bluuebunny/crossref_metadata_2025_split huggingface dataset:
Diverse IL-6 signalling modalities drive pathogenic T cell differentiation and graft-versus-host-disease after allotransplantation
Authors: Gartlan Kate H , Wilkinson Andrew , Chang Karshing , Kuns Rachel D , Henden Andrea , Minnie Simone A , Ensbey Kathleen S , Clouston Andrew , Zhang Ping , Koyama Motoko , Hidalgo Juan , Rose-John Stefan , Varelias Antiopi , Vuckovic Slavica , Hill Geoffrey R 
Published: January 2023

Abstract
Allogeneic stem cell transplantation (alloSCT) and graft-versus-host disease (GVHD) are characterized by systemic interleukin 6 (IL-6) dysregulation, which plays a significant role in shaping donor immune responses and T cell polarization. GVHD is a T cell-mediated disease and the severity and tissue distribution is heavily influenced by T cell-derived cytokines, therefore it is critical to understand the factors that drive T cell polarization in this context to inform therapeutic strategies. IL-6 has a unique receptor system composed of IL-6Ra and the signal transducing molecule gp130, in which signaling occurs via multiple pathways either directly (classical), indirectly via a soluble IL-6 receptor (trans), or presented via antigen presenting cells (cluster). We examined the influence of IL-6 signaling modalities on T cell polarization following allotransplantation, where we found specific targeting of these pathways modulates GVHD outcomes. Using donor grafts composed of IL-6Ra deficient T cells resulted in a profound loss of pathogenic Th17/Th22 differentiation and increased GVHD survival, demonstrating these populations are highly dependent upon classical IL-6 signaling post-transplant. Whilst targeting cluster signaling through IL-6Ra deficient DC had no effect on T cell cytokine responses, trans-signaling inhibition via soluble gp130-Fc resulted in severe skin GVHD. This effect was due to significant expansion of pathogenic donor Th22 and was prevented by donor IL-22 deficiency. These data demonstrate an important role for IL-6 trans signaling in regulating pathogenic T cell polarization pathways following allotransplantation and support IL-6 classical signaling as an important target for GVHD prevention.
Treg suppressor function within allografts is required for tolerance
Authors: Ossart Jason , Pena Andressa , Dai Hehua , Bauer Lynne , Williams Amanda , Camirand Geoffrey 
Published: January 2023

Abstract
The role of regulatory CD4+Foxp3+ T cells (Treg) suppressor function within allografts in tolerance remain unclear. To directly address this, we first used a model where immune reactions are restrained to the graft only. Effector T cells (Teff) alone, or with Treg (2–3×106 each) were transferred to B6 mice lacking secondary lymphoid organs (SLO−; splenectomized LTβR−/−) 5 days after islet allograft (Balb/c). Adoptive transfer of Teff alone induced acute rejection (10d MST). In contrast, Teff + Treg co-transfers significantly prolonged graft survival, with 65% of recipients surviving long-term (&gt;100d MST). Interestingly, Treg did not affect Teff proliferation or accumulation within allografts, but significantly reduced MHC-II expression on DCs and IFN-γ in Teff.
Secondly, given that S1PR1 signaling is necessary for lymphocyte exit from SLO, we prevented Treg migration to allografts during tolerance by removing S1PR1 expression in endogenous Treg specifically. Tamoxifen-fed B6.Foxp3EGFP-Cre/Ert2.S1PR1fl/fl (S1PR1KO-Treg) or B6.S1PR1fl/fl (WT-Treg) mice received Balb/c islets. Untreated mice rejected allografts acutely (18d MST). Interestingly, while anti-CD45RB treatment induced tolerance in WT-Treg control mice (95d MST), all S1PR1KO-Treg mice rejected their graft (28d MST; p&lt;0.05 vs WT-Treg). Also, these results correlated with a 25% decrease in Treg infiltrating islet allografts and with a 22% increase in Treg in lymph nodes on day 10 in S1PR1KO-Treg + anti-CD45RB compared to WT-Treg + anti-CD45RB recipients.
Overall, our data demonstrate that Treg can suppress rejection by Teff within allografts without prior activation in SLO, and suggest that Treg suppressor function within allografts is required for tolerance.
Tissue-Specific NK Cell Diversity in Solid Tumors
Authors: Dunbar Zerick T , Shanker Anil 
Published: January 2023

