bluuebunny/crossref_metadata_2025_split huggingface dataset:
Expansion and maintenance of long lived effector CD8 T cells is modulated by inflammation
Authors: Lucas Erin D , Huggins Matt , Thefaine Claire , Pierson Mark , Hamilton Sara 
Published: January 2023

Abstract
Long-lived effector T cells (LLEC) are a circulating population of KLRG1 and CX3CR1 expressing memory CD8 T cells. LLEC were first described by the Hamilton Lab and are highly effective at clearing systemic infections compared to other memory T cells. However, LLEC express lower levels of IL-7 and IL-15 receptor chains and are poorly proliferative in specific pathogen free (SPF) mice as compared to central and effector memory T cells, which leads to a slow attrition of their numbers. The factors supporting LLEC expansion and persistence are unknown, and the goal of this study is to understand what internal and external factors contribute to LLEC expansion and maintenance. Data from models of chronic inflammation suggest pro-inflammatory factors contribute to the expansion and/or survival of LLEC. Specifically, we have found that LLEC populations are increased in young mice that have been exposed to a diverse mixture of microbes through co-housing with pet-shop mice. This co-housing model chronically increases systemic inflammation, and LLEC specific for model antigens not only expand more, but also persist longer than in SPF mice. Similarly, we also find increased LLEC in aged mice, another model associated with chronic inflammation. In preliminary work, we find that LLEC division is increased during cohousing, and that IL-18R expression is elevated, further suggesting that inflammatory cytokines not typically associated with T cell persistence may support LLEC proliferation. Ongoing studies will determine which specific inflammatory markers support LLEC expansion and maintenance, whether these factors are detrimental to other memory subsets, and if this process can be altered to improve T cell responses to infection and vaccination.
Supported by grants from NIH (R01 AI143828).
Plasmodium infection elevates risk of severe secondary bacterial disease by altering the immunological landscape of the lung
Authors: Reed Jenna , Cornwall Douglas , Jacobs Nathan , Evavold Brian D , Lamb Tracey J 
Published: January 2023

Abstract
Lower respiratory infections cause an estimated two million sepsis deaths yearly, with &gt;50% attributed to the opportunistic pathogen Streptococcus pneumoniae. Sub-Saharan Africa, the site of most severe Streptococcus burden, is also ravaged by the malaria-causing Plasmodium parasite family, which afflicted 241 million people and killed 627,000 in 2020. Compared to healthy children, those with asymptomatic malaria have an elevated risk to severe bacterial infections, but the underlying mechanism(s) remain elusive. To fill this knowledge gap, we infected C57BL/6J mice with P. berghei NK65-NY either 7, 14, or 21 days prior to inoculation with S. pneumoniae 6304. Coinfected mice exhibited a 40–60% fatality rate, with death beginning 1–4 days post-bacterial infection. In contrast, singly infected groups survived the duration of the experiment. Coinfected mice exhibited equivalent parasitemia but significantly higher bacterial burden in the lung compared to mono-infections. Neutrophil recruitment and ROS production was altered in coinfected mice, which we predict is due to the formerly described effects of the Plasmodium metabolic product hemozoin. As such, we hypothesize that neutrophil control of S. pneumoniae is impaired. Since malaria has been shown to increase blood brain barrier permeability, we used Evans blue dye to test for a similar effect at the alveolar-capillary interface. Lung weight significantly increased as early as 6 days post-infection, while permeability trended towards a slight increase, peaking at day 9. Thus, we hypothesize that Plasmodium infection increases pathogenicity of Streptococcus infection due to a combination of impaired bacterial control and increased vascular leak in the lungs.
This work was supported by the NIH (ROl AI123425-01A1).
Host-targeted self-attenuated influenza virus as a potential therapeutic influenza vaccine
Authors: Ieremia Lanaiah M , Wen Ke , Wang Haiyan , Chen Yanping , Yang Huixiao , Zheng Zhichao , Yan Yongyong , Pena Adilene Realivazquez , Zeng Mingtao 
Published: January 2023

