bluuebunny/crossref_metadata_2025_split huggingface dataset:
The potential role of HLA-I associated adaptation in elevated risk of infection among HIV-1 STEP study vaccine recipients
Authors: Qin Kai , Carlson Jonathan , Boppana Sushma , Bansal Anju , Goepfert Paul 
Published: January 2023

Abstract
The biological mechanism(s) underlying the elevated risk of HIV-1 acquisition among vaccine recipients in Merck STEP study remain unclear. HIV-1 frequently adapts to CD8 T cell responses, leading to the accumulation of adapted epitopes (AE). Recently, we demonstrated that in chronic HIV infection, CD8 T responses skewed by HLA-I associated adaptation enhanced HIV trans-infection of CD4 T cells by dendritic cells (DCs). However, whether adaptation in HIV vaccines induces a pro-inflammatory status and aids in potentiating infection is unknown. Using in-vitro studies, we evaluated whether an enhanced CD4 T cell trans-infection mediated by AE-specific CD8 T cells could be a potential biological mechanism for the observed increased risk of HIV acquisition in the Step Study. We found that CD8 T cells responding to AE demonstrated a higher capacity in promoting DCs maturation than non-adapted epitope (NAE) (p=0.007). Additionally, DCs matured by AE-specific CD8 T cells, showed a higher trend in inducing HIV trans-infection of CD4 T cells in each of the four vaccinee samples tested. We also analyzed the potential in-vivo impact of adaptation on the rate of infection among vaccine recipients in the STEP study. Our data showed that vaccine recipients with high adaptation to the vaccine immunogen were at an increased risk of HIV acquisition compared to recipients with medium/low adaptation or placebos (HR=2.98, p=0.02, high (n=21), medium (n=639), low (n=192) and placebo (n=865)). Taken together, our results show that in STEP study, AE-specific CD8 T cells may have contributed to an enhanced risk of HIV-1 acquisition by promoting viral trans-infection of CD4 T cells by DC. These findings have important implications for future HIV vaccine design.
Generation of clinically-relevant vaccine vectors for use in schistosome-endemic areas
Authors: Norwood Stephanie K , Bement Zachary James , Shollenberger Lisa M 
Published: January 2023

Abstract
Schistosomiasis is a neglected tropical disease with 200–300 million people infected worldwide. Schistosome infections are immunomodulatory and induce a Th2 biased immune response. Chronic helminth infections, such as schistosomiasis, can cause Th1/Tc1 type vaccine failure. Schistosomiasis endemicity is geographically coincident for other diseases lacking effective vaccines (HIV, TB, and Malaria). New vaccine vectors are needed to counter the immune suppression displayed during this infection. Listeria monocytogenes has shown to function as an effective vaccine vector to induce cell-mediated immune responses. Wild-type Listeria expressing HIV-Gag was previously proven to be an effective bacterial vector, however there are safety concerns with using a pathogenic vaccine vector in humans. Therefore, it is important to develop a clinically relevant vaccine that instead uses an attenuated strain to overcome safety concerns. There are genetically attenuated strains of Listeria currently in use in human clinical trials as therapeutic cancer vaccine vectors. Here, we are cloning HIV-Gag into the attenuated strains to induce strong cell mediated immunity without risk of adverse effects from the pathogenic wild-type vector. We will confirm appropriate Gag protein expression from the novel vectors using standard methodology. Further, we will confirm the functionality of these vectors in naïve and schistosome-infected animals. Our aim is that these non-pathogenic vectors will be clinically relevant for use in schistosome-endemic areas.
Antibody therapeutics for emerging infectious diseases: Ebola virus and others
Authors: Dye John M , Chandran Kartik , Wec Anna , Walker Laura , Zeitlin Larry , Bornholdt Zachary , Herbert Andrew 
Published: January 2023

