bluuebunny/crossref_metadata_2025_split huggingface dataset:
Lentiviral-siRNA blockade of immunosuppressive genes increases DC vaccine efficacy. (131.42)
Authors: Lebson Lori , Rosenzweig Jason , Manicini Justin , Whartenby Katharine 
Published: January 2023

Abstract
Dendritic cells (DCs) play an essential role in the activation of T cells. Because of their potent activation capabilities, DCs have been used to stimulate anti-tumor responses via vaccines. While these vaccines have elicited immune responses, they have not been curative. In a tumor environment, suppressive factors can downregulate their ability to generate effective responses through modifying gene expression in DCs. We have been identifying such genes and proposed that blocking them in DCs would enhance their efficacy. To test our hypothesis, we silenced one immunosuppressive gene using a lenti viral vector siRNA construct. Our results demonstrate that blocking gene expression enhances activation and anti-tumor responses, leading to prolonged survival of tumor bearing mice. This approach may serve as a novel means to increase the efficacy of ex vivo developed DC vaccines.
Immunization with lentivector expressing HBV Ag markedly improves the tumor microenvironment and result in regression of established tumors (131.44)
Authors: He Yukai , Hong Yuan , Xiao Haiyan , Peng Yibing , Wang Guiqiang 
Published: January 2023

Abstract
One of the main reasons for the limited antitumor efficacy of current tumor vaccines is the failure of immune effector cells inside the tumor environment due to the infiltration of immune suppressor cells and immune dysfunction of effector cells. In this study, we investigated if the immune suppressive tumor environment can be improved by lentivector immunization. We found that lentivector immunization could not only stimulate potent T cell responses against HBV surface Ag and cause tumor influx of effector T cells, but also markedly changed the tumor microenvironment. Immunization with lentivector induces strong proliferation of CD4 and CD8 T effector cells but does not increase the proliferation of Treg in the vaccine draining LN. More importantly, lentivector immunization profoundly increase proliferation of the CD4 Teff cells in the tumor lesions, resulting in more CD4 Teff cells accumulation in the tumor lesions and decrease of Treg ratio in the tumor. Analysis of the tumor environment showed that lentivector immunization can skew immune suppressive tumor environment to Th1 and Tc1 like environment, with marked increase of T-bet, IFNγ, Perforin, and granzyme in the tumor lesions. Lentivector HBV-S-lv immunized mice were completely protected from HBV positive B16 tumor cells (B16-S) challenge. Furthermore, established B16-S tumor can be eradicated by lentivector immunization.
TSC2 regulates CD8+ T cell effector function and the generation of memory (132.14)
Authors: Pollizzi Kristen , Waickman Adam , Delgoffe Greg , Powell Jonathan 
Published: January 2023

Abstract
Recent studies implicate mTOR activity in the generation of memory CD8+ T cells. Tuberous Sclerosis Complex 2 gene (TSC2) is an upstream inhibitor of mTOR function. In this report, mice in which TSC2 is deleted in T cells were utilized to determine the role of TSC2 in controlling T cell function. TSC2-/- T cells demonstrate enhanced and constitutive mTOR Complex 1 (TORC1) activity. Using a recombinant vaccinia virus expressing OVA peptide (vac-OVA) as the primary stimulation we have found that loss of TSC2 promotes an enhanced effector response in CD8+ T cells in terms of killing ability and cytokine production 7 days after injection. However, when assessing function at 30 days post-infection the absolute number of OVA specific CD8+ T cells are reduced compared to wildtype controls. This correlated with diminished killing ability as shown by in-vivo CTL activity. Interestingly, when the TSC2-/- OVA specific CD8+ T cells were analyzed at day 30 for cytokine production, these cells secreted increased IFNγ and TNFα over their Wt counterparts. Together these data indicate that TSC2 deficiency impairs the effector to memory transition. However, the absence of TSC2 leads to enhanced CD8+ effector function during the primary immune response and in the remaining clonotypic CD8+ T cells that are present 30 days post infection. Overall, our findings implicate a novel and complex role for TSC2 signaling in the regulation of both CD8+ T cell effector function and memory generation.
Population level effects on Vγ2Vδ2 T cell frequencies in healthy adults: a difference in baseline values and resting phenotypes among Caucasian and African American donors (132.16)
Authors: Armstrong Cheryl , Cairo Cristiana , Cummings Jean-Saville , Deetz Carl , Davis Charles , Pauza C.David 
Published: January 2023

