bluuebunny/crossref_metadata_2025_split huggingface dataset:
The effect of neutrophils on limited hepatitis C virus vaccine immunogenicity in people who inject drugs
Authors: Roznik Katerina , Rousseau Kimberly E , Thompson Elizabeth A , Cox Andrea L 
Published: August 2023

Abstract
In the only efficacy trial of a hepatitis C virus (HCV) vaccine, the vector-based vaccine lowered peak HCV RNA levels in people who inject drugs (PWIDs) but did not prevent chronic HCV infection. The same vaccine regimen tested in healthy controls (HCs) resulted in a larger percentage of individuals responding to vaccination (100 vs. 78%) and a 3-fold higher magnitude of T cell responses compared to PWIDs. We evaluated baseline immune characteristics of PWIDs to understand the reduced responses, ruling out differences in baseline anti-vector immunity, subclinical HCV exposure, and increased Tregs. However, we found markedly elevated neutrophil counts in PWIDs at baseline compared to HCs. Elevated neutrophils were previously linked to nonresponse to hepatitis B vaccination in HCs, yet this phenomenon has been largely understudied. We analyzed baseline neutrophil phenotypes using flow cytometry and identified elevated PD-L1, CD11b, and CD63 on neutrophils from PWIDs compared to HCs. Stimulation of HC neutrophils with opioid receptor (OR) agonist DAMGO significantly upregulated PD-L1 and CD11b, an effect partially reversed with an OR antagonist. We also observed metabolic reprogramming of neutrophils in PWIDs compared to HCs, with higher expression of glycolytic machinery (GLUT1/HK2) and mitochondrial membrane protein VDAC1, indicative of increased mitochondrial mass and use of oxidative phosphorylation. Elevated baseline CD11b and CD63 suggest dysregulation and activation of neutrophils, with increased PD-L1 potentially contributing to poor T cell responses in PWIDs. Mechanistic studies of dysregulated neutrophil interactions with T cells may provide insights into vaccine-induced responses and future vaccine efforts.
Supported by NIH grant U19 AI159822.
Determining antigen-specific T cell phenotypes in cases of Hepatitis C virus clearance versus chronic infection
Authors: Stavrakis Georgia , Rousseau Kimberly , Thompson Elizabeth , Cox Andrea L 
Published: August 2023

Abstract
Globally, approximately 1.5 million new Hepatitis C virus (HCV) infections occur annually. A vaccine to prevent chronic HCV infection would limit HCV transmission and pathogenesis of HCV. To understand immunity to HCV infection in high risk populations, we prospectively followed HCV antibody negative high-risk people who inject drugs (PWIDs) in a protocol designed for monthly follow-up. We demonstrated previously that HCV reinfection in PWIDs is associated with a reduction in the magnitude and duration of viremia versus the initial infection and broadened cellular immune responses, consistent with protective immunity against chronic disease. We have characterized CD8+ HCV-specific T cells from the acute phase of HCV using high dimensional spectral flow cytometry to define the simultaneous expression of 25 intracellular metabolic and cell surface molecules. We found that HCV-specific CD8+ T cells from those who progress to chronic infection have a unique acute phase phenotype that clusters separately from those who later successfully control HCV. Acute phase CD8+ HCV-specific T cells from those who subsequently clear an initial HCV infection show increased expression of CD127 and CXCR3 and decreased expression of GLUT1, PD-1 and CD39 compared to acute phase T cells in those who later progress to chronic infection. Understanding HCV specific T cell phenotypes in successful control of HCV infection and reinfection will guide goals for vaccine induced immunity to prevent chronic HCV infection.
Supported by grants from the NIH (U19 AI159822)
TNFR2-targeting peptide for cancer immunotherapy
Authors: Chen Xin , Liao Ping , Chen Zhonghao 
Published: August 2023

