bluuebunny/crossref_metadata_2025_split huggingface dataset:
Contrasting CTL responses in protective and non-protective models of malaria (45.3)
Authors: Gupta Megha , Takagi Akihide , Richie Thomas L , Kappe Stefan H , Wang Ruobing 
Published: January 2023

Abstract
Protracted sterile protection conferred by a P. yoelii genetically attenuated parasite (PyGAP) vaccine was found to be completely dependent on CD8+ T lymphocytes. Immunization studies with perforin and IFN-γ knock out mice indicated that the protection was largely dependent on perforin as compared to IFN-γ. Both liver and spleen CD8+ T cells from PyGAP immunized mice induced massive apoptosis of liver stage (LS)-infected hepatocytes in vitro without the release of detectable IFN-γ and TNF-a, which directly correlated with GAP vaccine efficacy in vivo. Most importantly, we demonstrated that CD8+ T cells isolated from naïve mice that had survived wild-type P. yoelii sporozoite infection targeted mainly sporozoite-traversed and uninfected hepatocytes, revealing an immune evasion strategy that might have been used by wild-type parasites to subvert host immune responses. Differential processing of the parasite antigens in liver of immunized or naturally infected mice may be responsible for contrasting outcomes of their CTL responses. A controversial role of circumsporozoite protein (CSP) in host immune protection versus immune evasion against malaria is also highlighted by our findings.
This research was supported by SBRI innovation grant.
A study of mycobacterial lipid antigen-specific responses in a novel model of human group 1 CD1 (45.31)
Authors: Felio Kyrie , Nguyen Hanh , Dascher Christopher C , Choi Hak-Jong , Li Sha , Zimmer Michael I , Brenner Michael B , Wang Chyung-Ru 
Published: January 2023

Abstract
Group 1 CD1 presents mycobacterial lipid antigens to various T cell subsets. Although group 1 CD1-restricted responses have been implicated in anti-mycobacterial immunity in humans, little is known about their function, as mice lack group 1 CD1. To study group 1 CD1-restricted immune responses in vivo, we have generated human group 1 CD1-transgenic (hCD1Tg) mice. These mice express group 1 CD1 in a pattern similar to humans and support the development of group 1 CD1-restricted T cells, which can be induced both through infection with Mycobacterium tuberculosis (Mtb) and immunization with Mtb lipid antigens. hCD1Tg mice respond to Mtb epitopes recognized by human group 1 CD1-restricted T cells. The kinetics of these group 1 CD1-restricted responses resembles those of conventional T cells and is more rapid upon secondary exposure to antigen, but is distinct from those of group 2 CD1-restricted NKT cells. Furthermore, we find that the number of group 1 CD1-expressing antigen-presenting cells is altered over the course of Mtb infection, and correlates with the kinetics of group 1 CD1-restricted T cells. Taken together, our data demonstrate the viability of hCD1Tg mice as a model for the study of group 1 CD1-restricted T cells in anti-mycobacterial immunity. Such studies could lead to the development of new vaccines against Mtb. This work is supported by NIH grant NIH-A157460.
A systems biology approach on oral epithelial responses to periodontal pathogens (45.32)
Authors: Perumal Narayanan B , Kollipara Rahul Krishna , Janardhanam Srihari B , Srinivasan Mythily 
Published: January 2023

Abstract
Periodontitis, a chronic inflammatory disease, characterized by destruction of soft and hard tissues supporting the teeth affects ~20% in the US. The most widely implicated species in periodontitis are Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), and Fusobacterium nucleatum (Fn). We employ a systems biology approach with bioinformatic, genetic and biochemical methods to study differential responses of oral epithelial cells to specific pathogens. Using public domain microarray data, we compared gene expression profiles of cells infected with 4 different bacteria viz., Aa, Pg, Fn and Streptococcus gordonii (Sg). Various subsets of genes were differentially expressed (two-tailed T test) in these strains and clustered into 3 GO classifications: apoptosis, detection of external stimuli and cytokine activity. We have confirmed gene expression of some TLR genes (real time PCR) and cytokine genes (ELISA). Interestingly, apoptosis related genes were upregulated in cells stimulated with an oral commensal strain (Sg), but downregulated in cells exposed to pathogens (Pg/Aa). Also, many pro-inflammatory cytokines were upregulated in cells stimulated with Pg/Aa as compared to cells stimulated with Sg. In sum, our studies suggest that select periodontal pathogens enhance the survival of infected cells that secrete cytokines and mediate pathology.
PD-L1 expression in the CNS mitigates immune-mediated damage and morbidity at the cost of viral persistence (45.5)
Authors: Phares Timothy W , Atkinson Roscoe , Stohlman Stephen A , Bergmann Cornelia C 
Published: January 2023

