bluuebunny/crossref_metadata_2025_split huggingface dataset:
Dysregulation of complement regulator CD55 in the initiation of autoimmunity
Authors: Lee Iris , Ling Ivana , Pham Christine T , Kendall Peggy L 
Published: August 2023

Abstract
CD21 is a complement receptor involved in B cell tolerance. CD21 low age-associated B cells (ABCs) are an autoimmune-prone population of B cells that are expanded in autoimmunity. B cell receptor (BCR) and toll-like receptor 9 (TLR9) co-stimulation poise B cells for this autoimmunity-associated cell fate. We and others have discovered that BCR and TLR9 co-stimulation also causes downregulation of CD55, a complement regulatory protein that dampens complement activation by interfering with the production of complement fragments. We posit that CD55, by controlling the generation of complement fragments, regulates BCR-TLR9-triggered B cell responses in the context of autoimmunity. Congruent with this hypothesis, CD55 deficiency has been shown to cause exaggerated experimental encephalitis and disease in the MRL/lpr mouse model of lupus. Our overarching hypothesis is that CD55 regulates B cell responses in the context of autoimmune disease. For this study, we examined CD86 upregulation, proliferation, and cytokine production in B cells after knocking out CD55. We found that CD55 did not impact the CD86 upregulation or proliferation of B cells, but did induce a proinflammatory cytokine profile with increased IL-6 production. In addition, crossing CD55 deficiency onto anti-insulin B cells, a well-studied anergic model, showed that CD55 deficiency partially abrogated anergy-mediated tolerance, increasing the ability of these B cells to proliferate when stimulated through TLR9. These findings may have implications for the future use of complement inhibitors in the treatment of autoimmune disease.
Iris Lee holds a Dean’s Scholars Award from the Washington University Division of Physician-Scientists, which is funded by a Burroughs Wellcome Fund Physician-Scientist Institutional Award. The Kendall lab is supported by VA Merit I01BX002882 and R01 DK084246.
Interferon regulatory factor 3 disrupt the normal differentiation of epidermal keratinocyte by promoting the nuclear localization of interleukin-33
Authors: Jeong Na-Hee , Kim Sang-Hyun 
Published: August 2023

Abstract
Atopic dermatitis (AD) is a complex and multifactorial skin disease characterized by skin inflammation and intense pruritus. AD is often diagnosed based on symptoms, but a precise diagnosis and treatment are difficult due to various symptoms. This study aimed to develop a safe and effective therapeutic agent and identify a novel therapeutic target for AD. We verified the functions and mechanism of interferon regulatory factor3 (IRF3) as a novel therapeutic target of AD. IRF3, a transcription factor regulating various pro-inflammatory cytokines, including IL-33, was significantly increased in the skin lesions of AD mice and patients with AD. However, deficiency of IRF3 relieved AD symptoms and pathophysiological changes, including skin thickening (the dermis and epidermis thickening), immune cell infiltration, serum immunoglobulins level, and pro-inflammatory cytokines. In damaged AD tissues, IRF3 regulated the expression of cytokeratin 1 (KRT1) and 10 (KRT10). IRF3 also altered the expression of nuclear IL-33 in the epidermis and the extracellular IL-33 in the dermis of AD mouse tissues. In vitro experiments using human keratinocytes, knockdown or overexpression of IRF3 regulated the nuclear localization and the extracellular release of IL-33. Thus, IRF3 is an essential factor in the onset and exacerbation of AD by inducing skin barrier abnormalities as well as activation of the immune system. Collectively, our studies identify IRF3 as a positive regulator for IL-33, highlighting a novel therapeutic target for AD and suggesting a novel solution for treating intractable dermatitis.
Inflammatory conditions shape phenotypic and functional characteristics of lung-resident memory T cells
Authors: Schmidt Anna , Fuchs Jana , Lapuente Dennis , Hübner Julian , Antão Ana Isabel Vieira , Irrgang Pascal , Oltmanns Friederike , Paudyal Basudev , McNee Adam , Vatzia Eleni , Carr Veronica , Villanueva Sonia , Tchilian Elma , Tenbusch Matthias 
Published: August 2023

