bluuebunny/crossref_metadata_2025_split huggingface dataset:
Recipient LAG3 deficiency results in antibody-mediated rejection of mouse renal allografts
Authors: Nicosia Michael , Lee Juyeun , Fan Ran , Gorbacheva Victoria , Beavers Ashley , Dvorina Nina , Baldwin William , Fairchild Robert L , Min Booki , Valujskikh Anna 
Published: January 2023

Abstract
The functions of coinhibitory receptor lymphocyte activation gene-3 (LAG3) in T cells are well studied, however its role in humoral immune responses remains poorly characterized. The goal of this study was to test the role of recipient LAG3 in a mouse model of renal allograft rejection
Compared to WT animals, naïve B6.LAG3−/− mice have increased splenic cellularity and higher frequencies of CD44hi memory T cells, CXCR5hi follicular T cells, and B220+CD138+plasma cells yet do not develop spontaneous autoimmunity. Furthermore, naïve B6.LAG3−/−(H-2Db) mice have increased frequencies of memory T cells against H-2Dd, H-2Ds, H-2Dq and H-2Dk alloantigens. C3H (H-2Dk) kidney allografts were transplanted into B6.WT or B6.LAG3−/− recipients after bilateral nephrectomy. Whereas 4/4 WT recipients accepted C3H allografts for longer than 60 d, recipient LAG3 deficiency led to rapid allograft rejection (MST of 14 d, n=5) with serum creatinine levels of 0.1 and 1.35 mg/dl respectively at d14 posttransplant. Graft histology at rejection revealed minimal T cell infiltration, diffuse C4d staining, atrophic peritubular capillaries, endothelial swelling and edema characteristic of antibody mediated rejection (AMR). Compared to WT, LAG3−/− recipients had elevated frequencies of anti-donor IFNg producing T cells and increased levels of anti-donor MHC-II antibodies. Recipient CD8 T cell depletion did not alter the rejection kinetics in LAG3−/− recipients (MST of 16 d), while B cell depletion significantly extended C3H kidney allograft survival (MST of &gt;30 d).
These results suggest the predominant role of alloantibody rather than T cells in renal allograft injury and identify LAG3 is a potential therapeutic target for AMR prevention and treatment.
Formation and function of resident memory t cells in renal allografts
Authors: Daya Khodor Abou , Zhao Daqiang , Tieu Roger , Rammal Rayan , Oberbarnscheidt Martin H. , Lakkis Fadi G 
Published: January 2023

Abstract
The role of TRM in transplantation is unknown. In this study, we investigated the formation and function of TRM in a mouse kidney transplantation model.
(B6xBALB/c) F1.ova kidneys were transplanted to B6 recipients and 1m OT-I Teff cells transferred on day 2. Graft, blood, bone marrow, SLO, and liver were harvested 4 and 8 wks after transplantation. TRM were identified as CD44hiCD62LlowCD69+ CD103+/− cells after excluding in vivo labeled T cells. OT-I and polyclonal TRM were transcriptionally characterized using scRNAseq. TRM residency was tested by parabiosis and re-transplantation. To further establish a causal relationship between the TRM OT-Is and rejection we depleted them at wk 4 post adoptive transfer using anti-Thy1.1 (250 ug IV per day for 7d).
Mean serum creatinine (mg/dl) was significantly higher in allo vs syn group at wk 8 (0.7 vs 0.2, p&lt;0.05). Graft histology showed mixed acute and chronic rejection in the allo group. Flow analysis of allograft cells demonstrated TRM cells among OT-I and endogenous T cell populations at 4 & 8 wks. The OT-I population was exclusively TRM phenotype by flow and scRNAseq, rapidly produced IFNγ upon re-stimulation, and was not detected anywhere else. There was no significant difference in mean number of OT-Is between wk 4 and wk 8 (277K vs 234k, p=0.74). OT-I T cells could not be detected in the parabiont kidney graft or tissues or in the secondary host outside the re-transplanted kidney, indicating that the TRM are indeed resident in the graft and do not re-circulate. Depleting the OT-Is preserved kidney function at wk 8 compared to the non-depleted group (Cr= 0.7 vs 0.2, p&lt;0.05).
Our findings show that donor-specific TRM form in kidney allografts, are functional, and contribute to rejection.
The alarmin HMGB1 in cell cytoplasm can modulate the apoptosis to autophagy switch of pancreatic beta cells under stress
Authors: Chung Hyunwoo , Seo Sol , Hong Sung Ji , Choi So Won , Park Chung-Gyu 
Published: January 2023