Abstract
Interactions of programmed death 1 (PD-1) with its ligand (PD-L1) contribute to negative regulation of T cell function and dampened anti-viral responses. Following neurotropic coronavirus infection, PD-L1 is prominently upregulated on oligodendroglia, but only modestly on microglia and astrocytes. Concomitant expression of PD-1 on CNS infiltrating virus-specific CD8 T cells implicated PD-1:PD-L1 interactions in contributing to viral persistence in oligodendrocytes. This notion was confirmed by infection of PD-L1 deficient mice (B7-H1-/-). Virus is controlled more rapidly with significantly reduced numbers of infected oligodendrocytes in B7-H1-/- mice compared to wild-type. Accelerated control of infectious virus correlated with enhanced IFN-gamma production, critical to mediate anti-viral function in oligodendroglia. However, the lack of PD-L1 significantly increased disease severity and mortality. High Granzyme B levels and expression of TNF-alpha and iNOS were associated with the enhanced morbidity. Moreover, exacerbated disease in B7-H1-/- mice was coupled with greater axonal damage, while myelin loss was similar. The data demonstrate a significant contribution of PD-L1 in hindering viral clearance in glia cells, while protecting CNS tissue from immune-mediated damage. Supported by NIH NS18146.
Hyperactivation of CD8 T cells precedes exhaustion following viral infection (45.9)
Authors: Mackerness Kathryn J , Cox Maureen A , Stefanov Emily , Weaver Casey T , Harrington Laurie E , Zajac Allan J 
Published: January 2023

Abstract
Developing an in-depth understanding of the factors that regulate the induction, quality, and longevity of T cell responses is essential for devising rational strategies to promote anti-pathogen immunity. Cellular immune responses to many persistent viral infections are associated with the development of T cell exhaustion, which is characterized by the inability to produce anti-viral cytokines and proliferate. We have used IFN-γ-Thy1.1 cytokine reporter mice to dissect the initial activation events and functional traits of CD8 T cells as they respond following acute or persistent lymphocytic choriomeningitis virus (LCMV) infections. We show that a positive correlation exists between the expression of the Thy1.1 reporter molecule and IFN-γ production. Significantly, we show that expression of the Thy1.1 IFN-γ reporter molecule is elevated during the early stages of infections associated with the subsequent development of T cell exhaustion but is not as markedly upregulated following infections that become well controlled. This parallels the expression of PD-1, which is known to be upregulated on exhausted cells. Collectively, these data indicate that exhausted T cells, which are ineffective at eradicating persisting antigens, are derived from populations of cells that are initially hyperactivated. Thus, the blockade of this hyperactivation may represent a novel therapeutic approach to enhance cellular immunity against intracellular pathogens.
GATA-3 seals Notch-induced T cell commitment (46.1)
Authors: García-Ojeda Marcos E , Lemaître Fabrice , Richard-Le Goff Odile , Hendriks Rudi W , Cumano Ana , Di Santo James P 
Published: January 2023

Abstract
Transcription factors orchestrate T-lineage differentiation in the thymus. One early checkpoint involves Notch-1 stimulation of thymic immigrants that promotes T-cell commitment at the expense of the B-lineage program. The zinc-finger transcription factor GATA-3 is required for T cell specification, although its mechanism of action is unknown. Here we identify molecular targets of GATA-3 in early thymocytes and show that conditional ablation of GATA-3 expression in committed pro-T-cells unmasks a latent B-cell potential that is undetectable in normal T-cell precursors. Thus, GATA-3 delimits the developmental options in early thymocyte precursors by sealing T-cell fate induced by Notch signals.
M.E. García-Ojeda was funded by a Pasteur Foundation Fellowship. This work was supported by grants from the Institut Pasteur, the Inserm and the Ligue Nationale contre le Cancer. M.E. Garcia-Ojeda and F. Lemaître contributed equally to this work.
Ets1 is required for CD4 shut down during CD8 T cell differentiation in the thymus (46.11)
Authors: Zamisch Monica , Tian Linhua , Grenningloh Roland , Wildt Kathryn F , Ehlers Marc , Ho I-Cheng , Bosselut Remy 
Published: January 2023

