bluuebunny/crossref_metadata_2025_split huggingface dataset:
Novel mechanisms of anti-tumor action for therapeutic T-bet gene-modified dendritic cells (P4228)
Authors: Chen Lu , Taylor Jennifer , Storkus Walter 
Published: January 2023

Abstract
Clinical implementation of Type-1 polarized dendritic cell (DC) in vaccine formulations holds significant promise in the cancer therapy setting. We have recently demonstrated that DC engineered to express the T cell Type-1 transactivator protein T-bet (TBX21) promote robust Type-1 T cell-mediated immunity via a unique mechanism that is largely interleukin (IL)-12p70-independent. In the current study, we investigated the anti-tumor mechanisms of action associated with intratumoral administration of DC.Tbet developed from syngenic IL-12p35 -/-, IL-12p40 -/-, IFNγ -/- or β2m -/- mice. We observed that Type-1 anti-tumor immunity associated with slowed tumor growth was effectively developed in animals receiving DC.Tbet deficient in their ability to produce IL-12 family cytokines (i.e. IL-12, IL-23, IL-35) or IFN-γ. Surprisingly, we observed that the ability of DC.Tbet to present MHC class I/peptide complexes (i.e. lost in DC developed from β2m -/- mice) was not important for early priming of anti-tumor CD8+ T cells, but was essential for the sustained therapeutic efficacy of DC.Tbet (and treatment-induced anti-tumor CD8+ T cells) within the tumor microenvironment (TME). In order to best translate this cancer treatment modality into the clinic, we are currently evaluating how DC.Tbet conditions the TME for improved function of Ag-primed Type-1 T effector cells and how these APC collaborate with host DC to diversify the therapeutic T cell repertoire.
Adjuvant effect of a natural TLR4 ligand on dendritic cells (P4240)
Authors: Kim Hyung Sook , Ryu Hwa Sun , Lee Hong Kyung , Kim Yeon Jin , Kim Ji Sung , Park Yunsoo , Park Mi-jeong , Hong Jin Tae , Kim Youngsoo , Han Sang-Bae 
Published: January 2023

Abstract
Defect of dendritic cell (DC) maturation in tumor microenvironments is an important immunological problem limiting the success of cancer immunotherapy. In this study, we investigated the immunostimulatory effect of a Mori fructus polysaccharide (MFP) on DCs, a type of potent antigen presenting cells. MFP induced phenotypic maturation of DCs, as proven by the elevated expression of CD40, CD80/86, and MHC-I/II molecules. MFP induced functional maturation of DCs, in that MFP increased the expression of IL-12, IL-1β, TNF-α, and IFN-β, decreased antigen capture capacity, and enhanced allogenic T cell stimulation. MFP efficiently induced maturation of normal toll-like receptor4 (TLR4) DCs from C57BL/6 and C3H/HeN mice, but not mutated TLR4 DCs from C3H/HeJ mice, suggesting that TLR4 might be one of the membrane receptors of MFP. We showed that MFP increased phosphorylation of mitogen-activated protein kinase (MAPKs), and nuclear translocation of NF-κB p65 subunit, which are signaling molecules downstream from TLR4. These results indicate that MFP induced DC maturation through TLR4 signaling.
Effect of low level laser and Mesenchymal Stem Cells therapies on cigarette-induced lung inflammation in mice. (P4200)
Authors: Ligeiro de Oliveira Ana Paula , Silva Vanessa , Neves Vinicius , Vieira Rodolfo , Jazedje Tatiana , Pellati Mayra , Czeresnia Carlos , Perin Paulo , Maluf Mariângela , Nisenbaum Marcelo , Evangelista Lucila , Halpern Silvio , Zatz Mayana , Peron Jean Pierre 
Published: January 2023

