bluuebunny/crossref_metadata_2025_split huggingface dataset:
Inhibition of T cell Protein Tyrosine Phosphatase (TC-PTP) Enhances IL-18-dependent Hematopoietic Progenitor Expansion (111.57)
Authors: Bourdeau Annie , Trop Sebastien , Doody Karen , Dumont Daniel , Tremblay Michel 
Published: January 2023

Abstract
The clinical application of hematopoietic progenitor cell-based therapies for the treatment of hematological diseases is hindered by current protocols, which are cumbersome and have limited efficacy to expand the progenitor cell pool. We report that inhibition of T cell protein tyrosine phosphatase (TC-PTP), an enzyme involved in the regulation of cytokine signaling, through gene knockout results in a 9-fold expansion of functional hematopoietic progenitors in murine bone marrow (BM). This effect could be reproduced using a short (48 h) treatment with a pharmacological inhibitor of TC-PTP in murine BM, as well as in human BM, peripheral blood and cord blood. We also demonstrate that BM stem cells treated ex-vivo with a TC-PTP inhibitor can differentiate into functional progeny in vivo. We establish that the mechanism whereby inhibition of TC-PTP mediates its effects involves the IL-18 signaling pathway, leading to increased production of IL-12 and IFN-γ by progenitor cells. Together, our results reveal a previously unrecognized role for IL-18 in regulating progenitor cell expansion, and provide a novel and simple method to rapidly expand progenitor cells from a variety of sources using a pharmacological compound.
Notch Induces T Cell Receptor γδ Thymocytes to Differentiate along a Bipotent CD4 CD8 Double Positive Pathway (111.6)
Authors: Vandekerckhove Bart , Van Coppernolle Stefanie , Verstichel Greet , Snauwaert Sylvia , Langerak Anton , Kerre Tessa 
Published: January 2023

Abstract
It is well documented that in wildtype mice TCRγδ+ cells differentiate along a double negative (DN) pathway whereas TCRαβ+ cells differentiate along the double positive (DP) pathway, suggesting that the TCR itself induces lineage differentiation. Under experimental conditions and in genetically modified mice, however, evidence was presented suggesting that rather than TCR itself, TCR signal strength and notch determine lineage choice. In the human thymus, a “chimeric” DP TCRγδ+ cell population is present and constitutes a sizeable fraction of the γδ population. We asked the question whether these cells belong to the αβ or γδ lineage or whether these cells are bipotent. We found that TCRγδ DP cells are bipotent cells since strong TCR signals induces differentiation to TCRγδ cells, whereas Notch activation induces TCRαβ lineage differentiation. We furthermore could show that Notch signaling diverts TCRγδ DN cells to the DP pathway and induces strong proliferation. In line with these findings, TCRγδ+ acute lymphoblastic leukemias (ALL) with activating Notch1 mutations follow the DP differentiation pathway, whereas the DN ALL cells are devoid of these activating Notch1 mutations. We were able to confirm that also in vivo TCRγδ DP have rearranged TCRβ chains, actively rearrange the TCRα locus and delete the TCRδ locus (αβ lineage). Using TCRα rearrangements as a lineage marker, we could show that a subpopulation of mature TCRγδ cells is derived from DP cells.
Chitinase 3-like 1 enhances bacterial adhesion to colonic epithelial cells through the bacterial chitin-binding motif (112.9)
Authors: Mizoguchi Emiko , Lee In-Ah , Kamba Alan , Santhanam Radhika 
Published: January 2023

Abstract
Chitinase 3-like-1 (CHI3L1) is an inducible molecule in colonic epithelial cells (CECs), and the chitin-binding motif (CBM) in CHI3L1 regulates host-microbial interactions. To examine the existence of CBMs in E. coli, we used ChtBD3 as a search in the SMART database. We found that 11 genomes of E. coli strains contain a gene encodes for ChiA, which has multiple CBMs. There are 5 different amino acids (Q362K, E370K, V378A, V388E & E548V) in CBMs, and the E.coli are grouped into pathogenic- and non-pathogenic strains by substitution. To examine the role of CBMs in E. coli, we generated a deletion mutant (ΔchiA) of the Adherent Invasive E. coli (AIEC) LF82, and performed in vitro assays. LF82 ΔchiA has 50% less adhesion to CECs as compared to LF82. This phenotype is fully recovered when the mutant carries out pHSG575 with LF82 ΔchiA, but not withLF82 chiA plus 5 mutations or E. coli K12 chiA. To test the in vivo influence, B6 mice were treated 2% DSS to enhance the CHI3L1 on CECs and were orally inoculated with LF82 (WT or ΔchiA) for 15 days. In contrast to LF82, mice infected with LF82 ΔchiA showed less body weight loss, a lower clinical score, less stool bacteria and less cellular infiltration in the colon. A significantly increased number of E. coli adhered/invaded the colon in LF82-infected mice, as compared to LF82 ΔchiA-infected or control mice. In conclusion, CBMs of AIEC play a critical role in their adhesion to CHI3L1-expressing CECs during colitis.
HCV genotype 1a representative sequence elicits broad CD8+ T cell responses (113.10)
Authors: Burke Kelly , Munshaw Supriya , Osburn William , Levine Jordana , Liu Lin , Ray Stuart , Cox Andrea 
Published: January 2023

