bluuebunny/crossref_metadata_2025_split huggingface dataset:
Immunization with a mucosal, post-fusion F/G protein-based polyanhydride nanovaccine protects against BRSV infection in neonatal calves
Authors: Maina Teresia , Grego Elizabeth , Narasimhan Balaji , Sacco Randy E , McGill Jodi L 
Published: January 2023

Abstract
Human respiratory syncytial virus (HRSV) is a leading cause of severe acute lower respiratory tract (LRT) disease in young children worldwide. Currently, there is no approved vaccine for HRSV. Bovine RSV (BRSV) is closely related to HRSV and is the cause of severe acute LRT disease in young cattle. A neonatal calf model presents an opportunity to test HRSV vaccines and study a naturally susceptible host-pathogen interaction similar to HRSV infection. We tested the efficacy of a polyanhydride nanoparticle-vaccine (nanovaccine) encapsulating the post-fusion F and G proteins from BRSV and CpG adjuvant in neonatal calves with maternally-derived antibodies (MDA). A prime-boost immunization of neonatal calves with the BRSV post-fusion F/G CpG nanovaccine induced clinical protection in the presence of MDA. Vaccinated calves developed moderate or no clinical symptoms and were mostly protected from virus-associated lung pathology compared to unvaccinated controls. Calves that received a mucosal/systemic nanovaccine immunization had complete virological protection against BRSV infection in the lungs, which correlated with BRSV-specific immune responses in the bronchoalveolar lavage. Vaccination with the nanovaccine induced anamnestic BRSV-specific IgA titers in the lungs and systemic T cell responses with BRSV-specific CD4+ T cell proliferation and antigen-specific IL-17 and IFNγ secretion. Overall, the BRSV post-fusion F/G CpG nanovaccine is a promising candidate for further studies to protect neonatal calves and infants from RSV infection.
Polyfunctionality of T cell immunity in pre-immune and naïve individuals after COVID-19 mRNA vaccination
Authors: Moraes Vanessa S , Ross Ted M 
Published: January 2023

Abstract
Humans make SARS-CoV-2-specific antibodies, CD4+ T cells, and CD8+ T cells in response to infection. Neutralizing antibodies are associated with protective immunity against secondary infection and are the mechanism that can provide truly sterilizing immunity. While not necessarily providing sterilizing protection, T cell responses are critical for limiting disease severity and can limit the danger of antibody escape variants by providing for rapid clearance. Thus, studying antibody, memory B cell, CD4+ T cell, and CD8+ T cell memory to SARS-CoV-2 in an integrated manner is important for understanding mechanisms and durability of protective immunity against COVID-19. Here we add SARS-CoV-2-specific memory CD4+ T cells and CD8+ T cells to those efforts with a particular interest in assessing whether pre-existing cross-reactive T cell memory impacts vaccine-generated immunity. We monitored T cell responses in 70 healthy individuals vaccinated with Pfizer–BioNTech mRNA vaccine over 9 months. We found that vaccination induced the development of a sustained spike-specific CD4+ and CD8+ T cell response which persisted for at least 6 months. Vaccine-generated spike-specific memory CD4+ T cells were comparable in quantity and quality to convalescent cases 6 months post-boost, including the frequency of T follicular helper cells and IFNγ-producing cells. Subjects with pre-existing cross-reactive CD4+ T cell memory had increased CD4+ T cell and antibody responses to the vaccine, demonstrating the biological relevance of SARS-CoV-2–cross-reactive CD4+ T cells. These data show that vaccination elicits an immunologically competent and long-lived SARS-CoV-2-specific T cell population in naïve and pre-immune individuals.
Supported by National Institute of Allergy and Infectious Diseases (NIAID), a component of the U.S. National Institutes of Health (NIH), Department of Health and Human Services, contract 75N93019C00052
Clonal structure and epitope specificity of Ad5-nCoV vaccine-induced T cell response to SARS-CoV-2
Authors: Sheetikov Savely , Khmelevskaya Alexandra A , Zornikova Ksenia V , Zvyagin Ivan V , Shomuradova Alina S , Afanasiev Andrei , Dzutseva Vitalina , Efimov Grigory A 
Published: January 2023

