bluuebunny/crossref_metadata_2025_split huggingface dataset:
Humanized immune system mice: Improving the predictability of mouse models in immuno-oncology drug development (TUM3P.1044)
Authors: Hall Leon , Paz Keren , Kalbfliesh Megan , Pei Jiong , Baia Gilson 
Published: January 2023

Abstract
Mouse models are widely used in preclinical oncology research but species differences can limit efficacy predictions for clinical translation. Taconic Biosciences, Inc. is leveraging the super immunodeficient status of the CIEA NOD.Cg-Prkdcscid Il2rgtm1Sug/JicTac (NOG) mouse, which allows it so support the engraftment of human cells and tissues, to accelerate efficacy and safety testing of novel immunotherapies. We have optimized this model for efficient engraftment of the human immune system, and co-engraftment of patient derived xenografts (PDX). In a collaboration with Champions Oncology, the PDX engrafted humanized NOG mouse responded to immune checkpoint inhibition, by activating T cells which are able to infiltrate the tumor. An overview of Taconic's humanization program will be presented, including data showing the utility of humanized mice in PDX applications for immuno-oncology drug development, and recent advances in the development of next generation humanized mice
Characterization of the Anti-CD22 targeted therapy, moxetumomab pasudotox, for BCP-ALL (TUM3P.1053)
Authors: Kinjo Ichiko , Matlawska Ksenia , Winter Stuart , Burke Patricia , Wilson Bridget 
Published: January 2023

Abstract
Moxetumomab pasudotox (moxe, HA22, CAT-8015) is a recombinant, affinity optimized anti-CD22 immunotoxin. The demonstrated antitumor activity of moxe in patients with relapsed/refractory hairy cell leukemia supports its development in hematologic malignancies expressing CD22. Anti-leukemic activity has also been demonstrated in children with chemotherapy-refractory precursor B cell acute lymphoblastic leukemia (BCP-ALL) at various dose levels. Here, we explore potential mechanisms contributing to BCP-ALL sensitivity to moxe in vitro and in vivo. Binding, internalization and apoptosis studies were conducted using fluorescently-tagged moxe and two BCP-ALL cell lines (697, Nalm6), as well as blast cells from patients. Moxe cytotoxicity was only weakly correlated to CD22 expression levels, implicating additional processes after CD22-binding in the trafficking and processing of moxe also contribute to cytotoxicity. Studies in NSG mice engrafted with human BCP-ALL cell lines or patient blasts show that fluorescently-tagged moxe is taken up differentially by leukemia cells disseminated to different organs in the mouse (bone marrow, spleen, liver, brain). Studies were performed to evaluate the site-specific leukemia burden of cell lines and patient blasts in xenografted mice, and their correlation with in vitro uptake and cytoxicity assays. Our results suggest additional studies may be valuable to evaluate additional dosing strategies to maximize efficacy in these models.
Differential antibody induction to surface textured silica nanoparticle adjuvants (VAC3P.1055)
Authors: Tsirikis Peter , Wilson Kirsty , Kong Ying , Xiang Sue , Selomulya Cordelia , Plebanski Magdalena 
Published: January 2023