Abstract
There is compelling evidence that tumor necrosis factor receptor type II (TNFR2) is preferentially expressed by highly immunosuppressive CD4+Foxp3+ regulatory T cells (Tregs), especially those present in tumor-infiltrating lymphocytes (TILs). These findings led us and others to propose and experimentally proved that targeting TNFR2 with antibodies could induce anti-tumor immune responses. In this study, we identified a TNFR2-targeting peptide P20 through phage display. P20 had in vitro activity to block TNF-TNFR2 interaction and inhibit TNFR2-mediated activation/expansion of Treg cells. The in vivo effect of P20 on tumor growth was examined with mouse tumor models including CT26 colon cancer and OVA-EG7 lymphoma. The results showed that the treatment with P20 (i.p.) resulted in a marked inhibition of tumor growth, accompanied by a reduced number of Tregs and an increased number of IFN-γ+ CD8+ cytotoxic T lymphocytes in the tumor microenvironment. Furthermore, P20 could markedly enhance the efficacy of anti-PD-L1 in the inhibition of tumor growth in a syngeneic MC38 mouse colon cancer model. Thus, P20 may be a useful anti-tumor immunotherapeutic agent and merits further investigation. (Funded by FDCT 0099/2021/A2, UM MYRG2022-00260-ICMS, UM CPG2023-00025-ICMS)
There is compelling evidence that tumor necrosis factor receptor type II (TNFR2) is preferentially expressed by highly immunosuppressive CD4+Foxp3+ regulatory T cells (Tregs), especially those present in tumor-infiltrating lymphocytes (TILs). These findings led us and others to propose and experimentally proved that targeting TNFR2 with antibodies could induce anti-tumor immune responses. In this study, we identified a TNFR2-targeting peptide P20 through phage display. P20 had in vitro activity to block TNF-TNFR2 interaction and inhibit TNFR2-mediated activation/expansion of Treg cells. The in vivo effect of P20 on tumor growth was examined with mouse tumor models including CT26 colon cancer and OVA-EG7 lymphoma. The results showed that the treatment with P20 (i.p.) resulted in a marked inhibition of tumor growth, accompanied by a reduced number of Tregs and an increased number of IFN-γ+ CD8+ cytotoxic T lymphocytes in the tumor microenvironment. Furthermore, P20 could markedly enhance the efficacy of anti-PD-L1 in the inhibition of tumor growth in a syngeneic MC38 mouse colon cancer model. Thus, P20 may be a useful anti-tumor immunotherapeutic agent and merits further investigation
Blocking Respiratory Syncytial Virus infection utilizing decoy ICAM-1 receptor proteins
Authors: Geanes Eric S , McLennan Rebecca , Khanal Santosh , Bradley Todd 
Published: August 2023

Abstract
Respiratory syncytial virus (RSV) infections result in a significant number of hospitalizations for young children and the elderly each year, and current infection rates continue to rise, leading to a substantial public health burden. While numerous vaccine candidates are in clinical trials, there is currently no approved RSV vaccine to prevent infection or antiviral treatment after infection. There is an urgent need to find new therapeutics and strategies that reduce severe RSV disease that could improve health and save lives. The RSV fusion (F) protein is critical for binding host cells and mediating infection and is the target of most candidate vaccines. The RSV F protein has been shown to interact with several host-cell surface receptor proteins such as Intercellular Adhesion Molecule 1 (ICAM-1), NCL, EGFR, IGF1R, and infect respiratory epithelial cells. In this study, we confirmed ICAM-1 could bind RSV F protein and inhibit RSV infection via an RSV neutralization assay. Using RSV F protein as bait, we immunoprecipitated RSV F interacting proteins from A549 respiratory cells and performed mass spectrometry analysis to identify novel host proteins that bind RSV F protein. Finally, we engineered a mRNA-lipid nanoparticle (LNP) expressing soluble ICAM-1. The goal is to utilize the ICAM-1 mRNA-LNP as a passive intranasal vaccine that would result in over-expressed soluble ICAM-1 in respiratory mucosal surfaces to function as a decoy receptor that would reduce RSV infection. We are evaluating this therapeutic approach in preclinical models as a prophylactic or treatment after infection. ICAM-1 mRNA-LNP may provide a novel defensive strategy to reduce RSV infection or disease severity and augment future RSV vaccines that become approved.
Children’s Mercy Research Partners
Oral vaccination with live attenuated pathogenic bacteria protects immunocompromised hosts from lethal colitis
Authors: Wang Shuyu , Wan Fengyi 
Published: August 2023

