bluuebunny/crossref_metadata_2025_split huggingface dataset:
Low intensity vibration increases proliferation in human primary T cells: a non-invasive strategy to reduce “vein-to-vein” time in autologous cell therapies
Authors: Ashdown Christopher P , Chan Meilin , Rubin Clinton 
Published: August 2023

Abstract
Chimeric antigen receptor T cell therapy (CAR-T therapy), is revolutionizing cancer treatment. Nevertheless, autologous CAR-T therapy remains an inefficient process; CAR-T manufacturing is often prohibitively slow, requiring up to 4 weeks from white blood cell acquisition to infusion (“vein-to-vein” time). Our goal is to leverage the innate mechanosensitivity of human T cells through low intensity vibration (LIV) to promote proliferation and shorten the expansion phase of autologous CAR-T therapy. LIV is characterized as a low-magnitude (0.7g, where g=9.8ms−2) and high-frequency (30Hz) signal. LIV was delivered to human primary T cells in culture for two one hour bouts per day, with each bout separated by a 2h rest period. Following 5d, each group was analyzed for proliferation rates and activation status/exhaustion.
We found that subjecting T cells to LIV enhanced proliferation by 29% relative to sham controls (p&lt;0.004). This indicates a significant increase in proliferation over a relatively short period of time. Overstimulation of T cells can cause the development of a phenotype commonly known as “exhaustion”, which is characterized by decreased effector function. We used flow cytometry to examine the effect of LIV on T cell activation and exhaustion and found that there were no significant differences in surface receptors linked with T cell activation (CD25, CD62L, CD69) or T cell exhaustion (TIM-3, LAG-3, and PD-1) between control and LIV groups after 5d exposure to LIV.
These data suggest that LIV could be used to shorten the vein-to-vein time for CAR-T therapy. Furthermore, the lack of exhaustion is equally important because it suggests that LIV expanded cells are able to proliferate faster while maintaining their functionality.
Supported by grants from NIH REACH (U01) HL127522 and SUNY Technology Accelerator Fund (TAF), with additional support from the Snyder Family Fellowship (SBU Cancer Center).
CAR T-cell intrinsic PD1 Dominant Negative Receptor (PD1DNR) functions by both cell-intrinsic and extrinsic mechanisms
Authors: Amador Alfredo , Hou Zhaohua , Chintala Navin , Adusumilli Prasad S 
Published: August 2023

Abstract

Objectives
We have shown that chimeric antigen receptor (CAR) T cells with T-cell intrinsic PD1 dominant negative receptor (PD1DNR) prolong functional persistence. Herein, we further define T-cell intrinsic and extrinsic mechanistic functions of PD1DNR.

Methods
Human T cells were retrovirally transduced with mesothelin targeted, CD28 co-stimulated CARs– either M28z (M) or M28z with PD1DNR (MDNR). Cytotoxicity, proliferation, phenotype, cytokine and effector and inhibitory protein expression were assessed by chromium release, cell count, flow cytometry, olink and mass spectrometry, respectively. MDNR extrinsic activity was measured by the ELISA for soluble PD1 and assessing cytotoxic functions of M cells following co-culture with MDNR supernatant. CAR T-cell functional persistence was assessed in vitro by repeated antigen stimulation and in vivo by tumor treadmill test and mice survival.

Results
Following repeat antigen stimulation, MDNR cells demonstrate retained cytotoxicity, proliferation, accumulation, and enrichment of TNF-related cytokines (≥1× Log2), and overexpression of T-cell activation and proliferation proteins (GZMA, GNLY, IFNAR2, MFAP4). Soluble PD1 was 10-fold higher in MDNR supernatants. Co-culture of M cells with MDNR supernatant demonstrated higher cytotoxicity (p&lt;0.01). Mice treated with a single dose of MDNR show enhanced survival compared to M (median survival 68.5 vs. 27 days, P&lt;0.01). MDNR displayed also superior in-vivo functionality against advanced stage disease (median survival 62 vs. 12 days, P&lt;0.01).

