constructs with the C3d N- terminal peptide, were prepared to target T cells, and CD21+ splenic B cells and follicular dendritic cells for an increased response and memory B cell production. CD21 is the C3d receptor and MUC-4 is a tumor associated peptide. The immune response was measured using TF-Ag -BSA in enzyme immunoassays. The significance of antibody production to TF-Ag can be seen in the fact that passive immunotherapy with a monoclonal Ab to this Ag decreases lung metastasis and extends survival time of mice bearing breast cancer. The improved survival time is of particular interest because the McAb was not cytotoxic, and the enhanced survival was due to blocking metastasis. A successful vaccine would replicate this inhibition of metastasis and in addition provide cytotoxic effects to improve prognosis.
Tumor immunotherapy using chimeric receptor redirected allogeneic T cells across MHC barriers (131.39)
Authors: Marcus Assaf , Waks Tova , Eshhar Zelig 
Published: January 2023

Abstract
We present a strategy for adoptive immunotherapy using fully mismatched allogeneic chimeric receptor redirected T cells (allo-'T-bodies') in the absence of Graft vs. Host disease (GvHD). The strategy hinges on creating a therapeutic time-window using mild preconditioning which on one hand allows the allogeneic T-bodies enough time to effectively attack the tumor but on the other hand does not ablate the Host vs. Graft response (HvG) ensuring the eventual rejection of the allogeneic T cells thus preventing GvHD. We found that transfer of fully mismatched allogeneic T cells expressing a HER2/neu specific chimeric receptor following mild preconditioning roughly tripled the median survival of mice bearing pulmonary Renca-HER2 metastases with around 50% of mice surviving long term (&gt;200 days) in the absence of GvHD mortality. We show that allo-T-bodies provide comparable benefit to syngeneic T-bodies, and are far superior to non-specific allogeneic T cells, demonstrating that the response is indeed tumor-specific and not merely allo-specific. Taken together these data provide proof of principle for safe and effective allogeneic adoptive therapy allowing for the use of 'universal effector cells' as a standardized treatment for cancer.
Resveratrol enhances CD8+ T cell mediated effector responses by inducing type 3 deacetylase dependent mTOR kinase activity to promote T-bet expression (132.3)
Authors: Rao Rajesh , Li Qingsheng , Shrikant Protul 
Published: January 2023

Abstract
Caloric restriction (CR) augments cellular immunity and extends lifespan in mammals; however the mechanisms underpinning these observations remain unclear. The CR mimic; resveratrol, activates SIRT-1; a NAD+ dependent type III deacetylase, for promoting life-span. Herein, we show that resveratrol present during antigen stimulation of naïve CD8+ T (OT-I) cells enhances T-bet dependent type I effector maturation. Resveratrol mediated enhanced T-bet expression required Sirt-1-type III deacetylase activity dependent increases in mTOR kinase activity. In agreement, resveratrol administration enhanced viral immunization induced CD8+ T cell effector responses and restored the poor CD8+ responses observed in diet-induced obese mice. These results demonstrate that the CR-mimic resveratrol promotes effective CD8+ T cell responses in metabolically imbalanced hosts, which may lead to life-span extension. By delineating the role of Sirt-1; NAD+ dependent type III deacetylase activity for T-bet expression, we have identified a new pathway to regulate CD8+ T cell mediated host immunity.
A novel pro-survival and proliferation-inducing role for CCR4:TARC interaction in chronic lymphocytic leukemia (CLL) B cells (133.4)
Authors: Damle Rajendra , Temburni Sonal , Malavasi Fabio , Kaufman Matthew , Kolitz Jonathan , Allen Steven , Rai Kanti , Chiorazzi Nicholas 
Published: January 2023

