bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functionality of lung type II NKT cells developing in CD1d−/− animals
Authors: Pasquet Lise , Kato Shingo , Adams Tony , Sharrow Susan O , Davies-Hill Theresa , Jaffe Elaine S , Xia Zheng , Suzuki Motoshi , Kovalovski Damian , Berzofsky Jay A , Terabe Masaki 
Published: January 2023

Abstract
NKT cells are lymphocytes that recognize lipid antigens presented by CD1d. Type I NKT cells with semi-invariant TCR recognize α-galactosylceramide while many type II NKT cells with a more diverse TCR repertoire recognize sulfatide. We developed a stable sulfatide-loaded CD1d tetramer that allows the specific identification of type II NKT cells and found that they are enriched in the lung, a major site of tumor metastasis. An in-depth phenotypical analysis revealed that lung type II NKT cells were CD4 or CD8 single positive cells, like conventional T cells, whereas type I NKT cells were either CD4+CD8- or CD4-CD8-. We showed that type II NKT cells expressed markers of myeloid cells, c-Kit, CD11b and Ly6C, even though morphological analysis revealed lymphocytic morphology. In contrast to type I NKT cells, type II NKT existed in PLZF−/− and CD1d−/− mice. Thus, we asked the importance of CD1d in the in vivo and in vitro activation of type II NKT cells. In the steady state, lung type II NKT cells expressed granzyme A but not granzyme B or perforin. By injecting sulfatide in vivo, we observed that WT but not CD1d−/− lung type II NKT cells upregulated granzyme A and Ki67 expression. Using syngeneic WT bone marrow-derived dendritic cells (BMDC) as APCs, ex-vivo stimulation of WT but not CD1d−/− lung type II NKT cells with sulfatide induced the secretion of IFN-γ, IL-13 and IL-17. This was consistent with the expression of specific transcription factors, T-bet and RORγt, by lung type II NKT cells. In conclusion type II NKT cells still develop in CD1d−/− mice but are not functional. The ability to study responses of type II NKT cells in vitro can now help elucidate the mechanism by which these regulatory cells interact with other cells and modulate tumor immunity in the lung.
Immune microenvironment modulation via the interaction of human invariant natural killer T cell (iNKT cells) and Kupffer cells (KC), dendritic cells (DC) in colorectal liver metastasis
Authors: Feng Hao , Thasler Wolfgang 
Published: January 2023

Abstract

Aims
To investigate the immune microenvironment modulation via the interaction between human iNKT cells and KC, DC in colorectal liver metastasis.

Methods
Different subtypes of iNKT cells, KC and DC from the core of tumor (CT), tumor invasive margin (IM), and adjacent normal liver tissue (NL) were analyzed by BD LSRFortessa™, TMA, and immunofluorescence(n=138). Ex vivo 3D co-culture, Quansys Multiplex array, and Elisa array were performed.

Results
In metastasis, CXCR6+iNKT positively correlated with better prognosis. In addition, iNKT cells were enriched at IM, especially iNKT2. Intriguingly, although there were few iNKT cells in CT, the iNKT/Lymphocytes rate increased to ~5 folds and expressed elevated IL9, IFNγ, and FasL compared to NL and IM. This implied the critical role of iNKT cells in tumor control. Both DC and KC could drive iNKT activation, the proportion of BDCA-3+DCs significantly increased from NL to CT; though they expressed higher CXCL16 compared to other subtypes, the capability of activating iNKT cells via CD1d was less strong than plasmacytoid DC(pDC). pDCs showed significantly elevated α-GalCer: CD1d complex formation in an ex vivo iNKT cells activation assay, which implied its critical role of activating capability of pDC. However, pDCs, as well as BDCA-1+DCs, decreased from NL to CT. In this study, KC was found also activating iNKT cells. Interestingly, iNKT expressed Th2 cytokines when activated by KC, however, Th1 biased response could be found when activated with DC. In addition, KC subsets also differed with location. Furthermore, transmembrane CD1d density and intra/tmCXCL16 expression, Th1/2/17 biased immune responses of KC and DC were associated with the lipid metabolism preferences of these cells.
Group B Streptococcus infection induces M1-like activation in human placental and mouse decidual macrophages.
Authors: Sutton Jessica A , Rogers Lisa M. , Doster Ryan Steven , Gaddy Jennifer Angeline , Aronoff David M 
Published: January 2023

