bluuebunny/crossref_metadata_2025_split huggingface dataset:
TCR-T cells engineered to overexpress c-Jun have better functionality with improved tumor infiltration and persistence for treatment of Hepatocellular Carcinoma
Authors: Hussein Mohamed Shawky , He Yukai 
Published: January 2023

Abstract
Liver cancer is the sixth most common cancer in the world and the third highest cause of cancer death in the world, indicating the treatment gap and the urgent need for novel effective therapies. The majority (80–90%) of liver cancers are hepatocellular carcinoma (HCC), with a general 5-year survival rate is 20%. Adoptive cell therapy (ACT) targeting malignant tumors have been generally unsuccessful due to lack of T cell activation in vivo with poor expansion and short persistence of the tumor infiltrating T cells. We proposed that engineering T cells to overexpress c-Jun, a transcription factor required for T cell development and activation, would activate T cells, enhance their function, and anti-tumor efficacy. In this study, using T-cell receptor–engineered T-cell (TCR-T) against HCC-associated antigen α-fetoprotein (AFP), we demonstrated that human TCR-T cells engineered to overexpress c-Jun (TCR-JUN) have better expansion potential in culture with enhanced functional capacity against HepG2 tumor cells. Additionally, TCR-JUN cells were less apoptotic and more resistant to exhaustion after HepG2 tumor stimulation. In HCC xenograft tumor model, c-Jun overexpression enhanced TCR+ cells expansion and overall survival of the tumor bearing mice. Interestingly, the tumor infiltrating lymphocytes (TILs) of TCR-JUN cells demonstrated enhanced tumor infiltration, persistence, functionality, and were less exhausted. We conclude that engineering T cells to overexpress c-Jun could be a potential approach to enhance T cell cancer therapy.
This work is supported by NIH Grant R01CA235159 to Dr. Yukai He.
Intranasal adjuvant immunotherapy re-activates immunity and enhances survival in murine metastatic Lymphangioleiomyomatosis
Authors: Maisel Katharina 
Published: January 2023

Abstract
Lymphangioleiomyomatosis (LAM) is cystic lung disease affecting primarily women of age. Currently there is no cure for this disease and only modest progress has been made with the introduction of sirolimus as LAM treatment. Recently, we and others have identified that LAM may also induce immunosuppression, similar to cancer. Adjuvant-based therapies like toll-like receptor (TLR) agonists are a promising strategy to activate cancer immunity. We used a murine model of metastatic LAM, to test therapeutic efficacy of TLR agonists PolyI:C, MPLA, and CpG, administered intranasally 2x/week. We found that only CpG enhances survival in murine LAM and that this treatment synergized with anti-PD1 therapy, which we had previously found to enhance survival in murine LAM. We found that the inflammation induced by CpG must be carefully balanced with its beneficial effects – in early disease, 5 μg of CpG was more beneficial than 10 μg. However, in progressive disease, 10 μg did not enhance survival while 20 μg CpG did. Furthermore, we found that a single dose of CpG reduced regulatory T cells and migratory dendritic cells (mDCs) and increased ICOS on T cells in the lungs. CpG also increased mDCs in the lymph nodes, and these were reduced after CCR7 blockade. We also found that repeated administration of CpG reduced immune cell infiltration into the lungs, which suggests a tolerogenic effect that may contribute to the incomplete survival. Overall, our data suggest that CpG may be a new treatment for LAM, that its beneficial effects may be in part mediated by CCR7 dependent DC migration, and that dose and inflammation must be carefully monitored to optimize therapeutic efficacy.
Supported by the ALA Dalsemer Award and LAM Foundation Career Development Award.
Eicosanoid signaling as a therapeutic target in middle-aged mice with severe COVID-19
Authors: Zheng Jian , Wong Lok-Yin Roy , Wilhelmsen Kevin , Li Kun , Ortiz Miguel E , Schnicker Nicholas J , Thurman Andrew , Pezzulo Alejandro A , Szachowicz Peter J , Li Pengfei , Pan Ruangang , Klumpp Klaus , Aswad Fred , Rebo Justin , Narumiya Shuh , Murakami Makoto , Zuniga Sonia , Sola Isabel , Enjuanes Luis , Meyerholz David K , Fortney Kristen , McCray Paul B , Perlman Stanley 
Published: January 2023