Abstract
As a part of phase 3 clinical trial of a single-dose adenoviral COVID-19 vaccine, we evaluated the dynamics and clonal structure of T cell response to the SARS-CoV-2 Spike protein.
T cell response measured in 50 donors by IFNg ELISpot after Spike stimulation peaked on the 14th day (d14) after vaccination. Flow cytometry demonstrated that the response was skewed to CD4+ cells. To isolate Spike-specific T cells we performed short-term T cell expansion of d14 samples (n=17 donors) with the recombinant Spike, restimulated and FACS sorted IFNg+ cells.
T cell receptor (TCR) beta chain sequencing identified from 36 to 629 Spike-specific clones (median 186) per individual. CD4+ response was more clonal than CD8+ (median 116 and 17 respectively) but CD8+ occupied a larger share of the total TCR repertoire (median 0.25% for CD4+ and 0.52% for CD8+) as a result of the larger average clone size at d14 (3.4×10−5 for CD8+ vs. 6.6×10−6 for CD4+). Both the number and total share of Spike-specific clones in the TCR repertoire decreased at day 28 and further at day 180. At 6 months a median of 1 (0 to 18) of the initial Spike-specific clones were detectable in the peripheral blood. This number increased to 39 (9 to 105) as a result of T cell expansion.
For 14 donors we also performed rapid expansion of T cells sampled at d14 with 11 Spike-epitopes (9 MHC I and 2 MHC II) followed by sorting of activated cells and TCR sequencing. We identified on average 41 unique epitope-specific clones (2 to 141). They were mostly undetectable in the peripheral blood at 6 months (0 to 8 clones), but their numbers increased to the median of 7 clones (0 to 35) after cultivation with Spike.
Overall this demonstrates the induction of polyclonal and stable T cell response by a single injection of adenoviral vaccine.
The work was supported by the Russian Science Foundation grant 20-15-00395.
Antibody-dependent enhancement of SARS-CoV-2 spike binding to human ACE2 receptor by virus neutralizing antibodies
Authors: Magazine Nicholas J , Guan Yongjun , Yu Lei , Carossino Mariano , Zhang Tianyi , McGee Michael C , Huang Weishan 
Published: January 2023

Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its associated disease COVID-19 has caused over 275 million cases of illness and over 5 million deaths as of December 2021. The spike glycoproteins expressed on surface of the SARS-CoV-2 particle are the primary target of both vaccination and monoclonal antibody therapies. Widely reported evidence suggests that a subset of both naturally produced and synthetic antibodies (Abs) enhance infection, a phenomenon known as antibody-dependent enhancement (ADE). Recently, it has been demonstrated that the mechanism by which some Abs may elicit ADE is by enhancing the binding affinity of SARS-CoV-2 spike to its ligand - the human ACE2 receptor. Using in vitro spike-ACE2 binding assays, we identified two Abs which demonstrated ADE for spike/human ACE2 binding. Surprisingly, these binding-enhancing antibodies sufficiently neutralized SARS-CoV-2 pseudo-virus infection in human ACE2-knockin 293T cells, as well as live virus infection in Vero-E6 (non-human primate cell line) and Calu-3 (human cell line) cells in vitro. Furthermore, these receptor-binding enhancing antibodies significantly reduce but not enhance signs of SARS-CoV-2 infection in human ACE2-knockin mice, including viral replication and tissue pathology. Taken together, our data suggest a discrepancy between results of in vitro spike/receptor binding assay and virus neutralization assays for SARS-CoV-2 infection. Enhanced spike binding to host receptors is not a reliable measure of screening for ADE antibodies, but may be a mechanism by which virus neutralization is achieved in nature.
This work was supported in part by grants from the National Institutes of Health (R01 AI151139, R56 AI146226 and P20 GM130555). W. Huang also received research support from MegaRobo Technologies Corporation.
Differences between autoantibodies induced by SARS-CoV-2 infection and Pfizer-BioNTech SARS-CoV-2 vaccination
Authors: Geanes Eric S , Bradley Todd 
Published: January 2023

Abstract
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection can induce severe disease and lead to hospitalization and/or death in some subpopulations of patients. The underlying mechanisms of disease severity between individuals remain unclear. Recently, autoantibodies have been detected after severe SARS-CoV-2 infection, including anti-idiotypic IgM antibodies targeting ACE2, the receptor for SARS-CoV-2. In this study, we examined autoantibody differences within hospitalized patients with severe SARS-CoV-2 infection, individuals with previous SARS-CoV-2 infection (seropositive), and individuals with no previous exposure to SARS-CoV-2 (seronegative). An autoantibody multiplex panel was run on serum collected from seropositive and seronegative individuals before vaccination and at 7 weeks after two doses of vaccine. Anti-ACE2 antibodies were measured for these samples in addition to serum collected from hospitalized individuals with severe SARS-CoV-2 infection. Severe SARS-CoV-2 infection resulted in elevated anti-ACE2 IgG, IgA, and IgM antibodies compared to all vaccine groups. Seropositive samples resulted in significantly higher anti-ACE2 IgG after vaccination, and significantly higher IgA and IgM antibodies before vaccination compared to seronegative samples. ACE2 inhibition was measured for samples with measurable anti-ACE2 IgG to determine if anti-ACE2 autoantibodies were affecting ACE2 function. The findings of this study elucidate severe SARS-CoV-2 infection results in higher IgG, IgA, and IgM anti-ACE2 antibodies compared to non-hospitalized SARS-CoV-2 infection and SARS-CoV-2 naïve individuals and in turn may provide future novel identifiers for SARS-CoV-2 disease severity.
Supported by funds provided by the Children’s Mercy Research Institute
Optimizing a multi cancer antigen plasmid-based vaccine using an in situ prediction model.
Authors: Cecil Denise L , Drovetto Nicholas , Doan Ivy , Rodmaker Erin , Corulli Lauren , Sei Shizuko , Disis Mary L 
Published: January 2023