Abstract
Lung tissue-resident memory T cells (TRM) are critical for the local control of viral respiratory tract infections. However, the exact mechanisms of TRM stimulation and maintenance are not fully understood, rendering the induction of protective TRM responses by vaccination an ambitious goal.
Here, TRM populations induced by natural influenza A virus (IAV) infection or by mucosal immunization using adenoviral vector vaccines encoding the IAV antigens HA and NP as well as IL-1β as adjuvant were analyzed. Phenotypic and functional TRM responses were assessed in mice over 200 days via flow cytometry. Interestingly, long-lasting TRM responses with a less cytotoxic character were found scattered within the lung tissue after mucosal immunization. By contrast, TRM triggered by prior IAV infection surrounded bronchus-associated B cell clusters and their quantity waned over time more rapidly. Furthermore, single cell RNA sequencing revealed different gene expression profiles of NP-specific TRM depending on the priming condition. However, both TRM populations provided protection from heterologous challenge, resulting in an expansion of cognate antigen-specific TRM. Substantial differences in IAV-specific T-cell immunity after adenoviral vector immunization or prior infections could be confirmed in the Babraham pig model. Noticeably, increased TRM responses did not correlate with protection against an infection with a heterologous IAV strain, indicating potentially different correlates of protection in pig and mice.
In conclusion, mucosal immunization was associated with a prolonged tissue retention of functional TRM, opening up a promising line of enquiry for the design of future vaccines against respiratory viruses.
Main parts of this work were supported by grants from IZKF (advanced project A90). The translational part into the pig model was carried out at the Pirbright Institute, supported by grants from VetBioNet (grant agreement No 731014, Ref: VBN_20_49) and BBSRC.
Distinguished role of MerTk and Axl receptors-mediated efferocytosis in alveolar macrophages on pulmonary mucosal regulation
Authors: Pinto Kamila Guimaraes , Ferreira Antonia Correa , Leandro Monique , Rodrigues Leticia , da Silva Rigoni Thaís , Palermo Ester , Ferreira Jesuino Rafael Machado , Lopes Marcela , Vazquez Estefania , He Jianping , Kelsall Brian , Filardy Alessandra D’Almeida 
Published: August 2023

Abstract
Efferocytosis is paramount to regulating homeostasis and inflammation. The TAM receptor family (Tyro3, Axl and MerTk) mediates efferocytosis and simultaneously inhibits pro-inflammatory pathways. Here we investigated how MerTk and Axl regulate pulmonary mucosa homeostasis and inflammation, using silicosis as a study model. During homeostasis, lung cells and alveolar macrophages (AMs) from wild-type (WT) mice showed high expression of Axl, MerTk, and Gas6. Although we have found increased numbers of total airway cells, lower levels of TGF-b and IL-10, and higher levels of nitric oxide in the BALFs from both Axl−/− and MerTk−/− compared to WT mice, enhanced numbers of lung total cells, higher numbers of AMs, monocytes, and neutrophils were found in BALFs and lungs of MerTk−/− compared to Axl−/− or WT mice. We also found increased expression of CXCL1, CXCL2, TNF-a, and IL-6, and enhanced numbers of MHCII+AMs in the lungs of MerTk−/− compared to Axl−/− or WT mice. Furthermore, AMs from Axl−/− mice upregulated the expression of MerTk receptor. During silicosis, we found decreased pulmonary function, increased levels of TGF-b and CXCL1, as well as increased number of AMs and neutrophils in silicotic (SIL)-Axl mice compared to the SIL-MerTk or SIL-WT groups. In addition, SIL-Axl showed a reduced number of CD206+ AMs and an enhanced number of apoptotic, late apoptotic, and necrotic AMs compared to SIL-MerTk or SIL-WT groups. Finally, we found that both SIL-WT and SIL-MerTk upregulated mRNA levels for Axl compared to its PBS groups. Collectively, our data suggest that MerTk and Axl receptor-mediated efferocytosis are dedicated to regulating homeostasis and controlling inflammation in the lungs, respectively.
CNPq, CAPES and FAPERJ
Ionizing radiation induces lasting changes to the tissue resident memory T cell compartment
Authors: Hassert Mariah , Heidarian Mohammad , Badovinac Vladimir P , Harty John T 
Published: August 2023

