bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibiting fatty acid synthesis (FAS) can prevent and treat chronic graft versus host disease (cGVHD)
Authors: Aguilar Ethan George , Rhee Stephanie , Thangavelu Govindarajan , Loschi Michael , Paz Katelyn , Saha Asim , Ly Joanna , Sage Peter , Sharpe Arlene H , Maillard Ivan , Murphy William J , Serody Jonathan S , Jin Suji , Wagers Sara Bolivar , Nicholls Jemma , Hill Geoffrey , MacDonald Kelli PA , Compeer Ewoud , Dustin Michael , O’Connor Roddy , Sparwasser Tim , Blazar Bruce R 
Published: January 2023

Abstract
GVHD remains the primary complication of allogeneic hematopoietic stem cell transplantation. Considerable interest exists in understanding how metabolism affects immune cell function. We show targeting FAS represents an attractive strategy to ameliorate cGVHD. Acetyl-CoA carboxylase 1 (ACC1) catalyzes the first step of FAS, and can be pharmacologically inhibited with either Soraphen A or 5-(Tetradecyloxy)-2-furoic acid (TOFA). FAS is critical for T cell effector responses, a finding we confirmed using an in vivo multiorgan system cGVHD model with bronchiolitis obliterans, a model dependent on germinal centers (GC), IgG2c secretion, lung deposition and fibrosis. Compared to wild type T cells, cGVHD mice receiving CD4CreACC1fl/fl allogeneic T cells had reduced cGVHD severity assessed by pulmonary function test and immune phenotyping. As thymic regulatory T cell (tTreg) infusion can prevent and treat cGVHD, we examined tTreg FAS needs. Inhibiting FAS in sorted tTreg via drug (Soraphen A; TOFA) or genetic (FoxP3CreACC1fl/fl) strategies increased oxidative and glycolytic metabolism measured by Seahorse assay, that correlated with superior in vitro suppressor function. These findings were extended in vivo where transfer of FoxP3CreACC1fl/fl tTreg, at day 28 following disease establishment, decreased cGVHD severity. Daily treatment of cGVHD mice from day 28 to day 56 with TOFA reversed cGVHD. An in vitro assay in which T follicular regulatory cells suppress T follicular helper and GC B cell class switching showed that inhibiting tTreg FAS can increase their suppression of GC responses. These data provide evidence that inhibiting FAS can prevent or treat cGVHD and enhance Treg function, providing a new approach to ameliorate cGVHD.
Prophylactic antibiotic treatment exacerbates Graft vs. Host Disease-induced bone marrow failure and spleen hypoplasia
Authors: McDaniel Brianyell T , Enriquez Josue , Furr Kathryn , Grisham Matthew 
Published: January 2023

Abstract
Acute graft vs. host disease (aGVHD) is a major complication following hematopoietic stem cell transplantation. Preclinical and clinical studies suggest aGVHD may be potentiated by gut damage and translocation of intestinal microbiota following toxic, pre-transplant conditioning protocols. We recently reported that aGVHD-induced bone marrow (BM) failure and splenic hypoplasia develops in the absence of gut damage suggesting that intestinal bacteria may not be required for disease pathogenesis.

Objective
Determine whether prophylactic gut decontamination with broad spectrum antibiotics (Abx) affects the onset and/or severity of aGVHD-induced BM and spleen damage.

Methods
Syngeneic (Bl6) or allogeneic (Balb/c) CD4+CD25−T cells (5×106 cells) were injected into NK cell-depleted Bl6 RAG1−/− recipients. Prior to T cell transfer, RAG1−/− mice received water (ab libitum) containing aspartame (Asp) or an Abx cocktail containing Asp, neomycin and vancomycin for 7 days prior to and following T-cell transfer.