Abstract
Adult human γδ T cells are dominated by the Vγ2Vδ2 T cell receptor (TCR) which modulates MHC-unrestricted responses to non-peptidic antigens. This subset is so numerous that between 1 in 40 and 1 in 400 circulating CD3+ T cells will respond to stimulation with isoprenoid pyrophosphates. The normal frequency of Vγ2Vδ2+ cells reflects TCR selection and amplification by chronic antigen stimulation, but the regulatory mechanisms controlling population size have not been described. During clinical studies related to Vγ2Vδ2 T cells in cancer and AIDS, we identified a difference in the circulating populations between Caucasian and African American healthy volunteers. When expressed as the percentage of total lymphocytes, Caucasian donors had 5-times higher levels of Vγ2Vδ2 cells compared to African American donors and phenotypes characteristic of mature functional cells were also different among the groups, including CD56 expression that is linked to cytotoxicity. Analysis of the Vg2 repertoire showed a strikingly lower proportion of cells expressing the Vγ2-Jγ1.2 rearrangement. Our data suggest that distinct regulatory mechanisms, in addition to positive selection and chronic amplification, control Vγ2Vδ2 cell levels in blood and that further study of population level effects may uncover unique regulatory mechanisms that control this interesting population.
Multichromatic phenotypic analysis of peripheral lymphocytes in non-human primates (NHP) following total body irradiation (TBI) with 5% bone marrow shielding (BM5) (132.19)
Authors: Higgins Adam , Maciel Milton , Sztein Marcelo , MacVittie Thomas 
Published: January 2023

Abstract
High dose TBI of NHP leads to severe myelosuppression, neutropenia, thrombocytopenia, and immunosuppression, concurrent with an acute gastrointestinal syndrome. BM shielding mitigates these lethal effects by preserving a reservoir of unexposed hematopoietic progenitors. Rhesus macaques were exposed to either 10Gy (sub-lethal) or 11Gy (LD15 8-10 days) in the TBI/BM5 protocol and administered supportive care (fluids, antibiotics, transfusions). Peripheral blood was drawn over 150 days post irradiation (DPI) and lymphocyte subsets were assessed by flow cytometry. 10/11Gy TBI/BM5 caused a significant, early decrease in CD3+ T cells, sustained through 42 DPI, which reached baseline at 56 DPI. CD8+ T cells dominated T cell repopulation while CD4+ T cells showed a slower recovery. Most of the CD8+ T cells were CD62L+CD45RA+. CD20+ B cells reached 168% of baseline at 56 DPI and remained above baseline through 150 DPI. CD20+CD27+ memory B cell recovery kinetics were similar to B cells up to 77 DPI, with a sharp decrease at 90 DPI. CD3+ integrin α4β7+ (mucosal marker) cells recovered to 249% of baseline levels by 35 DPI and peaked at 400% of baseline at 120 DPI. Conversely, α4β7+ B cell recovery was slower, requiring 120 DPI to reach pre-irradiation levels. Functional assays will help understand the recovery pattern of lymphocyte subsets.
Various Sources, Types and Levels of Reactive Oxygen Species Make Distinct Contributions to Cell Cycle Progression And Survival (132.25)
Authors: Carey Gregory , Phatak Pornima , Koontz Jason 
Published: January 2023

Abstract
We examined several sources, types and roles of ROS in B cell lymphoma (BCL) survival. Initially following B cell receptor (BCR) crosslinking, hydrogen peroxide (H2O2), superoxide (O2•-) and cellular respiration were stimulated. Conversely, chronic BCR activation led to the loss of H2O2 and to massive O2•- induction, which were associated with tumor death. ROS scavengers reduced cellular ROS, but enhanced BCR-stimulated apoptosis, suggesting that ROS regulate BCL homeostasis positively. Next, we examined the RedOx- and activation-status of the survival protein, Akt after BCR-crosslinking. Interestingly, increased H2O2 production paralleled increased, reversible oxidation of Akt whereas the loss of H2O2 tone and massive O2•- induction paralleled irreversible Akt oxidation and its deactivation. Fractions of ROS were attributable to either NADPH oxidases, xanthine oxidase, the electron transport chain or glycolysis. Interestingly, however, it appeared that either particular ROS or distinct metabolic pathways differentially impacted the cell cycle. Thus, blockade of glycolytic respiration reduced ROS and arrested cells at Go/G1. In contrast, blocking oxidative respiration at complex I reduced a fraction of basal ROS and allowed the cells to exit Go/G1 and progress through S-phase, but to become arrested in G2/M. Together, these data suggest that the temporality, spatiality and magnitude of ROS production ultimately influence cell signaling and consequently, determine cell fate.
CXCL13 inhibition prevents bone metastasis in hormone-refractory prostate cancer (133.8)
Authors: Lillard James , Singh Rajesh , Sharma Praveen , Singh Shailesh 
Published: January 2023