Abstract
Little is known about optimization of a class II restricted plasmid-based vaccine targeting multiple cancer antigens. We set out to define parameters of an in situ prediction model in vaccine design for optimal immunogenicity. Using previously defined immunogenic class II epitopes from five cancer antigens, we evaluated the effects of using linkers between antigens and varying the antigen order. The construct with linkers was superior to the construct without linkers. The mice receiving the vaccine with linkers developed a greater magnitude (p&lt;0.0001), incidence and breadth IFN-g immune response. To determine the optimal antigen order, the sequences of all possible permutations were generated. Each sequence was evaluated in four publicly available algorithms to assess potential generation of immunity and physiochemical properties of the fusion protein. Of all parameters assessed, only predicted class II immunogenicity, in vivo half-life and percent of epitopes in helical secondary structure differed considerably among the constructs. We evaluated three constructs with similarly high class II immunogenicity; one with (A) long in vivo half-life and high percent in helices, one with (B) short half-life and high percent in helices or one with (C) short half-life and low percent in helices. Construct A generated a higher magnitude IFN-g response than Construct B or C (p&lt;0.006 for all). While there was 100% incidence of an immune response to all constructs, the breadth of the response for Construct A was greater than the other two constructs. Class II immunogenicity, in vivo half-life and the percent of the protein in a helical secondary structure can be used to discern the most immunogenic multi-antigen plasmid vaccine construct.
CD45RA expressing lymphocytes inhibit the outgrowth of Epstein-Barr Virus Specific T-cells from Patients with EBV+ Lymphoma
Authors: Sharma Sandhya , Woods Mae , Mehta Naren , Rooney Cliona 
Published: January 2023

Abstract
EBV antigen specific T-cells (EBVSTs) are difficult to expand from patients with EBV-positive lymphoma, perhaps because they circulate with low frequency or are rendered dysfunctional by the suppressive lymphoma microenvironment. As a result, we were unable to produce EBVSTs from ~25% of EBV+ lymphoma patients in our clinical trial (NCT01555892). To enrich memory T-cells prior to EBVST generation, we depleted the CD45RA+ subset of PBMCs, that includes naïve T-cells (TN), NK-cells, T-stem cells (TSCM) and terminally differentiated T-cells (TEMRA). CD45RA depletion (RAD) enhanced EBVST proliferation, antigen-specificity, polyfunctionality, cytotoxicity and tumor clearance in a murine model, and most importantly, enabled the outgrowth of EBVSTs from patient PBMCs that had previously failed to respond. This observation led to our hypothesis that bystander CD45RA+ cells inhibit the outgrowth of memory EBV antigen specific T-cells.
TCR-β chain sequencing showed that the clonotypes with greatest expansion in RAD-EBVSTs failed to expand or expanded poorly in EBVSTs from whole PBMCs (W-EBSVTs), reflecting a more selective clonal expansion in RAD-EBVSTs. TCR repertoire of W-EBVSTs was more similar to that of EBVSTs derived from CD45RA+ PBMCs that lacked detectable EBV specificity and to that of the original PBMCs. Subsets add back to RAD-PBMCs suggested that the CD3+ CD45RA+ subset inhibits the reactivation and clonal expansion of EBV-Ag specific memory T-cells. This may represent a physiological strategy to prevent the oligoclonal outgrowth of inappropriate memory T-cells during primary infection. This approach has been translated into our ongoing clinical trials using NCT01555892 and NCT04288726 using EBVST against lymphoma.
This work was funded and supported by CPRIT RP160283- Baylor College of Medicine Comprehensive Cancer Training Program, NIH-NCI P50 CA126752, American Society of Gene and Cell Therapy (ASGCT) Career Development Award, and Tessa Therapeutics.
Pre-existing biochemical entities predict severity of cytokine release syndrome in B-cell lymphoma patients treated with CD19-directed CAR T-cells
Authors: Gupta Neetu , Jalota Akansha , Hershberger Courtney E , Patel Manishkumar S , Mian Agrima , Faruqi Aiman , Khademi Gholamreza , Rotroff Daniel M , Hill Brian T 
Published: January 2023