Abstract
Coronavirus disease 2019 (COVID-19) is especially severe in aged populations. Resolution of the COVID-19 pandemic has been advanced by the recent development of SARS-CoV-2 vaccines, but vaccine efficacy is partly compromised by the recent emergence of SARS-CoV-2 variants with enhanced transmissibility. The emergence of these variants emphasizes the need for further development of anti-SARS-CoV-2 therapies, especially in aged populations. Here, we describe the isolation of a new set of highly virulent mouse-adapted viruses and use them to test a novel therapeutic drug useful in infections of aged animals. Similar to the human infection, aged mice infected with mouse-adapted SARS-CoV-2 develop more severe disease than young mice. In murine SARS, in which severity is also age-dependent, elevated levels of an eicosanoid, prostaglandin D2 (PGD2) and of a phospholipase, PLA2G2D, contributed to poor outcomes in aged mice. We now demonstrate that mRNA expression of PLA2G2D and PTGDR, a PGD2 receptor, and production of PGD2 increase in human PBMC-derived dendritic cells with aging and after SARS-CoV-2 infection. Using our virulent mouse-adapted SARS-CoV-2, we show that middle-aged mice lacking expression of PTGDR, or PLA2G2D are protected from severe disease. Further, treatment with a PTGDR antagonist, Asapiprant, protected aged mice from lethal infection. PTGDR antagonism is one of the first interventions in SARS-CoV-2-infected animals that specifically protects aged animals, suggesting that the PLA2G2D-PGD2/PTGDR pathway is a useful target for therapeutic interventions.
This work is supported in part by grants from the National Institutes of Health USA (NIH) (P01 AI060699 (SP, PBM) and RO1 AI129269 (SP) and BIOAGE Labs (SP). The Pathology Core, is partially supported by the Center for Gene Therapy for Cystic Fibrosis (NIH Grant P30 DK-54759), and the Cystic Fibrosis Foundation. PBM is supported by the Roy J. Carver Charitable Trust.
Chemokine CXCL10’s role in the immune response to SARS-CoV-2
Authors: Ivester Hannah , Morrison Holly , Finkielstein Carla , Duggal Nisha , Weger James , Allen Irving C 
Published: January 2023

Abstract
Since the beginning of the SARS-CoV-2 pandemic in early 2020, it was apparent that one of the most striking clinical features of the disease caused by the virus, COVID-19, was the lung damage that occurs. This lung damage is often the result of an immune response gone awry as the host attempts to clear the virus. This overactive immune response, the ‘cytokine storm,’ is responsible for much of the most severe cases of COVID-19 as well as the Multisystem Inflammatory Syndrome in pediatric patients. Since the start of the pandemic, a few key players have been identified in the progression of COVID-19 disease as well as the immune response to it. These include pro-inflammatory cytokines IL-6, TNF-alpha, and CXCL10. Many clinical reports have especially shown the importance of the chemokine CXCL10 as a prognostic biomarker with patients having higher levels of CXCL10 often having more severe outcomes. Recent studies have also shown an increase in CXCL10 and its associated pathways in animal models of COVID-19. Here we continue to provide evidence for the importance of the protein CXCL10 in SARS-CoV-2 immunity through whole genome transcriptomics studies in human and k18-hACE2 mice. Following these, further studies with a mouse-adapted SARS-CoV-2 virus and Cxcl10−/− mice also showed a decreased level of effectiveness in viral clearance in the absence of effective CXCL10. These data continue to show the importance of the chemokine CXCL10 in both disease severity and viral clearance of SARS-CoV-2 infection adding to the growing body of evidence related to SARS-CoV-2 pathobiology.
B cell convergence to distinct broadly reactive epitopes revealed by chimeric hemagglutinin vaccination
Authors: Guthmiller Jenna , Li Lei , Lan Linda Yu-Ling , Henry Carole , Stamper Christopher , Freyn Alec , Utset Henry , Han Julianna , Palese Peter , Coughlan Lynda , Ward Andrew , Krammer Florian , Wilson Patrick C 
Published: January 2023