Abstract
Chronic Obstructive Pulmonary Disease (COPD) has a high prevalence and economic and social cost. In this context, new therapeutic approaches are of great interest, and the use of Mesenchymal Stem Cells (MSCs) or low level laser therapy (LLL) may be innovative and promising approaches. Thus we investigated the effect of LLL (670 nm) and htMSCs (human tubal-derived Mesenchymal Stem Cells) in the treatment of cigarette-induced pulmonary inflammation in mice. C57Bl/6 mice were submitted to cigarette smoke for 75 days (2 times/day). Next, animals were treated with LLL (COPD+LLL group) or htMSCs (COPD+htMSCs group) 7 and 15 days before the experiment, on day 76th. Animals were sacrified for morphologic and functional analysis of the lung. Bronchoalveolar lavage (BAL) analysis showed that COPD+LLL and COPD+htMSCs groups had a significant decrease in neutrophils and lymphocytes when compared to COPD group. This was associated with reduced mucous secretion and collagen deposition. Moreover, LLL significantly reduced IL-6 and TNF-alpha, which was not observed aftetr htMSCs. In vitro experiments showed reduction in IL-6 secretion by bronchial epithelial cells (BEC) and TNF-alpha by pneumocytes type II (PII) exposed to cigarette extract at 2,5% and further irradiated with LLL. These results indicate that LLL and htMSCs therapies have anti-inflammatory effect on cigarette-induced lung inflammation reducing local inflammation and may be considered interesting therapeutic approaches.
Oral Salmonella expressing Colonization Factor Antigen I stimulates the rapid induction of Galectin-1 on induced Treg cells (P4204)
Authors: Jun Sangmu , Hoffman Carol , Hoyt Teri , Pascual David 
Published: January 2023

Abstract
Galectin-1 (Gal-1) is one of the 15 Galectin family members that bind to β-galactoside. Gal-1 binding to various cell surface glycoproteins leads to deletion of T effector functions. Gal-1 engagement to CD43 on APCs converts these cells to tolerogenic supporting the generation of Treg cells. Our previous work have shown that oral Salmonella-CFA/I strain H696 (expresses CFA/I fimbriae from enterotoxigenic E. coli) induces a highly suppressive Foxp3+ CD25+CD4+ Treg cells capable of suppressing the development of experimental autoimmune encephalomyelitis (EAE). Gal-1 expression was rapidly induced within 3 days by both Foxp3+ Treg cells obtained from Salmonella-CFA/I-immunized mice unlike mice vaccinated with isogenic Salmonella vaccine (strain H647) control. To assess the function of Gal-1, isolated Treg cells from H696- and H647-vaccinated mice were assessed for their ability to suppress OT-2 OVA-specific effector T cell proliferation following r estimulation with OVA peptide. Upon neutralization of Gal-1, Salmonella-CFA/I-induced Treg cells were unable to suppress OT-2 cell proliferation; similar neutralization of Salmonella vector-derived Treg cells had no effect. Kinetic analysis of Gal-1 expression by dendritic cells (DCs) following H696 imminization also showed early induction of Gal-1 suggesting the importance of Gal-1 to induce tolerogenic T cells and DCs. These results suggest Gal-1 is a major effector molecule for the Salmonella-CFA/I-induced Treg cells.
CD8 T cell exhaustion is influenced by genetic background (P6076)
Authors: Christiaansen Allison , Boggiatto Paola , Hartwig Stacey , Varga Steven 
Published: January 2023

Abstract
Lymphocyte choriomeningitis virus (LCMV) induces a robust CD8 T cell response allowing for viral clearance and limited disease following an acute LCMV Armstrong (Arm) infection in both C57BL/6 (B6) and BALB/c mice. In contrast, infection of B6 mice with LCMV Clone-13 (Cl-13) results in CD8 T cell exhaustion leading to the establishment of a chronic infection. We examined Cl-13 infection of BALB/c mice and observed a does-dependent increase in disease following infection. BALB/c mice infected with either Arm or a low dose of Cl-13 induced a robust NP118-126-specific CD8 T cell response whereas infection with a high dose of Cl-13 significantly blunted the induction of the NP118-126-specific CD8 T cell response. In addition, BALB/c mice infected with a high dose of Cl-13 failed to control the infection with prolonged virus replication in multiple peripheral organs. The more focused immunodominant CD8 T cell response elicited by LCMV in BALB/c versus B6 mice does not appear to account for the increased disease following high dose Cl-13 infection. Cl-13 infected B10.D2 mice exhibit an increased NP118-126-specific CD8 T cell response as compared to BALB/c infected mice. Furthermore, BALB.B mice exhibit a reduced magnitude of CD8 T cells specific to the three immunodominant epitopes as compared to B6 mice. Our results indicate that non-MHC genes influence the rate of CD8 T cell exhaustion and subsequent disease following a chronic Cl-13 infection.
SHP-1 deficiency does not restore the CD8+ T cell response to an LCMV viral variant (P6079)
Authors: Shorter Shayla , Evavold Brian 
Published: January 2023