Abstract
Vaccines designed to prevent or to treat hepatitis C viral infection must achieve maximum cross-reactivity against widely divergent circulating strains. Computer-generated sequences minimize genetic distance between circulating strains, and can be generated as a consensus sequence (most common amino acid at each position) or a representative sequence (derived with Bayesian and maximal likelihood phylogenetic tools). Broad recognition of such sequences in HCV has not been demonstrated. Consensus and representative sequences were generated from 390 full-length HCV genotype 1a polypeptide sequences. Sequence mutations in known epitopes were identified by longitudinal sequencing of HCV-infected subjects. Peptides encoding consensus, representative, and variant epitope sequences were tested for the capacity to expand CD8 T cells from HCV-infected subjects and to elicit cross-reactive responses by IFNg ELISpot. The representative and consensus sequences were identical for 9/11 epitopes examined. T cell lines generated against representative sequence peptides were generally cross-reactive to consensus sequence and natural sequence variants. Furthermore, representative sequence peptides generated more robust T cell responses than did natural sequence variant peptides. The broadest recognition of highly diverse circulating HCV strains was achieved using CD8+ T cells expanded with representative sequence HCV. These data support the use of representative sequence in HCV vaccine design.
Transcription factor expression profiling to characterize differentiation of antigen-specific CD8 T cells induced by a live attenuated viral vaccine. (113.18)
Authors: Billingsley James , Rajakumar Premeela , Salisch Nadine , Kuzmichev Yury , Hong Henoch , Connole Michelle , Reeves R. , Johnson R. 
Published: January 2023

Abstract
Live attenuated SIV (SIVΔnef) is the most effective approach to induce protection to pathogenic SIV challenge in macaques. Protective immunity, in part mediated by CD8 T cell responses, correlates with phenotypic maturation of SIV-specific T cells, but remains incompletely understood. Expression profiling of T cell transcription factors (TF) offers a novel approach to characterize antigen-specific T cell differentiation. Highly parallel qRT-PCR was used to characterize the expression of 21 TF in naïve, central, transitional, and effector memory T cell subsets, and in SIV-specific CD8 T cells obtained at times associated with lesser protection (wk 5) and greater protection (wk 20) following SIVΔnef vaccination. Unsupervised clustering organized samples from CD4 and CD8 T cells into groups concordant with cell surface phenotype. TF expression in SIV-specific CD8+ T cells segregated by time, with wk 20 cells exhibiting increased expression of TF associated with both maintenance of quiescence (TCF7, BAZF) and promotion of effector function (Eomes, T-Bet). Different expression profiles were observed in T cells specific for different SIV epitopes, consistent with different epitope escape kinetics. TF expression profiling suggests T cell responses correlated with protection may include characteristics of both memory and effector cells. TF expression profiling can provide data complementary to the analysis of memory cell differentiation based on classical phenotypic markers.
Vaccination of endangered Hawaiian geese (Nene, Branta sandvicensis) against West Nile Virus using a protein-based vaccine (WN-80E) (113.7)
Authors: Jarvi Susan , Hu Darcy , Misajon Kathleen , Coller Beth-Ann , Wong Teri , Lieberman Michael 
Published: January 2023