Abstract
Exposure to sublethal ionizing radiation can occur through various mechanisms, including purposeful medical exposure, nuclear accidents, and acts of nuclear terrorism, making the development of effective medical countermeasures a high public health priority. The induction of DNA damage through reactive oxygen species makes rapidly dividing cells, such as naïve T cells highly sensitive to radiation induced depletion. A previous study demonstrated that broadly, circulating memory T cells are more resistant to numerical loss following radiation exposure compared to naïve T cells. However, the effects of radiation on the more recently described memory T cell subset, tissue resident memory T cells (TRM) remain unknown. TRM have been identified as a critical T cell subset in protection from a number of infections including influenza, respiratory syncytial virus and malaria. In particular, lung TRM are critical for strain-transcending heterosubtypic immunity to influenza virus. In a murine model of influenza infection (IAV), we have found that sublethal thorax targeted radiation resulted in the rapid numerical decline of lung TRM without major alterations in circulating IAV-specific memory CD8+ T cells. The depletion of anti-IAV lung TRM was long-lasting, and resulted in the loss of heterosubtypic immunity to IAV. Interestingly, any surviving TRM display elevated granzyme B expression, indicating functional consequences for TRM following radiation exposure, which could have implications for immune-mediated pathology. Designing effective vaccination strategies to reinvigorate this critical T cell subset will be an important step in combating the immunological effects of radiation exposure.
1F32AI174382-01
Maternal beta-glucosylceramide induces the generation of IRF4+ dendritic cells in offspring of allergic mothers
Authors: Bloodworth Jeffrey C , Hoji Aki , Blankenship Ross , Cook-Mills Joan 
Published: August 2023

Abstract

Background
β-glucosylceramides (βGlcCers) are elevated in allergic mouse dams and are necessary and sufficient to induce allergy predisposition in offspring. βGlcCers increase dendritic cell (DC) numbers in the fetal liver and pup lungs. We hypothesize that these lung DCs predispose offspring to allergic lung responses. We also hypothesize that βGlcCers induce elevated pro-allergy IRF4+ DC numbers during fetal hematopoiesis.

Methods
Pregnant mice were injected with a mixture of βGlcCers known to predispose offspring to allergic asthma. DCs were assayed by flow cytometry. In another approach, lung DCs from pups of non-allergic dams or allergy predisposed pups born to allergic dams were transferred to lungs of pups of non-allergic dams. Recipient pups received suboptimal sensitization and airway allergen challenge.

Results
An increase in number of IRF4+ DCs was observed in fetal livers when dams received βGlcCers compared to fetal livers from vehicle-injected dams. Higher numbers of IRF4+ DCs were in the lungs of pups born to allergic dams compared to pups born to non-allergic dams. Transfer of CD45.1 lung DCs from pups born to allergic dams to lungs of CD45.2 pups born to non-allergic dams induced allergy predisposition in the recipients compared to lung DC transfers from pups born to non-allergic dams. Furthermore, elevated numbers of endogenous CD45.2 IRF4+ lung DCs were observed in allergen-challenged pups that received lung DCs from pups born to allergic dams.

Conclusions
These results suggest that βGlcCer induces increased numbers of pro-allergy IRF4+ DCs during fetal hematopoiesis. These results further suggest that lung DCs from pups born to allergic dams are induce allergen responsiveness and higher lung IRF4+ DC numbers.

Supported by grants from NIH (R01AI127695 and U01AI131337)
Iteratively stimulated T cells as tools to understand mechanisms of nonlymphoid recirculation
Authors: Künzli Marco , Soerens Andrew G , O’flanagan Stephen D , Joag Vineet , Quarnstrom Clare F , Scott Milcah , Masopust David 
Published: August 2023

Abstract
Memory T cell migration facilitates whole-body immunosurveillance and protection against recurrent infections and cancer. In the absence of inflammation, tissue resident memory T cells (TRM) patrol non-lymphoid tissues (NLT) while effector and central memory T cells (TEM and TCM) are mostly restricted to the blood and the blood/lymph system respectively. Memory T cells that retain the ability to recirculate through all compartments are rare and poorly characterized. Thus, the function of these cells as well as their non-lymphoid tissue entry and egress requirements under homeostatic conditions are not well defined.
Using a heterologous prime-boost immunization scheme and adoptive transfers, we have generated an abundant murine memory T cell population that has been exposed to &gt;50 viral infections over the last 10 years. We termed these cells iteratively stimulated T cells (ISTCs). Parabiosis experiments revealed that ISTCs acquired pan-migration properties: they can be found in blood, non-lymphoid tissues, lymph nodes, and thoracic duct lymph (TDL) of the naïve parabiont. ISTCs persist in NLTs like the lung which are usually associated with short-lived TRM populations, and mice harboring recirculating ISTCs are protected from lethal Influenza challenge. These data indicate that establishing recirculating memory T cells might provide a new strategy to generate long-lasting immunity in tissues like the lung. Single cell RNA-sequencing revealed tissue entry and egress target molecules that will be tested in in vivo CRISPR-Cas9 screens to elucidate mechanisms of T cell recirculation.
Supported by a grant from SNSF P2B-SP3_200187
Ablation of PD-1 Reduces the Stability and Lipid Metabolism of Regulatory T Cells in the Tumor Microenvironment
Authors: Kim Myeong Joon , Kim Kyungsoo , Park Hyo Jin , Kim Gil-Ran , Hong Kyeong Hee , Oh Ji Hoon , Son Jimin , Park Dong Jin , Kim Dahae , Choi Je-Min , Lee Insuk , Ha Sang-Jun 
Published: August 2023