Abstract
Viruses employ various mechanisms to evade recognition by host immune effectors and establish persistence in the host. We have previously shown that CD8+ T cells responding to a viral variant of lymphocytic choriomeningitis virus (LCMV) have high SHP-1 activity. Src homology region 2 domain-containing phosphatase-1 (SHP-1) is a non-receptor phosphatase found in hematopoietic cells and is thought to play a primarily negative regulatory role in many signaling pathways. SHP-1 exerts important negative regulation on proximal TCR signaling by blocking early T cell activation. Because of its role in dampening TCR signaling, we hypothesized that removal of SHP-1 in T cells would restore the response to the viral variant. Using CD4 Cre Fl SHP-1 mice, we evaluated the generation, activation and functionality of CD8+ T cells generated in response to the viral escape mutant. Loss of SHP-1 did not greatly affect response of the P14 TCR Tg cells line assessed in vitro. In vivo analysis of the response to the viral escape mutant did not demonstrate increased T cell numbers or functionality (IFNg secretion) in response to the viral escape mutant or the control chronic viral LCMV infection. Polyclonal T cell responses to the viral challenges were unaffected by SHP-1 deficient. These observations suggest that removal of a negative regulator may not have the expected therapeutic benefit of an enhanced T cell response.
Selective requirement of Ikaros zinc fingers in Treg and Th17 fate decision. (P1137)
Authors: Heller Jennifer , Schjerven Hilde , Qiu Ju , Lee Aileen , Smale Stephen , Zhou Liang 
Published: January 2023

Abstract
TGF-β is a common factor important for the differentiation of pro-inflammatory Th17 and anti-inflammatory inducible Treg cells. However, the precise molecular mechanisms underlying the fate decision of differentiating CD4+ T cells in the presence of TGF-β is poorly understood. Here, we show that distinctive N-terminal DNA-binding zinc fingers of Ikaros play essential roles in Treg and Th17 fate decision. Ikaros has a highly conserved DNA-binding domain near the N-terminus with four tandem zinc fingers. Zinc fingers 2 and 3 are required for stable binding to DNA, whereas fingers 1 and 4 appear to be important for differentially modulating binding properties to specific sites at target genes. Our data show that T cells lacking Ikaros zinc finger 4 but not 1 failed to differentiate into Foxp3+ Tregs upon TGF-β stimulation. Instead, TGF-β-skewed Ikaros zinc finger 4 mutant cells displayed aberrant upregulation of Th17-associated cytokines IL-17 and IL-22. IL-17 but not IL-22 upregulation is dependent on transcription factor RORγt. Aryl hydrocarbon receptor, an essential transcription factor required for IL-22 expression, was unexpectedly decreased. Together, our data uncover a novel selective requirement for Ikaros zinc fingers in the differentiation of Treg and Th17 cells and an intricate interplay among various transcription factors in programming Th17/Treg lineages. We are currently examining the role of Ikaros zinc finger 4 in infection and autoimmunity.
B-lymphocyte induced maturation protein 1 (Blimp-1) is required to limit the number of IL17A-producing CD4+ T cells in vivo. (P1139)
Authors: Bankoti Rashmi , Salehi Soofia , Benevides Luciana , Couse Michael , Silva Joao , Dhall Deepti , Meffre Eric , Targan Stephan , Martins Gislaine 
Published: January 2023

Abstract
Blimp-1 is a transcription factor required for embryonic development and immunity. Mice with T-cell-specific deletion of Blimp-1 (Blimp-1CKO) develop severe intestinal inflammation apparently mediated by CD4+Th cells, but little is known about Blimp-1’s role in regulating Th cell differentiation in vivo. Here we show that Blimp-1 restrains Th cell production of IL17 and IFNγ in vivo. Blimp-1CKO mice accumulate IL17 and IFNγ-producing TCRβ+CD4+cells in lymphoid organs and intestinal mucosa. Increased numbers of IL17-producing Th cells are also observed in WT and Blimp-1CKO-mixed BM chimeric mice containing WT regulatory T (treg) cells, suggesting a Treg-independent intrinsic role for Blimp-1 in constraining IL17production. Blimp-1-/-naïve CD4+Tcells are more prone to differentiate into IL17+IFNγ+ cells and cause severe colitis when transferred to Rag1-/- mice. In vitro-stimulated Blimp-1-/- CD4+T cells show increased expression of il17a and other Th17-related genes, suggesting that Blimp-1 could inhibit transcription of Th17 genes. Using ChIP assays we identified at least one site at the il17a gene that can be bound by Blimp-1 in Th cells, suggesting that Blimp-1 could function as a direct repressor of il17a. Single cell analysis reveals that Blimp-1 expression is selectively regulated during Th differentiation and directly correlates with suppression of Th17 traits in Th1cells. Collectively, these results establish a new role for Blimp-1 in regulating IL17 production in vivo
Effects of intracellular communication between T helper cells on Th2 differantiation (P1149)
Authors: Reich-Zeliger Shlomit , Antebi Yaron , Rimer Jacob , Hoberman-Arieli Tamar , Friedman Nir 
Published: January 2023