Abstract
We have recently reported, chemokine receptor CXCR5 and its ligand CXCL13 expression positively correlates with prostate tumor staging. We have also noted differential expression of CXCR5 by prostate cancer (PCa) cell lines. By virtue of the presence of elevated CXCL13 levels in patient serum, CXCL13 secretion by bone marrow endothelium, and expression of CXCR5 on the surface of PCa cells, we hypothesized that gradient-dependent CXCR5-CXCL13 interactions are responsible in part for PCa cell bone metastases and inhibition of this axis may prove beneficial for prevention of bone metastasis and tumor progression. Luciferase-expressing PC3 cells were injected into the right carotid artery or directly into the tibia of nude mice. Before and after the development of bone metastases, groups were treated with control or anti-CXCL13 antibodies every third day and monitored for tumor progression or regression by non-invasive bioluminescent and microCT in vivo imaging. We demonstrate CXCL13 is present in bone marrow of xenograft tumor-bearing mice following intra-cardiac /intra-tibial injection of PC3 cells. This hormone-refractory PCa cell line established skeletal (osteolytic) metastases and tumor growth, but CXCL13 blockade significantly delayed prostate tumor formation, osteolysis, and spread to bones as well as growth, than compared to mice treated with control antibody. In conclusion, our study shows that the CXCL13-CXCR5 axis supports PCa bone metastasis and growth.
Differential effects of Human Interferon-alpha subtypes on CD4 T cell function. (134.10)
Authors: Gold Doria , Hillyer Philippa , Raviv Nataly , Heuer Melissa , Chi Bo , Rabin Ronald 
Published: January 2023

Abstract
We compared the effects of each of the 12 IFN-α subtypes and IFN-β on CD4 T cell proliferation and cytokine expression in response to stimulation with SEB. Purified CD4 T cells or PBMC from healthy donors were treated with 4 doses of each IFN for 16 hours, and stimulated with SEB for either 4 hours or 4 days, to measure cytokine expression with flow cytometry or proliferation by 3H-thymidine uptake, respectively. While type I IFN did not affect the percentage of SEB-activated (CD40L+) CD4 T cells, they did enhance the percentage of CD40L+ cells that expressed IFN-γ, IL-2 and IL-4 in a dose-dependent manner. The greatest difference among the subtypes was at 25 U/mL (for example, the percent of IL-2+ cells in response to SEB alone, IFN-α1 and -α8 was 19.9, 23.4, and 26.2, respectively). Surprisingly, type I IFN decreased the level of expression of CD40L less than or equal to 50% of SEB alone, and this decrease highly correlated with the enhancement of cytokines. Proliferation was decreased by each IFN subtype with maximal differences among subtypes in roughly the same hierarchy as the effects on cytokine and CD40L expression. These data suggest that type I IFN enhance both types 1 and 2 CD4 T cell responses with differences among the subtypes at “physiological” levels. The decrease in CD40L expression may affect the level of B cell activation and DC-CD4 T cell cross-talk.
Genetic ablation of steroid receptor coactivator-3 promotes PPAR-β-mediated alternative activation of microglia in experimental autoimmune encephalomyelitis (135.1)
Authors: Zhang Yanyun , Xiao Yichuan 
Published: January 2023

