bluuebunny/crossref_metadata_2025_split huggingface dataset:
Sostdc1 regulates natural killer cell maturation and cytotoxicity
Authors: Millan Alberto Jovani , Elizaldi Sonny , Lee Eric , Murugesh Deepa , Loots Gabriela G. , Manilay Jennifer O. 
Published: January 2023

Abstract
Natural killer (NK) cells are specialized lymphocytes with innate ability to eliminate virally infected and cancerous cells, but the mechanisms that control NK cell development and cytotoxicity are incompletely understood. We identified novel roles for sclerostin domain containing-1 (Sostdc1) in NK cell development and function. Sostdc1-knockout (KO) mice display a progressive accumulation of transitional NK cells (CD27+CD11b+, tNK) with age, indicating a partial developmental block. The Ly49 repertoire on NK cells in KO mice is also changed. Lower frequencies (%) of KO splenic tNKs express inhibitory Ly49G2, but higher % of activating Ly49H+ and D+ cells. However, the % of Ly49I+, G2+, H+ and D+ populations were universally decreased at the most mature (CD27− CD11b+, mNK) stage. We hypothesized that the Ly49 repertoire in Sostdc1-KO mice would correlate with NK killing ability. We observed that KO NK cells are hyporesponsive against MHC-I-deficient cell targets in vitro and in vivo, despite higher CD107a surface levels and similar IFN-gamma expression to controls. Further investigation is required to determine Sostdc1’s role in NK cell cytotoxicity and self/non-self recognition. Sostdc1 is expressed in the bone periosteum, and we discovered Sostdc1 expression in NK cells. Consistent with this, reciprocal BM transplant experiments showed that Sostdc1 regulates NK cells through cell-extrinsic and cell-intrinsic manners. Transcriptomic profiling should identify if Sostdc1’s known roles in Wnt and BMP signaling are active, and reveal novel effects of Sostdc1 on NK cells. Taken together, these data support a role for Sostdc1 in the regulation of NK cell maturation, and NK cell cytotoxicity, and identify potential NK cell niches.
Antigen binding to B cells activates a metabolic program that in the absence of a second signal leads to mitochondrial dysfunction and cell death
Authors: Akkaya Munir , Roesler Alexander S. , Traba Javier , Miozzo Pietro , Akkaya Billur , Theall Brandon P. , Sohn Haewon , Pena Mirna , Sack Michael N. , Pierce Susan K. 
Published: January 2023

Abstract
Activation of B cells to proliferate and differentiate into antibody secreting cells is regulated to avoid unwanted antibody responses by two temporally distinct signals, the first provided by antigen binding to the B cell receptor (BCR) and the second by T helper cells. Despite the central role of these two signals in driving antibody responses, our understanding of the consequences of each signal is incomplete. Here using in vitro and in vivo models, we investigated the metabolic changes that accompany B cell activation and show that antigen binding to the BCR signals for rapid increases in both oxidative phosphorylation and glycolysis, however early events following antigen engagement including spreading and contraction of antigen bound BCR and internalization of the bound antigen depend solely on the energy produced by oxidative phosphorylation. Changes in gene expression, mitochondrial performance and glucose uptake follow the initial activation, preparing the cells to respond to a second signal. However, in the absence of a second signal B cells undergo progressive loss of mitochondria function and glycolytic capacity ultimately leading to apoptosis. Mitochondria dysfunction is a result of the gradual accumulation of intracellular calcium which leads to inefficient oxidative phosphorylation and increased ROS production and eventually swollen patches of mitochondria. This process can be avoided for approximately nine hours after B cell antigen binding by either T helper cells or by signaling through Toll-like receptor 9. Thus, antigen binding to B cells appears to activate a metabolic program that imposes a short time window in which B cells either receive a second signal and survive or alternatively face elimination.
Lack of TGFβR signaling confers more mature, activated, and educated phenotypes to NK cells leading to resistance to MCMV infection
Authors: Jang Youngsoon , Laouar Yasmina 
Published: January 2023