Results
Treatment of allogeneic mice with Abx reduced colonic bacterial load by more than 20-fold when compared to their Asp-treated counterparts. Abx treatment also resulted in large and significant reductions in BM- and spleen-residing T cells and myeloid cells as well as circulating erythrocytes, platelets and hematocrit when compared to Asp treated mice. These Abx-induced alterations were associated with significant increases (~4-fold) in serum IL-6 levels compared to Asp-treated mice.

Conclusions
Prophylactic Abx treatment exacerbates aGVHD-induced BM failure and spleen hypoplasia.
iNKT cell modulation of tumor microenvironment (TME) and antitumor immunity: the role of antitumorigenic and protumorigenic iNKT cells
Authors: Zhang Hui , Nguyen Jasmine , Pham Michael , Smith Anita 
Published: January 2023

Abstract
Preclinical and clinical data indicate that iNKT cells play important roles in antitumor immunity. High number of iNKT cells in the PBL of cancer patients usually correlates with a favorable prognosis. We found that compared to wild-type (WT) mice, depletion of iNKT cells did not affect tumor progression or metastasis in mice bearing melanoma or Lewis lung cancer; restoring iNKT cells by transferring thymocytes into iNKT KO mice dramatically enhanced antitumor immunity. The underlying mechanism is no known. Herein, we report progress on the mechanistic study. Thymocyte transfer only reconstituted iNKT cells in liver, spleen and blood, but not in LN of iNKT KO mice. This suggests that two functionally opposite iNKT subsets exist in the immune system, and thymocyte transfer only restored antitumorigenic iNKT, but not protumorigenic iNKT cells. Transferring IFNγ KO thymocytes diminished iNKT reconstitution-induced antitumor immunity; transferring IL-17A KO thymocytes enhanced iNKT reconstitution-enhanced antitumor immunity. Compared to WT mice or iNKT KO mice, iNKT reconstitution in iNKT KO mice up regulated the expression of XCL1, FLT3L and CCL5, down regulated IL-6, TGFβ1, TGFβ2, COX2 expression, increased IRF8+ cDC1 cells and Siglec F− DC, and decreased αSMA+ FAP+ cancer-associated fibroblasts (CAF) and MDSC in the tumor. Taken together, these results support that antitumorigenic iNKT cells produce IFNγ dominant cytokine and facilitate NK/cDC1 infiltration and activation in the tumor to induce antitumor immunity; protumorigenic iNKT cells induce CAF to produce TGFβ, which induces a myeloid cell-mediated inhibitory TME. The balance between antitumorigenic and protumorigenic iNKT cells shapes the TME and antitumor immunity.
LEF1 activates a central memory-like program in human NKT cells
Authors: Ngai Ho , Tian Gengwen , Courtney Amy N , Balzeau Julien , Liu Bin , Guo Linjie , Di Pierro Erica J , Metelitsa Leonid S 
Published: January 2023

Abstract
Vα24-invariant natural killer T cells (NKTs) possess potent antitumor activity that can be exploited for use in cancer immunotherapy. We recently demonstrated that the CD62L+ NKT subset persists longer and mediates more potent antitumor activity in vivo than its CD62L− counterpart; as such, CD62L+ NKTs are crucial to maximizing the therapeutic efficacy of NKTs. Key factors involved in transcriptional control of the CD62L+ central memory-like program in NKTs remain largely undefined. To address this gap, we performed comparative gene expression analysis in CD62L+ and CD62L− NKTs and found that LEF1, which encodes a mediator of Wnt/β-catenin signaling, was the top overexpressed gene in the positive subset. We also found that LEF1 protein expression was largely restricted to CD62L+ primary human NKTs and was associated with increased expression of Wnt target genes. While CRISPR-Cas9-mediated knockout of LEF1 expression did not impact CD62L+ cell numbers after initial ex vivo expansion, it did significantly reduce the frequency of these cells after antigenic restimulation. On the other hand, treatment with Wnt/β-catenin signaling activator TWS119 or Wnt3a ligand increased CD62L+ NKT cell frequency. Combined treatment of TWS119 with IL-21, which selectively protects the CD62L+ subset from apoptosis, resulted in significantly higher CD62L+ cell numbers than each single agent alone. These results identify LEF1 as a key transcriptional activator of the central memory-like program in CD62L+ NKTs and pave the way for targeted pharmacological enhancement of NKT cell-based cancer immunotherapy.
Distinct B cell signatures and tertiary lymphoid structures are driven by two etiol-ogies in head and neck cancer
Authors: Ruffin Ayana T , Cillo Anthony R , Tabib Tracey , Liu Angen , Onkar Sayali , Kunning Sheryl , Lampfield Caleb , Abecassis Irina , Qi Zengbiao , Soose Ryan , Duvvuri Umamaheswar , Kim Seungwon , Lafyatis Robert , Ferris Robert L , Vignali Dario , Bruno Tullia C 
Published: January 2023