Abstract
The importance of links between lipid metabolic regulation and immune functions is increasingly acknowledged, yet not enough. SRC-3 has been demonstrated to play important roles in the regulation of lipid metabolism. In the present study, the potential role of SRC-3 in EAE was examined by analyzing disease progression in SRC-3-/- mice. SRC-3-/- mice are lean and exhibit lower lipid deposition through modulating the expression of lipid metabolism genes by PPAR-β. Moreover, we found that SRC-3 deficiency significantly attenuated the disease severity of EAE along with decreased inflammatory infiltration and demyelination. However, these effects are not caused by inhibition of peripheral T cell response, but by up-regulated expression of PPAR-β in CNS, which induced an alternative activation state of microglia in SRC-3-/- mice. These alternatively activated microglia inhibited CNS inflammation through inhibition of the expression of pro-inflammatory cytokines and chemokines well as up-regulation of anti-inflammatory cytokines, and promoted myelin regeneration through increased accumulation of oligodendrocyte precursors (OPCs) in white matter and elevated expression of myelin genes in the spinal cords of SRC-3-/- mice. Our data uncover the mechanism by which PPAR-β promotes tissue repair in response to inflammatory injury in SRC-3-/- mice, which builds up important links between lipid metabolic regulation and immune functions.
IL-17 and IL-22 affected keratinocytes via IL-1β inducing NALP3-caspase-1 pathway (135.18)
Authors: Cho Kyung-Ah , Woo So-Youn , Lee Kyung Ho 
Published: January 2023

Abstract
Pathogenesis of inflammatory skin disease is involved in the interplay between immune cells and keratinocytes. Psoriasis, for example, can be triggered by dysregulation of T cell-mediated immune response and characterized by hyperplasia of the epidermis, infiltration of leukocytes into both the dermis and epidermis, and dilation and growth of blood vessels. In addition, chronic immune response of epidermal layer is involved in cytokine release from keratinocytes and IL-1β has been implicated in inflammatory skin disease. Th17 cells possess IL-1βreceptor and excess production of IL-1β in NALP3 mutation leads to augment Th17-dominant pathology. Th17 cells are induced by exposure of TGF-β, IL-6, IL-15 and IL-23 and Th17 cells are related to autoimmunity and inflammation by producing IL-17 and IL-22. To determine whether IL-17 or IL-22 may have an effect on IL-1β secretion from keratinocytes, we used human keratinocyte cell line, HaCaT, and capsase-1-deficient mice with the IL-17 or IL-22 stimulation. We found that under IL-17 and IL-22 treatment IL-1β was increased via NALP3 activation. These results suggest that cytokine from Th17 cells may potentiate IL-1β mediated skin inflammation and lead to phenotypic alteration of keratinocytes.
Profibrotic alternatively activated M2 macrophages are associated with increased severity of dilated cardiomyopathy in male BALB/c mice with cosackievirus B3-induced autoimmune myocarditis (135.20)
Authors: Coronado Michael , Frisancho-Kiss Sylvia , Bedja Djahida , Frisancho J. Augusto , Shin Jaewook , Gabrielson Kathleen , Fairweather DeLisa 
Published: January 2023

Abstract
Current research has shown heart disease as the leading killer in the US. Autoimmune myocarditis (AM) progresses to dilated cardiomyopathy (DCM) and occurs at a higher incidence in males. In order to study sex differences in AM we inoculated BALB/c mice with 103 PFU of coxsackievirus B3 with cardiac myosin ip to induce autoimmunity. At d10 post infection males develop severe myocarditis that progresses to DCM at d35. We investigated functional endpoints of DCM by echocardiography at d90 and found that males developed significantly more severe DCM. To investigate sex differences further we conducted a microarray comparing infected males and females. Approximately 500 genes were significantly up regulated in males. Genes increased with a &gt; 1.5 fold change fell into categories associated with Th2 responses, M2 macrophages (Macs), and extracellular matrix (ECM) remodeling. We showed previously that male cardiac CD11b+ GR1+ Macs express the mannose receptor (MR), TLR4, caspase-1 and IL-1β. This microarray confirmed that males have increased M2 markers including the MR, CD14 and IL-33. We also found upregulation of ECM remodeling genes including proteoglycan-4, timp-1 and tenascin-C indicating cardiac remodeling. Th2 responses are known to increase ECM remodeling and can drive differentiation of Macs to an M2 phenotype. Our data suggests that Th2 associated profibrotic M2 Macs may increase cardiac remodeling in males during AM resulting in DCM. Funded by NIH-HL-087033.
Maladaptive effector CD8+ T cells distinguish clinical subtypes of Juvenile Idiopathic Arthritis (135.36)
Authors: Dvergsten Jeffrey , Mueller Robert , Abedin Sameem , Michel Joshua , Rosenkranz Margalit , Vallejo Abbe 
Published: January 2023