Abstract
High mobility group box 1 (HMGB1) is an infamous alarmin which is known to be harmful to pancreatic beta cells and associated with diabetes mellitus pathogenesis and pancreatic islet graft failure. However, recent studies have indicated that cytosolic HMGB1 could function as a modulator to relieve cells from apoptotic stress by inducing autophagy. Particularly, pancreatic beta cells have been well-known to demonstrate the apoptosis to autophagy switch when exposed to hypoxia or lipoxiticy. In this study, we have investigated the beta cells under hypoxic and lipotoxic stress with CCK8 assay and flow cytometry while utilizing a small molecule inhibitor of HMGB1 which can suppress the cytosolic accumulation of HMGB1. The results demonstrated that the cytoplasmic HMGB1 blockade decreased the viability of mouse beta cell line MIN6 and primary islets. Moreover, the cytoplasmic HMGB1 blockade decreased the autophagic flux and increased the ratio of apoptotic MIN6 cells. Also, it was noteworthy that the MIN6 cells have shown high autophagic flux even in basal state. We believe this study linked the role of cytoplasmic HMGB1 in apoptosis to autophagy switch and the importance of autophagy in beta cells under apoptotic stress. We expect this study to open up a novel field of HMGB1 utilization within diabetes- and islet transplantation-related therapeutics, where the current field is mainly focused on the exhaustive blockade of HMGB1.
Autophagy Induced due to Moderate Endoplasmic Reticulum Stress in T Cells Enhances Immunotherapeutic Tumor Control
Authors: Chakraborty Paramita , Chatterjee Shilpak , Tran Danh T. , Pytel Dariusz , Kim Dosung , Nadig Satish N. , Atkinson Carl , Wang Hongjun , Diehl J. Alan , Gangaraju Vamsi , Palanisamy Viswanathan , Mehrotra Shikhar 
Published: January 2023

Abstract
A number of external or internal stimuli cause endoplasmic reticulum (ER) stress conditions. In response to ER stress, stressed cells activate well-orchestrated cellular signaling processes to resolve the stress or remove itself by triggering apoptosis. In the tumor microenvironment, induction of sustained ER stress is known to dampen the antitumor activity of T cells. However, how T cells respond to suboptimal ER stimuli is unknown. Using melanoma antigen gp100 reactive T cells, we found that T cells transiently activate protein kinase R-like endoplasmic reticulum kinase (PERK) dependent signaling network to ameliorate low level of ER stress. While transient PERK activation in T cell enhances stemness, promotes mitochondrial biogenesis, increases T cells anti-tumor immunity in B16 melanoma bearing mouse, overexpression of PERK triggers T cell dysfunctionality and death. Mechanistically, we discovered that transient activation of PERK decreases overall protein translation and induces protective autophagy to prevent cell damage. Conversely, knockdown of PERK abrogates autophagy activation, hampers mitochondrial biogenesis in response to suboptimal ER stress, which in turn compromises antitumor function. Furthermore, blocking autophagy in T cells hampers T cell anti-tumor activity. Lastly, using gp100 reactive LC3-GFP expressing T cells we demonstrated that, T cells that initiate autophagy due to suboptimal ER stress show better anti-tumor potential compared to those, that fail to enter into the process. Overall, these preclinical data highlight that, both transient ER stress and autophagy pathways are important for healthy T cell function and therapeutically, can be manipulated to regulate their antitumor potential.
Exploiting cancer cell metabolism with stimuli-responsive toll-like receptor agonists
Authors: Mancini Rock , Ryan Austin , Pulukri Anunay , Davaritouchaee Maryam , Abbasi Armina , Hendricksen Aaron , Opp Larissa , Burt Anthony , Nielsen Amy 
Published: January 2023

Abstract

Methods
We report stimuli-responsive Toll-Like Receptor (TLR) agonist prodrugs that rely upon the irregular metabolism (Warburg Effect and efflux potential) common among different types of cancers to activate immune cells. Beginning with Coley’s Toxins in the late 1800’s, TLR agonists have exhibited anticancer properties, and more recently have been shown to act synergistically with anti-CTLA-4 or anti-PD-L1 checkpoint blockades. Specifically, TLR7/8 agonists deplete MDSCs, activate CD8+ T-cells, M1 macrophages and pDCs; and enhance abscopal effects across animal models for melanoma, breast, and prostate cancers. Despite these profound effects, the severe systemic inflammatory toxicity elicited by TLR agonists presents a significant barrier to using them as drugs. To address this, we developed stimuli-responsive TLR agonist prodrugs targeted to cancer cell metabolism.