Abstract
Cytotoxic innate lymphoid cells, conventionally known as natural killer (NK) cells, play major roles in cancer immunity largely due to their natural cytolytic and immune regulatory functions. However, due to the extensive heterogeneity and plasticity seen among NK cells, their functions have yet to be fully harnessed in immunotherapy. The objective of this study is to elucidate NK cell diversity maturation, recruitment and effector function profiling in solid tumor microenvironments. We hypothesize that NK cells from different tissue locations display genetic and functional profiles that can predict NK effectiveness in solid tumor microenvironments. Here, we show the biological diversity that exists among NK cells from different locations pre- and post-interaction with solid breast cancer tumors, including heterogeneity of NK cell specific cluster of differentiation markers and gene expression perspectives based on flow cytometry, qPCR, RNA, and bioinformatics analyses. Understanding NK cells in the tumor microenvironment could help answer critical NK cell research questions and lead to advancements in cancer immunotherapy applications, ultimately helping to mitigate the cancer resources-results discrepancy.
Sympathetic nervous tone influences the development and function of natural killer cells
Authors: Nevin James Timothy , Moussa Marmar , Corwin William L. , Mandoiu Ion I. , Srivastava Pramod K. 
Published: January 2023

Abstract
Signals derived from the nervous system inform immune development and function in a variety of ways. For example, sympathetic nervous system (SNS) activity has been demonstrated to influence the function of innate lymphoid cells (ILCs) in the context of inflammation. Natural killer (NK) cells, a subset of ILCs, are important players in the anti-tumor immune response that possess the capability to respond to signals from the SNS via expression of adrenergic receptors (ARs) on their surface. Given the critical functions of NK cells in tumor immunity and their capacity to respond to SNS signals, we hypothesized that SNS input plays a key regulatory role in the NK-mediated anti-tumor response. Here, we demonstrate that the SNS delivers a critical maturation signal that promotes NK cell development and influences their cytotoxic capacity. Mice that lack functional SNS signaling after peripheral administration of 6-hydroxydopamine exhibit a higher proportion of poorly cytotoxic NK2 cells (CD11b−CD27+) and a lower proportion of potently cytotoxic NK1 cells (CD11b+CD27−) compared to controls. Furthermore, analysis of single-cell RNA sequencing establishes that SNS ablation disrupts normal NK development and that immature NK cells accumulate in SNS-ablated tumor-bearing mice. Together, these data demonstrate that tonic sympathetic nervous activity plays an important role in NK development. Further characterization of this role will improve our understanding of the consequences of neuroimmune interactions in cancer and inform the development of cancer immunotherapy.
LncRNA AK036396 inhibits maturation and accelerates immunosuppression of polymorphonuclear myeloid-derived suppressor cells by enhancing the stability of ficolin B
Authors: Tian Xinyu , Wang Shengjun 
Published: January 2023

Abstract
Long non-coding RNAs (lncRNAs) are emerging as crucial regulators of cell biology. However, the role of lncRNAs in the development and function of polymorphonuclear myeloid-derived suppressor cells (PMN-MDSCs) remains unclear. Here, we identified that the lncRNA AK036396 was highly expressed in PMN-MDSCs, and lncRNA AK036396 knockdown promoted the maturation and decreased the suppressive function of PMN-MDSCs. Ficolin B (Fcnb), whose expression could be used to reflect PMN-MDSC development, was the predicted target gene of lncRNA AK036396 based on microarray results. LncRNA AK036396 knockdown attenuated Fcnb protein stability in a manner dependent on the ubiquitin-proteasome system. Moreover, Fcnb inhibition downregulated the suppressive function of PMN-MDSCs. In addition, the expression of human M-ficolin, which is an orthologue of mouse Fcnb, was increased and positively correlated with arginase1 (Arg1) level in lung cancer patients. Based on these findings, lncRNA AK036396 can inhibit maturation and accelerate immunosuppression of PMN-MDSCs by enhancing Fcnb protein stability.
CCL2/CCR2 signaling protects against bladder cancer growth in a T cell dependent manner
Authors: Mukherjee Neelam , Ji Niannian , Shu Zhen-Ju , Curiel Tyler J , Svatek Robert S 
Published: January 2023