Abstract
B cells can regulate immune responses by presenting antigen, producing antigen-specific antibodies and immunomodulatory cytokines. Their role in the anti-tumor immune response is poorly understood. However, in many cancers including head and neck squamous cell carcinoma (HNSCC), intratumoral B cells correlate with better survival. HNSCC has two distinct etiologies (HPV−) and (HPV+) where patients who are HPV+ have increased B cell infiltration and respond better to therapy. We hypothesized that (1) intratumoral B cell phenotype is different between HPV+ and HPV− HNSCC (2) location within the tumor microenvironment (TME) is distinct and (3) antibodies produced by intratumoral B cell in HPV+ HNSCC are specific for viral antigens.
Using single-cell RNA sequencing and spectral flow cytometry, we observed that B cell signatures in HPV− HNSCC patients were predominantly memory B cells and plasma cells, while the signatures in HPV+ HNSCC were naïve and germinal center (GC) B cells. Further, we quantified B cells in tertiary lymphoid structures (TLS) using multispectral immunofluorescence, and the presence of GC-rich TLS were increased in HPV+ patients. In fact, GC-rich TLS within the tumor of HPV+ patients correlated with increased overall survival. Overall, high enrichment for GC B cells were positively associated with longer progression-free survival. Antibodies produced by intratumoral B cells from HPV+ patients were positive for HPV viral antigens. Ultimately, characterization of B cell phenotype and function in HNSCC is important for devising new therapeutic options for cancer patients. Specifically, therapeutics to enhance B cell responses in the TME should be prioritized as a compliment to T-cell mediated therapies.
Polyfunctional endogenous EBV-specific T cell response in nasopharyngeal carcinoma
Authors: Chen I-Yu , Lay Fang-Yun , Chia Jean-San , Liao Li-Jen , Chiu Yen-Ling 
Published: January 2023

Abstract
Nasopharyngeal carcinoma (NPC) is associated with Epstein-Barr virus (EBV) infection and can be treated by adoptive transfer of EBV-specific T cells. Although EBV-specific T cell immune response has been identified in NPC, the phenotype and functionality of EBV-specific, tissue-resident memory T cells has not been explored. Tumor-infiltrating lymphocytes (TILs) were isolated from newly-diagnosed NPC and two types of EBV-unrelated malignancies; oral squamous cell carcinoma (OSCC) and renal cell carcinoma (RCC) tissue samples. We found that endogenous EBV-specific T cell response can be detected in the all the tissue types, including NPC, OSCC and RCC, but were rare in peripheral blood. In all tissues, EBV-specific T cells in tumor tissues exhibit resident memory phenotype characterized by high CD69 and CD103 expression and also an exhaustion phenotype characterized by high CD39 and PD-1 expression. Unexpectedly, EBV-specific T cell frequency as well as response magnitude were not enhanced in NPC when compared to other tissue types. The lack of an enhanced EBV-specific T cell response magnitude in NPC tissue confirms the concept that enhancement of EBV-specific immunity by adoptive T cell therapy may improve the outcome of NPC.
A Novel Bifunctional Fusion Protein Comprising of TGF-βRII trap and IL15/IL-15Rα as an Immunotherapeutic against Cancer
Authors: Liu Bai , Chaturvedi Pallavi , Wang Meng , Zhu Xiaoyun , George Varghese , Revelo Kristine , Spanoudis Catherine , Kong Lin , Xing Jilan , Prendes Caitlin , You Lijing , Rhode Peter , Wong Hing C 
Published: January 2023