Abstract
Natural Killer (NK) cells are crucial in early resistance to murine cytomegalovirus (MCMV) infection. In B6 mice, the activating Ly49H receptor recognizes the viral m157 glycoprotein on infected cells. Balb/c mice lacking LY49H are more susceptible to MCMV infection than B6 mice. Strikingly, CD11cdnR mice that lack TGFβR signaling in NK cells are protected from MCMV infection in both strains. It is established that unlicensed NK cells that are not inhibited by self-MHC class I can control viral infection more than licensed NK cells. We found that CD11cdnR mice have increased frequency of unlicensed (LY49G2+) NK cells and slightly decreased frequency of licensed (LY49C/I+) NK cells in B6 strain. Moreover, TGFβ-resistant NK cells showed more activated phenotypes such as increased granzyme B protein expression and more KLRG1+ expression than control NK cells in both licensed and unlicensed cells. Interestingly, in CD11cdnR mice even unlicensed NK cells have a significant capacity to produce IFN-γ when triggered by anti-NK1.1, but not in control mice. Our data suggest that inactivation of TGFβR signaling might have a critical role in the licensing of NK cell and lead to the resistance against MCMV infection.
CRTAM shapes the microbiota and enhances the severity of gut infection.
Authors: Perez-Lopez Araceli , Nuccio Sean-Paul , Ushach Irina , Zlotnik Albert , Raffatellu Manuela 
Published: January 2023

Abstract
Gut lymphocytes and the microbiota establish a reciprocal relationship that tunes the host immune response. For example, the gut microbiota is required for T helper (Th)-17 cell differentiation; Th17 cytokines, in turn, stimulate the intestinal epithelium to produce antimicrobial proteins that control the gut microbiota. Residency and maintenance of T cells in the gut is controlled by Class I-restricted T cell-associated molecule (CRTAM). We thus evaluated whether CRTAM influences the gut microbiota composition. We found that CRTAM expression caused a shift in several members of the gut microbiota under homeostatic conditions. As the gut microbiota and T cells contribute to host defense against intestinal pathogens, we tested whether CRTAM plays a role during infection with the enteric pathogen Salmonella. We found lower levels of Salmonella colonization in the gut of Crtam−/− mice, along with lower expression of the Th17 cytokines Il17 and Il22 in the cecum. Salmonella-infected Crtam−/− mice exhibited fewer gut T cells and IL-17-producing cells. To investigate whether the altered gut microbiota composition of Crtam−/− mice influenced the host response to infection, we transplanted the fecal microbiota from WT or Crtam−/− littermate mice into germ-free mice, and then infected the mice with Salmonella. Relative to mice transplanted with “WT” fecal microbiota, mice transplanted with “Crtam−/−” microbiota exhibited lower numbers of T cells and IL-17-producing cells in the gut upon infection. Our results thus suggest an interplay between CRTAM expression and the gut microbiota, which ultimately impacts the mucosal immune system and the host response to enteric pathogens.
Absence of vacuolar ATPase in hematopoietic stem cells promotes breast tumor progression and pathogenesis
Authors: Sahoo Manoranjan , Katara Gajendra K. , Bilal Mahmood Y. , Kulshrestha Arpita , Ibrahim Safaa A. , Fleetwood Sara , Beaman Kenneth D. 
Published: January 2023