Abstract
Group B Streptococcus (GBS), a vaginal colonizing gram-positive bacterium, is a leading cause of neonatal death and sepsis. How GBS evades macrophage defenses in the gravid uterus is not yet known. In response to stimuli, macrophages are activated along a spectrum known as polarization ranging from classically-activated, pro-inflammatory “M1” to alternatively-activated anti-inflammatory “M2” macrophages. These activation states can contribute to disease pathogenesis. This study aims to define changes in macrophage activation in response to GBS infection. Human monocyte derived macrophages (MDMs) were infected with GBS or polarized in vitro with M1 (IFNγ/LPS) or M2 (IL-4 or IL-10) stimuli. Placental macrophages (PMs) were infected with GBS or treated with IFNγ/LPS. To mimic human ascending vaginal infection in vivo, C57BL/6 mice were inoculated vaginally with GBS at day E13.5 for 48h. Subsequently, decidual macrophages were isolated and analyzed for polarization markers via flow cytometry and tissues were used for histology. Resulting phenotypes were characterized using qRT-PCR, ELISA, and flow cytometry. In MDMs, GBS upregulated M1 genes (CCR7, CD80) and downregulated M2 genes (CD209, CD163). GBS increased the release of M1 cytokines (TNFα, IL-1β) in MDMs and PMs. PM surface expression of M1 markers (CCR7, CD80) increased while M2 markers (CD206, CD163, CD209) decreased. Mouse decidual macrophage surface expression of CD206 was downregulated while CD11c exhibited little change. Our results suggest that GBS may induce M1 activation in both human and mouse models. These studies provide new insights into the role of macrophage activation in GBS pathogenesis.
Studies supported by NIH NIAID Training Grant 5T32AI007281-27.
Cross-dressing and antigen transfer after DC targeting induce robust T cell response
Authors: Yao Chen , Hirai Toshiro , Igyarto Botond , Zurawski Sandy , Zurawski Gerard , Kaplan Daniel H 
Published: January 2023

Abstract
Cutaneous dendritic cells (DC) are critical in protecting host from environmental insults as well as maintaining skin homeostasis. Using mouse models that are lack certain subset of cutaneous DC, studies have showed the distinct function of different subsets of skin DC, However, the interaction between skin DC subsets is less known. By targeting antigen to Langerhans cells (LC), we showed that endogenous CD4+ T cell response is independent of MHC II expression on LC. We also showed that LC handed over antigen to CD11b+ dDC and DN dDC. At the same time, MHC II deficient LC acquired MHC-II in the lymph node through cross-dressing. We demonstrated unexpected collaboration between cutaneous DC subsets, and provided insight on DC targeted vaccine design.
STAT4 regulates CD8+Treg/Tfh cell axis and promotes atherogenesis in insulin-resistant Ldlr−/− mice
Authors: Galkina Elena , Taghavie-Moghadam Paresa L , Waseem Tayab C , Gleen Lindsey , Kaplan Mark H , Dobrian Anca D , Yang Yi , Nurieva Roza , Nadler Jerry L 
Published: January 2023