Abstract
Cytokines provide a main regulatory mechanism of immune responses, which allows for intercellular communication and enables cells to make collective decisions. Existing evidence for stochastiticity in expression of cytokines and their receptors raises the question: how does cytokine communication performs reliably in the face of noise? We study the effect of heterogeneity in IL4R expression level during Th2 differentiation as a model system. Naïve CD4 T cells were separated into different groups according to their IL4R expression levels and the different groups were cultured separately, under different IL4 concentrations driving their differentiation. Expression levels of Th2 markers, IL4 and GATA3, were measured at the single cell level. Our results reveal differences in the differentiation abilities of the different groups - higher initial receptor levels facilitate faster Th2 differentiation, at lower levels of IL4 as an input. Interestingly, we found that these differences in the differentiation abilities vanish when cells with high and low initial IL4R levels are co-cultured. These results suggest that intercellular interactions can decrease the potential heterogeneity in the population that results from noise in gene expression. These experiments provide an opportunity to evaluate the importance and roles played by intercellular communication in Th cell differentiation, and to assess the effects of stochasticity in protein levels on the differentiation process.
Analysis of factors that induce memory T cells. (P1157)
Authors: Soh Tetsuyoshi , Oki Yutaro , Uchiumi Fumiaki , Kishimoto Hidehiro 
Published: January 2023

Abstract
Naïve T cells recirculate between blood, lymph ducts and secondary lymphoid organs. Consequently, it should be different time that required for Ag-specific T cells, which are located in the various tissues, to populate and enter the draining lymph nodes in late time of the immune response. Late coming T cells were also primed in different environments and differentiated into memory T cells. We have investigated the kinetics of both OVA specific TCR Tg (OT-1 and OT-2) T cells stimulated earlier (early-stimulated T cells) and later in an immune response (late-stimulated T cells). To ascertain above studies, naïve T cells isolated from lymph nodes of OT-1/CD45.2+ mice and OT-2/CD45.2+ mice were injected i.v. into CD45.1+/CD45.2+ B6 host mice. For immunization of mice, ex-vivo expanded DCs that were induced from murine bone marrow cells by culturing with GM-CSF were incubated with OVA protein and LPS, and were injected s.c. into host mice. The early-stimulated T cells were developed by adoptive-transfer of Tg T cells into host mice before Ag-pulsed activated DCs were injected and the late-stimulated T cells were developed by adoptive-transfer of Tg T cells into mice 5-7 days after injection of Ag-pulsed activated DCs. The late-stimulated T cells divide slower and proliferate less than the early-stimulated T cells. These results suggest that the timing of recruitment of T cells into the draining lymph nodes affects T cells-fate that are influenced by different environments.
BCAP, a TLR/IL1R signaling adapter, intrinsically regulates differentiation of T helper cells (P1159)
Authors: Troutman Ty , Brewer Travis , Pasare Chandrashekhar 
Published: January 2023

Abstract
T helper cells utilize IL18 and IL1 signals provided by myeloid cells to achieve optimal Th1 and Th17 effector capacities. The IL1 and IL18 receptors belong to a family of receptors including Toll-like receptors (TLRs). The TLR/IL1R family share signaling components and (with the exception of TLR3) depend on the adapter MyD88 for engagement of downstream pathways. Signals from the receptor to the adapter are transmitted through homotypic interaction of TIR (Toll-Interleukin-1 receptor) homology domains found in all TLR/IL1R family members and their adapters. We have previously shown that BCAP (B-cell adapter for PI3K) contains a functional TIR domain enabling its participation in the TLR signaling pathway. Importantly, BCAP plays an obligate role in linking TLRs to activation of phosphoinositide 3-kinase (PI3K). Because the TLR/IL1R family share signaling components, we hypothesized that BCAP may regulate T cell differentiation through IL1R family signaling. Indeed, we found that BCAP intrinsically regulated differentiation of naïve T cells towards Th1 and Th17 effector lineages by participation in the IL1R family signaling pathways. Further, BCAP intrinsically regulated T cell proliferation and survival. Our continued efforts will further explore the molecular and transcriptional mechanisms by which BCAP regulates T cell activation. Significantly, this work will increase the understanding of key signaling pathways involved in disease and inflammation.
Role of BCL6 and PD-1 in CD4 memory T cell development (P1165)
Authors: Hollister Kristin , Wu Hao , Dent Alexander 
Published: January 2023