Abstract
Interleukin-15 (IL-15) is a promising cytokine for cancer therapy. However, it has shown limited antitumor efficacy as monotherapy in clinical trials to date. Removal of immunosuppression of the tumor microenvironment is considered a promising approach to enhance IL-15-mediated immune responses. TGF-β is a key component for creating an immunosuppressive tumor microenvironment. Therefore, we constructed a novel bifunctional fusion protein complex, designated HCW9218, comprising a soluble fusion of two TGFβRII, human tissue factor, and human IL-15, and a second soluble fusion of two TGFβRII and a sushi domain of IL-15Rα. HCW9218 activates IL-15R signaling and the dimeric TGFβRII functions as “trap” for all the three TGF-β isoforms. In healthy and tumor-bearing C57/BL6 mice, subcutaneously administrated HCW9218 was well tolerated with a long serum half-life and was able to stimulate and induce proliferation of CD8+ T cells and NK cells. Splenocytes from HCW9218-treated mice showed enhanced metabolic activity and cytotoxicity against Yac-1 target cells compared to untreated mouse splenocytes. In the syngeneic B16F10 melanoma mouse model, a single-dose of HCW9218 exhibited strong antitumor activity in combination with chemotherapy and TA99 antibody. HCW9218 also significantly increased immune cell infiltration into tumors. Mutagenesis studies demonstrated that both the TGFβRII and IL-15/IL-15Rα domains are essential for antitumor efficacy of HCW9218. Collectively, our preclinical data demonstrate that HCW9218 elicits potent immune responses with a remarkable safety profile and serves as a novel immunotherapeutic against cancer alone or in combination with therapeutic antibodies.
Antibiotic-mediated gut dysbiosis alters innate-like T cell frequency and phenotype in tumor-bearing mice
Authors: Wong Stephanie Wing-Yan , Ohashi Pamela S. , Sacher Adrian G. 
Published: January 2023

Abstract
Innate-like T cells bind to non-peptide antigens in a TCR-dependent manner and are not MHC-restricted. This subpopulation of T cells consists of mucosal associated invariant T cells (MAIT), natural killer T cells (NKT), and gamma delta (γδ) T cells. Innate-like T cells can recognize microbially-derived antigens and interact with gut commensals to help maintain intestinal homeostasis. Additionally, they are implicated in tumour immunity as producers of cytokines and cytotoxic molecules. Thus, innate-like T cells may serve as an intermediary between the gut microbiota and tumor development. Using syngeneic mouse tumour models and flow cytometric analysis, we aim to evaluate the relationships between gut microbiota modulation, tumour growth, checkpoint inhibitor response, and innate-like T cell phenotype. Gut dysbiosis was induced using broad spectrum oral antibiotics two weeks prior to subcutaneous injection of MC38 colon adenocarcinoma cells. Mice with palpable tumours were treated twice with anti-PD-L1 during the experiment. Compared to vehicle control, tumor size reduction in response to anti-PD-L1 was not significantly altered with antibiotic treatment. The following observations were noted in antibiotic-treated mice. The frequency of γδ T cells was decreased in the tumour, blood, and spleen. MAIT, NKT, and γδ T cells in the spleen had lower expression of tissue localization-associated markers, CD103 and CXCR6. MAIT cell activation state, characterized by CD25 and CD69 expression, was also elevated in the spleen. These findings suggest that changes to the immune landscape of tumor-bearing mice due to antibiotic-driven dysbiosis is characterized by alterations in innate-like T cell abundance and phenotype.
MHC genotype, immune repertoire, and tumor immunity all influence spontaneous cancer development and metastasis in spontaneous breast cancer models
Authors: Castro Sarah A , Pena Alvaro , Khazaie Khashayarsha , Pease Larry R 
Published: January 2023

