bluuebunny/crossref_metadata_2025_split huggingface dataset:
Unleashing ILC2-dependent anti-tumor immunity to melanoma
Authors: Belz Gabrielle T , Jacquelot Nicolas 
Published: February 2023

Abstract
Group 2 innate lymphoid cells (ILC2) are essential to maintain tissue homeostasis. In cancer, ILC2 can harbor both pro- and anti-tumorigenic functions but the mechanisms underpinning the execution of paradoxical outcomes is not known, nor how it affects clinical outcomes for tumors such as melanoma. We begin to unravel how ILC2 infiltrate human melanoma tumors and execute their anti-tumor potential through a variety of cytokines such granulocyte-macrophage colony-stimulating factor (GM-CSF) to coordinate the recruitment and activation of eosinophils to enhance the anti-tumor response. We identify cytokine-inhibitory receptor circuits that concurrently both promote and limit ILC2 function. Significantly, by understanding these hubs can break the negative impact of ILC2 through specific targetting to limit ILC2 promotion of tumor growth. Together, our results identified ILC2 as a critical immune cell type involved in melanoma immunity and revealed an as yet untapped synergistic approach to harness ILC2 function for anti-tumor immunotherapies.
IL-2 decreased PU.1 and PD-1 expression in NK-cells in NSCLC patients
Authors: Hohenberger Katja , Trufa Denis I , Geppert Carol-Immanuel , Rieker Ralf J , Sirbu Horia , Finotto Susetta 
Published: February 2023

Abstract
The transcription factor PU box binding-1 (PU.1), which is encoded by the SPI1 gene, is required for the development of most myeloid cell lineages as well as B- and T cells. Consequently, an alteration of the SPI1 gene and thus PU.1 can have severe impact on the immune system. However, the role of PU.1 in cancer is not yet fully understood.
In this study, we were looking for a hint on the influence of PU.1 expression on various cell types during non-small cell lung cancer (NSCLC). To start the investigation, we isolated peripheral blood mononuclear cells (PBMCs) from patients with NSCLC as well as from healthy controls. The PBMCs were cultured with different cytokines for 4 days and subsequently analyzed by flow cytometry. Furthermore, RNA was isolated from the PBMCs and qPCR was performed.
The qPCR results showed induced PU.1 mRNA levels in NSCLC patients in relation to GAPDH, compared to control patients. By flow cytometry, we could detect higher PU.1 expression in CD56+ Natural killer (NK) cells from lung cancer patients compared to the healthy donors. Moreover, NK-cells from lung cancer patients, in contrast to healthy controls, expressed the immunosuppressive protein Programmed Death 1 (PD-1). Furthermore, PBMCs from NSCLC patients that were cultured with aCD3/aCD28 antibodies, showed suppression of PU.1 as well as PD-1 in NK-cells, which was even more prominent after addition of IL-2.
These preliminary data suggest, that conditions inducing T cell activation like IL-2 and antiCD3/antiCD28 stimulation, can downregulate both the transcription factor PU.1 and PD-1 in NK-cells in tumor patients. Thus, improvement of current immunotherapy could use additional IL-2, which targets PU-1 as well as PD-1 in NK cells in lung cancer.
Tumor-resident memory CD8+ T cells have a pivotal role in cancer immunity and immunotherapy.
Authors: Corgnac Stéphanie , Mami-Chouaib Fathia 
Published: February 2023