Conclusions
PD1DNR expression in CAR T cells functions by both cell-intrinsic and extrinsic functions that are beneficial in solid tumors adoptive cell therapy.
LKB1 controls inflammatory potential through CRTC2-dependent epigenetic remodeling
Authors: Compton Shelby Elizabeth , DeCamp Lisa M , Kitchen-Goosen Susan M , Lau Kin , Dahabieh Michael S , Guak Hannah , VanderArk Alexandra , Vos Matthew , Williams Kelsey S , Shi Xiaobing , Rothbart Scott B , Krawczyk Connie M , Jones Russell G 
Published: August 2023

Abstract
Deregulated inflammation is a critical feature driving tumor progression in tumors harboring mutations in the Liver kinase B1 (LKB1), yet the mechanisms linking LKB1 mutations to deregulated inflammation remain undefined. Here, we identify deregulated signaling by CREB regulated transcription coactivator 2 (CRTC2) as an epigenetic driver of inflammatory potential downstream of LKB1 loss. We demonstrate that LKB1 mutations sensitize both transformed and non-transformed cells to diverse inflammatory stimuli, promoting heightened cytokine and chemokine production. LKB1 loss triggers elevated CRTC2-CREB signaling downstream of the salt inducible kinases (SIKs), increasing inflammatory gene expression in LKB1-deficient cells. Mechanistically, CRTC2 cooperates with the histone acetyltransferases CBP/p300 to deposit histone acetylation marks associated with active transcription (i.e., H3K27ac) at inflammatory gene loci, promoting cytokine expression. Together, our data reveal a previously undefined anti-inflammatory program, regulated by LKB1 and reinforced through CRTC2-dependent histone modification signaling, that links metabolic and epigenetic states to cell-intrinsic inflammatory potential.
Supported by the National Institute of General Medical Sciences (NIGMS, R35GM124736), the National Institute of Allergy and Infectious Diseases (NIAID, R21AI153997), the Paul G. Allen Frontiers Group Distinguished Investigator Program, the National Institute of Allergy and Infectious Diseases (NIAID, R01AI165722), and Van Andel Institute.
The effects of cannabinoid 1 receptor blockade on oxazolone-induced contact hypersensitivity mouse model
Authors: Han Ji Hye , Kim Wook 
Published: August 2023

Abstract
Contact hypersensitivity (CHS) is an experimental model of human allergic contact dermatitis, one of the representative skin diseases, where NLRP3 inflammasome plays a crucial role. Although the regulation of NLRP3 inflammasome activity by cannabinoid 1 receptor (CB1R) has been shown in metabolic diseases such as obesity and type 2 diabetes, the role of CB1R in CHS is unclear. Here, we investigate the putative role of CB1R in CHS by using a selective CB1R antagonist rimonabant. Pharmacological blockade of CB1R by rimonabant ameliorated oxazolone-induced CHS by suppressing hyperproliferation of keratinocytes and expression of pro-inflammatory cytokines in a TLR2/4-dependent manner. The inhibitory effect of rimonabant in oxazolone-induced CHS was not observed in NLRP3-deficient mice. These results suggest that CB1R could control TLR2/4 signaling pathway via NLRP3 inflammasome in CHS.
Supported by Basic Science Research Program through the National Research Foundation of Korea(NRF) funded by the Ministry of Education (NRF-2022R1I1A1A01071661)
Intercellular mitochondria transfer rescues aerobic respiration in metabolically compromised macrophages in vitro and in vivo
Authors: Brestoff Jonathan R , Giwa Rocky , Jia Wentong , Field Rachael L 
Published: August 2023