Abstract
The chemokine CCL2 (C-C motif ligand 2) is an important signaling axis underlying recruitment of pro-tumorigenic myeloid cells and is associated with worse outcomes in several tumor models. Currently, anti-CCL2 antibodies are being tested in cancer trials for solid tumors. To test CCL2 signaling in bladder carcinogenesis, wild type (WT) mice and mice deficient in CCL2 (CCL2KO) were given the urothelial carcinogen N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN). Surprisingly, tumor incidence was higher in CCL2KO mice demonstrating an unanticipated protective role for CCL2 signaling in bladder cancer (BC). We then orthotopically challenged WT, CCL2KO, and CCR2KO mice (lacking the major CCL2 receptor) with MB49 BC and confirmed the protective effect of CCL2/CCR2. WT mice had significantly more intratumoral T cells compared to CCL2KO mice suggesting that CCL2 is involved in recruiting T cells to the bladder. Further, depletion of T cells abolished this protective effect of CCL2. Adoptive transfer of CCR2+ T cells into CCR2KO mice restored protection against MB49. CCR2+ T cells were also more activated, functional and tumor specific compared to their CCR2− counterparts. We found that anti-CCL2 promotes BC growth highlighting a concern for use of anti-CCL2 in BC. Moreover, intravesical recombinant CCL2 (rCCL2) either alone or in combination with intravesical gemcitabine reduced bladder tumor and improved survival of mice with MB49 BC. We further developed a slow-release encapsulated nanoparticle formulation of rCCL2 which successfully treated MB49, C3H mice with MBT-2 BC and reduced urine hematuria in our first ever humanized model of orthotopic BC. Our results are rapidly translatable and identify a novel treatment strategy in BC.
Bifidobacterium improves the outcome of immune checkpoint blockade by modulating Treg cell function
Authors: Wang Feng 
Published: January 2023

Abstract
Immune checkpoint-blocking antibodies that attenuate immune tolerance have been used to effectively treat cancer, but they can also trigger severe immune-related adverse events. Previously, we found that Bifidobacterium could mitigate intestinal immunopathology in the context of CTLA-4 blockade in mice. Here, we examined the mechanism underlying this process. We found that Bifidobacterium altered the composition of the gut microbiota systematically in a regulatory T cell (Treg)-dependent manner. Moreover, this altered commensal community enhanced both the metabolic and suppressive functions of intestinal Treg cells, contributing to the amelioration of colitis during CTLA-4 blockade. Importantly, Bifidobacterium ameliorated gut immunopathology and boosted anti-melanoma immunity with combination therapy of anti-CTLA-4 and anti-PD-L1.
Emerging roles of Cxcl12/Cxcr4 signaling axis in breast cancer metastasis
Authors: Karagiannis George S , Rivera-Sanchez Luis , Duran Camille L , Oktay Maja H , Condeelis John S 
Published: January 2023

Abstract
The Cxcl12/Cxcr4 axis promotes breast cancer metastasis in mice and humans. Prior studies have linked Cxcl12/Cxcr4 to cancer cell homing, survival and proliferation at distant sites secreting Cxcl12, such as lungs and liver. However, the mechanism via which Cxcr4+ cancer cells escape primary tumors in the first place (which also highly express Cxcl12), remains elusive. Here, by measuring chemotactic gradients using immunofluorescence, we show that Cxcl12 gradients are concentrically expressed around cancer cell intravasation sites, and spatially associate with Cxcr4+ tumor cells. Without affecting the gradient itself, pharmacological inhibition of Cxcl12 suppresses the translocation of Cxcr4+ cancer cells to intravasation sites. Paradoxically, tumor cell-specific deletion of Cxcr4 results in a suboptimal prometastatic response. We theorized that Cxcr4+ tumor-associated macrophages (TAMs), previously shown to accompany migratory tumor cells, could instead read the Cxcl12 chemotactic gradient, thus partially rescuing the prometastatic phenotype. Indeed, clodronate-mediated TAM depletion abrogates Cxcr4+ cancer cell translocation to intravasation sites and subsequent dissemination to secondary sites. We also found that breast cancer endothelium is lined by Cxcl12+Pdgfrb+ stromal cells; however, Cxcl12 gradients originating from intravasation sites are specifically tied to a subset of Cxcl12High perivascular TAMs. As such, pharmacological inhibition of Pdgfrb depletes Pdgfrb+Cxcl12+ stromal cells, but does not affect Cxcl12+ TAM-dependent translocation of Cxcr4+ tumor cells to intravasation sites. Overall, our data support a new model for the implication of Cxcl12/Cxcr4 in the early stages of metastasis.
Preclinical evaluation of efficacy and cytokine release of bispecific T cell engagers in humanized NSG mice
Authors: Yao Li-Chin , Cai Danying , Aliperta Roberta , Colthart Allison , Meier Werner , Cheng Mingshan , Keck James G 
Published: January 2023