Abstract
The metabolic syndrome support atherosclerosis, but the exact mechanisms for accelerated atherogenesis remain unclear. While the pro-inflammatory role of signal transducer and activator of transcription 4 (STAT4) in atherosclerosis and diet-induced insulin resistance (IR) was recently established, an impact of STAT4 on atherogenesis in conditions of IR is not known. To study the role of STAT4 in IR-accelerated atherosclerosis we generated Stat4−/−Ldlr−/− mice that were fed a diabetogenic diet with added cholesterol (DDC). DDC fed Stat4−/− Ldlr−/−mice demonstrated improved glucose tolerance, insulin sensitivity, and a 36% reduction in atherosclerosis compared with Ldlr−/−controls. Interestingly, we detected a reduction in T follicular helper (Tfh) and plasma B cells, but a sharp elevation in CD8+ Tregs in spleens and aortas of Stat4−/−Ldlr−/−versus Ldlr−/−mice. Similarly, STAT4 deficiency supported CD8+ Treg differentiation in vitro. Additionally, Stat4-deficient CD8+Tregs suppressed Tfh and germinal center B cell development upon immunization with KLH indicating an important role for STAT4 in CD8+ Treg functions in vivo. Effects of STAT4 deficiency were not only restricted to T cells, but had a significant impact on macrophage phenotype. Stat4−/−Ldlr−/− macrophages displayed decreased IFNγ and MHC-II expression. Conditioned media from Stat4−/−Ldlr−/−MFs supported CD8+ Treg but not Tfh cell differentiation in TGFβ-dependent manner, suggesting a role for MF-specific STAT4 in Th cell differentiation. These new findings suggest a novel mechanism by which STAT4 supports atherosclerosis in obese, IR Ldlr−/− mice via increases in MF activation, and modulation of the Tfh/ CD8+ Treg axis systemically and within the aorta.
Comparative analysis of stress on faculty and students based on salivary IgA
Authors: Redmond Sarah , Biggs Erick , Blevins Kasey , Brimer Kelsey , Brown Cameron , Bryan Sandra , Burger Haley , George Anna , Hunt Mckenzie , Jannise Caitlin , Kesterson Melissa , Kwan Anthony , Marshall Jacob , Miller Kenzie , Mohamed Attia , Rajashekara Arpitha Mysore , Pagan Rachael , Peters Mary , Saenz Allison , Steeves Gregory , Stowers Samuel , Welch Nate , Williams Alexis , Williams Devonn , Wingfield Kyle , Zimmermann Krista 
Published: January 2023

Abstract
The goal of this project is to test students and faculties stress levels in their saliva compared between the beginning of this semester and the end of the semester. IgA levels are influenced by many physiological factors, including the hormones associated with stress. The findings conclude that there is significance between IgA levels in a student or faculty based on hours since they had left class and hours until they have class. Students and faculty showed a correlation between IgA and time since being in class or work (p-value = 0.03), however no significance of the duration of the expected rest period (p-value = 0.71). The significance of this finding is that students and faculty tend to be more stressed after being in class/work based on IgA levels in their saliva.
The expressions of 164 histone modification enzymes in seven groups are modulated in respond to pathogenic processes of metabolic diseases, tumors and regulatory T cell differentiation
Authors: SHAO YING , Chernaya Valeria , Wang Hong , Yang Xiao-Feng 
Published: January 2023

Abstract
To determine whether the expression of histone modification enzymes is regulated in physiological and pathological conditions, we took an experimental database mining approach pioneered in our labs to determine a panoramic expression profile of 164 enzymes in 19 human and 17 murine tissues. We have made the following significant findings: 1) Histone enzymes are differentially expressed in cardiovascular, immune and other tissues; 2) Our new “pyramid model” showed that heart and T cells are among a few tissues/cell types, in which enzymes mediating histone acetylation/deacetylation, histone methylation/demethylation are in the highest varieties; and 3) Histone enzymes are more downregulated than upregulated in metabolic diseases, and regulatory T cell polarization/ differentiation, but not in tumors. These results have demonstrated that histone modification enzyme expressions are modulated in different pathophysiological conditions as well as in the processes of T cell subset conversion, differentiation and polarization, which makes upregulated histone enzymes being potential novel therapeutic targets in suppressing disease activities and Treg functions.
Cytotoxic CD8+ interleukin (IL)-17+ T-cells play a role in hypertension-specific inflammatory responses associated with ovariectomy in Dahl rats
Authors: Pai Amrita V , West Crystal A , Souza Aline , Ji Hong , Wu Xie , Sandberg Kathryn 
Published: January 2023