Abstract
Influenza is still one of leading causes of morbidity and mortality around the world, despite of annual influenza immunization in many countries. New strategies are urgently needed for developing effective vaccines and therapeutics against influenza viruses. In this study, we have constructed new recombinant influenza viruses, named host-targeted self-attenuated influenza viruses (SAIVs), which can express functional mammalian species-specific artificial microRNAs (amiRNAs). The expression of these amiRNAs can inhibit expression of some host factors critical for influenza replication, and therefore the resultant recombinant influenza viruses are replication restricted and attenuated in the host cells. One of these SAIVs, which can express an amiRNA that inhibits expression of the host cellular Cdc2-like kinase 1 (CLK1), was produced in embryonic chicken eggs and evaluated in a mouse model of influenza infection. It elicited robust antibody responses against influenza virus and demonstrated significantly protective efficacy against lethal infection with wild type influenza virus H1N1 PR8 after single dose of intranasal vaccination. Additionally, post-exposure treatment with this CLK1-targeted SAIV showed therapeutic effect against PR8 lethal-dose infection. Our research finding provided a proof of concept that the new host targeted self-attenuated influenza virus can be further developed to a therapeutic vaccine for prophylactic and therapeutic use against influenza.
Supported by a research grant from National Institute of Allergy and Infectious Diseases (AI133207).
Role of inflammation in the pulmonary hemorrhage induced by a snake venom metalloproteinase
Authors: Castro Ana Cristina , Gutiérrez José María , Escalante Teresa , Rucavado Alexandra 
Published: January 2023

Abstract
Snakebite envenoming (a neglected tropical disease that affects millions of people worldwide every year) is characterized by a complex pathophysiology. In cases of viperid envenomings, local and systemic hemorrhage is one of the major systemic effects that manifest in patients. This work focuses on studying pulmonary hemorrhage caused by a type P-III snake venom metalloproteinase (SVMP) extracted from Crotalus simus venom, and the extent to which inflammation affects the outcome of this hemorrhage. In order to determine the role of inflammation in pulmonary hemorrhage, we created a new automated image analysis methodology using the CellProfiler platform to quantitatively assess hemorrhage under different conditions, in conjunction with a statistical analysis program to process and analyze said image analysis results. With this toolset, we subsequently analyzed hemorrhage under the effects of treatment with three anti-inflammatory agents (a COX inhibitor, a TNF-ɑ inhibitor, and anti-neutrophil antibodies) in combination with the toxin on a murine model. Our findings conclude that inhibiting inflammation reduced lung hemorrhage significantly, which shows promise as a potential novel avenue for the treatment of snake envenomings. Moreover, we propose new hypotheses for how the toxin may enter lung capillaries and subsequently cause basement membrane damage which eventually leads to lung hemorrhage.
This study was supported by Vicerrectori?a de Investigación, Universidad de Costa Rica (Project 741-B4-524), and by a fund for support of graduate studies of the Sistema de Estudios de Posgrado (Universidad de Costa Rica)
Imaging Mass Cytometry identifies structural and cellular composition of the mouse tissue microenvironment
Authors: Lowrie Kerry , Raza Qanber , Cohen Michael , Kala Smriti , Awong Geneve , Quong Andrew , Loh Christina 
Published: January 2023

Abstract
Imaging Mass Cytometry™ (IMC™) is a vital tool to deeply characterize the complexity and diversity of any tissue without disrupting spatial context. The Hyperion™ Imaging System utilizes IMC, based on CyTOF® technology, to assess up to 40 individual structural and functional markers in tissues, providing unprecedented insight into the organization and function of tissue microenvironment. We have previously demonstrated the application of IMC in combination with Maxpar® panel kits on human tissues. Here, we showcase Maxpar OnDemand Antibodies for IMC application including 11 new highly relevant markers to construe cellular and molecular composition of mouse tissues.
We analyzed a normal mouse tissue microarray using IMC spatial proteomic analysis. Tissues were stained with a 20-marker panel designed to highlight tissue architecture and major immune lineage markers. We generated a detailed spatial map of the diverse tissue architecture and successfully identified immune, epithelial, and stromal cell populations in various mouse tissues. Additionally, we classified the activation state of immune cell populations, adhesion state of epithelial cells, and molecular composition of the extracellular matrix.
This work demonstrates the capability of IMC to identify subcellular localization of cellular and structural markers in the mouse tissue microenvironment. Future studies utilizing IMC in combination with Maxpar OnDemand Antibodies will enable in-depth phenotypic characterization of the tissue microenvironment in various mouse models of development and disease, and thus provide the basis for the use of high-multiplex imaging in preclinical investigations.
For Research Use Only. Not for use in diagnostic procedures.
Robust human regulatory T cell expansion with fusion proteins HCW9302 and HCW9213 circumvents need for magnetic-bead or feeder cell approaches for adoptive cell therapy
Authors: Wong Hing C , Shrestha Niraj , George Varghese , Dee Michael J , Chaturvedi Pallavi , Zhu Xiaoyun , Spoundis Catherine , Liu Bai , Kong Lin , Echeverri Christian A , You Lijing , Egan Jack O , Jiao Jin-An , Rhode Peter R 
Published: January 2023