Abstract
Nanoparticles have been shown to be a potent adjuvant in vaccine design. Previous studies demonstrated that carboxylated 40-50 nm polystyrene nanoparticles (PSNPs) with covalently bound antigen offer a new class of vaccine with only two elements (antigen and particle) but with no added inflammatory stimuli. Herein, we report on a deconstructed two-part vaccine whereby the adjuvant and protein carrier are co-administered to elicit potent antibody responses in vivo. Recent works indicate that particle shape can also influence the immune response. As such, we investigate the influence of surface morphology using smooth and rough surfaced silica nanoparticle adjuvants, showing smooth nanoparticles as the more potent antibody inducers. MTT assay confirms that both silica nanoparticle adjuvant and polystyrene protein carrier display excellent biocompatibility with over 95% viability with COS-7 cells. Preliminary in vitro dendritic cell (DC) culture experiments indicate that silica nanoparticle adjuvants, similarly to PSNPs, do not induce the up-regulation of maturation markers, CD80, CD86 and MHC II on CD11c+ gated DCs. This suggests the particle carrier itself is non-inflammatory, a required feature for our vaccine approach. Ongoing studies are focusing on the cytokines induced in vitro and in vivo by these novel constructs. The outcomes from this study provide key design criteria in the development of novel nanoparticle immunotherapeutics for the treatment of disease.
Cell-mediated immune responses to ZOSTAVAX© are characterized by a boost in polyfunctional central memory T cells (VAC4P.1115)
Authors: Sei Janet , Cox Kara , Dubey Sheri , Antonello Joseph , Vora Kalpit 
Published: January 2023

Abstract
Herpes Zoster (HZ) is a debilitating disease caused by reactivation of latent varicella-zoster virus (VZV). ZOSTAVAX is approved for the prevention of HZ and its complications in individuals 50 years and older in many countries around the world. The mechanistic immune correlates of protection and VZV antigens responsible for induction of protective CMI are not defined. To screen for dominant VZV antigens, whole blood from subjects pre-and-post ZOSTAVAX vaccination were assessed for production of IFN-ϒ, IL-2 and, TNF-α. VZV antigens eliciting high cytokine levels were used to stimulate PBMC and intracellular cytokine staining performed. We show that the VZV antigens that led to a boost in the frequency of polyfunctional CD4+ T cells (TCD4+) were IE63&gt;IE62&gt;gB&gt;ORF9&gt;gE. Polyfunctional TCD4+ produced higher levels of cytokines than monofunctional cells, and a boost in expression of cytokines was observed, post-ZOSTAVAX. Phenotypic characterization of polyfunctional TCD4+ demonstrated that they are TCM cells. Notably, we detected ORF9-specific CD8+ T cells in 52% of subjects tested, however whether these responses are cross-functional or VZV-specific needs to be determined. Overall, these studies outline the CMI that may correlate with ZOSTAVAX protection.
Inhibition of GSK3beta leads to increased survival, proliferation and memory phenotype generation of GBM-specific CAR T cells (VAC5P.1125)
Authors: Sengupta Sadhak , Guha Prajna , Katz Steven , Sampath Prakash 
Published: January 2023

Abstract
Application of chimeric antigen receptors (CAR), where an antibody or ligand-binding domain is fused with the zeta signaling chain of the T cell receptor, is a novel and alternative strategy to bypass the failure of cytotoxic immune response induction by most tumor cells. Although CARs have shown impressive preclinical efficacy, translational success in solid tumors like glioblastoma (GBM) is often restricted by the limited viability of the T cells either due to the absence of IL2 in the central nervous system and/or because of mandated chemotherapy. Although multiple doses of T cell transfer and systemic administration of IL2 are effective for short time periods, unregulated overdose of T cells and IL2 result in immune toxicity. Inhibition of glycogen synthase kinase 3 beta (GSK3β) in GBM-specific IL13CAR T cells increased IL2-independent survival and proliferation of the CAR-positive T cells upon activation with IL13Rα2-expressing GBM tumor cells or soluble target antigen, when compared to bystander CAR-negative T cells or untreated IL13CAR T cells. GSK3β-inhibited IL13CAR T cells also showed increased IL7R and CD45RO expression compared to the bystanders as well as untreated IL13CAR T cells indicating a possible development of memory population. Exposure to target antigen increased cytotoxic efficacy and tumor-killing ability specifically in GSK3β-inhibited IL13CAR T cells. These observations suggest an innovative advancement in anti-GBM CAR T cell-mediated immunotherapy.
Critical role of ITK in mouse γ-herpes virus 68 infection as a model for human immunodeficiency (VIR1P.1140)
Authors: Cho Hyoung-Soo , Shin HyunMu , Berg Leslie 
Published: January 2023