Abstract
Vacuolar ATPase (V-ATPase) is expressed on surface of most cancer cells where it acidifies the extracellular milieu and promotes metastasis. We have previously demonstrated that the a2 isoform of V-ATPase (a2V) of mammary epithelial cells, plays a key role during breast tumor progression and metastasis. However, the role of a2V in hematopoietic stem cells (HSC) during breast tumor progression is not known. Therefore, to understand the role of a2V in HSC during breast tumor progression, we conditionally knocked out a2V (cKO) from the HSC of mice. We observed that deletion of a2V leads to a significant reduction of CD4+ and CD8+ T cells in the spleen of the a2V cKO mice compared to control mice. We hypothesized that the reduction of helper CD4+ cells, or cytotoxic CD8+ T cells would result in larger and faster growing tumors in the a2V-cKO mice. After implantation of a syngeneic tumor cell line (E0771) in the breast tissue of mice, we observed that lack of a2V leads to larger and faster growing breast tumors compared to controls. Further investigation of the tumor microenvironment revealed that there was a significant reduction in percentage of CD4+ and CD8+ T cells in the a2V-cKO mice compared to control mice. Targeted RNAseq of the tumor microenvironment demonstrated that out of 499 genes those were analyzed, only 3 genes were significantly upregulated while 144 genes were significantly downregulated in the a2V cKO mice. The 144 downregulated genes included genes for pro-inflammatory cytokines, T cell specific genes, death receptors and pro-apoptotic genes. Collectively, these results demonstrate that deletion of a2V from the HSC leads to a decrease in CD4+ and CD8+ T cells in the periphery, which results in a faster growing and larger breast tumor.
Arginase-1 is neither constitutively expressed in nor require for myeloid-derived suppressor cell (MDSC)-mediated inhibition of T cell proliferation
Authors: Kidder Koby S. , Bian Zhen , Shi Lei , Liu Yuan 
Published: January 2023

Abstract
Although previous reports suggest that tumor-induced myeloid-derived suppressor cells (MDSC) inhibit T cell proliferation by L-arginine depletion through arginase-1 activity, we herein show that arginase-1 is neither inherently expressed in MDSC nor required for MDSC-mediated inhibition of T cell proliferation. Employing Percoll density gradient centrifugation, large expansions of MDSC in the bone marrow of tumor-bearing mice were isolated and exhibit potent inhibition on T cell proliferation induced via TCR-ligation, Concanavalin A, PMA plus ionomycin, or IL-2. These MDSC, though suppressive toward T cell proliferation, do not hinder TCR-induced IFNγ secretion or granzyme B expression. Despite effectively inhibiting T cell proliferation, both G- and M-MDSC exhibit no arginase-1 expression; however, arginase-1 can be induced by exposure to TCR-activated T cells or their culture medium, but not by T cells activated by other means or growing tumor cells. Western blot analysis revealed that TCR-activated T cells confer arginase-1 expression in MDSC by secreting cytokines that manifest as two distinct signaling-relay axes, IL-6-to-IL-4 or GM-CSF/IL-4-to-IL-10. Specifically, IL-6 signaling increases IL-4R on MDSC, enabling IL-4 to induce arginase-1 expression; similarly, GM-CSF in concert with IL-4 induces IL-10R on MDSC, allowing IL-10-mediated induction. Surprisingly, our data demonstrate that the induction of arginase-1 is not conducive, per se, for MDSC-mediated inhibition of T cell proliferation, as neither supplementation of L-arginine nor arginase-1 inhibitor rescue T cell proliferation. Instead, direct cell contact is required for the inhibition and is not dampened by PD-L1 blockade or SIRPα deficiency.
Tumor-resident macrophages modulate intestinal barrier function through sialylation of Mucin 1 in IBD and colitis-associated cancer.
Authors: Cascio Sandra , Kvorjak Michael , Hashash Jana Al , Sriram Raahul , Hartman Douglas , Binion David , Finn Olivera J. 
Published: January 2023