Results
Our prodrugs are inactive until conversion by complementary stimuli selected from α-mannosidase, β-galactosidase, or β-glucuronidase with concomitant efflux via MDR1 and comparable processes. We observe conversion and efflux in several cancer types, including B16 melanoma, 4T1 breast, and TRAMP prostate cancers, resulting in immunogenicity as measured by activation of primary BMDCs. In-vivo, prodrugs do not induce systemic inflammation and perform differently than parent TLR agonist in a 4T1 neoadjuvant resection metastasis model.

Conclusion
These results suggest that stimuli-responsive TLR agonist prodrugs target the action of TLR agonists to cancer cell metabolism and mitigate systemic inflammatory effects relative to the parent TLR agonist.
Innate immune populations contribute to effective immune responses against B lymphoblastic leukemia cells
Authors: Hunter Rae , Henry Curtis J. , Porter Christopher C. 
Published: January 2023

Abstract
Pediatric patients diagnosed with acute lymphoblastic leukemia (ALL) have a 15–20% mortality rate, in spite of current advances in therapies, including immunotherapies. Despite the use of immunotherapies in patients with relapsed or refractory ALL, relapse rates post-treatment remains over 50 percent. These observations highlight our lack of understanding of immune evasion mechanisms used by leukemia cells.
Our group has previously published that IL-12 promotes a very effective immune response to B-cell ALL cells in vivo. While T cells are required to maintain remission, we observed prolonged survival in IL-12 treated Rag1−/− mice with leukemia as compared to untreated mice, suggesting a major role of the innate immune response in ALL cells. Depletion of NK cells with anti-NK1.1 did not abrogate the elimination of an immunogenic model of leukemia, in which calcineurin has been knocked down by shRNA. However, depletion of myeloid cells from immunocompetent mice using colony-stimulating factor-1 receptor (CSF1R) depleting antibodies promoted the progression of this leukemia, which was eliminated in all isotype treated, immune-competent recipients, indicating myeloid cells play a critical role in the immune response to ALL cells. Ongoing studies suggest dendritic cells (DCs), specifically CD11b+CD11c+ DCs, are reduced in mice transplanted with parental relative to immunogenic, Cn-deficient ALL cells. Furthermore, macrophages cultured in vitro with parental cells expressed significantly lower surface levels of CD80, CD40 and CD86 compared to levels found on the surface of cells cultured with Cn-deficient leukemia cells. Overall, our data suggest that ALL cells may compromise the T-cell priming capacity of innate immune cells.
Tumor-associated myeloid cells provide critical support for T-ALL in vivo
Authors: Lyu Aram , Triplett Todd A. , Godfrey Wesley H. , Nam Seo Hee , Ames Rachel Y. , Hu Zicheng , Sarang Adviti , Selden Hilary J. , Lee Chang-Han , Georgiou George , Horton Terzah M. , Ehrlich Lauren I. R. 
Published: January 2023

Abstract
T cell acute lymphoblastic leukemia (T-ALL) is a malignancy of immature T cells. Despite harboring multiple genetic alterations that drive disease, primary T-ALL cells require exogenous signals to survive. We previously reported that tumor-associated myeloid cells support survival of primary mouse T-ALL cells in vitro. However, the contribution of myeloid cells to T-ALL progression in vivo remains unclear. Here, we depleted phagocytic myeloid cells from leukemic mice, resulting in a significant reduction in tumor burden in multiple organs and prolonged survival. T-ALL burden also diminished following myeloid depletion in lymphodeficient mice, indicating that adaptive immunity was not required to eliminate T-ALL cells, thus suggesting tumor-associated myeloid cells could directly support T-ALL growth. Consistent with this possibility, multiple subsets of tumor-associated myeloid cells support survival of mouse and primary patient T-ALL cells in vitro. Acute myeloid depletion reduced activation of several pathways regulating cell survival in T-ALL cells, implicating these pathways in promoting T-ALL progression in vivo. Computational analysis of published datasets shows an association between enriched macrophage gene signatures and poor prognosis in T-ALL patients, further implicating a pro-tumor role for myeloid cells. Together, our results indicate that an altered myeloid compartment directly promotes T-ALL progression at divergent T-ALL sites in vivo.
A novel Cullin 4A/BCereblon E3 Ligase Modulator, FPFT-2216, offers protective effect against Myelodysplastic syndrome
Authors: Hailu Yohannes Gemechu , Kishimoto Tadamitsu 
Published: January 2023