Abstract
Developing medical countermeasures against emerging infectious diseases remains a challenging task. Ebola virus, Crimean CongoHemorrhagic Fever Virus, and Hantaviruses are ever growing threats on the global landscape. A consortium of worldwide researchers has developed a platform for novel monoclonal antibody discovery. This platform is predicated on the understanding that harvesting antibody sequences from convalescent survivors of previous outbreaks may lead to the most effective treatments for future outbreaks. Utilizing flow cytometric sorting of antigen-specific memory B cells for the elucidation of heavy and light chain sequences, novel biologics were identified and generated for evaluation. Specifically, a cocktail of two monoclonal antibodies was discovered and developed for Sudan virus, Ebolavirus, and Bundibugyo virus. Previously, published antivirals have been virus-specific with no cross reactivity against additional species of virus beyond the original agent. This cocktail of monoclonals neutralized and provided efficacy against infection with each of these viruses in animal models (rodent and non-human primate studies). This cocktail is currently being moved into advanced development and phase I clinical trials. More generally, this successful discovery platform is currently being utilized for developing novel monoclonal treatments for alphaviruses, hantaviruses, arenaviruses and other filoviruses. New data on the development of biologics against these agents will be discussed as well.
Bacillus Calmette-Guérin (BCG) vaccine reprograms human neonatal metabolism in vivo and in vitro
Authors: Arce Joann Diray , Jensen Kristoffer Jarlov , Angelidou Asimenia , Conti Maria Giulia , Kelly Rachel S. , Pettengill Matthew , Liu Mark , van Haren Simon D. , Benn Christine Stabell , Ozonoff Al , Steen Hanno , Lasky-Su Jessica , Levy Ofer 
Published: January 2023

Abstract
Immunization has significantly reduced infectious morbidity and mortality, but vaccine optimization and development is limited by incomplete understanding of how early life vaccines engage the distinct infant immune system to induce protection. We used plasma metabolomics to gain immunometabolic insight into the effects of Bacille Calmette-Guérin (BCG) which prevents disseminated TB and induces heterologous immunity to unrelated pathogens. For in vivo studies, low-birth weight newborns in Guinea-Bissau received a neonatal dose of Bacille Calmette-Guérin (early BCG) at birth or a catch-up after blood collection at four weeks post-birth (delayed BCG). For in vitro studies, human neonatal cord blood samples were stimulated for 18 hours with either vehicle (saline) or BCG. In vivo and in vitro plasma samples were profiled for global untargeted metabolomics and complex lipid panel lipidomics.
BCG-induced perturbation of neonatal metabolic pathways was observed in vivo and confirmed in vitro. Lipid biomarkers of early BCG-induced response identified increased free fatty acids (FFA), and decreased lysoPC (LPC), lysoPE (LPE), monoacylglycerol (MAG), phosphatidylcholine (PC), phosphoethanolamine (PE) and triacylglycerol (TAG). BCG identified decrease on plasma levels of LPC, LPE, PC and MAG in vitro. Nucleotide purine and pyrimidine pathways are perturbed both in vivo and in vitro. Neonatal BCG vaccination alters the early life metabolism, in particular lipids implicated in immune response. This suggests that BCG-induced alterations in lipid pathways may contribute to its protective effect. Further studies may identify biomarkers of vaccine immunogenicity to inform development and optimization of early life vaccines.
Arenavirus vectors for massive tumor self-antigen-specific CD8 T cell attack
Authors: Pinschewer Daniel D. , Bonilla Weldy V. , Kallert Sandra , Kirchhammer Nicole , Marx Anna-Friederike , Krzyzaniak Magdalena , Schmidt Sarah , Raguz Josipa , Berka Ursula , Günther Stephan , Magnus Carsten , Orlinger Klaus , Zippelius Alfred 
Published: January 2023

Abstract
Therapeutic vaccination regimens inducing high frequencies of clinically effective tumor-targeting CD8 T cells represent an unmet need in cancer immunotherapy.
We exploit cutting-edge viral reverse genetics to harness the immunostimulatory properties of the arenaviruses Pichinde virus and lymphocytic choriomeningitis virus for therapeutic cancer vaccination. When administering to mice life-replicating vector formats of these two viruses, both delivering the same cancer testis self-antigen in a heterologous prime-boost regimen, tumor-specific CD8 T cell responses reached up to 50% of the circulating CD8 T cell pool. This massive CD8 T cell attack eliminated established solid tumors in a significant proportion of animals, accompanied by complete protection against tumor re-challenge. The magnitude of CD8 T cell responses was driven by alarmin signals and depended on the combined use of two genealogically distantly related arenaviruses. Combinations of closely related vectors or the repeated administration of the same vector were not inhibited by vector-neutralizing antibodies, but CD8 T cell immunodominance hierarchies favored vector backbone-targeted responses at the expense of self-reactive T cell specificities.
These findings establish a powerful arenavirus-based tumor immunotherapy regimen exploiting key innate signaling pathways and reshuffling immunodominance hierarchies to break tumor self-directed tolerance
Pharmacological inhibition of protein kinase C-eta inactivates the contact-dependent suppressive activity of regulatory T cells (Tregs): A novel approach for cancer immunotherapy?
Authors: Xie Jiji , Liu Hsin-Yu , Pedros Christophe , Kong Kok-Fai , Altman Amnon 
Published: January 2023