Abstract
During the latest steps of thymocyte differentiation, T cell precursors that have rearranged TCRbeta and TCRalpha genes and express both CD4 and CD8 (double positive, DP) are selected into MHC II-restricted CD4 T cells, or MHC I-restricted CD8 T cells. Several transcription factors regulate this developmental transition: Gata3 and Zbtb7b promote the generation of CD4 cells, whereas Runx3 is important for CD8 T cell development by promoting the cessation of CD4 expression. The transcription factor Ets1 was reported to contribute to CD8 cell development, but its function in this process remains unclear. The present study has tested the hypothesis that Ets1 participates in CD8 cell differentiation by enabling Runx3 function. Expression of the P14 MHC I-restricted transgenic TCR in Ets1-deficient mice resulted in the generation of a population of CD8 lineage cells that failed to downregulate CD4, suggesting that Ets1 is required for the down-regulation of CD4. Transient transfection analyses failed to reveal any Ets1-Runx3 cooperation on CD4 expression. Rather, we found that Ets1 is necessary for the proper expression of Runx3 during CD8 thymocyte differentiation, and enforced expression of Runx3 restores CD4 shut-down in Ets1-/- MHC I-restricted thymocytes. These findings identify Ets1 as a key player during CD8-lineage differentiation and show that it acts at least in part by promoting Runx3 expression.
Impaired development of human Th1 cells in patients with deficient expression of STAT4 (46.14)
Authors: Chang Hua-Chen , Han Ling , Goswami Ritobrata , Nguyen Evelyn Thi , Pelloso David , Robertson Michael J. , Kaplan Mark H. 
Published: January 2023

Abstract
IL-12 activates STAT4, which is a critical regulator of inflammation and Th1 lineage development in murine systems. The requirement for STAT4 in the generation of human Th1 cells has not been examined thoroughly. Expression of the Th1 signature markers IFNγ, IL-12Rβ2, TNFα, and CXCR3 is greatly reduced in Th1 cultures of CD4 T cells isolated from Non-Hodgkin's Lymphoma patients following autologous stem cell transplant, which have acquired STAT4 deficiency, compared to control Th1 cultures. T-bet expression is not affected whereas IL-4 and IL-5 production is increased in patient Th1 cultures. STAT4 deficiency does not result in defects in the development of Th2 cells. Reconstitution of STAT4 in patient T cells allowed recovery of Th1 signature markers IFNγ, IL-12Rβ2, and CXCR3. In contrast, ectopic expression of IL-12Rβ2 in patient Th1 cells, which was also decreased with acquired STAT4-deficiency, did not rescue STAT4 expression, though it did increase IFNγ production to levels intermediate between control and patient samples. These results demonstrate that as in murine systems, STAT4 is required for optimal human Th1 lineage development.
Bcl6 is required for the development of T follicular helper cells marked by downregulation of P-selectin glycoprotein ligand-1 (46.7)
Authors: Poholek Amanda C , Hernandez Sairy , Chandele Anmol , Dong Xuemei , Odegard Jared , Kaech Susan M , Dent Alexander L , Craft Joe 
Published: January 2023

Abstract
Follicular helper T cells (TFH), defined by expression of the surface markers CXCR5, PD-1 and ICOS, are located in germinal centers where they provide help for B cell maturation through the secretion of IL-21. The transcriptional repressor Bcl6 is upregulated in TFH cells; however, the role that it plays in their development is unknown. We describe downregulation of P-selectin glycoprotein ligand-1 (PSGL1) as a new surface marker for TFH cells, and show that Bcl6 is required for the development of TFH cells that are PSGL1lo. Activated antigen-specific CD4 PSGL1lo T cells are enriched for expression of CXCR5 and PD-1, and highly upregulate IL-21 and Bcl6. Over-expression of Bcl6 leads to an increase in PSGL1lo TFH cells; however, it cannot rescue TFH cell development in the absence of ICOS. Consistent with others, we find B cells are required for full development of TFH cells, but not for downregulation of PSGL1, suggesting there are multiple steps in development of the subset. This data demonstrates that Bcl6, independent of ICOS signaling, is critical for the development of this lineage.
Fas induced apoptosis and Restimulation Induced Cell Death (RICD) is restricted to human effector memory CD4+ T subsets (46.8)
Authors: Ramaswamy Madhu , Cruz Anthony C , Siegel Richard M 
Published: January 2023