Abstract
Potentially severe immune-related adverse events (irAEs) develop in relapsed/refractory large B cell lymphoma (R/R LBCL) patients during treatment with anti-CD19 CAR T-cell therapy. Therefore, prediction and better management of toxicities is critically required to improve patient outcomes. Cytokine release syndrome (CRS), one of the most notable acute irAEs, is the result of an inflammatory response of activated CAR T-cells and myeloid cells. Interestingly, high levels of glucose and low levels of certain amino acids are associated with pro-inflammatory immune activation in infectious and autoimmune diseases, but such associations have not been investigated in CAR T-cell therapy. Therefore, we employed mass spectrometry-based untargeted and targeted metabolomics analysis to identify pre-treatment host biological metabolites that predict rapid onset and severe CRS in 41 R/R LBCL patients treated with Axicabtagene Ciloleucel or Tisagenlecleucel. Analysis of plasma metabolites revealed significant associations of CRS with several metabolic classes, including carbohydrates, lipid, amino acids, dipeptides and nucleotides. Interestingly, high levels of the carbohydrates, glucose, mannose and 1,5-anhydroglucitol were observed in patients who experienced more rapid onset of CRS. In contrast, patients with greater severity of CRS had lower plasma abundance of the amino acids, glycine, proline and glutamate. Our data suggest that specific pre-existing plasma metabolites have the potential to serve as predictive biomarkers of CRS in patients undergoing anti-CD19 CAR T-cell therapy, and may be used for risk stratification and clinical management of irAEs in advance of treatment.
Promoted checkpoint inhibitor-based tumor immunotherapy by small-molecule prodrug hydrogelator
Authors: Xu Dongqing , Wan Fengyi 
Published: January 2023

Abstract
Cancer immunotherapy with immune checkpoint inhibitors (ICIs) has shown great promise in harnessing host immune system to combat cancer. That said, only a portion of patients has benefit directly from the anti–programmed cell death protein 1 (anti-PD1) therapy, whereas the others with an immunosuppressive tumor microenvironment (TME) achieve poor clinical outcomes. Hence, promoting an immune-stimulating TME that favors tumor-infiltrating lymphocytes is an important means to maximize the cancer immunotherapeutic potentials. Small-molecule prodrugs conferred intratumoral delivery of ICI agents have emerged as novel local delivery approaches to enhance antitumor immunity for cancer therapy. Our recent study revealed a combined chemoimmunotherapy could serve as a reservoir for extended release of DNA-damaging agent camptothecin and ICI agent anti-PD1 antibody, thus eliciting robust and durable systemic antitumor immunity in mice. The underlying mechanisms on how this carrier-free therapeutic system boosts immune sensing of immunogenic tumors and priming of CD8+ T cells against tumor antigens and the impact of microbiota will be discussed. Together, these results provide important insights into the use of small-molecule prodrugs as both chemotherapeutic and carrier, to awaken and enhance antitumor immune system for improved cancer therapy.
Supported by NIH grant R01CA244350
Effect of Standard of Care Treatment on Tumor Membrane Vesicle Vaccine for Triple-Negative Breast Cancer
Authors: Monterroza Lenore , Munoz Luis , Bommireddy Ramireddy , Samaranayake Sarah , Pack Christopher , Selvaraj Periasamy 
Published: January 2023