Abstract
Regulatory T cells (Treg cells) are responsible for immune homeostasis and highly express PD-1 in the tumor microenvironment (TME). However, the role of PD-1 in tumor-infiltrating (TI) Treg cells remains controversial. Indeed, PD-1 expression in Treg cells was even higher than in CD8+ T cells and conventional CD4+ T cells. Here, we identified that conditional deletion of PD-1 in Treg cells delayed tumor growth by reducing TI Treg cell pool and amplifying the functionality of TI CD8+ and CD4+ T cells. In Pdcd1fl/flFoxp3eGFP-Cre-ERT2(+/−) mice, in which both PD-1WT and PD-1KO Treg cells coexisted, TI PD-1KO Treg cells exhibited the impaired proliferative and suppressive capacity compared to TI PD-1WT Treg cells. Additionally, exTreg cells, which lost their Foxp3 expression, were more abundant in PD-1KO Treg cells than PD-1WT Treg cells. In TC-1 lung cancer, PD-1 antibody therapy was effective in reducing TI Treg cell pool. Single-cell analysis identified that PD-1 signaling promoted various pathways related to lipid metabolism, proliferation, and suppression in TI Treg cells. Single-cell TCR sequencing revealed that the clonal expansion of TI Treg cells was enriched in PD-1WT Treg cells compared to PD-1KO Treg cells. We also showed that conditional deletion of PD-1 in Treg cells reduced lipid uptake and mitochondrial mass of Treg cells in TME. These results suggest that PD-1 ablation or blockade can enhance antitumor immunity by exacerbating Treg cell stability and metabolic fitness in the TME.
This study was supported by National Research Foundation of Korea (NRF) grants funded by the Korean government (MSIT) (2017R1A5A1014560, 2019M3A9B6065221 to S-.J.H.; 2018R1A5A2025079, 2019M3A9B6065192 to I.L.). This study was also supported by the Korean Health Technology R&D Project (HV20C0144, HN21C1410 to S-.J.H.) through the Korean Health Industry Development Institute (KHIDI) funded by the Ministry of Health & Welfare. The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Improving anti-tumor CD8+ T cell function through manipulating glutamine metabolism
Authors: Fisher Emilie L , Chi Channing , Suguira Ayaka , Madden Matthew , Rathmell Jeffrey C 
Published: August 2023

Abstract
Immunotherapies that bolster the anti-tumor effects of cytotoxic CD8 T lymphocytes (CTLs) have improved outcomes for many, yet most patients fail to achieve durable responses. Investigations into strategies to further enhance the cancer killing capacity of these cells are crucial in the effort to improve immunotherapies. T cells radically alter their metabolism upon activation, and prior studies have shown pan-inhibition of glutamine metabolism (glutaminolysis) using the inhibitor 6-Diazo-5-oxo-norleucine (DON) enhances the function of CTLs in the tumor microenvironment. However, DON’s toxicity combined with this drug’s lack of specificity demonstrate a need for a more directed glutaminolysis targeting approach. I utilized a targeted glutaminolysis CRISPR library to investigate the role of individual enzymes targeted by DON. To interrogate the importance of these enzymes specifically in anti-tumor CTLs, I developed a model in which CRISPR-library edited antigen-specific CTLs were adoptively transferred into tumor-bearing mice. After 7 days of in vivo selection, next generation sequencing was performed on isolated CTLs to evaluate the survival advantage or disadvantage of these gene knockouts. While multiple DON targets resulted in decreased CTL fitness, deletion of the gene encoding for glutamine synthetase (GS) – which converts glutamate to glutamine – conferred an advantage to CTLs. Upon inhibition of GS, cells may increase cellular levels of glutamate, and preliminary data suggest this results in a decrease in glycolysis and increase in mitochondrial respiration capacity. Lastly, in vitro data demonstrate increased inflammatory cytokine production of CTLs upon genetic knockout or pharmacologic inhibition of GS.
Supported by grants from NIH (T32 GM007347, R01 CA217987)
Modulating T Cell Metabolism to Improve CAR-T Cell Therapy success
Authors: Wu Meng-Han , McCalab Megan , Pham-Danis Catherine , Kohler Mark Eric , Rincon Mercedes 
Published: August 2023