Abstract
Our earlier work has revealed a novel, Treg-intrinsic signaling pathway consisting of a CTLA4 and PKC-eta complex, which is required for Treg function to suppress immune responses, including tumor-specific immunity, in a contact-dependent manner, but is dispensable for suppressions mediated by suppressive cytokines (Nat. Immunol. 2014; J. Clin. Invest. Insight. 2017). Furthermore, we found PKC-eta to be dispensable for the activation and effector function of mouse and human CD8+ T cells, implying that PKC-eta is a promising cancer immunotherapy target, whose pharmacological inhibition would inhibit the suppressive activity of tumor-specific Tregs without impairing function of tumor-specific cytotoxic T cells. We therefore initiated, in collaboration with Takeda Oncology, the testing of two lead small molecule compounds previously reported to selectively inhibit the catalytic activity of Ca2+-independent PKC subfamily members, including PKC-eta. Using in vitro assays, we titrated these compounds and established a range of compound concentrations that significantly inhibited the in vitro contact-dependent suppressive activity of human Tregs, while only minimally inhibiting the activation and proliferation of conventional human T cells. Furthermore, they did not inhibit the production of IL-10 by stimulated human Tregs, consistent with our hypothesis that the CTLA4-PKC-eta signaling pathway is selectively required for contact-dependent suppression. Such compounds could potentially serve as a novel form of CTLA4-based checkpoint blockade for cancer immunotherapy and, at the same time, minimize immune-related adverse effects associated with clinically used anti-CTLA4 antibodies.
Exploiting tumor necrosis with novel immunotherapies
Authors: Franks S. Elizabeth , Solocinski Kristen , Schlom Jeffrey , Hodge James 
Published: January 2023

Abstract
Tumor necrosis prior to therapy historically correlates with aggressive tumor phenotypes. However, necrosis in response to chemotherapy leads to increased progression-free and overall survival in several cancer types. Chemotherapy with docetaxel results in inhibition of mitosis, leading to necrosis. To exploit the therapeutic potential of docetaxel, a bifunctional molecule consisting of a mAb specific for histone H1 and two molecules of IL-12 (NHS-IL-12), was engineered to deliver rIL-12 to the tumor microenvironment (TME) through specific targeting of necrotic regions in the tumor. IL-12 is a proinflammatory cytokine that increases lytic capacity of T and NK cells. Although promising preclinically, rIL-12 when delivered systemically leads to severe toxicity. We hypothesize inducing necrosis leads to immunogenic modulation that can be exploited with targeted therapies resulting in increased tumor cell killing.
Here, we show pathologist confirmed increase in tumor necrosis post-delivery of docetaxel, leading to improved retention of NHS-IL-12 in the TME. Significant anti-tumor activity is observed when combining docetaxel and NHS-IL-12, but there is elevated PD-L1 expression on tumor cells and serum TGFb levels, which correlate with poor prognosis. M7824 is a fusion protein containing a mAb against PD-L1 linked to the extracellular domain of TGFbR2, sequestering TGFb. Following the strategic addition of M7824, TGFb levels in mice receiving the triple combination had significantly reduced TGFb levels. Additionally, the greatest anti-tumor activity and increase in survival was observed in cohorts receiving docetaxel, NHS-IL-12 and M7824. Data presented herein have aided in the rationale design of a clinical trial at NCI.
Investigating the pathogenesis of chronic chikungunya virus infection
Authors: Locke Marissa Christine , Young Alissa , Cook Lindsey E. , Monte Kristen , Lenschow Deborah 
Published: January 2023