Abstract
Metastatic triple-negative breast cancer (TNBC) has a poor prognosis primarily due to metastatic potential and resistance to currently available therapies. Recent studies suggest that TNBC may be a viable target for immunotherapy due to the presence of high levels of infiltrating lymphocytes. However, many TNBC patients do not respond due to high intra-tumoral and inter-patient heterogeneity. Utilizing a novel protein transfer method, we generate a personalized vaccine based on tumor membrane vesicles (TMVs) derived from whole tumor tissues modified with glycolipid-anchored forms of the immunostimulatory molecules (GPI-ISMs) B7-1 and IL-12. In prior studies we observed that simultaneous delivery of TMVs along with biological adjuvants enhanced CD8 T cell immunity. Herein, we investigated the impact of standard-of-care (SOC) chemotherapy on the resulting quality of the TMV vaccine. Mice were injected with 4T1 TNBC cells subcutaneously and treated with three rounds of SOC drugs starting at day 15. Chemotherapies affected tumor growth to varying degrees. Mice were euthanized and tumors were harvested for TMV preparation. Results show that depending on the treatment, the yield and particle size of TMVs was impacted. While SOC drugs affected some surface markers on TMVs, expression of CD24 and CD11b was unchanged. Further, there was reduced uptake of TMVs by dendritic cells prepared from cisplatin and 5-fluorouracil treated tumors. GPI-ISMs were equally incorporated by protein transfer onto TMVs despite SOC treatments. In summary, our studies in the 4T1 model show that a TMV vaccine can be successfully prepared after chemotherapy, but the impact on TMV yield and size justifies further investigation.
Supported by R01 CA202763
Extracellular CIRP Induces Novel Nectin-2+ (CD112+) Neutrophils to Promote Th1 Differentiation in Sepsis
Authors: Murata Kensuke , Murao Atsushi , Aziz Monowar , Wang Ping 
Published: January 2023

Abstract
Neutrophil heterogeneity represents different subtypes, states, phenotypes, and functionality of neutrophils implicated in sepsis pathobiology. Extracellular cold-inducible RNA-binding protein (eCIRP) is a damage-associated molecular pattern that promotes inflammation and alters neutrophil phenotype and function through TLR4. Nectin-2 or CD112 is an Ig-like superfamily member. CD112 serves as the ligand for DNAM-1 (CD226), which induces Th1 differentiation in naive CD4+ T cells. Th1 cells produce IFN-γ to fuel inflammation. CD112 is expressed mainly on APCs, but its expression in neutrophils is unknown. We hypothesize that eCIRP induces CD112 expression in neutrophils, promoting Th1 differentiation in sepsis. Incubation of neutrophils with recombinant murine (rm)CIRP significantly increased the gene and protein expression of CD112 in neutrophils. Anti-TLR4 Ab-treated neutrophils significantly decreased CD112+ neutrophils compared with controls upon rmCIRP stimulation. After 4 h of rmCIRP injection in mice, CD112+ neutrophils were significantly increased in the blood and spleen. At 20 h after cecal ligation and puncture–induced sepsis, CD112+ neutrophils were also significantly increased. Blood and splenic CD112+ neutrophils in septic CIRP−/− mice were much lower than in septic wild-type mice. Coculture of naive CD4 T cells with rmCIRP-treated (CD112+) neutrophils significantly increased IFN-γ-producing Th1 cells compared with coculture with PBS-treated neutrophils. CD112 Ab significantly attenuated Th1 differentiation induced by rmCIRP-treated neutrophils. Thus, eCIRP increases CD112 expression in neutrophils via TLR4 to promote Th1 differentiation in sepsis. Targeting eCIRP may attenuate sepsis by reducing Th1-promoting CD112+ neutrophils.
TBK1 Isoform Inhibits Grass Carp Reovirus Infection by Targeting the Degradation of Viral Nonstructural Proteins NS80 and NS38
Authors: Zhang Jie , Chang Ming Xian 
Published: January 2023

Abstract
TANK-binding kinase 1 (TBK1) undergoes alternative splicing, and the previously reported TBK1 isoforms are negative regulators of RIG-I–like receptor–mediated type I IFN production. Although a study has suggested that grass carp TBK1 has an opposite effect at high- and low-titer of grass carp reovirus (GCRV) infection, the functions of grass carp TBK1 isoforms in GCRV infection remain unclear. In this study, we show that a TBK1 isoform from grass carp (Ctenopharyngodon idellus) named as gcTBK1_tv3, which has a 1-aa difference with zebrafish TBK1_tv3, inhibits the replication and infection of GCRV both at high and low titers of infection in C. idellus kidney cells. gcTBK1_tv3 can colocalize and interact with the NS80 and NS38 proteins of GCRV. Furthermore, gcTBK1_tv3 specifically degrades the NS80 and NS38 proteins of GCRV through the ubiquitin-proteasome pathway. Mechanistically, gcTBK1_tv3 promotes the degradation of NS80 or NS38 for K48-linked ubiquitination by targeting the Lys503 residue of NS80 or Lys328 residue of NS38, respectively, which ultimately impairs the production of cytoplasmic viral inclusion bodies and limits GCRV replication and infection. Taken together, our findings provide insight into the function of TBK1 isoform in the antiviral immune response and demonstrate that TBK1 isoform can target the nonstructural proteins of GCRV for impairing the formation of viral inclusion bodies.