Abstract
20 to 30% of breast cancers overexpress ERRB2, encoding the human epidermal growth factor-2 receptor (HER2). Overexpression of HER2 is associated with poorer prognosis, particularly in metastatic disease. Although HER2 therapeutics exists many patients eventually develop uncontrolled tumor growth. The BALB/c-neuT mouse has been a popular model for investigating HER2-positive breast cancer. We have found that immune responsiveness and MHC genotype are important factors in spontaneous tumor development and tumor rejection in the neuT model. To assess the influence of immune response potential on tumor biology we introduced the skg ZAP-70 mutation or the H-2b MHC haplotype into the neuT model. Spontaneous tumors appeared with decreased frequency and metastasis or increased frequency and metastasis, respectively, in comparison to the BALB/c model. H-2b tumors were readily transplantable into the breast fat pads of syngeneic BALB.B-neuT hosts which developed systemic immune abnormalities very similar to those observed in BALB/c - neuT or BALB/c WT mice bearing neuT tumors. However, BALB.B WT hosts initially accepted the grafts, but subsequently rejected them after they were established. Adoptive transfer of splenocytes from BALB.B WT mice which had rejected tumor conferred resistance to the tumor grafts, as well as to spontaneous tumors developing in BALB.B neuT littermates. The strong H-2b-mediated antitumor response in WT mice indicates efficient antigen presentation, and presumably leads to strong tolerance in neuT+ animals. The importance of an intact T cell signaling and MHC genotype indicate a critical role for T cell-mediated immunity and tolerance in the biology of the spontaneously developing tumor and metastasis.
Reactive oxygen species and L-type calcium channels mediates hypoxia-dependent CCL2 secretion in mast cells
Authors: Castillo-Arellano Jorge Ivan , Ramírez-Moreno Itzel Guadalupe , Ibarra-Sánchez Alfredo , Jiménez-Estrada Manuel , Blank Ulrich , Gonzalez-Espinosa Claudia 
Published: January 2023

Abstract
Mast cells (MCs) are well-known cells for their important role in initiating allergic response through of cross-linking of FcɛRI receptor with IgE-Ag complexes. As participants on inflammatory reactions, they are exposed to environmental conditions such as hypoxia, which on MC-dependent cytokine secretion has not been described.
Here, we show that bone marrow derived MCs (BMMCs) exposed to hypoxia, produce important amounts of CCL2 mRNA and protein, that is maximal secreted between 12 and 24 hours. CCL2 production induced by hypoxia was sensitive to the antioxidant trolox and L-type Ca2+ channel blocker nifedipine.
In order to analyze the possible mechanism involved on hypoxia-induced CCL2 production, we found that this conditions caused an increase on reactive oxygen species (ROS) that was maximal after one hour. We also observed an increase on [Ca2+]i two hours after hypoxia, which was sensitive to trolox. With the aim to confirm this mechanism, BMMCs were treated with H2O2 and as expected, H2O2 caused an increase on ROS and [Ca2+]i in BMMCs. Ca2+ increase produced by H2O2 was sensitive to nifedipine. CCL2 mRNA accumulation and CCL2 protein secretion induced by H2O2 was sensitive to trolox and nifedipine too.
When trolox and nifedipine were used in the B16-F1 murine model of melanoma, a close relationship between ROS, Ca2+ rise and CCL2 production was found, since those compounds were able to prevent pimonidazole-positive areas and CCL2 accumulation inside tumors. Our data suggest that conditions prevalent in tumor microenvironment contribute to the generation of ROS and L-type Ca2+ channel-dependent Ca2+ entry in MCs, causing the secretion of the potent immunosuppressor chemokine CCL2.
Metastatic ovarian cancer shapes the abdominal immune microenvironment by manipulating IL20-IL20RA signaling pathway
Authors: Xiang Rong , Li Jia , Wang Longong , Shi Yi 
Published: January 2023