Abstract
Mitochondria evolved through an ancient endosymbiotic process and are essential for eukaryotic cellular metabolism. An emerging body of literature indicates that some cells export mitochondria for delivery to other cells, a process termed intercellular mitochondria transfer. Recently, we demonstrated that adipocytes transfer mitochondria to macrophages in vivo and that this process maintains white and brown adipose tissue homeostasis (Brestoff et al., Cell Metab, 2021; Rosina et al., Cell Metab, 2022; Borcherding/Jia et al., Cell Metab, 2022). It is believed that macrophages degrade exogenous mitochondria as a form of licensed mitophagy that supports adipocyte mitochondria quality control, however it is unclear whether externally-derived mitochondria are able to respire in macrophages once captured. We administered purified mitochondria to wildtype (WT) or NDUFS4 KO mice, which lack an enzyme required for mitochondrial Complex I activity. Although WT peritoneal macrophages quickly degraded exogenous mitochondria, KO peritoneal macrophages captured, retained, and utilized exogenous mitochondria to rescue cell-intrinsic defects in aerobic respiration. Similarly, healthy BV2 microglial cells quickly degraded captured mitochondria in vitro, but BV2 microglial cells exposed to rotenone and Antimycin A captured and utilized exogenous WT mitochondria to rescue the toxin-induced defects in aerobic respiration. Purified mitochondria from NDUFS4 KO mice failed to rescue aerobic respiration in toxin-stressed BV2 cells. These data suggest that macrophages with defective mitochondrial metabolism can obtain and utilize exogenous, cell-free mitochondria from their environment to overcome a metabolic crisis.
Supported by a grant from the Burroughs Wellcome Fund (CAMS #1019648) to JRB.
A novel role of EphA4/Stat6/Mertk signaling in cleaning up the brain after trauma
Authors: Soliman Eman , Gershenson Ilana , Basso Erwin Gudenschwager , Leonard John , Mills Jatia , Ju Jing , Tartaglino Chiara , Elhassanny Mohamed , Chen Michael , Wang Xia , Theus Michelle 
Published: August 2023

Abstract
Traumatic brain injury (TBI) is a major cause of disabilities and deaths from acute injuries in the US and worldwide. Neuroinflammation and neuronal apoptosis are hallmarks of TBI; however, the mechanism of clearance of apoptotic debris by efferocytosis remains ill-defined. Our previous studies demonstrated that EphA4 receptor in peripheral-derived macrophage (PDM) mediates the pro-inflammatory response after injury. Here, we aimed to evaluate whether PDM-specific EphA4 restricts efferocytosis following TBI. We generated wild-type (WT) and EphA4-knockout (KO), GFP+ bone marrow (BM) chimeric mice and performed Controlled cortical impact injury. We observed that KO mice displayed improved motor function, cerebral blood flow, reduced cortical cell death as measured by TUNEL, and enhanced efferocytosis compared to WT mice. The percentage of GFP+/Iba1− expressing PDMs that engulf NeuN+ or TUNEL+ cells was significantly higher in KO mice (p&lt;0.05, n=5). We further performed an in vitro efferocytosis assay using pHrodo-labeled apoptotic Jurkat cells co-cultured with GFP+ WT and KO BM-derived macrophages. We found a significant increase in efferocytosis efficiency in KO compared to WT cells (p&lt;0.05, n=4). In addition, mRNA isolated from KO cells show increased expression of the efferocytosis receptor Mertk and its ligands Gas6 and Pros1 and phospho-array analysis shows increased p-Stat6, a key regulator of Mertk/Gas6. Interestingly, the use of Stat6 and Mertk inhibitors attenuated efferocytosis effects in KO macrophages in-vitro. Our findings implicate the EphA4/Stat6/Mertk axis as a novel regulator of apoptotic debris clearance that may be targeted to augment resolution of inflammation following TBI.
National Institute of Neurological Disorders and Stroke of the National Institutes of Health, NS096281, NS119540, and NS121103 (MHT)
CD8α identifies human NK cells with altered proliferation, metabolism, and IL-15 signaling
Authors: Cubitt Celia , Wong Pamela , Tran Jennifer , Foltz Jennifer Ann , Marin Nancy D , Marsala Lynne , Foster Mark , Schappe Timothy , Berrien-Elliott Melissa M , Fehniger Todd A 
Published: August 2023