Abstract
CD8+ resident memory T (TRM) cells expressing the CD103 integrin frequently accumulate in human lung tumors and are associated with a favorable prognosis. However, the contribution of this T-cell subset to anti-tumor immunity and to the response to immune checkpoint blockade (ICB) immunotherapy has not been established. Using quantitative multiplex immunofluorescence staining on a cohort of non-small-cell lung cancer (NSCLC) patients treated with PD-1-targeted immunotherapy, we show that an increased density of CD103+CD8+ cells in tumors is associated with greatly improved outcome. CD103+CD8+ cells co-expressed CD49a and CD69, and display a molecular profile characterized by expression of PD-1 and CD39 exhaustion markers, and T-bet, AhR and phosphorylated-STAT-3 transcriptions factors, which confer them a Tc1/Tc17 profile. Moreover, CD103+CD8+ tumor TRM cells, but not CD103−CD8+ (non-TRM) tumor-infiltrating counterparts, demonstrate enhanced proliferation and cytotoxicity toward autologous cancer cells, and display oligoclonal expansion of specific TCRβ clonotypes. Human lung tumors are also infiltrated by CD103+CD4+ T cells likely suggesting a role for this resident population in orchestrating the antitumor T-cell response. Moreover, data obtained in pre-clinical mouse tumor models demonstrate that the absence of CD103 on CD8+ T cells impact the antitumor immune responses. These results explain why CD103+CD8+ tumor-reactive TRM cells are associated with better outcome in ICB-treated patients.
Elucidating the roles of PIK3IP1/TrIP in PI3-kinase regulation, T cell differentiation and T cell function in vivo
Authors: Murter Benjamin M , banerjee hridesh , Szymczak-Workman Andrea , Kane Lawrence P 
Published: February 2023

Abstract

Background:
The cell surface protein TrIP (Transmembrane Inhibitor of PI3K, gene name: Pik3ip1) functions to downregulate PI3K signaling in T cells and can act as a negative regulator of T cell immune responses. This negative immune regulation was recently reported to promote anti-tumor immunity, implicating TrIP as a potential immunotherapy target. Additionally, TrIP was shown to regulate Th1 vs. Th17 cell fate decisions, suggesting a role in Th17-driven autoimmune disease.

Methods:
Using a conditional TrIP knockout mouse model we developed, we have begun to interrogate how TrIP expression regulates the effector function of CD8+ T cells and its impact on tumor immune landscape and overall tumor burden. In parallel, we have assessed the effects of TrIP conditional knockout in T cells on progression of experimental autoimmune encephalomyelitis (EAE) in vivo, and also analyzed the effects on Th17 skewing in vitro.

Results:
We have found that TrIP knockout in CD8+ T cells leads to lower tumor burden compared to WT counterparts. We have also found that the absence of TrIP expression in T cells leads to lower clinical scores and slower disease progression of EAE.
Conclusions/Future Directions: We describe preliminary data showing that the absence of TrIP expression on CD8 T cells promotes anti-tumor immunity and have begun to explore its effects on the tumor microenvironment. Expanding on this work, we are developing a P14-transgenic TCR system for use with gp33-expressing tumor lines to investigate potential differences in T cell trafficking to the tumor. Our preliminary data also demonstrates that TrIP knockout T cells lead to a resistance to EAE, prompting further exploration of the role of TrIP in this disease model and in multiple sclerosis.
Genetically engineered CD4 T cells as a novel immunotherapy against drug resistant Klebsiella pneumoniae infections.
Authors: Hoffmann Joseph P , Iwanaga Naoki , Yang Haoran , Chen Kong , Norton Elizabeth B. , Rangel-Moreno Javier , Kolls Jay K , McCombs Janet E. 
Published: February 2023