Abstract
IL-2-inducible T cell kinase (ITK) is a Tec family tyrosine kinase predominantly expressed in T cells. In the absence of ITK, the magnitude of T cell receptor (TCR) signaling is greatly reduced, but not abolished, leading to an overall decrease in TCR signaling strength. Recently, various mutations in ITK were identified in patients suffering from a fatal lymphoproliferation due to a failure to control Epstein-Barr virus (EBV) infection. Interestingly, Itk-/- mice have been shown to mount protective responses to several acute virus infections. Thus, we hypothesized that ITK plays an essential role in long-term anti-viral immunity against latent γ-herpes virus infection. To study this, we infected WT and Itk-/- mice with mouse γ-herpes virus 68 (MHV68). Despite a reduced response at D7 post-infection, Itk-/- mice ultimately controlled MHV68 replication in the spleen by D14. After the initial response, both WT and Itk-/- mice showed no difference in their viral titers in the spleen at later time points. However, Itk-/- mice developed severe gastrointestinal pathology following LPS-mediated reactivation of latent MHV68, accompanied by a high viral titer in the intestine. Furthermore, the number of lymphocytes in the intestinal epithelium and lamina propria in naïve Itk-/- mice was substantially less than WT mice. Overall, these data suggest that ITK is essential for anti-viral immunity against γ-herpes virus infection in the gut following reactivation from latency.
Recruitment of GL7+ B cells to the inflamed CNS prior to peripheral germinal center formation (VIR1P.1151)
Authors: DiSano Krista , Phares Timothy , Bergmann Cornelia 
Published: January 2023

Abstract
Central nervous system (CNS) inflammation induced by viral infection and autoimmune disease results in the accumulation of various B cell subsets, including antibody secreting cells (ASC). However, temporal dynamics and precursor relationships of B cell subsets between the periphery and the inflamed CNS remain unclear. Encephalomyelitis induced by gliatropic coronavirus JHMV resolves into a persistent CNS infection that is controlled by local humoral immunity. Prior to the accumulation of essential CD138+ ASC, CD138- B cells traffic to the CNS by day 5 post infection (p.i.). Analysis of these early accumulating B cells by flow cytometry shows they comprise several phenotypes, which typically reside within the cervical lymph node (CLN), including naïve, activated, germinal center (GC)-like (GL7+), and unswitched memory B cells. B cell activation within the CLN occurs by day 7 p.i., but GC formation is not robust until day 14 p.i.. Unexpectedly, GL7+ B cells are detected within the CNS prior to GC formation. Moreover, in contrast to naïve/early activated B cells, which decline in number, GL7+ B cells increase differentially in brain and spinal cord throughout persistence. Maintenance of more differentiated B cell subsets within the CNS suggests either selective ongoing recruitment from the periphery or preferential local CNS maintenance. The lack of evidence for ectopic follicles suggests activated B cells, including GC-like B cells, can be recruited from the CLN.
The role of KIR and HLA polymorphism in HIV acquisition: association and in vitro functional studies (VIR1P.1159)
Authors: erlich henry , Wolinsky Steven , Penugonda Sudhir , Hayes Elizabeth , Nichols Robert , Saeteurn Koy , Duggal Priya , Maljkovic Berry Irina , Reddy Susheel , Swantek Peter , Ladner Martha , Lin Lin , Braun Jonathan , Trachtenberg Elizabeth 
Published: January 2023