Abstract
Pervasive T cell clonality is a feature of Juvenile Idiopathic Arthritis (JIA), but the role of oligoclonal T cells in this chronic disease of childhood is poorly understood. We propose that T cell clonality in JIA is indicative of replicative pressure leading to accumulation of terminally differentiated cells. Hence, we examined the T cell repertoire of children with confirmed diagnosis of JIA and their age-matched healthy counterparts. Results showed that naive T cells of children with JIA had significantly lower telomere-FISH fluorescence indicating a long replicative history. Multicolor cytometry analyses also showed that patients had over abundance of CD8+CD31+CD28null T cells in blood and synovial fluid irrespective of disease duration and medication. Frequency of these unusual T cells was significantly higher among patients with the more severe, polyarticular form of JIA disease than those with oligoarticular disease. Furthermore, these T cells expressed high levels of γH2AX and pRB suggesting senescence. In proliferation assays using CFSE, these T cells remained CFSEhi but were CD69hiCD25lo/neg, results consistent with senescence and not quiescence. In receptor crosslinking assays, ligation of CD31 alone sufficiently induced tyrosine phosphorylation, cytolysis, and production of IFNγ, TNFα, and IL-10. These data indicate premature immune senescence in JIA, with CD8+CD31+CD28nullH2AX+ T cells as maladaptive effectors, and are potential targets for therapy.
Toll-like receptor 9 is alternatively cleaved to generate a soluble form of the receptor that downregulates response to CpG-DNA (136.25)
Authors: Chockalingam Annapoorani , Brooks James , Cameron Jody , Leifer Cynthia 
Published: January 2023

Abstract
Proper regulation of nucleic acid sensing Toll-like receptors (TLRs), including TLR9, is critical to prevent autoimmune disease and promote host defense. The objective of our research is to elucidate the molecular mechanisms that control TLR9 signaling. Localization of TLR9 in endolysosomes is one regulatory mechanism that facilitates recognition of CpG-DNA and prevents response to self DNA. Restricted localization of TLR9 controls access to proteases, including cathepsins B and L and asparagine endopeptidase, which generate an active 80 kDa (p80) form of the receptor. Here we use mass spectrometry analysis to identify a 100 kDa soluble form of TLR9 (sTLR9, amino acids 1-740). The proteolytic cleavage of sTLR9 is distinct from that of p80 since it is not blocked by Z-FA-fmk or E-64 protease inhibitors, or by bafilomycin A1 or chloroquine. These data suggest that generation of sTLR9 does not occur in acidic endosomes. We show that the generation of sTLR9 requires cathepsin-S, a protease inhibitor active at neutral pH. Most importantly we show that the sTLR9 binds to CpG-DNA and down regulates CpG-DNA induced TLR9 signaling. Balance between proteolytic cleavage occurring at two distinct sites on TLR9 ectodomain is likely a major factor that modulates host innate immune responses to microbial infection and prevents development of autoimmune disease.
Inflammasome-dependent IL-1β production is critical for complete Freund’s adjuvant-induced helper T cell polarization (136.44)
Authors: Shenderov Kevin , Barber Daniel , Mayer Katrin , Jankovic Dragana , White Sandy , Caspar Pat , Hieny Sara , Trinchieri Giorgio , Besra Gurdyal , Cerundolo Vincenzo , Sher Alan 
Published: January 2023

Abstract
The innate immune system recognizes pathogens/antigens via several families of pattern recognition receptors (PRRs). Often, several PRRs will be triggered, resulting in an innate immune response via a complex integration of signals. However, the importance of different PRR signaling pathways in determining adaptive responses such as helper T cell polarization is not fully understood. We studied the role of two pathways—TLRs and the inflammasome—in promoting helper T cell polarization in response to complete Freund’s adjuvant (CFA). Mice immunized with ovalbumin (OVA) in CFA developed an OVA-specific Th1/Th17 response that was dependent on the signaling adaptor MyD88. TLRs were only partially required for this response, whereas interleukin 1 (IL-1) receptor signaling and the inflammasome were critical. We found that heat-killed M. tuberculosis H37Ra, a component of CFA, induces mature IL-1β production in vitro in a process that requires potassium efflux and reactive oxygen species and is mediated by the NLRP3 receptor. We further showed that the major inflammasome-triggering component of H37Ra is the peptidoglycan. However, the peptidoglycan-sensing receptors NOD1 and NOD2 were not needed for this response. Our data suggest that NOD1/2-independent recognition of mycobacterial peptidoglycan by the NLRP3 inflammasome induces IL-1β production and may play an important role in CFA-induced helper T cell responses. This work was supported by the NIAID Intramural Research Program.