Abstract
With mortality rates ranging from 30% to 70%, carbapenem-resistant Klebsiella pneumoniae (Kp) is a significant public health threat. With growing incidence of drug resistance, it is imperative to develop alternative strategies to combat these severe infections. One novel immunotherapy is adoptive T cell transfer therapy using T cells genetically modified to express a T cell receptor (TCR) specific to an antigen of interest. Though successful against cancers and viral infections, this strategy has yet to be utilized for bacterial infections. Our study’s objective was to validate adoptive transfer as an immunotherapy against Kp infections using CD4 T cells from immunized mice. Further, we sought to identify TCRs specific to Kp epitopes for the generation of engineered Kp specific T cells. CD45.1 WT mice were intratracheally vaccinated with outer membrane protein X (OmpX) adjuvanted with heat-labile enterotoxin A1 twice at three weeks apart. Lung and splenic CD4 T cells from immunized mice were transferred into CD45.2 WT mice, followed by a challenge with Kp ST258 C4 strain. Immunized CD4 T cells were sequenced by single cell TCRseq to identify Kp specific TCR sequences. OmpX immunization resulted in a robust population of Kp specific Th17 cells in the lungs. Adoptive transfer of lung, but not splenic T cells from immunized mice significantly reduced bacterial burdens in challenged mice. Additionally, we identified 7 enriched TCR sequences that may be Omp specific. We have confirmed the efficacy of adoptive T cell transfer therapy against Kp infections using T cells from immunized mice. Moving forward we will validate our TCR sequences as Kp specific to genetically engineer both primary and cell line T cells for future therapeutic studies.
The role of monocytes in UVB-mediated acute skin damage in lupus prone mice
Authors: Maz Mitra Padideh , Shi Hong , Wolf-Fortune Sonya , Estadt Shannon N , Kahlenberg J Michelle 
Published: February 2023

Abstract
Systemic lupus erythematosus (SLE) is a heterogeneous autoimmune disorder with manifestations including skin, joint, and kidney inflammation, that is characterized by production of autoantibodies and sensitivity to ultraviolet (UV) light. The precise mechanisms governing UVB-mediated inflammation are incompletely understood; thus, our study objective is to delineate specific innate immune cell populations contributing to UV-mediated inflammation in lupus-prone mice. To that end, lupus-prone NZM mice, lupus-prone iNZM mice (knockout of alpha chain of type I interferon receptor), and wild type Balb/c mice were treated with 100mJ of UV light daily for 5 days. 24 hours after the final dose, skin samples were harvested from each, and CyTOF analysis was performed, revealing skewing of monocyte and macrophage populations in the skin of lupus prone versus wild type mice in a type I IFN dependent fashion. Subsequently, RNA sequencing was performed on skin samples of UV-irradiated NZM and Balb/c mice, which showed upregulation of known inflammatory monocyte chemoattractant genes in lupus prone mice after UV exposure compared to wild type mice. We further characterized monocyte populations recruited to the skin of NZM, iNZM, and Balb/c mice 24 and 72 hours after a single 300mJ dose of UV light by flow cytometry. Our data show a type I IFN dependent, transient predominance of inflammatory Ly6c hi monocytes in lupus prone versus wild type mice at 24 hours after UV exposure. By 72 hours after UV exposure, proportions of inflammatory monocytes in the lupus prone mice decrease. Future studies will be directed at understanding how these inflammatory monocytes differentiate in the context of a type I IFN rich environment and how this contributes to disease.
STAT4 in Conventional Dendritic Cells Regulates Pathogenic T Cell Differentiation and Inflammation in Experimental Autoimmune Encephalomyelitis
Authors: Alakhras Nada Salem , Kaplan Mark H 
Published: February 2023

Abstract
Multiple sclerosis (MS) is an inflammatory demyelinating disease affecting the central nervous system (CNS). Signal transducer and activator of transcription 4 (STAT4) is a major regulator of inflammatory in protective immunity and autoimmunity. STAT4 is activated downstream of two cytokines known to mediate the pathogenic T cell differentiation in autoimmune diseases, IL-12 and IL-23. In experimental autoimmune encephalomyelitis (EAE), an animal model of MS, germ-line deletion of STAT4 in mice results in resistance to the development of EAE. In this study, we investigated the contribution of innate myeloid cells to the phenotype observed in STAT4−/− mice in EAE. Using conditional mutant mice, we immunized mice deficient in STAT4 in myeloid cells and found that mice deficient in STAT4 in CD11c+ cells, but not Lyz2 + cells, are resistant to disease. STAT4fl/flCD11cCre mice are resistant to EAE induction and have inefficient differentiation of the pathogenic IL-17A, GM-CSF, and IFN-γ secreting CD4+ T cells. More importantly, STAT4fl/flCD11cCre mice have a significant decrease of migrated MHCIIhigh+ CD11chigh+CD26+ conventional dendritic cells (cDCs) to the CNS, suggesting a critical role of STAT4 on cDC migration to the CNS. Overall, our results identify a functional requirement of STAT4 in cDCs during EAE induction and propose a potential therapeutic target for autoimmune neuroinflammation diseases.
Card9 promotes pathogenic neutrophil responses that induce Th17-mediated autoimmune arthritis and spondylitis
Authors: Napier Ruth J , Vance Emily E , Lee Ellen J , Rosenzweig Holly L 
Published: February 2023