Abstract
Bispecific T cell engagers (BiTEs) are a growing class of cancer therapy that redirects effector T cells against target-expressing tumors. Mouse models to assess efficacy and cytokine release associated with T cell engagement are needed. JAX humanized NOD scid gamma (Hu-NSG™) mice are engrafted with cord blood-derived CD34+ HSC and have shown robust T cell reconstitution. In this report we evaluated two novel BiTEs, EGFR-CD3 and BCMA-CD3, in Hu-NSG™ mice implanted with MDA-MB-231 human breast cancer cells and NCI-H929 multiple myeloma, respectively. Following the initial dosing, we measured human cytokines and analyzed T cell activation by flow cytometry. Clinical observations and body weight change were also monitored. Treatment with EGFR-CD3 for 2 weeks resulted in complete tumor regression when the dosing was initiated around 100 mm3. 50% of the treated-tumors remained nonpalpable for 10 weeks after dosing ceased. EGFR-CD3 also inhibited tumor growth in mice bearing tumors of ~400 mm3 average in volume. In both cases human IFN-γ, IL-6, and IL-10 levels were elevated from the first dose before returning to the baseline and the frequency of CD69+ cells in T cell subsets increased after the treatment. BCMA-CD3 treatment also induced T cell activation and elevated levels of human IFN-γ, TNF-a, IL-6, and IL-10. NCI-H929 tumor growth inhibition by BCMA-CD3 showed donor to donor variability. We observed that significant growth inhibition in one donor while the other donor did not respond. Together, our Hu-NSG™ platform enables preclinical evaluation of drug efficacy and cytokine release induced by BiTEs and has the potential to be applied to other immunotherapies.
Therapeutic targeting of CD38 enhances temozolomide activity in glioblastoma cells and modulates their tumor microenvironment by eliciting an enhanced CD8+ T cell response
Authors: Manna Alak , Aulakh Sonikpreet , Chanan-Khan Asher , Paulus Aneel 
Published: January 2023

Abstract
Glioblastoma (GBM) is an aggressive primary malignant brain tumor with poor prognosis. Surgical resection followed by chemoradiotherapy (temozolomide [TMZ] + radiation) results in a median survival of only 12-15 months. Recently, immunotherapy with targeted antibodies, therapeutic vaccination and adoptive cell therapy offers a promising option for treating GBM. Herein we observed that CD38, a catalytic receptor and regulator of NAD homeostasis, is expressed on GBM cells and their microenvironment in primary patient-derived GBM tissues. Blacher et al have shown that blocking CD38 enzymatic functions results in reduced GBM cell growth. This evidence prompted us to investigate the antitumor and immune engaging effects of a clinical grade anti CD38 therapeutic monoclonal antibody (Daratumumab/DARA) in preclinical models of GBM. We demonstrated that DARA significantly enhanced the direct apoptotic effect of TMZ and induced ADCC/CDC of GBM cells, irrespective of the presence of microglia. In an in vitro 3D simulated GBM + T cell coculture system, DARA induced activation (CD69/GITR upregulation/decreased PD1) and enhanced proliferation of CD8+ T cells/cTL activity against GBM cells. DARA also resulted in increased IFNγ and decreased IL-10; suggesting an improved antitumor cytokine milieu and ensued in downregulated inflammatory cytokine associated JAK2/STAT3 signaling pathway. Finally, we tested TMZ+DARA in a human GBM xenograft in vivo model and noted significantly prolonged survival of mice [TMZ+DARA vs. vehicle: 98.13±1.88 vs. 63.63±3.77 days, p&lt;0.001]. Altogether, these observations led us to investigate the clinical activity of DARA+ TMZ in GBM patients in a phase I/II clinical trial.
Outlining key inflammation associated parameters during early phase of an experimental gram-negative sepsis model in rhesus macaques (Macaca mulatta)
Authors: Rosado-Franco Jose J , Ramos-Benitez Marcos J , Parodi Laura M , Rosario Derick , Compo Nicole , Giavedoni Luis D , Espino Ana M 
Published: January 2023