Abstract
CLL is a lymphoproliferative disorder characterized by an accumulation of CD5+ B cells. Somatic mutation of Ig V genes and cell surface CD38 expression by CLL cells stratifies cases into clinically distinct subgroups. CLL cells are likely sustained by the stimulation via BCR, TLR, and cytokine/chemokine:ligand interactions. Here we report on CCR4 expression and function in CLL. Cases with high numbers of CD38+ cells (≥30%) showed significantly higher percentages of CCR4-expressing cells (52.8 ± 5.2; p&lt; 0.01) than counterparts with low numbers of CD38+ cells (&lt;30%) (33.7 ± 5.3). Among CD38+ clones, those with mutated Ig V genes exhibited more CCR4-expressing cells than those with unmutated Ig V genes (p&lt; 0.01). CCR4 molecules were functional as defined by proximal events (significant increases in phosphorylation of Btk and STAT5 in response to varying doses of TARC, specific decreases in surface CCR4 expression, and, migratory responses to TARC). Among its downstream effects, TARC alone rescued 25-60% of CCR4+ CLL cells from apoptosis. Simultaneously TARC induced proliferation of CLL cells in a dose-dependent manner. BCR crosslinking, TLR-9 agonists, T cell help, and CD38:CD31 interactions increased density of CCR4, likely facilitating ligand-mediated effects. Taken together, these findings suggest an important and novel role for CCR4:TARC interaction in promoting CLL cell survival and proliferation downstream of several triggering molecules.
IL-27 promotes Tc1 polarization of human CD8 T lymphocytes and enhances their proliferation and effector functions (134.2)
Authors: Arbour Nathalie , Schneider Raphael , Yaneva Teodora , Beauseigle Diane 
Published: January 2023

Abstract
IL-27 has been shown to exhibit both pro- and anti-inflammatory properties. This cytokine favors naïve CD4 T cell differentiation into Th1 cells to the detriment of Th17 and Th2 skewing. Moreover, IL-27 promotes proliferation and cytotoxic functions of mouse CD8 T cells, but no data is available for human cells. We investigated the impact of IL-27 on human CD8 T cells. Surface IL-27 Receptor (IL-27R), composed of TCCR (also known as WSX-1) and gp130, is detectable on subsets of peripheral blood T cells. More CD8 than CD4 T cells are IL-27R positive and these proportions increase upon polyclonal activation. IL-27 activates rapid STAT1 and STAT3 signaling and transcription of SOCS1 and SOCS3 mRNA in CD8 T cells. Addition of IL-27 to anti-CD3 activated CD8 T cells leads to a significant augmentation of proliferation, and IFN-gamma and granzyme B production. Furthermore, purified naïve (CD45RA) but not memory (CD45RO) CD8 T cells are susceptible to those IL-27-enhancing effects. Moreover, anti-CD3 activated CD8 T cells express higher levels of T-bet upon IL-27 addition. Thus, our results support the capacity of IL-27 to drive naïve CD8 T cell polarization toward a Tc1 phenotype via STAT phosphorylation and induction of T-bet expression. Finally, the presence of IL-27 in the environmental milieu of naïve human CD8 T cells being activate through their TCR enhances the proliferation, IFN-gamma and granzyme B production thus contributes to generate more efficient effector cells.
Enhanced liver regeneration in IL10 deficient mice after partial hepatectomy via activating hepatocyte STAT3 and stimulating inflammatory response (134.6)
Authors: Yin Shi , Wang Hua , Park Ogyi , Wei Wei , Shen Jilong , Gao Bin 
Published: January 2023

Abstract
A series of evidence show that pro-inflammatory cytokines, including tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), play a critical role in the initiation and progress of liver regeneration. Relatively little is known regarding the role of anti-inflammatory cytokine IL-10 in liver regeneration after partial hepatectomy (PHx). Here, we examined the role of IL-10 in liver regeneration using a model of PHx in several strains of genetically modified mice. IL-10 -/- mice, and IL10-/-STAT3hep-/- double knockout mice in which IL-10 gene was globally disrupted and STAT3 gene was deleted specifically in hepatocytes; and their littermate controls underwent 70% PHx. Liver regeneration was evaluated by BrdU incorporation. After PHx, expression of IL-10 mRNA was significantly elevated after PHx. Compared to wild type mice, IL-10-/- mice had enhanced liver regeneration as determined by measuring hepatocyte proliferation and mitosis with the peak of hepatocyte proliferation occurring 48 hours post PHx. This increased liver regeneration in IL-10-/- mice was associated with elevated serum levels of TNF-alpha and IL-6, and enhanced activation of the IL-6 downstream signal STAT3 in the liver. An additional deletion of STAT3 in hepatocytes reduced liver regeneration in IL-10-/- mice. These findings suggest that IL-10 may negatively regulate liver regeneration via limiting inflammatory cytokine responses and subsequently tempering hepatic STAT3 activation.
Increased autoimmune disease activity in MRL-lpr mice following influenza infection. (135.21)
Authors: Crowe Sherry , James Judith , Edwards Erica , Anderson Jourdan 
Published: January 2023