Abstract
Polymorphism in the genes that encode the HLA antigens and the KIR receptors on NK cells influence both adaptive and innate immunity. We analyzed the role of HLA and KIR polymorphisms in HIV acquisition by comparing 405 HIV+ and 523 HIV- men in the Chicago MACS cohort. We found no significant association with any HLA allele, in contrast to the well established strongly protective effect of HLA-B*57:01 for HIV disease progression. The inhibitory KIR ligands are HLA class I epitopes and fall into the groups: HLA-C1,C2, HLA-Bw4, and HLA-A3/A11. Association analyses of individual KIR genes in the presence/absence of their cognate HLA ligand revealed that one KIR/HLA ligand genotype combination was protective for HIV acquisition. The HLA ligand genotype C1/C1 was protective compared to C1/C2 and C2/C2 but only with genotype KIR2DL3+/KIR2DL2- (OR = 1.76, p = 0.007). This KIR genotype marks homozygosity for the centromeric KIR A haplotype, which is more inhibitory, has fewer genes, and less diverse than KIR B haplotypes. Since C2 is the ligand for the KIR2DL1 receptor, and C1 is the ligand for the KIR2DL3 receptor, the protective association observed with C1/C1 homozygotes suggests that a lack of signaling via the strongly inhibitory KIR2DL1 receptor in addition to signaling via the KIR2DL3 receptor is protective. In vitro assays of NK cells from KIR2DL3 homozygous individuals demonstrate that C1/C1 vs C1/C2 and C2/C2 (C2+) genotypes are more active in terms of cytokine release.
Ly6C+ monocyte efferocytosis and cross-presentation of cell-associated antigen
Authors: Larson Sandy R , Thomas Stacey , Atif Shaikh M , Gibbings Sophie , Prabagar Miglena , Henson Peter M , Jakubzick Claudia 
Published: January 2023

Abstract
Recently it was shown that circulating Ly6C+ monocytes traffic from tissue to the draining lymph nodes (LNs) with minimal alteration in their overall phenotype. Furthermore, in the steady state, Ly6C+ monocytes are as abundant as classical dendritic cells (DCs) within the draining LNs, and even more abundant during inflammation. However, little is known about the functional roles of constitutively trafficking Ly6C+ monocytes. In this study we investigated whether Ly6C+ monocytes can efferocytose (acquire dying cells) and cross-present cell-associated antigen, a functional property specially attributed to Batf3+ DCs. We demonstrated that Ly6C+ monocytes intrinsically efferocytose and cross-present cell-associated antigen to CD8+ T cells. In addition, efferocytosis was enhanced upon direct activation of the Ly6C+ monocytes through its corresponding TLRs, TLR4 and TLR7. However, only ligation of TLR7, and not TLR4, enhanced cross-presentation by Ly6C+ monocytes. Overall, this study outlines two functional roles, among others, that Ly6C+ monocytes have during an adaptive immune response.
Loss of phagocytic and antigen cross-presenting capacity in aging DCs is associated with mitochondrial dysfunction
Authors: Janssen Edith , Thacker Robert , Shehata Hesham M , Lages Celine Silva , Hennies Cassandra , Lehn Maria , Chougnet Claire 
Published: January 2023

Abstract
Impaired functionality of dendritic cells (DCs) significantly contributes to decreased adaptive immune responses in aged hosts. The expression of MHC-peptide on the DC surface is the critical first step in T cell priming, but few studies have addressed the effect of aging on antigen (Ag) acquisition, processing, and presentation by DCs. Here we show that aged murine DCs were less efficient in the cross-presentation of cell-associated Ag and subsequently in the cross-priming of CD8+ T cells than their young counterparts.
The decreased cross-presentation was associated with a reduction in the frequency of CD8αDCs and merocytic (CD8α-CD11b-)DCs that could endocytose cell-associated Ag, as well as the number and the size of the endocytosed particles in the DC that did internalize cell-associated materials.
Mechanistically, phagocytic capacity has been associated with mitochondrial activity and membrane potential (Δψm). Aged DCs exhibited profound signs of mitochondrial dysfunction, illustrated by lower Δψm, reduced ATP turnover and coupling efficiency, decreased baseline oxidative phosphorylation (OXPHOS), and greater proton leak and ROS production. Mimicking the aged metabolic phenotype in young DCs by pharmacologic manipulation indicated that the reductions in Δψm and ATP impeded the phagocytic capacity while ROS interfered with a later step in the cross-presentation process. Conversely, in vitro scavenging of ROS partially restored cross-presentation by aged DCs.
Together, these data suggest that improvement of aged DC functionality might be feasible in the elderly, by targeting metabolic dysfunction, or its downstream sequelae, thereby opening new avenues for enhancing vaccine efficiency in this population.
Potential interaction of WT and Mutant TNF-Receptor 1 and Pyruvate Kinase M2 in TNF-Receptor-Associated Periodic Fever Syndrome (TRAPS)
Authors: Ploch Michelle , Pelletier Martin , Billingham Leah , Siegel Richard M 
Published: January 2023