Abstract
Sepsis is a life-threatening syndrome characterized by metabolic and systemic dysregulation in the presence of an invading pathogen, such as bacteria or fungi. In the initial stages of this syndrome, the body, in an attempt to eradicate the pathogen, secretes high levels of pro-inflammatory cytokines and chemokines that leads to an anti-inflammatory state, which leads to organ dysfunction and death. Despite the efforts to find targeted therapeutics to treat septic patients, physicians rely on the administration of antibiotics and fluids. Hence, the development of novel targeted therapeutics to treat septic patients at early stages of sepsis is highly important to save the lives of these patients. The aim of this study was to identify inflammation markers during the early phase of sepsis in rhesus macaque. Four rhesus macaque were given an intravenous dose of 1010 CFU/kg of E. coli. Blood samples were collected before, 30 minutes, 2, 4, 6 and 8 hours after E. coli infusion. Physiological parameters, bacteremia, endotoxemia, C-reactive protein (CRP), procalcitonin (PCT), and cytokines and chemokines were determined for each animal at each time point. Bacteremia was present in all animals from 30 minutes to 3 hours after E. coli infusion. Endotoxin, CRP and PCT levels remained detectable during the whole experimental window. Signature cytokines and chemokines such as TNFα, MIP1α, and MIP1β peaked about 2 hours after E. coli infusion and decreased thereafter. Plasma IL6, IL12p40, IFNγ, and IL1Ra, ITAC, MIG, IP10 and MCP1, remained at detectable levels after 4 hours of E. coli infusion. This non-human primate model could be useful for the assessment of new therapeutics aiming to suppress key inflammatory markers throughout sepsis early phases.
Glucose is required for IFN-γ secretion but not IFN-γ mRNA expression by peripheral T cells
Authors: Loving Crystal L , Bond Zahra F 
Published: January 2023

Abstract
T cells play a key role in adaptive immunity, proliferating and producing effector molecules upon activation. T cell effector function is regulated by changes in their metabolism, which is necessary for ensuring availability of biosynthetic precursors for expansion, gene transcription, and protein translation. Impaired or altered immune T cell function often coincides with cellular metabolic impairment or shifts, and may be a cause for defective immune responses. For example, peripheral porcine T cells have increased glycolytic capacity following Arterivirus infection, but fail to produce IFN-γ protein in response to antigen stimulation. To understand the mechanism by which porcine T cell effector function is controlled by metabolic substrate availability a series of in vitro studies with varying culture conditions were performed. Specifically, peripheral blood mononuclear cells were stimulated with phorbol 12-myristate 13-acetate plus ionomycin in the presence or absence of glucose for varying amounts of time, and IFN-γ mRNA expression and IFN-γ protein secretion assessed. Glucose was required at the time of stimulation for both IFN-γ mRNA expression and translation/secretion. If glucose was removed 24h after stimulation, IFN-γ mRNA and protein were still produced, though protein levels were reduced. However, if glucose was not readily available upon stimulation, IFN-γ mRNA was transcribed, but not secreted. L-glutamine was supplied in all assays. Collectively, glucose availability to T cells can impact effector function and may be a factor in impaired responses during infection.
Cross-species gene expression analysis of human, canine and murine natural killer cells suggests convergence of activated dog and human transcriptomic profiles
Authors: Gingrich Alicia A , Reiter Taylor , Judge Sean J , Sturgill Ian , Monjazeb Arta M. , Murphy William J , Brown C Titus , Canter Robert J. 
Published: January 2023

Abstract
Key species differences exist between human and murine natural killer (NK) cells. Although outbred dogs are a link for translational NK studies, better characterization of dog NK cells is needed. We used RNAseq to profile resting and activated NK cells from mice, dogs and humans. Cells were isolated based on putative NK cell populations from healthy humans (CD56+CD3−), beagles (CD5dim) and C57BL/6J mice (NK1.1+CD3−) and sequenced from steady-state or following exposure to irradiated human feeder cells (K562 clone 9) with 100 IU/mL rhIL-2 × 14 days (human and dog) or 1000 IU/mL rhIL-2 × 7 days. Purity of expanded human and dog NK cells was ~ 90%, while for mouse it was ~ 70%. We performed differential gene expression (DGE) across species and conditions using ~7000 1:1 orthologous genes. DGE revealed distinct transcriptional profiles for each species (FDR &lt;0.05). Hierarchical clustering demonstrated mouse NK cells as an outgroup compared to dog and human. PCA showed within-species spatial clustering of resting NK cells. After activation, variance between dog and human NK cells decreased, variance between human and mouse NK cells increased (PC1 40%, PC2 28%). Species differed in expression of key genes after activation, including PRF1, GZMA, SPP1, IL2RB, CTSD and FCER1G, which showed greater increases in abundance counts in mice than dogs and humans. In this first transcriptomic sequencing of dog NK cells, we show distinct gene profiles of resting cells across the most commonly used mouse, dog, and human NK populations, with convergence of dog and human NK cells upon stimulation under these standard conditions. By defining the cross-species comparative transcriptional profiles of putative NK cells, these data fill a gap in translational NK studies.