Abstract
Enteropathogenic Escherichia coli(EPEC) and enterohemorrhagic E. coli(EHEC) are significant causative agents for diarrheal diseases globally. Vaccination has been deemed as the most effective strategy for preventing EPEC and EHEC caused foodborne illnesses; however, FDA has not yet licensed any vaccine against EPEC or EHEC. Of note, live attenuated bacterial vaccine strategy has been scarcely investigated for against EPEC or EHEC infection, in particular in immunocompromised populations where the infections are associated with severe symptoms and outcomes. Infection with Citrobacter rodentium(CR), the murine equivalent of EPEC and EHEC, in mice has been widely used to develop novel vaccine strategies and evaluate vaccination efficacy. CR infection in immunocompromised mice such as interleukin-22 knockout (Il22−/−) mice leads to severe symptoms and mortality, which mirrors the severe outcomes of EPEC and EHEC infections in immunocompromised populations. We explored whether oral inoculation with live CR mutants with rationally attenuated virulence could protect lethal infection in susceptible Il22−/− mice. We showed that oral administration of live attenuated CR strains elicited efficient systemic and humoral immunity against variant CR virulence proteins, hence protecting otherwise lethal CR infection, in Il22−/− mice. Our findings provide proof of concept evidence for live attenuated vaccination strategy to prevent EPEC/EHEC infections in immunocompromised hosts associated with more severe symptoms and lethality, and reveal novel CR-specific antibody subtypes and virulence determinants that are critical for live attenuated vaccination offered immune protection.
Vaccine antigen archiving by lymphatic endothelial cells boosts archived antigen-specific CD8 T cell memory responses during a pathogenic challenge
Authors: Doan Thu A , Sheridan Ryan , Forward Tadg , Tamburini Beth 
Published: August 2023

Abstract
Numerous studies have shown viral antigens can persist in lymph nodes after the resolution of the infection. We demonstrated that lymphatic endothelial cells (LEC), which comprise the lymphatic vasculature necessary for antigen drainage from the tissue, are the predominant cell type required for the persistence of both vaccine and viral antigens within the lymph node. We termed this process antigen archiving due to the ability of LEC to actively archive antigens to which an immune response has occurred. This process involves antigen acquisition, retention, and exchange of the antigen between LECs and dendritic cells (DC), resulting in the presentation of the archived antigens to archived antigen-specific CD8 memory T cells. In this study, we evaluated the effect of LEC archived antigen on archived antigen-specific memory T cells during a pathogenic challenge. We predicted that a pathogenic challenge would result in the release of the archived antigen and subsequent archived antigen presentation. Following a pathogenic challenge, we found a significant increase in archived antigen-specific endogenous or transferred memory CD8 T cells, compared to bystander activation seen by memory T cells of an unrelated specificity. The phenotype of the archived antigen-specific memory CD8 T cells was more effector-like and led to a significant increase in protection against Listeria monocytogenes (LM)-expressing the archived antigen, in the local area. In conclusion, these findings suggest that archived antigen may be released during a pathogenic challenge. As such, the archived antigen may be acquired by migratory DCs as they traffic through the lymphatics in order to present the archived antigen to memory CD8 T cells acting as an effective boost.
NIH R01 AI121209
The mature c-terminal domain of filamentous hemagglutinin is sufficient to provide protection against Bordetella pertussis in the murine nasal cavity
Authors: Pyles Gage Michael , Weaver Kelly , Witt William , Huckaby Annalisa , Mateu-Borras Margalida , Damron Fredrick , Barbier Mariette 
Published: August 2023

Abstract
Bordetella pertussis is the causative agent of whooping cough. The transition from whole cell (wP) to acellular pertussis vaccines (aPs) triggered an increase in pertussis incidence despite wide-spread vaccine coverage. aPs contain protein antigens purified from B. pertussis: pertactin, fimbriae, filamentous hemagglutinin (FHA), and pertussis toxin (PT). Antibodies are critical for protection against B. pertussis, and have been demonstrated to trigger opsonization, block bacterial adhesion, and neutralize disease symptoms. Following aP-vaccination, however, antibodies target a diminished number of functional B cell epitopes as the antigens are chemically denatured during aP manufacturing. The effects of denaturation on PT are well documented, but this has not been studied for other vaccine antigens, such as FHA. Additionally, FHA is a large antigen, and most of the protein does not play a role in bacterial adhesion. To direct antibody responses toward non-denatured, functionally relevant portions of FHA we aimed to test the Mature C-terminal Domain (MCD) of FHA as vaccine antigen. Protection provided by MCD was compared to several vaccine preparations, including aP and native FHA. We observed that aP and wP vaccines lead to the induction of antibodies that recognize FHA, MCD and B. pertussis. Using a CD-1 murine model of vaccination and challenge we demonstrated that MCD is immunogenic and sufficient to decrease bacterial burden in the murine airway, providing ~85% reduction in B. pertussis present in the nasal cavity. Overall, our data supports the use of MCD as a vaccine antigen for protection against B. pertussis. Ongoing studies focus on the selection of the adjuvants and route of administration to optimize vaccine efficacy.
Mariette Barbier is funded by NIH grant 5R01AI141671-04. Fredrick Damron is funded by NIH grant 5R01AI137155-04. The Vaccine Development Center Research Challenge Grant (no. HEPC.dsr.18.6) from the Division of Science and Research, WV Higher Education Policy Commission also contributed support.
Mucosal chemokine adjuvant supports heterologous protection against Delta and Omicron VOCs in the context of a SARS-CoV-2 wild-type DNA vaccine
Authors: Gary Ebony Nicole , Tursi Nicholas J , Warner Bryce M , Parzych Elizabeth M , Ali Ali R , Frase Drew , Moffat Estella , Embury-Hyatt Carissa , Smith Trevor R F , Broderick Kate E , Humeau Laurent , KobasA Darwyn , Patel Ami , Kulp Daniel W , Weiner David B 
Published: August 2023