Abstract

Background and Objective
Oophorectomy is associated with increased body weight (BW) gain and incidence of hypertension (HT) compared with age-matched women. Increased BW gain and HT coincide with increased inflammatory biomarkers. To separate immune responses due to HT from those due to BW gain, we compared T-cell subpopulations in normotensive Dahl (DR) and hypertensive Dahl (DS) rats after ovariectomy.

Methods
Six week old DS and DR rats were ovariectomized (Ovx) or had sham surgery (Sham). Mean arterial pressure (MAP) was measured in anesthetized rats after 4 and 10 months (mo) acutely by indwelling vascular catheter. Splenic T-cells were assessed by flow cytometry at 10 mo.

Results
Ovariectomy increased the MAP in DS but not DR rats at 4 mo [MAP (mmHg): DS-Ovx, 143±1.0 vs DS-Sham, 129±2.9; p=0.0002 & DR-Ovx, 123±4.4 vs DR-Sham, 126±1.3; ns n=10–12/group]. At 10 mo, the MAP remained higher in the ovariectomized DS compared to DR rats [MAP (mmHg): DS-Ovx, 190±12 vs DR-Ovx, 114±1.2; n=5–8/group; p=0.002]. At 10 mo, ovariectomy increased BW to a similar extent in DS and DR rats [BW gain (g): DS, 179±13 vs DR, 202±14; ns]. The frequency of CD8+IL17a+ T-cells increased in the ovariectomized DS but not DR rats [Frequency (%) at 10 mo: DS-Ovx, 9.6±2 vs DR-Ovx, 1.3±0.4; n=5–8/group; p&lt;0.0001].

Conclusion
Cytotoxic CD8+ IL-17a+ T-cells may play a role in the HT-specific inflammatory response in ovariectomized DS rats.
T Lymphocyte Immunophenotyping: 14-Color Flow Cytometry Panel Design Using the Attune NxT Flow Cytometer and Super Bright Fluorescent Dyes
Authors: Bradford Jolene A , Zhang Elizabeth Yan , Smith Wesley , Punj Manik , Buller Gayle , Hoenes Candice , Sardina Patricia , Melquist Penny , Oram Shane 
Published: January 2023

Abstract
The Invitrogen™ eBioscience™ Super Bright polymer dyes represent a suite of bright fluorophores excited by the violet laser (405 nm). Optimized for use in flow cytometry, the Super Bright dyes allow for expanded use of violet laser excitation and promote streamlined multicolor panel design. Due to their inherent brightness, detection of cell populations with low abundance targets is possible. A 14-color panel used to characterize human T-cell lymphocytes is presented, with information on differentiation profiles, activation or exhaustion status, and co-stimulation activity. Both cryopreserved human peripheral blood mononuclear cells (PBMC) and fresh blood collected from a normal donor was used. Acquisition and analysis was performed using the Attune™ NxT Flow Cytometer four laser system, which has the ability to detect multiple violet-excited fluorophores. The results show well characterized cell populations, with expected changes between unstimulated and stimulated PBMC ex vivo. Correction for spectral overlap was performed using standard instrument auto-compensation procedures. The unique acoustic focusing technology of Attune NxT Flow Cytometer enables rapid and accurate detection with sensitivity at high flow rates without compromise of data quality, as demonstrated with this 14-color data set. The Attune NxT cytometer is designed to accommodate a wide variety of experimental conditions and combined with the Super Bright dyes provides expanded choice for panel design. For research use only. Not for use in diagnostic procedures.
T Cell Activation and the Role of Ubiquitin Ligases
Authors: Lu Xiaolong , Singh Rajesh , Song Changcheng , Mason Mark 
Published: January 2023