Abstract
NK cells are cytotoxic innate lymphoid cells that can differentiate into memory-like (ML) NK cells following IL-12/15/18 stimulation. Previous work identified that CD8α expression on donor ML NK cells was negatively associated with outcome for cellular therapy of AML. However, the role of CD8α on human NK cells remains poorly studied, and murine NK cells are CD8α-precluding study in mice. We found that CD8α-NK cells had greater proliferation and survival in vitro and in vivo. Interestingly, we observed that CD8α expression was dynamic, and a subset of CD8α-NK cells upregulated CD8α (“new” CD8+) after IL-15 stimulation. Notably, the “new” CD8+ NK cells were the most proliferative in vitro and in vivo, and had significantly greater signaling following IL-15 stimulation. Strikingly, we also found that “new” CD8+ NK cells had significantly higher glucose uptake, nutrient receptor expression, and an enhanced capacity for glycolysis and oxidative phosphorylation. These results indicate that recent CD8α acquisition marks the ability of NK cells to respond to IL-15 signals to engage in the robust metabolic activity required for proliferation. We also investigated the role of CD8α in NK cell activation, given that CD8α can bind HLA and associate with Lck. In the absence of CD8α, NK responses (CD107a, IFNg, TNF) following activating receptor ligation were enhanced for several receptors, suggesting an intrinsic role for CD8α in inhibiting NK cell activation. Together, these results identify a novel association of CD8α with treatment outcome for NK cellular therapy and identify a previously unappreciated role of CD8α on human NK cell proliferation, metabolism, and response through activating receptors.
Supported by grants from NIAID F30AI16131
“The role of macrophage scavenger receptors in engineered biomaterial scaffold remodeling”
Authors: Faust Mondreakest T , Sadtler Kaitlyn , Destefano Sabrina , Fertil Daphna , Josyula Aditya 
Published: August 2023

Abstract
CD206, also known as mannose receptor C type 1, is used frequently as a marker to identify the M2 polarized macrophages in the investigation of immune responses to biomaterials. M2 macrophages help with tissue repair, inflammation reduction, building immune tolerance, and defense against excessive inflammation. CD206 is found on the surface of M2 macrophages and dendritic cells, and functions as a pattern recognition receptor for a variety of connective tissues in health, including pulmonary tuberculosis, liver fibrosis, and others. Previous research has shown a role of CD206 in internalization of collagen fragments in tumors. In this study, we aim to determine whether CD206 is more than just an M2 marker in response to biomaterials. We have found that adaptive immune cells as well as cross-presenting dendritic cells are critical for CD206 induction in biomaterial-treated muscle injury. In Batf3−/− mice, where CD206 expression is lost, we find hemosiderin deposition suggesting defects in phagocytosis. In future studies, we will use CD206-deficient to directly evaluate its role in scaffold remodeling at the site of injury.
TNF receptors orchestrate differential neutrophil responses critical to immunity against Staphylococcus aureus skin infection
Authors: Youn Christine , Pontaza Cristina , Wang Yu , Dikeman Dustin A , Joyce Daniel P , Alphonse Martin P , Nolan Sabrina J , Ahmadi Michael , Young Jeremy , Tocaj Aron , Garza Luis A , Miller Lloyd S , Archer Nathan K 
Published: August 2023