Abstract
Regulatory T cells (CD4+CD25+FoxP3+) (Tregs) are a subset of CD4 T cells that suppress the activities of other immune cells and have applications in the treatment of autoimmune and inflammatory diseases. Their use as an adoptive cell therapy has been limited by the practicality of expanding and purifying clinically sufficient numbers of cells. HCW9213 and HCW9302 are fusion proteins based on HCW Biologics’ TOBI™ technology platform, consisting of anti-CD3/anti-CD28 antibody domains and IL-2 domains, respectively. When used in combination, these fusion proteins were capable of expanding human Treg cells in vitro without the use of anti-CD3/CD28 magnetic beads and/or feeder cells, improving the overall yield, process efficiency and overcoming regulatory hurdles in manufacturing. Tregs generated with these molecules displayed similar phenotypes and suppressive cytokine production as Tregs expanded with recombinant human IL-2. Using a proprietary anti-CD39 antibody to isolate CD39+ Tregs, we have also been able to generate a Treg population with twice the suppressive activity against CD4+ T responder cells as traditional CD4+CD25+CD127lo Tregs. Thus, using its novel fusion proteins, HCW Biologics has been able to develop a superior Treg cell product ideal for the use in adoptive cell transfer. Additionally, this Treg platform can potentially be further optimized with addition of disease-targeted chimeric antigen receptors (CAR).
Characterizing arterial lesions in cardiac allograft vasculopathy rejected grafts using NanoString GeoMx digital spatial profiling
Authors: Nevarez-Mejia Jessica , Pickering Harry C , Sosa Rebecca A , Lau Ryan P , Fishbein Gregory A , Baldwin William M , Fairchild Robert L , Reed Elaine F 
Published: January 2023

Abstract
Cardiac allograft vasculopathy (CAV) is the leading cause of graft failure following heart transplantation. Donor specific antibodies (DSA) contribute to CAV by triggering vessel inflammation and endothelial cell injury. This results in thickening of the arterial intima and vessel occlusion. In this study, we aim to define the innate and adaptive immune cells mediating inflammation and vessel neointima formation using CAV+DSA+ rejected cardiac explants (N=3) and GeoMx digital spatial profiling (DSP). Arterial regions containing macrophages (CD68+CD163+) within vessels (CD34+) were selected for whole genome sequencing and a 76-protein panel. Arteries were scored as containing ‘low’ or ‘high’ neointima based on H&E. A total of 41 proteins including markers of inflammation and apoptosis were found to be expressed in all arteries. The expression of CD8, CD56, CD20, CD127, and CD11c exhibited a significantly positive correlation with RNA counterparts. A total of 15 differentially expressed proteins (e.g., T-cell makers: CD8, CD45RO, and CD127), and high expression of genes related to inflammation (ITGAX), and apoptosis (TP63), along with pathways for antigen presentation were elevated in arteries with low neointima. High neointima arteries showed higher SMA protein expression and increased genes related to cell growth (LECT2), complement regulation (CFHR3) and anti-inflammatory factors (FNDC4). Enriched pathways in high neointimal arteries included platelet activation/degranulation and cell migration. Our results provide insight into the mechanisms mediating CAV progression seen by inflammatory profiles in arteries with low neointima followed by pro-fibrotic/reparative phenotypes in high neointima arteries.
This work is funded by the Ruth L. Kirschstein National Research Service Award (NRSA) T32HL069766 (UCLA Vascular Biology Training grant), the Eugene V. Cota Robles Fellowship and NIH Grant 441450 ER 29762
Loss of IFNgR1 signaling glioblastoma drives resistance to CAR T cell binding avidity and cytotoxicity due to lower downstream expression of ICAM-1
Authors: Larson Rebecca , Kann Michael , Bailey Stefanie , Haradhvala Nicholas , Stewart Kai , Bouffard Amanda , Scarfo Irene , Leick Mark , Berger Trisha R , Jan Max , Joung Julia , Ouspenskaia Tamara , Law Travis , Regev Aviv , Getz Gad , Maus Marcela 
Published: January 2023