Abstract
Caspase recruitment domain-containing protein 9 (Card9) promotes anti-fungal immunity following C-type lectin receptor Dectin-1 activation by b-glucans. CARD9 has also been implicated in the pathogenesis of T cell-mediated inflammatory diseases including rheumatoid arthritis and ankylosing spondylitis, but how Card9 controls non-infectious inflammation is poorly understood. Using genetically susceptible SKG mice that develop b-glucan (zymosan)-induced, Th17-mediated autoimmune arthritis and spondylitis we found that Card9-deficiency (Card9−/− SKG mice) conferred complete protection from disease. Card9 promoted arthritis via a Dectin-1-mediated mechanism, as the Dectin-1-specific ligand (curdlan) induced disease and bacterial/fungal ligands specific for Dectin-2 or Mincle did not. At 8 weeks post-zymosan Th17 cells and neutrophils were dramatically reduced in Card9−/− SKG mouse joints, indicating an interplay between Card9, neutrophils and Th17 cells in promoting arthritis. Importantly, pre-arthritic Card9−/− SKG mice had reduced neutrophils in lavage fluid 4 hours post-zymosan compared to SKG mice, but no changes in dendritic cells or macrophages. Lavage fluid neutrophils from Card9−/− SKG mice had similar Dectin-1 expression to SKG mice, yet decreased activation (retention of CD62L) and degranulation of primary (CD63) and secondary (CD66b). Akin to Card9−/− SKG mice, neutrophil-deplete SKG mice had decreased Th17 cells and delayed arthritis onset, supporting a pathogenic function for neutrophils in induction of arthritis. These data suggest that Card9 functions downstream of Dectin-1 to promote pathogenic neutrophil responses necessary to induce Th17-mediated arthritis and spondylitis.
PD-L1 on macrophages protects from severe collagen induced arthritis
Authors: Wood Megan Kay , Talor Monica V , Won Taejoon , Cihakova Daniela 
Published: February 2023

Abstract
Immune related adverse events linked to severe inflammatory arthritis have been identified in 10–43% of patients receiving immune checkpoint inhibitor therapy, aCTLA-4, aPD-1 and aPD-L1, indicating the importance of immune inhibitory pathways in maintaining homeostasis in the synovium. We found that immune checkpoint, PD-L1, is primarily expressed in the joint compared to other immune checkpoints. Furthermore, blocking of PD-L1 during collagen induced arthritis (CIA) resulted in more severe arthritis. Mice synovial macrophages have the highest PD-L1 expression level at homeostasis and macrophage PD-L1 expression is further increased during CIA. Similarly, in human synovial fluid, macrophages have the highest frequency of PD-L1 expression. We hypothesize that the expression of PD-L1 by macrophages is essential for synovial homeostasis and regulation of joint inflammation. We selectively deleted PD-L1 in macrophages using the cre/flox system and induced CIA analyzing disease severity using flow cytometry. LyzcrePD-L1fl/fl showed more severe CIA with a greater change in paw thickness, higher disease and histology score and a higher frequency of CD4 and CD8 T cells in the synovium. Interestingly, female LyzcrePD-L1fl/fl mice have higher arthritis severity to LyzcrePD-L1fl/fl males. This indicates the importance of sex specific PD-L1 expression by synovial macrophages during CIA. Our project has high translational potential since joint specific PD-L1 expression could be modulated for therapeutic purposes in humans with rheumatoid arthritis or check point inhibitors induced arthritis.
Elucidating the role of IL-27 in Sjögren disease-like autoimmunity in the non-obese diabetic mouse model
Authors: Debreceni Ivy L , Chen Yi-Guang , Lieberman Scott M 
Published: February 2023