Abstract
Protein ubiquitylation has emerged as a key regulatory mechanism in innate and adaptive immune systems. E3 Ligases such as Cbl play fundamental roles in immune regulation. These enzymes are expressed in all leukocytes and are key checkpoints in maintaining the balance between activating and inhibitory signaling to immune cells, including T cells, NK cells, B cells and different types of myeloid cells. Thus, targeting ubiquitin ligases is a novel strategy to treat human diseases such as cancer and autoimmunity. However, the detailed mechanisms and functions of ubiquitin ligases in human cell is still largely unclear. Several ubiquitylation substrates of Cbl have been identifed in immune cells, e.g., Tyro3, Zap70, p85 and LAT. Therapeutic modulation of ligases is expected to alter the ubiquitylation of their substrate proteins, and documentation of such changes is critical to establishing the mechanism of action of therapeutic agents. Current methods to study poly-ubiquitin on substrates are cumbersome and inefficient. Here we describe a powerful technology, UbiQuant S, that allows selective and quantitative analysis of poly-ubiquitylated proteins in a cellular, high throughput format. We are able to track the changes in ubiquitylation status of Cbl targets upon T-cell activation, which provides valuable information on how Cbl and other ligases regulate T cell signaling elements. This technology also provides a powerful cellular platform for validating small molecule cancer immunotherapy agents and other checkpoint inhibitors.
Vaccinia virus preferentially infects primary human memory B cells
Authors: Shepherd Nicole , Lan Jie , Li Wei , Yu Qigui 
Published: January 2023

Abstract
Vaccinia virus (VACV) has shown an extreme preference for binding to and infection of primary human antigen-presenting cells including monocytes, macrophages, and dendritic cells in the peripheral blood. Although VACV infection in monocytes, macrophages, and dendritic cells has been examined, few studies have evaluated VACV infection in B cells. In this study, we characterized VACV binding to and infection of primary human B cells. Flow cytometric analysis revealed that B cells isolated from human peripheral blood mononuclear cells exhibited two distinct populations of B cells, referred to as Bsmall and Blarge. Phenotypic analysis demonstrated that the Bsmall compartment was more quiescent compared to the Blarge population and enriched with CD138-positive plasma cells. VACV preferentially entered into and infected memory B cells. Especially, VACV preferentially infected ex vivo CXCR5+ B cells, particularly CXCR5+HLA-DRhigh B cells. Assessment of VACV gene expression by RT-PCR in total B cells to evaluate permissiveness of VACV infection in B cells revealed early and intermediate viral gene expression, but little to no late viral gene expression, suggesting that VACV infection was aborted at the late viral gene expression stage in unstimulated B cells. Additionally, preliminary data demonstrated enhanced VACV binding to and infection of B cells stimulated by B cell receptor crosslinking and TLR-9 agonist treatment. With the risk of smallpox or monkeypox being used as bioterrorism agents and the multitude of adverse reactions to the current vaccine, understanding VACV infection in B cells is important for developing a better smallpox vaccine.
Suppression of IFN-β production by Epstein-Barr virus lytic transactivator Zta
Authors: Wong Nok Hei Mickey , Yuen Kit-San , Chan Chi-Ping , Jin Dong-Yan 
Published: January 2023