Abstract
Mucin 1 (MUC1) is a transmembrane glycoprotein overexpressed and hypoglycosylated in chronic inflammatory conditions and human adenocarcinoma compared to healthy tissues. We have reported that AOM/DSS-treated human MUC1.Tg mice showed higher tumor incidence and greater inflammation when compared to wild-type (WT) mice. Moreover, high expression levels of pro-inflammatory cytokines, including TNF-α and IL-6, were found in MUC1+ inflamed colon tissues and exogenous TNF-α promoted the transcriptional activity of MUC1 as well as over-expression of its hypoglycosylated form. As macrophages are the major source of TNF-α and IL-6, we analyzed infiltrating type 1 (M1) and type 2 (M2) macrophages into the inflamed and tumorigenic colon tissues. Immunofluorescence assay indicated increased presence of macrophages, and in particular CD206+ (M2), in inflamed colon tissues of MUC1.Tg mice compared to WT mice. In addition, immunostaining of macrophages in colon tissues from IBD patients showed expression of hypoglycosylated MUC1 and the presence of macrophages expressing CD163+, a human M2 marker. Interestingly, in vitro co-culture model system indicated that M1 and M2 polarized macrophages, induced different glycoforms of MUC1 in HT-29 colon cancer cells. In addition, macrophage-secreted factors modulated glycosyltrasferase enzymes, sush as ST6GalNAc1 and ST3GAL1, involved in O-glycan sialylation. We are currently dissecting the molecular basis by which macrophages induced hypoglycosylated and tumor forms of MUC1 during inflammation and colitis-associated cancer (CAC). Our findings will elucidate novel functions of tumor-associated macrophages in altering the intestinal barrier function in IBD and CAC.
A novel mechanism of innate immune dysfunction in untreated Chronic Lymphocytic Leukemia.
Authors: Manso Bryce A. , Zhang Henan , Mikkelson Molly G. , Gwin Kimberly A. , Secreto Charla R. , Ding Wei , Parikh Sameer A. , Kay Neil E. , Medina Kay L. 
Published: January 2023

Abstract
The consequences of immune dysfunction in B-Chronic Lymphocytic Leukemia (CLL) likely relate to the incidence of serious recurrent infections and second malignancies that plague CLL patients. The known immune abnormalities are not able to consistently explain these complications and it is not easy to correct their immune status. Here, we report bone marrow (BM) hematopoietic dysfunction in early and late stage untreated CLL patients. We found reduced functional capacity of hematopoietic progenitors in BM using colony forming unit assays and flow cytometry revealed significant reductions in frequencies of hematopoietic stem and progenitor cell (HSPC) populations. Consistent with the reduction in HSPCs, monocytes and natural killer cells in BM were reduced and the reduction in BM was recapitulated in blood. Finally, we report imbalances in protein levels of the transcriptional regulators HIF-1α, PU.1, and GATA-2 in CLL patient BM, providing molecular insight into the basis of HSPC dysfunction. Importantly, PU.1 and GATA-2 were rapidly increased when healthy HSPCs were exposed in vitro to TNFα, a cytokine constitutively produced by CLL B cells. Together, these findings reveal BM hematopoietic dysfunction in untreated CLL patients, adding to the complex immunodeficiency state of these patients and the reduced capacity of the innate immune system.
IL-36γ promotes a local immune response via the formation of tertiary lymphoid structures in colorectal carcinoma.
Authors: Weinstein Aliyah M. , Chen Lu , Giraldo Nicolas , Petitprez Florent , Lacroix Laetitia , Brzana Emily A. , Fridman Wolf H. , Sautes-Fridman Catherine , Storkus Walter J. 
Published: January 2023