Abstract
Chikungunya virus (CHIKV) is an arthritogenic alphavirus that acutely causes fever as well as severe joint and muscle pain. Chronic musculoskeletal pain persists in a substantial fraction of patients for months to years after infection. While replicating virus is not detected in joint-associated tissues of patients nor in various animal models at convalescent time points, viral RNA is detected months after acute infection.
To identify the cells that might contribute to chronic pathogenesis, we developed a recombinant CHIKV that expresses Cre recombinase (CHIKV-3′-Cre). CHIKV-3′-Cre infection of tdTomato reporter mice resulted in a population of tdTomato+ cells that persisted for at least 112 days. Immunofluorescence and flow cytometric profiling 28 days post infection revealed that these tdTomato+ cells were predominantly myofibers and fibroblasts as well as a small population of macrophages. Further, tdTomato+ cells are enriched for CHIKV RNA. Interestingly, the population of surviving cells was dynamic over time with macrophages making up a high percentage of the cells early and then tapering off.
We applied this system to determine the differential roles of the type I interferon (IFN) subtypes during chronic CHIKV infection. Results using IFN blocking antibodies revealed that mice lacking the IFNαs, but not IFNβ, show an increase in chronic RNA levels and the number of fibroblasts that survive CHIKV infection in distal sites, indicating that type I IFNs have divergent roles during chronic CHIKV disease. Ongoing studies will explore the differential functions of IFNαs and IFNβ, will determine why the surviving cells and viral RNA are not cleared by the immune system, and will elucidate their contribution to chronic inflammation.
CD4+ T cell mediated HLA class II cross-restriction in HIV controllers
Authors: Gras Stephanie 
Published: January 2023

Abstract
Rare individuals, termed HIV controllers, spontaneously control HIV infection by mounting efficient T cell responses against the virus. Protective CD4+ T cell responses from HIV controllers involve high affinity public T cell receptors (TCRs) recognizing an immunodominant capsid epitope (Gag293) presented by a remarkably broad array of HLA class II molecules. Here we determine the structures of a prototypical public TCR bound to HLAD-R1, –DR11, and –DR15 molecules presenting the Gag293 epitope. TCR recognition was driven by contacts with the Gag293 epitope, a feature that underpinned the extensive HLA cross-restriction. These high affinity TCRs promoted mature immunological synapse formation and cytotoxic capacity in both CD4+ and CD8+ T cells. The public TCRs suppressed HIV replication in multiple genetic backgrounds ex vivo, emphasizing the functional advantage conferred by broad HLA class II cross-restriction.
Serine phosphorylation of the STAT1 transactivation domain promotes autoreactive B cell and systemic autoimmunity development
Authors: Fike Adam J , Chodisetti Sathi Babu , Domeier Phillip P , Singh Harinder , Schell Stephanie L , Mockus Taryn E , Choi Nicholas M , Corradetti Chelsea , Kawasawa Yuka Imamura , Hou Baidong , Atkins Hannah M , Caricchio Roberto , Decker Thomas , Lukacher Aron E , Olsen Nancy J , Rahman Ziaur S.M. 
Published: January 2023

Abstract
Although STAT1 tyrosine-701 phosphorylation (STAT1-pY701) is indispensable for STAT1 function, the requirement for STAT1 serine-727 phosphorylation (STAT1-pS727) during autoimmune and anti-pathogen responses remains unclear. Here we report that STAT1-pS727 promotes autoimmune antibody-forming cell (AFC) and germinal center (GC) responses, driving systemic lupus erythematosus (SLE) development. STAT1-pS727, however, is not required for GC and antibody responses to foreign-antigens including pathogens or gut microbiota. STAT1-pS727 plays an important B cell-intrinsic role in driving autoimmunity. Transcriptomic analysis of B cells from TLR7-accelerated SLE-prone mice reveals STAT1-pS727-mediated gene regulation of cellular pathways known to be involved in autoimmune GC and AFC responses. Mechanistically, TLR7 activation in B cells induces STAT1-pS727 and during autoimmune responses TLR7 signaling converges with IFNγ-STAT1 signaling in B cells by recruiting STAT1 into the MyD88 signaling complex. Together, we identify previously unappreciated differential regulation of autoimmune and anti-pathogen responses by STAT1-pS727, and implicate STAT1-pS727 as a therapeutic target for SLE.
Food allergens could promote the Th2 mediated autoantibody development in an autoimmune skin disease bullous pemphigoid
Authors: Qian Ye , Lin Lan , Moran Timothy Paul , Yang Jinsheng , Liu Zhi , Culton Donna Aline 
Published: January 2023