Abstract
Emerging variants of concern (VOCs) and waning vaccine-induced immunity challenge SARS-CoV-2 control. Directing vaccine-induced humoral and cell-mediated responses to mucosal surfaces could enhance vaccine efficacy and potentially reduce transmission. We immunized mice with DNA plasmids expressing a wild-type SARS-CoV-2 spike protein (pS) alone, or with the DNA-encoded mucosal chemokine adjuvant cutaneous T cell-attracting chemokine (pCTACK). Mice co-immunized with pS and pCTACK exhibited increased spike specific IFNy T cell responses compared to those receiving the pS alone. While both groups showed similar levels of spike receptor binding domain (RBD)-specific IgG in their serum, in the mucosa, co-immunization with pCTACK enhanced RBD-specific IgA, and cecal extracts from pCTACK co-immunized animals neutralized pseudotyped viruses. When ACE2 transgenic mice were immunized, rested for 3 months, and challenged with a heterologous SARS-CoV-2 Delta variant, co-immunization with pCTACK supported 100% survival while immunization with wild-type pS antigen alone supported 60% survival. Enhanced protection from morbidity was also observed as 100% of pCTACK co-immunized animals did not exhibit infection-induced weight loss. When animals were immunized similarly and challenged with Omicron BA.2, pCTACK co-delivery resulted insignificantly decreased viral loads compared to pS-only immunization. These studies demonstrate that mucosal chemokine adjuvants can direct vaccine-induced responses to specific immunological sites with significant impact on heterologous challenge. These data have significant implications for the design of improved vaccines targeting SARS-CoV-2.
This work was supported by NIH NCI award T32 CA09171 (ENG) and NIH NIAID award T32-AI-055400 (EMP) and The Wistar Institute coronavirus discovery fund with additional support from CEPI/Inovio Pharmaceuticals.
Acute and chronic viral infection generate brain-resident memory T cells with distinct phenotypic, functional, spatial, and metabolic profiles
Authors: Mix Madison R , Epping Lecia L , Hancox Lisa S , Merrill Ronald A , Taylor Eric B , Badovinac Vladimir P , Harty John T 
Published: August 2023

Abstract
The rising incidence of post-viral neurological sequelae represents a substantial burden on global public health. Therefore, there is an urgent need to investigate alterations in brain immune composition after acute and chronic viral infection. While virus-specific tissue-resident memory (TRM) CD8+ T cells are observed in human health and neurological disease, these populations remain understudied in the brain. Here, we leveraged the comparative model system of lymphocytic choriomeningitis virus (LCMV) strain Armstrong vs. Clone 13 infection in mice to generate brain TRM with quiescent and exhausted profiles respectively. We then interrogated the 1) phenotype, 2) functionality, 3) localization, and 4) metabolism of anti-viral TRM in the brain. Importantly, brain TRM exhibit higher expression of exhaustion markers (PD-1, LAG-3, TIM-3, TOX) compared to T circulating memory (TCIRCM) cells in the same host. Furthermore, brain TRM in chronically infected hosts are functionally impaired in cytokine production and cytolytic capacity. Anti-viral brain TRM seed distinct neuroanatomical sites, such as the cerebrospinal fluid, parenchyma, and choroid plexus, often adopting aggregate-like structures in solid compartments. Compared to TCIRCM, quiescent and exhausted brain TRM also demonstrate higher expression of surface amino acid transporters with disparate amino acid uptake. These findings are contextualized by whole brain metabolomic profiling indicating global perturbations in amino acid metabolism post-acute and chronic viral infection. Altogether, these data highlight brain-intrinsic memory T cell signatures and offer broad insight into how brain TRM may influence post-viral neurological outcomes.
Supported by grants from the NIH (AI042767, AI114543, AI167847, 5T32GM007337) and the University of Iowa Post-Comprehensive Research Fellowship.
Rhesus macaque cytomegalovirus deploys viral Fcγ receptors to evade antibody-mediated cellular immunity
Authors: Otero Claire E , Petkova Sofia , Taher Husam , Scheef Elizabeth , Moström Matilda , Sprehe Lesli , Kendall Savannah , Papen Courtney , Früh Klaus , Kaur Amitinder , Permar Sallie , Hengel Hartmut , Malouli Daniel , Kolb Philipp 
Published: August 2023