Abstract
Tertiary lymphoid structures (TLS) form at sites of chronic inflammation, and locally prime the immune system independently of secondary lymphoid organs. The intratumoral presence of TLS has been associated with a positive prognostic outcome for patients with solid tumors, including colorectal carcinoma. Thus, the therapeutic induction of TLS would be hypothesized to confer clinical benefit to patients.
Indeed, the therapeutic introduction of the IL-1 family cytokine IL-36 gamma into the tumor microenvironment (TME) of murine MC38 colon carcinoma leads to delayed tumor progression and the development of TLS at 5 days post-treatment. The TLS present with characteristics of secondary lymphoid organs, including PNAd+ high endothelial venules (HEV) surrounded by CD3+ T cells and CD11c+ dendritic cells, but are considered as “non-classical” due to the absence of a defined B cell zone. The formation of TLS is dependent upon the adaptive immune response, as IL-36 gamma-treated tumors grown in Rag2−/− animals fail to present with PNAd+ vasculature within the tumor. Thus, crosstalk between the immune system and cells of the vasculature is key to the efficacy of the IL-36 gamma-induced anti-tumor immune response.
In human colorectal carcinoma, IL-36 gamma was also observed to promote the local anti-tumor immune response. Expression of IL-36 gamma by vascular endothelial cells of HEV was associated with an increased density of CD20+ B cells in TLS. Specific IL-36 gamma production by CD68+ macrophages was associated with an increased infiltration of CD4+ central memory T cells into the TME. Therefore, IL-36 gamma is a pro-inflammatory mediator involved in the anti-tumor immune response in both murine and human colorectal cancer.
Phytohemagglutinin activates CD8 T cells in a model of the tumor microenvironment.
Authors: Riggs James Edward , Lomakova Yelizavet , Goldman Naomi , Somerville John 
Published: January 2023

Abstract
Tumors often include a high proportion of immune modulatory cells and molecules that restrain the anti-cancer response. Activation of T cells to eliminate cancer cells within the immune suppressive tumor microenvironment (TME) remains a challenge. We have shown that C57BL/6J peritoneal cavity (PerC) cell culture models features of macrophage (Mθ)-rich TMEs as TCR ligation fails to activate T cells unless IFNγ is neutralized or iNOS is inhibited. We tested other forms of T cell activation and found phytohemagglutinin (PHA) distinctive in the ability to markedly expand CD8 T cells in the TME model. Neither IFNγ neutralization or iNOS inhibition were required for this response. IFNγR−/− PerC cells revealed that the PHA response is IFNg-dependent, yet this mitogen triggered less IFNγ production than TCR ligation. PHA-FITC staining revealed strong binding to Mfs and PerC cell addition to normally quiescent spleen (SP) cell cultures led to a marked increase in T cell expansion. The stimulatory capacity of PHA might reflect an ability to bind Mθs with CD8 T cells. Intriguingly, PerC Mf expression of the T cell inhibitory PD-L1 molecule was lower after PHA stimulation than that seen following TCR ligation. If PHA can increase CD8 T cell activation within the TME without increasing PD-L1 expression it might serve as an antitumor immune stimulant.
Brightfield and Fluorescence Imaging using 3D PrimeSurface® Ultra-Low Attachment Microplates
Authors: Dang Anju , Larson Brad 
Published: January 2023

Abstract
Culturing cells in three-dimensions (3D) has become a well established approach as it is more representative of the in vivo environment than traditional two-dimensional (2D) cultures. Allowing cells to interact with each other in a spheroid creates a micro-environment which mimics in vivo tissue and is a better model for examining the effect of drugs in cancer. Developing uniform spheroids becomes especially important as it forms the basis for robust and reliable assays. S-BIO PrimeSurface® cultureware are ultra low attachment (ULA) dishes and plates that promote scaffold free, self assembly spheroid formation. The plates are pre-coated with a proprietary hydrophilic polymer that enables spontaneous spheroid formation of uniform size. PrimeSurface 96 and 384 ULA plates have good optical clarity making them highly suitable for brightfield and fluorescent imaging. Imaging technologies such as the BioTek Cytation™ 5 allows researchers to study not only spheroid proliferation through brightfield imaging, but also phenotypic events such as hypoxia, apoptosis, or necrosis induction through the use of fluorescent probes and fluorescence imaging. Incorporation of z-stacking and projection techniques in the Gen5™ Microplate Reader and Imager Software create in-focus images of spheroidal cells, allowing accurate, robust, and repeatable determination of the effect of test molecules or conditions. In this app note, we present data generated with BioTek Cytation 5 using PrimeSurface ULA plates to develop simple robust spheroid assays for brightfield and fluorescence imaging.
Tumor transcriptome analysis and immune infiltrate profiling suggests a role for the IFNL3/IFNL4 genomic locus in liver cancer
Authors: Onabajo Olusegun O. , Wang Fang , Banday Rouf , Prokunina-Olsson Ludmila 
Published: January 2023