Abstract
Though CAR T cell therapy has had success treating hematologic malignancies, these treatments have shown only modest efficacy in solid tumors. Little is known about the necessary molecular components required for CAR T cell cytotoxicity, especially in the context of solid tumors. We investigated the role of IFNgR signaling on solid tumors and discovered across tumor types (including glioblastoma, pancreatic, ovarian and lung) loss of IFNgR1 signaling resulted lower CAR T cell cytotoxicity in vitro and that this was irrespective of target antigen (including endogenous EGFR, IL13Ra2, mesothelin, and exogenous CD19). We also validated this in orthotopic xenograft models of glioblastoma and pancreatic cancer in vivo. CAR T cells exposed to wildtype (WT) or IFNgR1 KO tumor had similar transcriptional profiles, but the tumor cells had different signatures in response to CAR T cell co-culture. Further investigation showed marked upregulation of cell adhesion pathways in WT cells compared to IFNgR1 KO. We utilized microscopy with acoustic force and discovered CAR T cells had lower cell binding avidity to cells lacking IFNgR1 compared to WT cells. Following CAR T cell exposure, Intercellular Adhesion Molecule 1 (ICAM-1) was strongly upregulated at both the transcriptional and protein levels in WT cells but not IFNgR1 KO cells. We overexpressed ICAM-1 on IFNgR1 KO tumor cells and observed that CAR T cell binding avidity and cytotoxicity was restored to that of WT levels. This work highlights the importance of CAR T cell binding avidity and adhesion for optimal cytotoxicity. Better understanding of CAR T cell function will inform future construct design for successful therapies against solid tumors.
RCL was supported by T32 GM007306, T32 AI007529, and the Richard N. Cross Fund. ML was supported by T32 2T32CA071345-21A1. SRB was supported by T32CA009216-38. NJH was supported by the Landry Cancer Biology Fellowship. JJ is supported by a NIH F31 fellowship (1F31-MH117886). GG was partially funded by the Paul C. Zamecnik Chair in Oncology at the Massachusetts General Hospital Cancer Center and NIH R01CA 252940. MVM and this work is supported by the Damon Runyon Cancer Research Foundation, Stand Up to Cancer, NIH R01CA 252940, R01CA238268, and R01CA249062.
ZBP1 is a crucial regulator for tumor necroptosis during tumor development.
Authors: Baik Jin Young , Choksi Swati 
Published: January 2023

Abstract
Tumor necrosis happens commonly in advanced solid tumors. We reported that necroptosis plays a major role in tumor necrosis. By studying TNF-induced necroptosis, it is now known that receptor interacting protein kinase 1 and 3 (RIPK1 and RIPK3) and the mixed lineage kinase domain-like (MLKL) constitute the core of the necroptosis machinery. Two other proteins, Z-DNA-binding protein 1 (ZBP1) and TIR-domain-containing adapter-inducing interferon-β (TRIF) are known to function upstream of RIPK3 and interact with RIPK3 to mediate necroptosis in response to viral infection or TLR signaling respectively. Our study shows that it is the ZBP1, and not RIPK1 nor TRIF, that mediates tumor necroptosis during tumor development in preclinical cancer models. We found that ZBP1 expression is dramatically elevated in the later stages of both the MVT-1 breast cancer model and the MMTV-PyMT, a genetically engineered mouse model (GEMM) of breast cancer, when necrosis happens. Next, we generated CRISPR ZBP1 knockout (ZBP1 KO) MVT-1 cells to examine whether ZBP1 is needed for tumor necroptosis during MVT-1 mammary tumor development. ZBP1 deletion blocks tumor necroptosis during primary tumor development, more importantly, we observed decreased lung metastasis comparing to wildtype. We showed that glucose deprivation (GD) triggers ZBP1-depedent necroptosis in tumor cells. GD causes mitochondrial DNA (mtDNA) release to the cytoplasm and the binding of mtDNA to ZBP1 to activate MLKL in a BCL-2 family protein, NOXA-dependent manner. Therefore, our study reveals ZBP1 as the key regulator of tumor necroptosis and provides a potential drug target for controlling tumor metastasis.
Abrogating regulatory T cells overcomes tumor-specific T cell exhaustion and prevents metastatic pancreatic cancer
Authors: Schmiechen Zoe C , Burrack Adam L , Miller Ebony , Rollins Meagan , Wang Iris X , Cruz Eduardo , Patterson Michael , Stromnes Ingunn 
Published: January 2023