Abstract
Sjögren’s is an autoimmune disease targeting lacrimal and salivary glands leading to debilitating ocular and oral complications. Lack of understanding of the immunopathogenic mechanisms of disease has hindered development of effective therapies. Non-obese diabetic (NOD) mice spontaneously develop gland inflammation with features similar to humans providing a model to study immune signals that drive this autoimmune process. IL-27 is elevated in serum and saliva of Sjögren’s patients and is required for disease development in NOD mice. However, the mechanisms by which IL-27 contributes to Sjögren’s is not fully elucidated. Here, we define the requirements of IL-27 signaling in effector T cell subsets in the context of lacrimal gland autoimmunity.
IL-27 is a heterodimeric cytokine (EBI3 + p28) and binds a heterodimeric receptor (gp130 + IL-27Rα). IL-27Rα is expressed on CD8 and CD4 T cells, and both T cell subsets play pathogenic roles in lacrimal gland disease development. We utilized IL-27 KO and IL-27Rα KO NOD mice and an adoptive transfer model, to determine which T cell subsets required intact IL-27 signaling to mediate lacrimal gland inflammation in NOD-SCID recipient mice. Neither CD4 nor CD8 T cells sort-purified from IL-27Rα KO donors transferred disease. In contrast, while CD4 effector T cells from IL-27 KO NOD mice transferred disease, IL-27 KO CD8 T cells did not. Together these data demonstrate a T cell-intrinsic requirement for IL-27 signaling in both effector T cells subsets and suggest that CD8 T cells may also produce IL-27 to drive lacrimal gland inflammation. Flow cytometric and gene expression analyses to further define the immunological pathways driven by IL-27 in the context of lacrimal gland autoimmunity are underway.
TNF plays a crucial role in inflammation by signaling via T cell TNFR2
Authors: Alam Muhammad S 
Published: February 2023

Abstract
TNF, produced largely by T and innate immune cells, is potently pro-inflammatory, as are cytokines such as IFN-γ and IL-17 produced by Th1 and Th17 cells, respectively. Analysis of single-cell RNA-Seq datasets from multiple sclerosis (MS) patients identified CSF-infiltrating CD4+ T cells that had inflammatory gene signatures including high levels of TNF. This prompted us to ask if TNF is upstream of Th skewing toward inflammatory phenotypes. We found that exposure of naive human or mouse CD4+ T cells to TNF and TGF-β generated Th17 cells that express low levels of IL-17 (RORγt+IL-17lo) and high levels of inflammatory markers, independently of IL-6 and STAT3. This was mediated by the non-death TNF receptor TNFR2, which was also found to contribute to the generation of inflammatory Th1 cells. Single-cell RNA-seq of CNS-infiltrating CD4+ T cells in mouse experimental autoimmune encephalomyelitis (EAE) found an inflammatory gene expression profile similar to MS. Notably, TNFR2-deficient CD4+ T cells produced fewer inflammatory mediators and were less pathogenic in EAE and colitis. IL-1β, a Th17-skewing cytokine, induced TNF and pro-inflammatory GM-CSF in both human and mouse T cells, which was prevented by disruption of TNFR2 signaling, demonstrating IL-1β can function indirectly via the production of TNF. Thus, TNF is not just an effector cytokine but also an initiator of inflammatory Th differentiation, raising the possibility that targeting TNFR2 may diminish the generation of pathogenic Th cells in diseases in which TNF, IFN-γ, and IL-17 contribute.
Single-cell clonotype tracing of tissue Th17 cells during CNS autoimmunity reveals the conversion of intestinal homeostatic cells to pathogenic cells
Authors: Schnell Alexandra , Huang Linglin , Singer Meromit , Singaraju Anvita , Bollhagen Alina , Thakore Pratiksha I , Dionne Danielle , Delorey Toni M , Pawlak Mathias , Irizarry Rafael , Regev Aviv , Kuchroo Vijay K 
Published: February 2023