Abstract
Seasonal influenza virus vaccines provide modest protection against endemic viruses but lack effectiveness against pandemic-threat viruses. Therefore, new immunogens and vaccine platforms that induce antibodies against broadly protective epitopes are needed to provide robust protection against seasonal and novel influenza viruses. In a phase 1 clinical trial, the chimeric hemagglutinin (cHA) immunogen induced antibody responses against the conserved HA stalk domain. However, the landscape of the B cell specificities and subsets induced by this vaccine remain undetermined. Using single cell RNA-sequencing and repertoire sequencing, we identified that formulation of the cHA vaccine differentially induced B cell responses against distinct epitopes. Notably, those participants that received an inactivated vaccine with a squalene-based adjuvant had a greater proportion of B cells targeting two conserved neutralizing epitopes of the stalk domain: the central stalk and anchor epitopes. Moreover, participants that received an inactivated vaccine with adjuvant had distinct transcriptional B cell subsets, including an increase in atypical memory B cells. B cell clonal overlap between acute and memory timepoints was substantial, with several multi-donor clones that targeted distinct broadly protective epitopes. In addition, we identified stalk targeting germline antibodies that could neutralize recent pandemic H1N1 viruses. Altogether, our study reveals that the inactivated cHA vaccine with an oil-in-water adjuvant promotes B cell activation and maturation against broadly protective epitopes.
Supported by National Institute of Allergy and Infectious Diseases (NIAID) National Institutes of Health grant numbers K99AI159136 (JJG), U19AI082724 (PCW), U19AI109946 (PCW), U19AI057266 (PCW), P01AI097092 (PP), R01AI145870-01 (PP), R21AI146529 (LC), and T32AI007244-36 (JH), the NIAID Centers of Excellence for Influenza Research and Surveillance (CEIRS) grant number HHSN272201400005C (PCW), HHSN272201400008C (LC, FK, PP), and the NIAD Centers of Excellence for Influenza Research and Response (CEIRR) grant number 75N93019R00028 (PCW, FK, PP). This work was also partially supported by the NIAID Collaborative Influenza Vaccine Innovation Centers (CIVIC; 75N93019C00051, FK, PP, ABW, PCW).
Identifying maternal humoral immune responses associated with control of viremia after primary maternal CMV infection in a rhesus macaque model
Authors: Otero Claire E , Nelson Cody S , Scheef Elizabeth , Sprehe Lesli , Mostrom Matilda , Malouli Daniel , Fruh Klaus , Chan Cliburn , Kaur Amitinder , Permar Sallie R 
Published: January 2023

Abstract
Congenital cytomegalovirus (cCMV) is the most common in utero infection and causes major neurodevelopmental deficits, but there remains no licensed vaccine to prevent cCMV. Little is known about maternal immune responses that can prevent placental CMV transmission, which could guide rational design of an effective vaccine. Using the rhesus macaque (RM) model of primary RM CMV (RhCMV) infection during pregnancy, we established that pre-existing RhCMV-neutralizing IgG protected against cCMV, even in the setting of CD4+ T cell depletion, where vertical transmission occurs consistently in RhCMV-seronegative RMs. However, the antibody (Ab) functions mediating this protection have not been fully defined. To identify humoral immune correlates of containment of viremia and protection against cCMV we used samples from previous studies in this model to measure Ab binding to whole RhCMV virus and key glycoproteins, neutralization, Ab dependent cellular phagocytosis (ADCP), and cytotoxicity (ADCC). Immunocompetent RMs developed both RhCMV-specific neutralizing and Fc-mediated effector functions (ADCP, ADCC) 2–4 weeks post infection. However, Ab responses were not statistically distinct between transmitters and non-transmitters during this period. Maternal viremia outperformed maternal Ab responses as a predictor of placental transmission. Therefore, we correlated each Ab response with maternal plasma viral load at day 21 post infection. Interestingly, IgG binding to gB and the pentamer, immunodominant viral glycoproteins involved in viral entry, displayed significant inverse relationships with maternal viremia (n=15, Spearman r=−0.71, p=0.003 for both), highlighting these as critical targets for vaccine design.
Supported by grants from NIH (NIAID P01-AI129859, NCI T32-CA009111)
Impact of pre-existing immunity on the development of de novo virus-specific TRM following live attenuated influenza vaccination
Authors: Lobby Jenna L , Danzy Shamika , Lowen Anice , Kohlmeier Jacob E 
Published: January 2023