Abstract
Cytomegalovirus (CMV) is a top priority in vaccine development due to the high burden of disease in immunocompromised populations and congenitally infected infants. Evidence suggests that CMV-specific Fc-mediated antibody effector responses, such as antibody dependent cellular phagocytosis and cytotoxicity, are key for prevention of multiple CMV disease outcomes. However, human CMV expresses three Fcγ receptors that have been shown in vitro to synergistically interfere with anti-CMV IgG engagement of host Fcγ receptors (FcγRs). We recently identified three functionally homologous viral Fcγ receptors (vFcγRs) in rhesus macaque (RM) CMV (RhCMV), which allow us to validate the function of vFcγRs in vivo. Using a full-length (FL) RhCMV backbone (Taher, et al. 2020), we produced a mutant RhCMV lacking all identified vFcγRs (ΔvFcγR) and compared infection of CMV-seronegative male RMs with ΔvFcγR RhCMV (n=4) to FL RhCMV (n=3). RMs infected with ΔvFcγR and FL RhCMV experienced similar peak DNAemia levels 1 week post infection but ΔvFcγR RhCMV DNAemia was contained by week 3 while FL RhCMV remained detectable in plasma through week 9. The observed early control of infection is concurrent with the development of RhCMV-specific IgG responses which peak 3–4 weeks post infection in both groups. The kinetics of RhCMV-specific binding and neutralizing antibody responses were similar between the two groups of RMs, suggesting that anti-RhCMV Fc-mediated antibody effector responses may have been more effective against the RhCMV deleted of vFcγRs. Thus, vFcγRs are promising candidates for vaccine immunogens to improve Fc mediated effector responses and efficacy to reduce incidence of CMV-associated birth defects and transplant complications.
Supported by grants from NIH P01-AI129859 and P01-AI129859_S1, NIH T32-CA009111, National CMV Foundation ECRA
T-bet controls the maintenance and differentiation potential of flu-induced effector memory B cells
Authors: Risley Christopher A , Schultz Michael D , Allie S Rameeza , Peel Jessica N , Liu Shanrun , Fucile Chris , Scharer Christopher D , Rosenberg Alex , Boss Jeremy M , Randall Troy D , Lund Frances E 
Published: August 2023

Abstract
Although the transcription factor (TF) T-bet is expressed by antigen-experienced human B cells, including the extrafollicular IgDnegCD27neg memory-like DN2 cells and the IgDnegCD27+ effector memory cells (eBmem), it is unknown whether T-bet expression influences the development, maintenance, or function of Bmem cells. Using oligo-labeled recombinant influenza nucleoprotein (NP) tetramers and paired single cell RNA and V(D)J sequencing, we characterized NP-specific Bmem in the draining LN of flu-infected memory mice. We identified 7 transcriptionally distinct clusters of mature non-replicating β Bmem cells. While none of the Bmem clusters expressed TFs required for plasma cell (PC) lineage commitment, one cluster, containing clonally-expanded somatically-mutated Bmem cells, expressed significantly higher levels of the T-bet gene, Tbx21, and was transcriptionally very similar to human T-bet+ eBmem. The NP+ Tbx21+ Bmem upregulated gene networks associated with metabolic reprogramming, protein synthesis and the mTOR-dependent unfolded protein response, suggesting that these cells might represent Bmem that are metabolically poised for rapid PC differentiation. Using constitutive and inducible models to delete Tbx21 specifically in B cells we showed that the rapid differentiation potential of Bmem is controlled by T-bet and that T-bet is required for the development and persistence of NP+ eBmem subsets in LN and lung. Thus T-bet not only marks Bmem with effector potential but also regulates the persistence and function of flu-induced NP+ Bmem that are transcriptionally poised to provide rapid humoral protection from infection.
Supported by a grant from the NIH (R01 AI110508)