Abstract
Pancreatic ductal adenocarcinoma (PDA) is the 4th leading cause of cancer related deaths and has a dismal 5-year survival rate of 10 percent. PDA lethality is attributed to late diagnosis, early metastasis, and therapeutic resistance. Metastasis can occur before the development of histologically detectable tumors and is a leading cause of cancer-related deaths. We identified that pancreatic tumor cells derived from mice that resist immunotherapy (e.g., tumor escape variants, TEV) and re-implanted into the pancreas of syngeneic and immunocompetent mice rapidly metastasize, reflecting the pathogenesis of human PDA. We show that TEVs retain the targeted tumor antigen, and despite a defect in IFNgamma-inducible MHC class I upregulation, TEVs remain sensitive to tumor antigen specific T cell-mediated lysis in vitro, suggesting TEVs may confer unique qualities in vivo to resist T cell killing. Using a peptide:MHC tetramer to identify the tumor-specific CD8 T cells, we identified that intratumoral T cells in EV tumors have increased Granzyme B production and a reduction in prototypical exhaustion markers PD1, Lag3, and Tox, perhaps due to reduced MHC class I signaling. Notably, primary tumors from TEVs were significantly enriched for Foxp3+ Tregs as compared to parental tumors. Using a genetic model, we identified that Treg depletion resulted in a drastic reduction in tumor burden and metastasis and improved tumor-specific T cell function in TEV tumors. Tumor-cell intrinsic changes driving Treg accumulation in TEVs will be discussed. In summary, Tregs are key drivers of both T cell exhaustion and immunosuppression in pancreatic cancer and may prove a valuable clinical target for tumors that evade immune checkpoint blockade.
Supported by grants from NIH (R01 CA249393)
Tumor immunity is enhanced in mice expressing the pro-autoimmune allele of tyrosine phosphatase Ptpn22, but not in Ptpn22 knock-out mice
Authors: Orozco Robin C , Marquardt Kristi , Mowen Kerri A , Sherman Linda A 
Published: January 2023

Abstract
The 1858C&gt;T allele of the tyrosine phosphatase PTPN22 (causing amino acid substitution R620W in encoded protein Lyp) is present in 5–10% of the North American population and is strongly associated with numerous autoimmune diseases. Although much research has been done to define how this allele potentiates autoimmunity, the influence PTPN22 and its pro-autoimmune allele has in tumor immunity is poorly defined. To interrogate the role this allele has during the anti-tumor immune response, we used CRISPR/Cas9 to generate mice in which the ortholog of Lyp, PEP, is mutated at position 619 to produce the relevant pro-autoimmune mutation (PEP-619WW) or lack Ptpn22 expression (PEP-null). Using the B16 and other tumor models, we tested the hypothesis that pleiotropic effects of the PTPN22 pro-autoimmune allele enhances tumor immunity. Results of this study show that PEP-619WW mice, but not PEP-null mice, resist tumor growth as compared with wildtype mice. Consistent with these results, tumors from PEP-619WW mice have more CD45 infiltrates containing; more activated CD8 T cell and CD4 T cells, more cDC1 cells, and less MDSCs than WT animals. Interestingly, the tumor infiltrating PEP-619WW cDC1s have lower PD-L1 expression compared to cDC1s from PEP-WT mice. Furthermore, using single-cell RNA sequencing we show that intra-tumoral myeloid cells from PEP-619WW mice have a more cytokine responsive transcriptional signature compared to PEP-WT cells. Our results suggest that a pro-autoimmune allele can be beneficial by promoting a strong anti-tumor immune response and may have a protective effect in this disease. Also, these data highlight an important difference between the PEP-null and PEP-619WW murine models during the tumor response.
Supported by grants from NIH (UO1 AI130842 and T32 AI007354 27).