Abstract
Thalidomide is an immunomodulatory drug (IMiD) having a therapeutic action in hematological malignancies and inflammatory diseases. Despite the therapeutic benefits, thalidomide is known for its high toxicity and responsible for a number of children born with anomalies when taken by pregnant women. Thalidomide’s high toxicity resulted in the development of more powerful and less toxic analogs.
The mouse is resistant to IMiDs effect and for this reason, we developed mice carrying human cereblon to study IMiDs mechanism. We investigated the potential IMiDs effect on T cell stimulation/suppression. We found that T-lymphocytes from humanized cereblon mice treated with a novel IMiD, FPFT-2216, result in higher production of IL-22, IL-2 and IL-9 whereas wild type mice were resistant. Furthermore, IMiDs showed potential degradative effects towards Cereblon substrates like Aiolos and interferon regulatory factor 4 among humanized cereblon mice but not in wild type. These IMiD-mediated T-cell stimulations might augment anti-tumor immunity.
Next, we assessed the potential therapeutic effects of FPFT-2216 in Myelodysplastic syndrome (MDS). NSG mice were engrafted with MDS patient derived cell line, MDS-L, and treated by DMSO, or FPFT-2216. Mice treated with FPFT-2216 showed a longer survival rate and less infiltration of MDS-L into their bone marrow as compared to their control. Moreover, In vitro experiments confirmed that FPFT-2216 offers a significant anti-MDS effect. In conclusion, FPFT-2216’s strong preclinical effect in MDS-L cell provides a hint that it can help in treating MDS patients and it supports further safety-check in humanized cereblon mice, other primates and clinical trials.
Conflict of interest: None
Modified culture protocol for differentiation of human promyelocytic leukaemia PLB-985 cell-line into mature neutrophil-like granulocytes
Authors: Shehu Shaayau , Edwards Steven W. , Wright Helen L. 
Published: January 2023

Abstract
Neutrophils are considered to be mature and terminally differentiated. They are short-lived, lack proliferation capacity and are impossible to transfect in vitro to express exogenous genes or proteins. These properties have made their ex vivo experimental manipulation and biochemical studies of their signal activation mechanisms and genetic makeup, challenging. Establishment of cell line models capable of differentiating along the myeloid lineage into mature neutrophil-like granulocytes, with similar morphological and functional properties to blood neutrophils would, therefore, be an important tool to enable transfection of key genes and determine the effects of such manipulations on neutrophil functions. PLB-985 is a recently established cell line that has been demonstrated to show greater levels of differentiation into granulocyte in the presence of appropriate differentiation-inducing agents, but the efficiency of differentiation reported so far was very low, and the differentiated cells only partly resembled mature blood neutrophils. This study developed a modified differentiation protocol and an optimised culture conditions that induced the PLB-985 cell line to differentiate into mature, neutrophil-like granulocytes, Terminally-differentiated, neutrophil-like phenotypes of PLB-985 cells that resembled mature blood neutrophil morphology, and which had an appreciably delayed apoptosis have been consistently cultured. This was evident by the acquisition of multi-lobed nucleus, granulated cytoplasm and delayed apoptosis. The differentiated PLB-985 cells may, therefore, provide an excellent neutrophil-model system for in vitro study of neutrophil differentiation, signalling and functions.
Neutral ceramidase regulates ferroptosis in myeloid-derived suppressor cells
Authors: zhu huabin , Lu Chunwan , Yang Dafeng , Redd Priscilla S , Ibrahim Mohammed L. , Klement John D. , Smith Alyssa D. , Albers Thomas , Lebedyeva Iryna , Coant Nicolas , Hannun Yusuf , Liu Kebin 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSCs) are key targets in cancer immunotherapy. Neutral ceramidase (nCDase), a key enzyme that hydrolyzes ceramide into sphingosine, is highly expressed in colon carcinoma and regulates apoptosis and autophagy in colon carcinoma cells. We report here that nCDase is also highly expressed in tumor-infiltrating immune cells in mouse and human colon carcinoma in vivo. To target nCDase, we performed molecular docking based on human nCDase structure aganist a virtual chemical library. In vitro enzymatic inhibition analysis of top hits identified nCDC06 as a potent nCDase inhibitor. Chemical and NMR analysis determined nCDC06 structure and molecular weight (325.1 Dalton). nCDC06 has an IC50 of 10.16 to 25.91 mM for human nCDase and 18.6–30.2 mM for mouse nCDase. nCDC06 induces MDSC cell death in a dose-dependent mechanism. Inhibition of ferroptosis decreased nCDC06-induced MDSC cell death. Analysis of ultracellular structure, ROS accumulation, and glutathione revealed that nCDC06 suppresses MDSCs at least in part through a ferroptosis-dependent mechanism. Gene expression profiling revealed that nCDC06 activates the p53 pathway in MDSCs. Furthermore, nCDC06 therapy significantly inhibited tumor growth in an experimental lung metastasis mouse model. We determined that nCDase regulates MDSC ferroptosis and nCDC06 is potent in activating the p53 pathway to induce MDSC ferroptosis to suppress tumor growth in vivo.
Metabolic barriers imposed on tumor associated macrophages
Authors: Roy Sohini , Young Kim 
Published: January 2023