Abstract
Live attenuated influenza vaccine (LAIV) elicits both humoral and cellular immune memory in children, but its efficacy is limited in adults. We hypothesize that pre-existing immunity from past infections and/or immunizations prevents the attenuated vaccine from establishing an immune response. To determine if we can overcome this limitation by increasing the antigenic distance of the vaccine strain from previous circulating seasonal strains, we generated a series of drifted LAIVs with successive mutations in the HA protein, allowing for increasing levels of escape from pre-existing antibody. We also inserted a CD8+ T cell epitope from the Sendai virus nucleoprotein (SeV-NP) as a readout for generation of a de novo TRM response following immunization. Surprisingly, we were unable to identify SeV-NP+ CD8+ TRM following LAIV immunization in PR8-immune mice, even with LAIV strains that can fully escape pre-existing antibody. As these data suggested a role for cell-mediated immunity in limiting LAIV efficacy, we investigated several scenarios to assess the impact of pre-existing LAIV-specific TRM in the upper and lower respiratory tract. Ultimately, we found that deletion of the immunodominant influenza NP366–374 epitope was sufficient to escape pre-existing CD8+ TRM and to establish de novo CD8+ TRM. Combined, these studies demonstrate that both pre-existing humoral and cellular immunity can limit the effectiveness of LAIV, thereby informing future design of vaccine vectors against respiratory pathogens.
Supported by grants from NIH (F31 HL156639-01, R35 HL150803)
Requirement for robust T and B cell responses for immune protection against Powassan virus (POWV) in virus-like particle (VLP) vaccination.
Authors: Stone E Taylor , Hassert Mariah , Geerling Elizabeth , Wagner Colleen , Brien James , Ebel Gregory , Hirsch Alec , German Cody , Smith Jessica , Pinto Amelia K 
Published: January 2023

Abstract
Powassan virus (POWV) is a tick-borne pathogen for which humans are an incidental host. POWV infection can be fatal or result in long-term neurological sequelae; however, there are no approved vaccinations for POWV. Integral to efficacious vaccine development is the identification of correlates of immune protection, which we accomplished in this study by utilizing a murine model of POWV infection. Using POWV lethal and sub-lethal challenge models, we show that 1) robust B and T cell responses are necessary for immune protection, 2) POWV lethality can be attributed to both viral- and host-mediated drivers of disease and 3) knowledge of the immune correlates of protection against POWV can be applied in a virus-like particle (VLP)-based vaccination approach that provides protection from lethal POWV challenge. We also provide evidence that POWV-experienced CD8+ T cells may potentiate immune-mediated pathogenesis during infection. Identification of beneficial and detrimental aspects of POWV-immune protection is significant as it will aid in the rational design of POWV vaccines.
This research was supported in part by the National Institutes of Health, National Institute of Allergy and Infectious Diseases (NIAID): 5R01AI152192-02, 5R01AI137424-03 and F31AI152460-01. This research was also supported in part by the Department of Defense (Award#: PR192269). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Dysregulation of CD8+ T cell function in the setting of altered cytokine signaling due to inborn errors of immunity
Authors: Campos Duran Jose S , Conrey Peyton E , Sayed Samir , Di Chao , Gonzalez Elena N , Romberg Neil D , Bergerson Jenna R E , Holland Steve M , Freeman Alexandra F , Su Helen C , Leiding Jennifer W , Forbes Lisa R , Vogel Tiphanie P , Henrickson Sarah E 
Published: January 2023

Abstract
A key outcome of chronic antigenic stimulation in the setting of sustained inflammation is the development of CD8+ T cell exhaustion. However, the relative roles of inflammation and antigen stimulation on T cell dysfunction remain unclear. Inborn errors of immunity (IEIs) provide an opportunity to study the effect of genetic alterations in signaling pathways on immune function. Here, we focus on an IEI resulting from genetically amplified cytokine signaling: gain-of-function mutations in STAT3 (STAT3 GOF). STAT3 signals downstream of multiple inflammatory cytokines (e.g., IL-6 and IL-27) and we used STAT3 GOF as a genetic mimic of chronic inflammation. To assess how amplified STAT3 signaling impacts immune cell subset distribution and activation status, PBMCs from STAT3 GOF patients and age-matched healthy controls were profiled via mass cytometry (CyTOF). This revealed altered states of CD8+ T cell activation including increased expression of immunoregulatory receptors (e.g., PD-1 and CD39). STAT3 GOF CD8+ T cells also showed impaired cytokine production and reduced proliferative capacity. Bulk RNA sequencing of naïve STAT3 GOF CD8+ T cells identified an exhausted-like transcriptional profile. Quantitation of metabolic function shows alterations in glucose use and oxidative phosphorylation in STAT3 GOF. Our studies suggest that inflammation alone may directly impair CD8+ T cell function. Understanding the mechanisms by which inflammation impacts T cell function may identify components of T cell dysregulation that can be targeted therapeutically in these rare patients, as well as those with more common inflammatory diseases (e.g., obesity) with shared aspects of T cell dysfunction.
Supported by CHOP Research Institute, The Primary Immune Deficiency Treatment Consortium (PIDTC) and Immune Deficiency Foundation (IDF).
Association of elevated CD151+ T cell frequencies with impaired immune function
Authors: Perez Mildred , Lowman Kelsey , Moylan David C , Tidwell Christopher M , Schroeder Andrew , Duverger Alexandra , Wagner Frederic H , Erdmann Nathan , Goepfert Paul , Hu Hui , Sabbaj Steffanie , Kutsch Olaf 
Published: January 2023