Abstract
As the leading cause of skin and soft tissue infections, Staphylococcus aureus is a major health burden and a continued threat to human health due to the emergence of antibiotic-resistance. Through this current study, we aimed to better characterize the protective immune mechanisms against S. aureus skin infection to identify novel targets for the development of host-directed immunotherapeutics as alternatives to antibiotics. Tumor necrosis factor (TNF) is a master regulatory cytokine whose inhibition is associated with the increased risk of S. aureus skin colonization and infection in humans. Despite the clinical implications, the contribution of TNF and its cognate receptors, TNFR1 and TNFR2, to immunity against S. aureus skin infection is unclear. Utilizing an in vivo mouse model of S. aureus intradermal infection, we determined that immune cell-derived TNF engages with both TNFR1 and TNFR2 to promote protection against S. aureus in the skin. Neutrophil-intrinsic TNFR1 and TNFR2 signaling directed differential antimicrobial mechanisms against S. aureus skin infections. Mechanistically, TNFR1 mediated neutrophil recruitment to the skin, whereas TNFR2 directed neutrophil activation (e.g.-NOX2 activation, ROS production, and PAD4-dependent neutrophil extracellular trap (NET) formation) and prevented systemic dissemination of skin infection. Collectively, our findings revealed non-redundant functions and cellular mechanisms mediated by TNFR1 and TNFR2 in neutrophils for immunity against S. aureus.
This study was funded in part by grants R01AR073665 (NKA), R01AR069502 (NKA), R01AI111205 (NKA), and R01AI146177 (NKA) and from the United States National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS) and National Institute of Allergy and Infectious Diseases (NIAID).
SARS-CoV-2 Envelope Protein Tolerizes Macrophage Response to Secondary Inflammatory Stimuli
Authors: Pierce Stephen H , Mclennan Rebecca , Khanal Santosh , Menden Heather , Sampath Vankatesh , Bradley Todd 
Published: August 2023

Abstract
Exposure of innate immune cells to bacteria or viruses may result in epigenetic rewiring that either enhances or weakens innate immune responses to a secondary challenge. One primary mechanism of innate immune cell function is through toll-like receptors (TLRs) that sense a variety of bacterial and viral molecules and cause immune activation. SARS-CoV-2 Envelope protein (E protein) has been shown to mediate innate immune activation, through engagement of TLR2. In this study, we determined the secretome of chemokines, cytokines and growth factors from monocytes in response to E protein stimulation and compared this response with other viral antigens and TLR agonists. Next, we stimulated monocytes with E protein followed by secondary stimulation with lipopolysaccharide (LPS) on monocyte-derived macrophages (MDM) 1 week later. Reduced expression of CCL3, CCL4, CCL5, CXCL10, TNF-A, and IL-12 upon secondary LPS or autologous E protein stimulation were statistically reflecting a more tolerant MDM state that is imprinted after E protein exposure. Finally, we demonstrated that immunizing neonatal mice with SARS-CoV-2 E protein induced proinflammatory cytokine secretion and lung tissue inflammation pathology and demonstrated a long-term impact on immune cell function and secondary response to LPS. Thus, E protein induces macrophage proinflammatory networks that lead to tissue inflammation, but also induce a more tolerant state after return to a resting state that could impact the ability to respond to secondary infection. Further understanding of the molecular pathways that drive this immune activation and tolerance could identify treatment targets to restore immunity after SARS-CoV-2 infection.
Understanding the role of CD22 on Macrophage and Dendritic Cell Function
Authors: Zastrow Alexi , Friedman David J , Crotts Sydney B , Rajcula Matthew , Hammer Brady , Elissa Mai , Shapiro Virginia Smith 
Published: August 2023

Abstract
At the terminal position of many glycan chains are unique sugars known as sialic acids. Sialic acids bind sialic acid immunoglobin-like lectins (Siglecs). Our lab is interested in the role of CD22 (Siglec-2), which binds to α2,6-linked sialic acid generated by ST6GalI. CD22 can associate with itself or other molecules on the same cells in a “cis” configuration or bind in “trans” with ligands on other cells. CD22 contains four ITIMs within its cytoplasmic tail and is primarily known to function as an inhibitory co-receptor of the B cell receptor (BCR). CD22-ligand interactions restrain CD22 and BCR association which dampens BCR signaling. While CD22 is usually characterized as a B cell specific protein, we found that CD22 is highly expressed in hybrid macrophages (CD11b+ CD11c+ F4/80+) and expression decreases with activation, implying that CD22 may modulate myeloid cell responses. Using novel macrophage and dendritic cell-specific CD22 cKO mouse models, we will explore how CD22 impacts myeloid cell activation. We propose that CD22 impacts myeloid cell function and modulating this interaction may improve immune responses.
Inflammation-induced development and migration of a novel monocyte-like dendritic cell population that regulates antiviral CD8 T cell immunity
Authors: Oliva Kimberly , Reagin Katie , Klonowski Kimberly D 
Published: August 2023