Abstract
Interferon β (IFN-β) production is an innate antiviral immune response against virus infection and lytic virus production. Several molecular mechanisms, including the sensing of cytosolic double-stranded DNA (dsDNA) by cyclic GMP-AMP synthase (cGAS), have been described to induce IFN-β production. Epstein-Barr virus (EBV) is a γ-herpesvirus that infects most of the population in the world and is associated with the development of various lymphoid and epithelial malignancies in a subset of people. In this study, we identified and characterized EBV lytic transactivator Zta as a potent suppressor of IFN-β production. EBV lytic infection was shown to suppress IFN-β production. To identify the viral proteins that mediate this suppression, a functional screen was performed using a dual-luciferase reporter system and an expression library of all EBV proteins. In this screen EBV lytic transactivator Zta was found to be capable of potently suppressing IFN-β production induced by different stimuli of interferon regulatory factor 3 (IRF3) activation. Zta is previously known to directly interact with and suppress IRF7, but its impact on IRF3 and the mode of action have not been demonstrated. We showed that Zta prevented the activation of IRF3 and IFN-β promoter by cGAS and STING (stimulator of interferon genes). Mechanistically, Zta suppressed the phosphorylation of IRF3 and its upstream interacting partner TANK-binding kinase 1 (TBK1), leading to the inhibition of IRF3 activity and nuclear translocation. Therefore, Zta may help EBV to evade host immunity during lytic infection by circumventing IRF3 activation and thus IFN-β production. Our findings revealed a new mechanism for EBV evasion of innate immunity.
Roles of Reactive Oxygen Species in Modulation of Thymocyte Subpopulations during Thymic Atrophy
Authors: Majumdar Shamik , Raghavan Abinaya , Nandi Dipankar 
Published: January 2023

Abstract
The thymus is the site of T cell development, selection and maturation. Precursor cells migrate from the bone marrow to the thymus, where they undergo sequential development followed by selection to give rise to mature and functional T cells. The cell surface expressions of various markers define distinct stages of development and maturation of thymocytes. With the help of these markers and multicolour flow cytometry, we have standardized staining procedures to study different thymocyte subsets during atrophy. We have also standardized an acute and lethal model of oral Salmonella Typhimurium infection in BALB/c mice, wherein drastic reduction of thymic cellularity is observed. Using multicolour flow cytometry, we investigated the changes occurring in thymocyte subsets in this model of infection-induced thymic atrophy. In addition, thymic atrophy was also induced by administration of LPS, the chemotherapeutic drug etoposide and the synthetic glucocorticoid, dexamethasone. Extensive thymic atrophy occurred across the different groups and thymocyte subsets were studied in a comprehensive manner. CD4 and CD8 profile on total thymocytes revealed CD4+CD8+ cells to be the most sensitive across all the modes of thymic atrophy studied. Subsequent detailed analysis of the thymic subpopulations revealed distinct changes during thymic atrophy. The roles of reactive oxygen species (ROS) in modulation of thymocyte subsets in the above mentioned conditions were studied using the ROS quencher, N-acetyl cysteine. We further intend to understand the mechanisms involved in changes occurring in the thymocyte subsets that may aid in development of candidate therapeutic molecules with thymopoietic potential.
Epigenetic regulator CXXC5 recruits DNA demethylase Tet2 to regulate TLR7/9-elicited IFN response in pDCs
Authors: Xiao Hui , Ma Shixin 
Published: January 2023

Abstract
TLR7/9 signals are capable of mounting massive IFN response in pDCs immediately after viral infection, yet the involvement of epigenetic regulation in this process has not been documented. Through immune gene profiling, we found that the zinc finger CXXC family epigenetic regulator CXXC5 is specifically expressed in pDCs, but not in cDCs or macrophages. Interestingly, CXXC5-deficient pDCs exhibited impaired IFN-I response to TLR7/9 ligands and RNA/DNA viruses. Furthermore, we uncovered that CXXC5-deficiency resulted in aberrant methylation of CpG DNA islands (CGIs) of a subset of genes including Irf7 in the steady-state pDCs, leading to diminished expression of this group of genes in pDCs. Mechanistically, CXXC5 seemed to be responsible for the recruitment of DNA demethylase Tet2 to maintain the hypomethylation of these CGIs in pDCs, a process coincided with histone remodeling and active transcription in these regions. Upon challenged by viruses, CXXC5-deficient mice were unable to launch robust IFN-I response at the early stage, and became highly susceptible to HSV and VSV infection. Together, our results not only identified CXXC5 as a novel regulator of TLR7/9-induced IFN-I response in host defense, but also provided new insight into the understanding of epigenetic regulation of pDC function.