Abstract
At homeostasis, most Th17 cells are found in the lamina propria of the intestine, where they contribute to tissue homeostasis by inhibiting microbiota from tissue invasion and promote barrier functions. Recent studies in humans and mice have implicated intestinal Th17 cells in extra-intestinal autoimmune diseases, including multiple sclerosis, but the mechanism in which intestinal Th17 cells mediate these dichotomous functions remains unknown. Here, we combined single-cell RNA- and TCR-seq (scRNA/TCR-seq) with fate mapping of tissue Th17 cells to profile over 84,000 tissue Th17 cells and characterize their heterogeneity, plasticity, and migration at homeostasis and during CNS autoimmunity. We discovered a homeostatic Th17 cell population, that is induced by the intestinal microbiota, is present in both lymphoid organs and the intestine, and expresses IL-17. Joint scRNA- and TCR-seq showed that during EAE this homeostatic population gives rise to a pathogenic Th17 cell population, that migrates specifically through the draining lymph nodes and the spleen to the CNS, and highly expresses GM-CSF and IFNγ, but expresses little or low levels of IL-17. This conversion of homeostatic Th17 cells into encephalitogenic Th17 cells depended on IL-23R signaling. Our work characterizes Th17 heterogeneity, plasticity, and migration during homeostasis and CNS autoimmune disease, identifies a mechanism by which intestinal Th17 cells influence the generation of an autoimmune disease-inducing population, and provides further evidence for a direct relationship between the IL-17-producing and GM-CSF- and IFNγ- producing pathogenic T cells.
STAT3 gain-of-function variants and T cell-mediated inflammation
Authors: Toth Kelsey A , Saucier Nermina , Agbigbe Tejiri , Schmitt Erica , Keppel Molly P , Klechevsky Eynav , Kim Brian , Cooper Megan A 
Published: February 2023

Abstract
STAT3 is a transcription factor that plays a role in the proliferation, survival, and function of many different cell types. In T cells, STAT3 is required for the polarization and function of Th17 cells and can inhibit development of peripheral Tregs. STAT3 polymorphisms have been linked to several common autoimmune disorders where the balance between Th17 and Tregs has been disrupted. Patients with germline gain-of-function variants in STAT3 present with a syndrome of early-onset poly-autoimmunity. We generated two mouse models of STAT3 GOF by CRISPR/Cas9. In these models, mice have a single point mutation, p.G421R or p.T716M, which are two of the exact amino acid substitutions identified in patients. Heterozygous STAT3 GOF mice do not show spontaneous disease or constitutive activation of STAT3 and instead display delayed STAT3 de-phosphorylation following IL-6 stimulation. We hypothesized that STAT3 GOF would enhance Th17 responses and found increased Th17 polarization from naïve STAT3 GOF T cells in vitro. To test whether this increased disease susceptibility in vivo, we treated WT and STAT3 GOF mice with topical imiquimod. Compared to WT, STAT3 GOF mice have more severe disease as demonstrated by ear swelling and epidermal thickness, as well as increased numbers of Th17 cells in peripheral organs at day 7 and in the skin at day 14. No difference was observed in IL-17+ γδ T cell populations in these organs at either time point. STAT3 GOF mice crossed to Rag 1 −/− or IL-22−/− show decreased disease severity, indicating that this response may be mediated by adaptive lymphocytes and IL-22 signaling. These data indicate that STAT3 GOF can exacerbate disease by enhancing Th17 responses that may drive autoimmunity.