Abstract
We recently discovered a CD11c+MHC-II+CD103+CD11b+ monocyte-like dendritic cell (moDC) population that colocalizes with CD8 T cells in the draining lymph node (dLN) early after respiratory infection. Importantly, depletion of these cells enhances the formation of CD8+ tissue resident memory T cells (TRM) in the lung. Given the importance of TRMafter respiratory challenge, we aimed to understand the origin and migration of these moDCs. To this end, we intranasally (i.n.) infected C57Bl/6 mice with influenza virus A HK-×31 (×31) or with vesicular stomatitis virus (VSV), the latter which allows us to compare with identical systemic infection. moDCs were detected in the bone marrow (BM) ~2 days post infection, one day prior to their peak recovery from the dLN. BM moDCs phenotypically resembled dLN moDCs, including expression of the inflammatory monocyte-associated markers CCR2 and Ly6C. Adoptive transfer of BM Ly6C+ monocytes into congenically mismatched recipients showed a meaningful percentage of moDCs were monocyte-donor derived. Additionally, type I IFNs, which can induce monocyte differentiation and are crucial in early antiviral immunity, were required for moDC development. We showed that moDCs are decreased in the BM, and consequently the dLN of Ifnar1−/− mice compared to WT mice following respiratory infection. Lastly, moDCs were detected in the BM after intravenous (i.v.) VSV but not in the dLN. This could be attributed to the increased expression of CCL2, MIP-1β, and IL-12p40 in the dLN of i.n. VSV or ×31 infected mice compared to naïve or i.v. VSV infected mice, where inflammation is dispersed. Overall, we provide potential targets in the moDC development and migration process that could enhance antiviral CD8 T cell immunity.
Supported by grant from NIH (R21 AI131093)
Surfactant lipids inhibit PI3K-dependent signaling pathways induced by IL-4 in alveolar macrophages
Authors: Casals Cristina , García-Fojeda Belen , Tenreiro Paula , Minutti Carlos M 
Published: August 2023

Abstract
Alveolar macrophages (AMs) are less able to respond to IL-4 in vivo than macrophages from the peritoneal cavity, due to a still-unknown factor of the lung environment. The aim of this study is to investigate whether surfactant lipids, which are continuously endocytosed by AMs, could influence IL-4-mediated alternative activation and proliferation of AMs. To that end, AMs were preincubated with surfactant lipids and stimulated with IL-4 in the presence or absence of surfactant protein SP-A, an amplifier of IL-4 actions. We found that alveolar lipids reduced IL-4- and IL-4+SP-A-dependent arginase activity, the expression of genes associated with alternative activation, and proliferation of AMs. Mechanistically, endocytosed lipids decreased IL-4- and IL-4+SP-A-induced activation of the PI3K-Akt-mTORC1 signaling axis, but not the IL-4-dependent STAT6 axis. Lipid-dependent inhibition of the Akt/mTORC1 signaling axis is consistent with reduced IL-4+SP-A-driven glycolysis and mitochondrial respiration as well as decreased ATP citrate lyase expression and histone acetylation stimulated by IL-4. We conclude that surfactant lipids inhibit PI3K-dependent signaling pathways and suggest that decrease of surfactant lipids in chronic lung diseases might augment IL-4-dependent fibrotic responses.
This research is funded by the Spanish Ministry of Science and Innovation through Grant PID2021-123044OB-I00 to C. Casals