Abstract
Chimeric antigen receptor (CAR)-T cell therapy is a promising treatment for lymphoblastic leukemia and lymphoma. Despite initial response to CAR-T cell therapy, about 40% of patients relapse. Modulating CAR-T cell metabolism (immunometabolism) is being considering as a novel approach to improve CAR-T cell therapy efficacy. We have identified MCJ (DnaJC15) as a novel target to enhance CAR-T cell mitochondrial metabolism. MCJ is an endogenous negative regulator of Complex I of electron transport chain. The absence of MCJ in MCJ KO CD8 cells increases Complex I activity, mitochondrial membrane potential, and mitochondrial respiration without affecting glycolysis or ROS production. Lack of MCJ also enhances ATP-dependent effector functions of CD8 cells, including cytokine secretion and cytotoxicity. Using the B16 melanoma cell model, we have shown that antigen-specific MCJ KO CD8 cells are superior in eliminating melanoma cells through enhanced effector function in vitro and in vivo. Importantly, we have generated MCJ-deficient CD8 CAR-T cells against CD19+ B cell leukemia and shown that they exhibit superior killing capacity through their increase cytotoxicity activity in vitro. MCJ KO CAR-T cells have higher mitochondrial maximal respiration and spare respiratory capacity. Furthermore, in vivo MCJ KO CAR-T cells have superior efficacy than WT CAR-T cells, and increase overall survival of the leukemia-bearing mice. Together, these data demonstrate MCJ as a novel target to enhance CAR-T cell therapy efficacy. To show the translational relevance of our studies and moving forward to the clinic, we are currently investigating whether silencing MCJ in human CD8 CAR-T cells also improve their efficacy.
R21 AI110016 R21 AI149187 R56 AI148434 R01 CA260909 R21 CA223389
Caloric restriction induces macrophage reprogramming and improved epithelial ovarian cancer survival
Authors: Singh Harshit , Udumula Mary Priyanka , Hijaz Miriana , Munkarah Adnan , Giri Shailendra , Rattan Ramandeep 
Published: August 2023

Abstract
Calorie restriction (CR) without malnutrition is a dietary intervention where energy intake is reduced by 15 – 40%. The benefits of CR in improving various diseases and extending lifespan has led its investigation in cancer. We have reported 30% CR to limit epithelial ovarian cancer (EOC) progression. Here, we highlight the ability of CR to induce immuno-metabolic reprogramming of tumor associated macrophages and improve EOC survival. Female C57/B6 mice were injected intraperitoneally with 8 × 106 mouse EOC ID8 (p53+/+, p53−/−, or ID8 p53−/−,BRCA1−/−) cells and fed ad libitum (RD) or subjected to 30% CR. Immune profiling was done by flow cytometry. Metabolic phenotype was determined by XF seahorse analyzer. Macrophage marker were measured by qPCR and western blot. Mice on 30% CR exhibited reduced tumor progression and increased overall survival compared to RD mice in all ID8 models as indicated by median survival; ID8p53+/+ (65 vs 95 days), ID8p53−/− (50.5 vs 76.5 days), and ID8p53−/−,BRCA1−/− (44 vs 53 days). Immune profiling showed increase in antitumor associated pro-inflammatory macrophages (F4/80+CD38+) and decrease in protumor anti-inflammatory (F4/80+CD206+) macrophages, increasing the pro / anti-inflammatory ratio. Pro-inflammatory macrophages showed increased intracellular iNOS. The phenotype change in CR mice was accompanied with reprogrammed energy metabolism indicated by increased glycolysis, reflective of pro-inflammatory macrophages. Decrease in anti-inflammatory macrophages resulted in robust anti-tumor T cell response. A 30% CR reprograms macrophage metabolism and promotes their pro-inflammatory anti-tumor phenotype, restores antitumor immunity resulting in reduced EOC growth and improved survival
R01 B11209 to RR Henry Ford Cancer institute Post Doctorate Fellowship to HS