Abstract
Follicular T helper (TFH) cells have been shown to play an indispensable role within the germinal center (GC). To study the effects of TFH cell loss, our lab has developed a mouse line wherein BCL6, the master transcription factor behind TFH cell differentiation, is conditionally deleted in specific cell subsets (BCL6fl/fl). When mated to Cre-CD4 mice, loss of BCL6 specifically in CD4 T cells resulted in a complete absence of GCs. Data from control mice confirmed the correlation between BCL6 and PD-1 expression, with the highest levels of both found in TFH cells. Further analysis showed the few CD4+ CXCR5+ ICOS+ PD-1hi cells found in BCL6fl/fl Cre-CD4 mice have higher rates of apoptosis than those in control mice. Because previous studies have shown that Bcl6 regulates T cell memory, we investigated whether there were any effects on CD4 T cell memory development and found BCL6fl/fl Cre-CD4 mice have an altered memory cell phenotype, with higher levels of central memory cells and lower effector memory cells than control mice. Furthermore, memory cells in these mice show significantly lower rates of apoptosis as well as surface expression of PD-1. Taken together, this data suggests a complex regulatory network involving BCL6 and PD-1 expression, which effects not only TFH cell differentiation, but the development of CD4 T cells into effector versus central memory cells as well.
Bioenergetics of stomatin-like protein-2-deficient T cells reveal a regulatory role for this protein in immunometabolism (P1173)
Authors: Mitsopoulos Panagiotis , St-Pierre Julie , Dunn Stanley , Madrenas Joaquín 
Published: January 2023

Abstract
Stomatin-like protein (SLP)-2 is a mainly mitochondrial protein identified in proteome analysis of glycolipid-enriched membrane microdomains of mammalian cells, and modulates T cell activation. To study SLP-2 function we generated T cell-specific SLP-2 knockout (SLP-2 T-K/O) mice since conventional SLP-2 knockout mice were embryonic lethal at the pre-implantation stage. These mice showed reduced CD4+ T cell responses and decreased respiratory complex I and II+III activities. Our mechanistic studies suggest SLP-2 recruits prohibitins to cardiolipin (CL)-enriched domains in the mitochondrial inner membrane to form specialized CL-enriched membrane microdomains, required for optimal cellular respiration. We hypothesized SLP-2 functions in this manner to regulate the bioenergetics of T cells. Seahorse XF analysis revealed that although SLP-2 T-K/O cells showed unaltered total cellular and mitochondrial oxygen consumption rates (OCRs), they had significantly greater non-mitochondrial OCR, mitochondrial OCR uncoupled from ATP synthesis, and greater reliance on glycolysis (71% of mice). Surprisingly, SLP-2 T-K/O cells exhibited greater spare respiratory capacity, a phenomenon not due to differential sensitivities of cells to uncoupling agents. This new immunometabolic phenotype suggests SLP-2 may regulate assembly of functional respiratory supercomplexes. Bioenergetic and biochemical analyses of mitochondria are underway to establish the regulatory role of SLP-2 in T cell metabolism.
Th17 cells differentiated without TGF-β undergo FasL-dependent AICD (P1183)
Authors: Zhang Jimin , Ren Guangwen , Shi Yufang 
Published: January 2023

Abstract
Interleukin (IL)-17-producing T helper (Th17) cells can be differentiated under the combination of cytokines: IL-6, a proinflammatory cytokine, and transforming growth factor (TGF) beta, an antiinflammatory cytokine. We previously reported that TGF-beta is acting indirectly for Th17 cell differentiation by blocking expression of the transcription factors signal transducer and activator of transcription (STAT) 4 and GATA-3, thus preventing Th1 and Th2 cell differentiation. From T cells derived from Stat-6(-/-)T-bet(-/-) mice, which are unable to generate Th1 and Th2 cells, IL-6 alone can induce robust differentiation of Th17 cells. Here we report that Th17 cells derived from Stat-6(-/-)T-bet(-/-) mice, which were differentiated without TGF-β, underwent FasL-dependent activation-induced cell death (AICD). FasL was up-regulated upon TCR reactivation of those Th17 cells and the addition of FasL blocking antibodies abolished those Fas induced Th17 cell death. These features were comparable to wild type Th17 cells differentiated under the traditional combination condition of IL-6 and TGF-β. In all, these data demonstrate that Th17 cells differentiated without TGF-β undergo FasL-dependent AICD in a similar way to those wild type Th17 cells differentiated with TGF-β, indicating that Th17 cells derived from these two different conditions share common characteristics in AICD.