Abstract
FasL, a member of the TNF super family and its receptor Fas, are critical for peripheral tolerance of T cells via a mechanism of TCR-induced apoptosis of activated cells, termed RICD. Studies of RICD have shown that human CD4+T cells require TCR crosslinking to maximally undergo apoptosis when stimulated with Fas. However, the relative sensitivity of different CD4+ T cells subsets to Fas stimulation have not been addressed.
We found that essentially all cells sensitive to Fas stimulation derive from human memory CD4+ T cells and within the memory T cell pool, 'effector' cells lacking CCR7 and CD27 were consistently the most sensitive to Fas-induced apoptosis. Fas resistant 'central memory' as well as Fas-sensitive 'effector' cells had similar levels of Fas, induced FasL as well as pro-survival Bcl2/BCLxL levels. Biochemical studies indicate a highly efficient formation of the death-inducing signaling complex (DISC) in memory and effector T cells, which correlates with their apoptotic response. Finally, in ALPS patients with Fas mutations, resistance to Fas apoptosis was found predominantly in the effector memory subsets.
These results suggest for the first time that within a heterogeneous population of CD4+ T cells, effector memory T cells are specifically programmed for sensitivity to Fas-induced apoptosis compared with activated naïve T cells.
Differential sensitivity of Th1, Th2 and Th17 cells to Fas-mediated apoptosis (47.15)
Authors: FANG Yujiang , YU Shiguang , MULLEN Helen 
Published: January 2023

Abstract
In response to T cell receptor crosslinking, naïve CD4+ T cells can differentiate into three major subsets, Th1, Th2 and Th17 cells. Th17 cells play an important role in the pathogenesis of autoimmune diseases, but little is known about the regulation of apoptosis in Th17 cells. This study was undertaken to directly determine and compare the sensitivity of in vitro polarized Th1, Th2 and Th17 cells to Fas-mediated apoptosis and to determine if the anti-apoptotic molecule FLIP could inhibit apoptosis of each of the three subsets. Using several different methods, the order of sensitivity of T cell subsets to Fas-mediated apoptosis is: Th1&gt;Th17&gt;Th2. The greater sensitivity of Th17 cells compared to Th2 cells correlated with their higher expression of FasL. The decreased sensitivity of Th17 compared to Th1 cells correlated with higher expression of FLIP by Th17 cells. Transgenic overexpression of FLIP in T cells protected all three subsets from Fas-mediated apoptosis. Using splenocytes from IFNγ-/- mice, Fas-mediated apoptosis of T cells activated under Th1 polarization-inducing conditions were shown to be dependent on IFN-γ, while Fas-mediated apoptosis of Th17 and Th2 cells was IFN-γ independent. These findings provide new knowledge for understanding how survival of different subsets of T cells is regulated. This information might be useful for the design of new strategies to treat autoimmune diseases (NIH DK35527).
A European inter-laboratory pre-validation of in vitro assays to evaluate immunotoxicity (47.18)
Authors: Vandebriel Rob , Kleensang Andre , Koeper Lydia-Mareen , Biola-Vidamment Armelle , Fiechter Danielle , Carfi Maria , Gribaldo Laura , Corsini Emanuela , Pallardy Marc , Pieters Raymond , Vohr Hans-Werner , Van Loveren Henk 
Published: January 2023

Abstract
The new European Union REACH (Registration, Evaluation and Authorisation of new and existing CHemicals) policy will increase the use of laboratory animals if alternative methods are not available. The immune system is a target for many chemicals and drugs, with potential adverse effects on human health. We performed an inter-laboratory pre-validation of a set of in vitro assays to detect immunosuppressive capacity.
Human PBMC and murine and rat splenocytes were used. Six immunosuppressive compounds (benzo(a)pyrene, cyclophosphamide, methotrexate, rapamycin, dexamethasone, and urethane) and one non-immunosuppressive compound (D-mannitol) were tested. Basal cytotoxicity was assessed by measuring LDH-release. Human PBMC were stimulated with anti-CD3/anti-CD28, murine splenocytes with anti-CD3/anti-CD28, Con A, LPS, and anti-CD40/IL-4, and rat splenocytes with Con A and LPS. Cell proliferation and IFN-γ and TNF-α production were measured.
Five out of 6 immunosuppressive compounds could be identified as such, and the non-immunotoxic one came out as negative. This suggests that in vitro assays are able to detect immunosuppressive capacity. The sensitivity of the various species/stimulant/endpoint combinations ranged between 38 and 61%, while the specificity ranged between 67 and 100%. This suggests that from these results a choice for specific species/stimulant/endpoint combinations cannot be made.
Simultaneous detection of human IL-17, FoxP3 and other Th cytokines and surface markers by multi-color flow cytometry (47.4)
Authors: Li Li , Wu Xiaowei , Boyce Christopher , Wei Aili , Rohrer Jurg 
Published: January 2023