Abstract
HCV infection and cirrhosis are among the strongest risk factors for developing liver cancer. Impaired HCV clearance is strongly associated with a dG allele of a genetic variant, rs368234815-TT/dG; this allele creates an open reading frame for interferon lambda 4 (IFNL4). The dG allele has also been linked to reduced risk of cirrhosis. Despite strong association of HCV and cirrhosis with liver cancer, no link between rs368234815-TT/dG and liver cancer has been reported. To evaluate potential association of rs368234815-TT/dG with liver cancer, we explored resources of The Cancer Genome Atlas (TCGA). We used a genetic variant rs12980275-A/G located downstream of IFNL3 gene as a TCGA-genotyped proxy for rs368234815-TT/dG. Using multivariate statistical models, we observed that liver cancer patients homozygous for rs12980275-G allele (corresponds to rs368234815-dG allele) had less cirrhosis but reduced survival. Tumor transcriptome analysis showed association of rs12980275-G allele with immunosuppressed tumor microenvironment in non-cirrhotic patients. By deconvoluting tumor infiltrating immune cell signatures, we observed lower levels of CD8+ and follicular helper T cells, but increased M1 macrophages in non-cirrhotic patients with rs12980275-G allele. Transcriptome analysis also identified metallothioneins (MTs), which are involved in removal of reactive oxygen species in the liver, to be differentially expressed according to rs12980275 genotypes. Using hepatoma cell lines, we show that MTs are induced by IFNL4. Our results suggest an association of IFNL3/IFNL4 locus with both an immunosuppressed tumor microenvironment in liver cancer, and increased expression of MTs which could contribute to reduced fibrosis.
Identification of novel α-fetoprotein-specific T cell receptors to redirect human T cells for hepatocellular carcinoma immunotherapy
Authors: He Yukai , Zhu Wei He/Wei , Peng Yibing , Wang Lan , Hong Yuan , Jiang Xiaotao , Li Qi , Liu Heping , Huang Lei , Wu Juan , Celis Esteban , Merchen Todd , Kruse Edward 
Published: January 2023

Abstract
Hepatocellular carcinoma (HCC) is the major form of liver cancer for which there is no effective therapy. Genetic modification with T cell receptors (TCR) specific for HCC-associated antigens, such as α-fetoprotein (AFP), can potentially redirect human T cells to specifically recognize and kill HCC tumor cells to achieve antitumor effects. In this study, by using lentivector and peptide immunization, we identified a population of CD8-T cells in HLA-A2 transgenic AAD mice that recognized AFP158 epitope on human HCC cells. Adoptive transfer of the AFP158-specific CD8-T cells from immunized AAD mice eradicated HepG2 tumor xenografts as large as 2cm in diameter in immunocompromised NSG mice. We then established T cell hybridoma clones from the AFP158-specific mouse CD8-T cells and identified three sets of paired TCR genes out of 5 hybridomas. Expression of the murine TCR genes redirected primary human T cells to bind HLA-A2/AFP158 tetramer. The TCR gene-engineered human T cells (TCR-T) specifically recognized HLA-A2+AFP+ HepG2 tumor cells and produced effector cytokines. Importantly, the TCR-T cells could specifically kill HLA-A2+AFP+ HepG2 tumor cells without significant toxicity to normal hepatocytes in vitro. Adoptive transfer of the AFP-specific human TCR-T cells could eradicate HepG2 tumors in NSG mice.

Conclusion
We have identified novel AFP-specific murine TCR genes that can redirect human T cells to specifically recognize and kill HCC tumor cells, and those AFP158-specific TCRs have a great potential to engineer a patient’s autologous T cells to treat HCC tumors.