Abstract
Bullous pemphigoid (BP) is the most common autoimmune skin blistering disease and is characterized by the presence of autoantibodies (autoAbs) against skin hemidesmosomal adhesion molecules. The predominant antigenic target is the non-collagenous domain 16 (NC16A) of BP180. AutoAbs against NC16A lead to loss of adhesion and subsequent subepidermal blistering. It is unclear what triggers the development of autoAbs. Food allergens are known to induce T helper 2 (Th2) antibody responses and our previous studies have shown the possible role of environmental antigens/allergens in promoting autoAb development. The goal of this study is to investigate whether Th2 mediated immune responses against food allergens drive the autoAb development in BP. BP serum antibodies were tested against a panel of common environmental and food allergens. Anti-NC16A IgE and IgG4 monoclonal antibodies (mAbs) and their naïve counterparts were generated from a BP patient to determine their specificity or cross-reactivity. We found that antibodies from BP patients recognized food allergens, such as walnut, but not other types of allergens, such as insects, epithelia, fungi, or pollens. The levels of serum IgE and IgG4, but not IgG1, against selected food allergens were significantly higher than controls. The anti-NC16A IgE and IgG4 mAbs cross-reacted to food allergens, and their naïve mAb counterparts had reduced reactivity to NC16A, but still reacted strongly to food allergens. In addition, mice immunized with walnut allergen produced anti-NC16A antibodies. Our results indicate that Th2-associated autoAbs from BP subjects might be linked to antibody responses toward food allergens and may provide a target for the management and treatment of BP.
Autoreactive B cells have a specific metabolic response during humoral responses
Authors: Choi Seung-Chul , Li Wei , Zhang Xiaojuan , Teng Xiangyu , Morel Laurence 
Published: January 2023

Abstract
Germinal centers (GC) provide a unique microenvironment for B-cell affinity maturation and class-switching recombination to occur. We compared for the first time the metabolism of B cells between lupus-prone B6.Sle1.Sle2.Sle3 (TC) mice and B6 controls at steady state, as well as during T cell-dependent (TD) and T cell-independent (TI) immunizations. B cells from TC mice showed an elevated glycolysis and mitochondrial oxidative metabolism, which was normalized by inhibiting glycolysis with in vivo 2-deoxy-D-glucose (2DG) treatment. 2DG greatly reduced the production of TI-antigen-specific antibodies, but showed minimal effect with TD-antigens. In contrast, glutaminolysis inhibition with 6-Diazo-5-oxo-L-norleucine (DON) had a greater effect on TD than TI Ag-specific antibodies in both strains. Interestingly, 2DG, but not DON reduced TI-Ag-specific IgM in TC mice, whereas both 2DG and DON prevented Ag-specific IgM production in B6 mice. Thus, autoreactive and control B cells have different intrinsic metabolic requirements. Autoreactive B cells are more glycolytic, which mirrors our previous results showing that autoreactive TFH cells have opposite glucose and glutamine requirements. The requirement for glutamine in TD-responses could be due to a direct effect on GC B cells, or an indirect effect through TFH cells. Overall, these results predict that targeting glucose metabolism provides an effective therapeutic approach for systemic autoimmunity by eliminating both autoreactive TFH and B cells, although TI-responses may be reduced in lupus-prone mice.
Deletion of von-Hippel Lindau in Dmp1-expressing cells impairs B cell development
Authors: Chicana Betsabel , Burns Christian , Spencer Joel , Manilay Jennifer 
Published: January 2023

Abstract
Our objective is to characterize how altered bone homeostasis influences B lymphocyte development. The von-Hippel Lindau protein (VHL) regulates hypoxia-inducible factor degradation and is involved in responses to low oxygen environments. Osteoblasts (OBs) support B cell development and mature into osteocytes (OCYs). We utilized Dmp1-Cre;Vhl conditional knockout mice (cKO), in which Vhl is deleted in OBs and OCYs. The cKO display dysregulated bone growth, high bone density, smaller bone marrow (BM) cavity volume, and an overall decrease in BM cellularity compared to wild-type (WT) mice. B cell progenitors is significantly increased in frequency and numbers whereas Hardy Fractions B, C, D, E and F were significantly decreased in the BM of cKO mice. Analysis of BM B cells 16 weeks after whole BM transplantation, showed a significant reduction in Fractions B-C and later Fractions in the WT→cKO mice, while B cell development was normal in the cKO→WT chimeras, indicating a cell-extrinsic effect of Vhl on B cells within the cKO BM microenvironment. We hypothesize that B cell development is not supported due to reduction of key niche cells and decreased production of IL7, SCF, CXCL12, and Flt3. To define the defective niche cells, we will use in vitro B cell development assays in which WT B cells will be co-cultured with sorted cKO OBs, MSCs, and OCYs. B cell apoptosis, proliferation and turnover rates will be measured. We have begun preliminary studies using intravital and ex vivo imaging of the cKO BM to identify changes in oxygen tension, blood vessel type and structure, the presence and location of stromal cells. These studies have the potential to reveal novel molecular mechanisms by which Vhl in Dmp1 expressing cells may serve as a B cell niche.