Abstract
Naïve CD4+ T helper cells can be induced to differentiate towards at least four types of committed helper T cells, namely T helper 1 (Th1), Th2, Th17 and regulatory T (Treg) cells according to the local cytokine milieu. The committed cells are characterized by the expression of specific transcription factors and by the cytokines released. Th17 cells are acknowledged to be instrumental in the response against microbial infection, but are also very efficient inducers of tissue inflammation and crucial initiators of organ specific autoimmunity. FoxP3+ Treg cells are necessary and sufficient to prevent autoimmunity throughout the life span of an individual. Immune and autoimmune responses are regulated by a fine balance between effector and regulatory T cells. There is a reciprocal relationship between pathogenic Th17 cells and Treg cells. Therefore the need to simultaneously detect multiple transcription factors and cytokines to define these populations is becoming increasingly important. In this report, we describe the simultaneous detection of IL-17 and FoxP3 as well as other Th cytokines (IFN-gamma and IL-4) and cell surface markers (CD3, CD4, CD25, CCR4, CCR6, CD45RA) using optimized buffer systems and protocols with multi-color flow cytometry.
Regulation of effector CD4+ T cell functions by Tbet (48.13)
Authors: Yeh Wen-I , Harrington Laurie E. 
Published: January 2023

Abstract
Many chronic inflammatory diseases, including inflammatory bowel disease and multiple sclerosis, are reported as Th1 mediated disorders; however, recent data from animal models suggests that Th17 cells are responsible for disease. Interestingly, IFNγ does not appear necessary for disease induction, but the key Th1-associated transcription factor Tbet is required. Therefore, we investigated the relationship between Tbet expression and effector CD4 T cell functions. Colitis induced by adoptive transfer of IFNγ KO CD4 T cells resulted in rapid disease and surprisingly over 60% of CD4 T cells expressed Tbet. We next examined the kinetics and functions of Tbet in vitro during Th1 and Th17 polarization. Tbet was upregulated within 24 hrs of stimulation under Th1 conditions, but remained low in Th17 cells. Moreover, Th17 differentiation was unaffected by Tbet deficiency. In order to define molecules regulated by Tbet, we examined gene expression in WT and Tbet KO CD4 T cells by microarray analysis. Consistent with our earlier results, Th1 associated cytokines were reduced but Th17 and Th2 associated genes were elevated in Tbet KO CD4 T cells. Notably, both the mRNA and protein levels of IL-10, an inhibitory cytokine, are increased in Tbet KO CD4 T cells suggesting a potential role for IL-10 in vivo. In conclusion, we propose that Tbet is critical in maintaining pathogenic Th1 cells and suppressing IL-10 expression.
Cellular Damage Determination in Newborn Balb/c Mice Skin Through Ro60, Hsp27, 60, 70 and 90 expression. (48.32)
Authors: Aguilera Galaviz Luis Alejandro , Olarte Saucedo Maricela , García Cabral Fernando , Lopez Luna Maria Argelia , Sanchez Rodriguez Sergio Hugo 
Published: January 2023

Abstract
Systemic Lupus Erythematosus is an autoimmune disease, it could be multi-systemic and characterized by auto-antibodies, especially antinuclear (anti-Ro, anti-La, anti-DNA), damage is mediated by immune complexes. The injuries in the skin of patients with SLE are aggravated by sun exposition (UV light and heat), altering the cellular expression and distribution of Ro60 antigen. In order to understand the damage mechanism involved in skin of patients with LES by environmental stressors, we developed a murine model focused in Ro60 and the heat shock proteins family (Hsp) expression. Objective: Determinate the cellular damage of the skin of newborn Balb/c mice, which was measured through the expression of the Ro60, Hsp27, 60, 70 and 90. Methods: Balb/c mice were exposed to different physical and chemical stressors like UV irradiation, heat (40ºC), H2O2, ethanol, HgCl2, camptotecine and cicloheximide drugs, in presence or absence of caspase 3 inhibitor. Skin extracts were obtained and the proteins Hsp27, 60, 70 and 90 were characterized and immunodetected by PAGE-SDS and Western blot using ECL, densytometric analysis was performed. Results: The Ro60 expression is modified with physical and chemical stressors, also it is increased mainly with UV light and heat, and decreased with caspase 3 ihnibitor. Hsp27, Hsp70, Hsp90 was similar to Ro60. The over expression of those proteins during the stress have the same profile. However using caspase 3 inhibitors before the stress, a down regulation are observed.