Abstract
Tumor Necrosis Factor (TNF) is a trimeric cytokine induces innate inflammatory responses or cell death through TNFR1. Extracellular mutations in TNFR1 are associated with Receptor-Associated Periodic Syndrome (TRAPS), an autosomal dominant autoinflammatory disorder characterized by recurring fevers, dermal and synovial inflammation and elevated risk of amyloidosis. Previous work by our group has demonstrated that a TRAPS-like mutation in TNFR1 leads to accumulation of the receptor in the ER, leading to abnormal MAPK activation and correlated with reduced glucose metabolism. To better understand how the mutant TNFR1 signals to induce these abnormal cellular phenotypes, we immunoprecipitated the wild-type and TRAPS-associated mutant receptors and performed mass spectroscopy to identify associated proteins. Both WT and Mutant were able to bind the known adapter protein TRADD, but not the death domain (DD) deleted mutant. In addition, we also identified a novel association of WT and mutant TNFR1 with pyruvate kinase M2 (PKM2). Pyruvate kinase is the rate limiting enzyme in glycolysis and PKM2, and has been shown to be more activated in cancer cells, leading to abnormal metabolism known as the Warburg effect where cells undergo aerobic glycolysis. In addition, other studies suggest a link between activated PKM2 and induction of inflammatory responses, increasing TNF and IL1-b production. To confirm this interaction, we utilized colocalization microscopy techniques and IP/Western blot analysis which inferred a link between PKM2 and mutant TNFR1 more than the wild-type. Association between PKM2 TNFR1 may be a novel link between TNFR1 signaling and metabolism and explain some of the metabolic abnormalities in TRAPS cells.
Superoxide deficiency protects against Coxsackievirus-induced Type 1 diabetes through dampened macrophage anti-viral responses
Authors: Burg Ashley R , Das Shaonli , Padgett Lindsey E , Tse Hubert Ming 
Published: January 2023

Abstract
Viral infections are highly-suspected environmental triggers of autoimmune Type 1 diabetes (T1D), but the mechanism remains unclear. Many innate immune pro-inflammatory anti-viral factors, including IFN-α/β, TNF-α, CXCL10 and CCL5, are pathogenic in T1D. We recently demonstrated a crucial role for reactive oxygen species (ROS) production in modulating these factors, as Non-Obese Diabetic (NOD) mice deficient in NADPH oxidase (NOX)-derived superoxide (NOD.Ncf1m1J) are highly protected against T1D, partly due to impaired inflammatory macrophage responses. Therefore, we hypothesize that NOD.Ncf1m1J mice will be protected from virus-induced diabetes due to dampened pro-inflammatory anti-viral macrophage responses. Upon infection with Coxsackie B3 (CB3), a suspected diabetogenic virus, NOX-derived superoxide is rapidly generated by NOD macrophages, but is significantly decreased in NOD.Ncf1m1J macrophages. Loss of redox-dependent pro-inflammatory cytokine and chemokine synthesis significantly protected NOD.Ncf1m1J mice from virus accelerated T1D, as infected NOD.Ncf1m1J pancreata revealed significant decreases in Tnf, Cxcl10 and Ccl5 mRNA accumulation. This corresponded with 2.5-fold decreases in TNF-α- and IL-1β-producing pancreas-infiltrating macrophages, despite having no significant defect in CB3 viral clearance. Mechanistically, we determined that MDA5, a viral RNA sensor, was regulated by ROS. CB3-infected NOD.Ncf1m1J macrophages displayed a decrease in MDA5 expression, but was rescued by addition of exogenous ROS. Therefore, our results provide evidence that ROS can synergistically influence anti-viral responses to promote autoimmune diabetes in response to diabetogenic viral infections.
NKG2D-H60a interaction via homotypic T cell contact delays NOD diabetes development.
Authors: Trembath Andrew P , Sharma Neekun , Matocha Kimberly Nicole , Markiewicz Mary A 
Published: January 2023