Abstract
Macrophages comprise a broad spectrum of heterogeneous cell population with diverse functions in immune responses in pathological conditions including cancer, inflammation and infection. Macrophages are extremely plastic in nature and highly specialized in sensing their microenvironment which governs their functional phenotype. Simplistically, macrophages can be categorized into classical inflammatory or M1 type and anti-inflammatory or M2 type. While the M1 macrophages play a crucial role in activating immune responses and release a wide array of pro-inflammatory and cytotoxic molecules, M2 macrophages facilitate resolution of inflammation by secreting immunosuppressive anti-inflammatory factors, phagocytosing and clearing apoptotic debris and promoting sustainability of regulatory T cells while suppressing effector T cells. Emerging evidences indicate an intimate connection between a robust switch in metabolic adaptations and different classes of macrophages. Tumor associated macrophages (TAM) are abundantly present in tumor microenvironments and can account for up to 50% of myeloid infiltrate in some types of human solid malignancies and contribute to disease progression, therapy resistance and clinical outcome. However, the metabolic circuitries in TAM underlying their ability to sustain tumor evolution and influence their own functional polarization is complicated and less characterized in many tumor types. In the current study, using multi-prong approaches like bulk myeloid as well as single cell RNA sequencing as well as functional assays, we highlight the metabolic pattern of TAMs in head and neck cancer and melanoma and propose immunometabolic strategies to harness TAM for controlling tumor growth.
Dendritic cells dysfunction in tumor-draining lymph nodes
Authors: Bandola-Simon Joanna Marta , Wong Nathan , Cam Maggie , Roche Paul A 
Published: January 2023

Abstract
An effective anti-tumor response requires activation of both CD4 helper T cells (through MHC-II molecules) and CD8 cytotoxic T cells (through MHC-I molecules). Activation of tumor-specific T cells depends on tumor antigen presentation by DCs. However, it is known that the tumor microenvironment largely suppresses immune responses. The focus of our study is to characterize the functional status of DCs in tumor-draining lymph nodes and to identify the mechanisms that lead to defective presentation of tumor antigens. We transplanted tumors into the flank of mice and analyzed DCs from both tumor-draining lymph node (TDLN) and non-draining lymph nodes (NDLN) as our model system. Cross-presentation of OVA to CD8 T cells was not suppressed in TDLN in comparison to the NDLN either in vivo and ex vivo. However, both in vivo and ex vivo CD4 T cell activation by DCs from TDLN was defective. Subset analysis revealed that both migratory and resident DCs in TDLN show a less mature phenotype as compared to their counterparts isolated from NDLN. Functional assays revealed defects in antigen uptake and processing in DCs from TDLN. We also identified differences in the proportion of MHC-II molecules containing the Invariant chain CLIP peptide in DCs from TDLN in comparison to NDLN. These features of DC function, supported by RNA-Seq analysis, contribute to defective antigen presentation to CD4 T cells and may explain the dysfunction of DCs in tumor-bearing mice. Further analysis should enable moving towards “correcting” DC defects that would allow CD4 T cell priming to provide help to anti-tumor CD8 T cells.