Abstract
Influenza virus infection induces IFNα and TNFγ, which have been linked to increased severity of select autoimmune diseases. Influenza also stimulates TRL3 and TLR7, which are associated with increased kidney damage in mouse models of lupus. Therefore it seems likely that influenza infection of individuals with systemic lupus erythematosus, or lupus, would accelerate autoimmune disease progression. We used the MRL-lpr mouse model of spontaneous lupus-like disease to examine the impact of influenza infection on end stage organ damage. Influenza infection of MRL-lpr mice resulted in significant peri-vascular cellular infiltrates in the kidneys, composed primarily of CD4+ T cells. The infected mice also had accelerated proteinuria and lymphadenopathy of the cervical and mediastinal lymph nodes. Infection also resulted in a transient but significant increase in the concentration of anti-cardiolipin antibodies, but no changes in other autoimmune specificities. Experiments are underway to determine if this end-stage organ damage is dependent upon virus replication or if exposure to viral proteins, such as following vaccination, would result in increased kidney damage. This model will provide a unique system to dissect the immune-pathogen interaction in an autoimmune disease setting. These data will also provide valuable information on how infection impacts autoimmune disease development and progression.
CCR6-dependent recruitment of dendritic cells to the central nervous system during autoimmune encephalomyelitis (135.26)
Authors: Wu Gregory , Archambault Angela , Debes Gudrun , Laufer Terri 
Published: January 2023

Abstract
The antigen presentation requirements during experimental autoimmune encephalomyelitis (EAE), a model for the human disease multiple sclerosis (MS), are incompletely known. Dendritic cells (DCs) are specialized antigen presenting cells that critically participate in neuro-inflammation. Previously, we found that DCs alone are sufficient as antigen presenting cells for the induction of active EAE. We hypothesized that DCs arising from the periphery are specifically recruited to the central nervous system (CNS) during EAE. We utilized radiation bone marrow chimeras to address the role of peripheral DCs during EAE. We found that bone marrow-derived DCs accumulate within the CNS during disease. Further, limiting expression of MHCII to radiation-sensitive DCs was sufficient to support wild-type levels of disease. Using mixtures of CCR6-deficient and wild-type bone marrow as donors for radiation chimeras, we determined the proportion of each that arose in the CNS during EAE. These mixed bone marrow chimeras were equally susceptible to EAE compared to chimeric mice with entirely wild-type bone marrow. However, the fraction of DCs lacking CCR6 was reduced compared to CCR6-expressing DCs. Thus, in a competitive environment, CCR6-deficient DCs are inefficiently recruited to the CNS during EAE. These results suggest a potential mechanism for the entry of DCs during the development of neuro-inflammation.
Anti-neutrophil autoantibodies in tissue damage in systemic lupus erythematosus (135.31)
Authors: Yu Yangsheng , Fan Run , Zhang Yan , Yue Yinshi , Gould Karen , Thiele Geoffrey , Klassen Lynell , Fassina Giorgio , Kimberly Robert , Zhang Zhixin , Su Kaihong 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is a complex autoimmune disease in which abnormal immune responses attack multiple organs, leading to life-threatening complications. Although the pathogenic role of autoantibodies in SLE has been well established, the most disease-relevant reactivity of autoantibodies and their use as disease progression biomarkers are not fully understood. Using a single-cell PCR approach, we generated a large panel of recombinant antibodies from B cells of SLE patients. We found that a high frequency of SLE-derived antibodies (~ 20%) bind to and activate neutrophils as assayed by degranulation, oxidative burst, calcium influx, and tyrosine phosphorylation. One of the antibodies, S4Ab4, also induces neutrophil extracellular trap (NET) formation and increases cell death to human vein endothelial cells. Using mass spectrometry, we found that S4Ab4 reacts with two neutrophil proteins: lactoferrin and nucleotide-binding oligomerization domains 27 (NOD27). While lactoferrin is a known neutrophil autoantigen, NOD27, a family member of NOD-like pattern recognition receptors, might be a novel autoantigen in SLE. Furthermore, S4Ab4 antibody can cause proteinuria and kidney and lung manifestations when administered into NZB/W F1 lupus-prone mice. Our results show that SLE-derived autoantibodies can directly bind to and activate neutrophils. These anti-neutrophil autoantibodies may play important roles in the disease progression and tissue damage in SLE.