Abstract
While West Nile Virus (WNV) has not yet been reported in Hawaii, eventual introduction appears unavoidable. Nene are endemic, endangered Hawaiian geese, and are susceptible to WNV. The goal of this study is to demonstrate that a vaccine developed against WNV for humans (WN-80E) is also highly immunogenic in Nene, and does not produce adverse biological effects. Six captive, non-breeding Nene were immunized with two 10 µg doses (four weeks apart) of the WN-80E recombinant protein adjuvanted with Montanide ISA720. Two Nene were similarly injected with mock preparation as controls. Blood samples were collected prior to the first dose, at two weeks and six months after the second dose. Detailed observations were noted for 48 hours post injection, followed by daily observations of health status to monitor for adverse biological effects. WNV-specific antibody titers were determined by an endpoint ELISA. An unpaired t-test demonstrated significantly higher geometric mean titers for immunized vs. control groups at two weeks post dose two (4129 and 100, respectively, p = 0.0096) and at 6 months post dose two (246 and 63, respectively, p = 0.0023). No swelling was reported at any time during the study at the site of injection. No serious adverse biological effects were observed directly due to the immunization. The vaccine containing the WN-80E and Montanide ISA720 adjuvant appears to be safe and effective in Nene, and should be considered for widespread use in Hawaii.
Critical role of the inflammasome during central nervous system Staphylococcus aureus infection (114.1)
Authors: Hanamsagar Richa , Kielian Tammy 
Published: January 2023

Abstract
Staphylococcus aureus (S. aureus) infection in the central nervous system (CNS) can lead to abscess formation typified by widespread inflammation and necrosis. A key cytokine driving inflammation during this process is IL-1β, which is processed into its active form by the inflammasome. Our recent studies demonstrated that the NOD-like receptor 3 (NLRP3) inflammasome and its adapter protein ASC are required for IL-1β secretion by microglia, the resident immune cells of the CNS. To understand the role of NLRP3 inflammasome activation during brain abscess formation, C57BL/6 wild type (WT), ASC knockout (KO), and NLRP3 KO mice received intracerebral infections with 1000 cfu S. aureus. Interestingly, ASC KO mice succumbed to infection within 24 h, yet bacterial burdens were identical to WT animals. Inflammatory mediators including CXCL1, CXCL10, CCL2, and IL-6 were significantly reduced in ASC KO mice, suggesting that ASC loss may impact processes extending beyond IL-1β cleavage. To extend the survival of ASC KO mice, animals were infected with a 1-log lower inoculum, revealing significant reductions in neutrophil and CD4+ T cell infiltrates compared to WT animals. In contrast, NLRP3 loss had minimal impact on brain abscess development. Collectively, these results indicate that ASC is involved in diverse pathways affecting inflammatory mediator expression and immune cell infiltration into the CNS, which may occur independently of its classical inflammasome actions.
HMGB1 facilitates inflammasome-dependent IL-1β release (114.2)
Authors: Keyel Peter , Vernon Philip , Chu Jessica , Lotze Michael , Salter Russell 
Published: January 2023

Abstract
HMGB1 is a potent mediator of late-phase shock, endotoxin toxicity and other inflammatory processes. HMGB1 is released passively from necrotic cells, and also following inflammasome activation in macrophages and dendritic cells. We hypothesized that caspase-1 activated nucleases liberate HMGB1 from the nucleus and allow for subsequent cellular release. To test this hypothesis, we measured inflammasome-mediated HMGB1 release in the presence of various nuclease inhibitors. Inhibitors of DNase γ blocked inflammasome-activated HMGB1 release. Surprisingly, DNase γ inhibitors also blocked IL-1β release, suggesting that nuclear proteins play a role in nonconventional cytokine secretion. To determine if HMGB1 regulates IL-1β release, we tested its intracellular association with IL-1β and the ability of HMGB1-deficient cells to secrete IL-1β. We found that HMGB1 co-immunoprecipitated with IL-1β in cells, indicating an interaction prior to IL-1β release. Furthermore, genetic disruption of HMGB1 in dendritic cells reduced their ability to secrete IL-1β in response to inflammasome activation. Taken together, these data suggest that HMGB1 and other nuclear proteins regulate IL-1β release through an inflammasome dependent process.
PKR is required for NLRP3 inflammasome activation and HMGB1 release (114.7)
Authors: Lu Ben , Nakamura Takahisa , Inouye Karen , Valdes-Ferrer Sergio , Olofsson Peder , Li Jianhua , Wang Haichao , Erlandsson-Harris Helena , Kalb Thomas , Andersson Ulf , Yang Huan , Chavan Sangeeta , Hotamisligil Gökhan , Tracey Kevin 
Published: January 2023