Abstract
Type I diabetes is an autoimmune disorder mediated by T cells that destroy the insulin-secreting β-islet cells in the pancreas. Both human and murine studies implicate the immune receptor natural killer group 2 member D (NKG2D) in enhancing type 1 diabetes development. However, a mechanism for NKG2D influence in type 1 diabetes development is not clear. Utilizing NKG2D-deficient non-obese diabetic (NOD) mice, we demonstrate that NKG2D expression affects diabetes development via multiple mechanisms in multiple anatomical locations, with both pro- and anti-diabetic effects. First, we found NKG2D influences NOD diabetes development via alteration in the microbiota composition. When we eliminated this microbiome effect, NKG2D-deficient mice had enhanced diabetes development, demonstrating a protective role for NKG2D in this disease. Our in vitro studies suggest this protection is mediated via a previously undescribed interaction between NKG2D and the NKG2D ligand H60a, both expressed by activated NOD CD8+ T cells, during diabetogenic cytotoxic T cell (CTL) generation. We show that during CD8+ T cell differentiation into CTLs, NKG2D-H60a interaction generally decreases the subsequent CTL effector cytokine response. Taken together, these findings demonstrate there is a protective role for NKG2D during diabetes development that is likely mediated by NKG2D-ligand interaction during diabetogenic CTL differentiation.
Loss of intrasplenic 33D1+DC migration impairs alloimmune responses against RBC transfusion
Authors: Liu Dong , Calabro Samuele , Gallman Antonia , Nascimento Manuela , Chen Pei , Krishnaswamy Kumar , Uthaman Gowthaman , Xu Lan , Xu Lan , Williams Adam , Eisenbarth Stephanie C 
Published: January 2023

Abstract
Although the migration of tissue DCs from periphery to draining LNs is the crucial step for the induction of T cell responses, it is not clear whether the same holds true within the spleen. Here we use red blood cell (RBC) transfusion model to specifically study intrasplenic DC migration in the induction of alloantibodies to blood group antigens present on the donor RBCs. We observed that mice deficient in DOCK8 (dedicator of cytokinesis 8), a guanine nucleotide exchange factor that regulates the activity of Rho GTPase family members, fail to produce alloantibodies following transfusion. Two major splenic conventional DC subsets, the Batf3-dependent CD8+ XCR1+ subset and the IRF-4-dependent CD11b+33D1+ subset, prime CD4+ and CD8+ T cells, respectively. Our data demonstrate that only bridging channel 33D1+ DCs migrate to the T cell zone to initiate CD4+ T cell responses during RBC transfusion. Using in vivo antibody labeling method, we are able to differentiate marginal zone/red pulp dendritic cells from T cell zone/white pulp dendritic cells. In vivo labeling reveals that the failure of generating alloantibodies to RBCs in DOCK8 deficient mice is due to impaired 33D1+ DC migration. Interestingly, although the XCR1+ DCs in DOCK8 deficient mice migrated normally, they played no role in alloimmune responses. Taken together, our data demonstrate that subset specific migration of DCs within spleen is critical for initiation of CD4 T cell response and generation of alloantibodies. As equivalent populations of DC subsets exist in humans, our work suggests that blocking certain DC subset migration therapeutically during RBC transfusion could potentially prevent the harmful sensitization that occurs in patients who require lifelong RBC support.