Abstract
The disseminated of Ovarian cancer (OC) cells shape the tumor abdominal immune environment (TAIE) for survival. However, the key genes and mechanisms mediating the reprogramming of TAIE by OC remain largely unexplored. Here, through a genome-wide in vivo knockout screening by CRISPR/Cas9, we identified IL-20-IL20RA as a key checkpoint preventing the abdominal metastasis of OC cells. In OC patients, the level of IL20RA dramatically decreases in metastatic OC cells when compared with paired OC specimen in the primary sites. Ectopic expression of IL20RA in highly metastatic OC cells greatly suppressed the abdominal metastasis and reduced the formation of ascites. In contrast, silencing IL20RA could increase the metastasis of OC cells. We further examined IL02RA mediated crosstalk between the disseminated cancer cells and the TAIE in both in vivo and in vitro. Silencing IL20RA in OC cells dramatically decreased the M1 macrophages and increased the M2 subtype, while the populations of T and B lymphocytes in ascites were not affected. Further mechanistic studies revealed that the OC cells greatly induced the mesothelial cells in peritoneum to produce IL20RA ligands, i.e., IL-20 and IL-24, which in turn activated the IL20RA downstream signaling in OC cells to trigger the production and maturation of IL-18. IL-18, when secreted into the microenvironment, was able to promote the polarization of macrophages into M1-subtype to kill the cancer cells. Thus, we identified a novel immune checkpoint IL-20/IL20RA that had to be overcome during the metastasis of OC cells to avert an anti-tumor TAIE, which has the potential to be developed as a novel immunotherapy method for OC.
IL-15 deficient colon carcinomas have decreased cytolytic lymphocytes and skewed myeloid cell populations
Authors: Schluns Kimberly S , Chou Chih-Chien , Carrero Rosa M Santana , Davis Richard Eric , Hegde Shweta M 
Published: January 2023

Abstract
Loss of IL-15 expression by colon tumors strongly correlates with classification as microsatellite stable, decreased tumor infiltrating lymphocytes (TILs), increased metastasis, and poor responses to immunotherapy. To better understand how IL-15 expression regulates immune cells in the tumor microenvironment (TME) and identify the importance of IL-15 expressed by tumor cells, we developed a mouse model system to examine this by generating MC-38 tumor cells deficient in IL-15 using CRISPR/Cas9 system. After one round of IL-15 deletion, MC-38 cells that lack one IL-15 allele (i.e IL-15+/− MC-38 cells) were identified and expressed decreased levels of soluble IL-15 complexes. Surprisingly, deletion of one IL-15 allele was sufficient to affect tumor growth as IL-15+/− MC-38 tumors grew faster than Wt MC-38 tumors. IL-15+/− MC-38 tumors had decreased CD8 and NK TILs and increased CD11b+Ly6G+ and CD11b+Ly6ChiLy6G− myeloid cells than Wt tumors. Gene expression analysis of myeloid cells showed that blocking IL-15 in tumors increased expression of CD206, ARG1, and TDO2 and decreased IL-12β and IL-6. A second round of deletion led to complete deletion of IL-15 in MC-38 cells (i.e. IL-15−/− MC-38) and upon implantation developed tumors with further decreases in CD8 and NK TILs. In mice lacking IL-15Rα in the intestinal epithelium, AOM/DSS-induced colon tumors also harbored decreased numbers of CD8 TILs. These results demonstrate that even partially decreasing levels of IL-15 in the TME negatively impacts the numbers of CD8 and NK TILs and skews myeloid cells towards a pro-tumor landscape. Overall, these models that can be used to identify the mechanisms contributing to the poor therapeutic responses observed in human colon carcinomas.