Abstract
We recently reported that in humans the tetraspanin CD151 marks phenotypically distinct T cell subsets. The frequency of CD151+ T cells differs between T cell lineages and in between memory T cell populations. CD151+ T cells are hyper-proliferative and hyper-activated, as shown by phospho-proteomic analysis. We found that CD151 is an activation marker that is upregulated following TCR/CD3 activation, but at baseline, the presence of CD151 was associated with the loss of CD28 expression, a sign of T cell senescence. We hypothesized that the presence of CD151+ T cells is associated with immune hyperactivation and in extension, impaired immune surveillance. Unfortunately, we found that not even basic human CD151 T cell biology is reproduced in mouse models and thus had to resort to the study of pathological human conditions to elucidate the role(s) of CD151+ T cells. As cancer development requires immune escape, cancer diagnosis could be used as an indicator of impaired immune surveillance. Benefitting from the availability of longitudinal patient sample and data collections from people living with HIV (PLWH) we found that PLWH that developed cancer all had CD8+CD151+ T cell frequencies outside a normal reference range. CD8+CD151+ T cell frequency increase was not cancer-specific and could be detected years before the cancer diagnosis (0.5–12 years), indicating that the increase was not part of an anti-tumor response. Other established immune markers showed no correlation with the development of cancer. The findings associate elevated CD8+CD151+ T cell frequencies with impaired immune surveillance and suggest that elevated CD8+CD151+ T cell frequencies, at least in PLWH can be used as prognostic marker for cancer development.
Supported by grants from NIH R01-AI122842, NIH R33-AI116188, NIH R33-AI133679
Comprehensive analysis of the durability and the quality of humoral and T cell responses in SARS-CoV-2 vaccinated and infected subjects.
Authors: Woloszczuk Kyra , Connors Jennifer R , Cusimano Gina , Bell Matthew , Furukawa Sara , Melnyk Nataliya , Lin Edward , Venkateswaran Kodumudi , Bernui Mariana , Kutzler Michele , Cairns Charles , Haddad Elias K 
Published: January 2023

Abstract
Understanding adaptive immune memory to SARS-CoV-2 is relevant for the development of vaccines and new immunotherapies. Specifically, understanding the interactions between memory B cells and T-cell subsets can shed light on persistent immune memory and its effects on antibody durability and antibody affinity. Studying the frequencies of SARS-CoV-2 memory B cells and antigen-specific T-cells 3–6 months following natural infection or vaccination would aid us in vaccine development and vaccination in infected subjects. In this study, we analyzed these responses in individuals who were hospitalized with COVID-19 (n=20), individuals who were vaccinated after COVID hospitalization (n=20), and uninfected individuals (n=40) with two doses of the Pfizer or Moderna SARS-CoV-2 vaccine. We hypothesize individuals who were vaccinated after hospitalization will have the highest frequency of memory T follicular helper cells, memory B cells and antibody durability. To accomplish this, we will analyze anti-SARS-CoV-2 antibodies, including 7 RBD variants as well as 65-cytokines in all patients via Luminex and examine frequencies of memory T follicular helper cells, persistent memory B-cells specific to SARS-CoV-2 using flow cytometry. All immune responses will be analyzed 3–6 months after hospitalization or after a second vaccination. These results show the differences in durability and quality of humoral and T cell responses between vaccinated and infected subjects.
Supported by grants from NIH supplement #U19AI128910-04S1