Abstract
The NLR family pyrin domain-containing 3 (NLRP3) inflammasome, an intracellular oligomeric protein complex, regulates release of caspase-1 activation-dependent cytokines, including IL-1β, IL-18, and high-mobility group box 1 (HMGB1). During the course of studying HMGB1 release mechanisms from macrophages, we unexpectedly observed that double-stranded RNA dependent protein kinase (PKR) is required for NLRP3 inflammasome activation. PKR inactivation by genetic deletion or pharmacological inhibition blocked NLRP3 inflammasome activation in response to double-stranded RNA, ATP, monosodium urate, adjuvant aluminum, and live E coli. Overexpression of PKR significantly enhanced the reconstituted NLRP3 inflammasome-induced caspase-1 activation and IL-1β maturation; whereas blocking PKR significantly inhibited the reconstituted NLRP3 inflammasome activation. PKR physically interacts with NLRP3, and PKR autophosphorylation is required for efficient PKR and NLRP3 interaction. Importantly, PKR autophosphorylation in a cell free system with recombinant NLRP3, ASC and pro-casapse-1 reconstitutes inflammasome activity. Together, these results indicate that PKR is required for inflammasome activation, and that it should be possible to target this molecule to inhibit inflammasome activity during inflammation.
Cytosolic K+ and extracellular ATP as regulators of NLRP3 inflammasome activation and the IL-1β secretion response of macrophages to crystalline stimuli. (114.9)
Authors: Katsnelson Michael , Smith Corey , Dubyak George 
Published: January 2023

Abstract
The crystalline particulate Alum has widely been used as an adjuvant in vaccines. Alum and other crystalline particulates also stimulate NLRP3/caspase-1 inflammasome assembly and secretion of mature IL-1β via poorly understood mechanisms that involve decreases in cytosolic [K+]. This study characterized mechanisms for K+ efflux in LPS-primed murine bone marrow-derived macrophages (BMDM) stimulated with either Imject Alum or the Na+, K+-ATPase inhibitor ouabain. Treatment with Alum for 3 h induced robust secretion of IL-1β that was similar in the absence or presence of antagonists of the P2X7 receptor, an ATP-gated K+ efflux channel known to regulate NLRP3 inflammasome assembly. However, the ATP-scavenging enzyme apyrase markedly reduced this IL-1β secretion, suggesting that NLRP3 activation and IL-1β release induced by Alum crystals is at least partially dependent on a P2X receptor subtype(s) other than P2X7. Current experiments are testing possible involvement of P2X4 receptor channels. Treatment of LPS-primed BMDMs with ouabain for 6 h also induced significant release of IL-1β. We hypothesize that ouabain causes NLRP3 activation and IL-1β secretion by inhibiting the steady-state Na+, K+ATPase activity which counteracts the slow efflux of cytosolic K+ via leak/rectifier-type K+ channels.
CD4+ and CD8+ T cell β antigen receptors have different and predictable V and J gene usage and CDR3 lengths (115.10)
Authors: Robins Harlan , Emerson Ryan , Sherwood Anna , Desmarais Cindy 
Published: January 2023

Abstract
CD8+ and CD4+ T lymphocytes have different and complimentary roles: CD8+ (cytotoxic) T cells directly kill cells presenting non-self epitopes while CD4+ (helper) T cells regulate the immune response to a particular antigen. These T cells recognize antigens by binding to epitope MHC protein complexes, with CD8+ T cells binding class I and CD4+ T cells binding class II MHC protein complexes. While it’s known that the CD8 and CD4 proteins determine which MHC class the T cells bind, it is unknown if the antigen receptor sequences vary between the two populations. The diversity of possible receptors is huge and until recently this diversity precluded the possibility of capturing the antigen receptor repertoire. Adaptive Biotechnologies has developed a novel method that amplifies rearranged T cell receptor β (TCRB) CDR3 sequences and uses high throughput sequencing to sequence millions of TCRB CDR3 chains. To determine if the two populations of T cells have unique TCRB profiles, PBMC from 19 unrelated individuals were sorted 4 ways into CD4+ and CD8+ naïve and memory T cells. V and J gene usage and CDR3 sequence length could distinguish the TCRB CDR3 sequences amplified from CD4+ and CD8+ T cells. These sequence differences can be exploited and used to estimate the proportion of CD4+ to CD8+ T cells in a mixed population of T cells. These data indicate a biological difference between the antigen receptors that bind class I versus class II MHC protein complexes.
Pregnancy alters T-cell development in the mouse thymus (115.11)
Authors: Jaiswal Mukesh , Mallers Timothy , Chaouat Gerard , Gilman-Sachs Alice , Beaman Kenneth 
Published: January 2023

Abstract
The syngeneic and allogeneic mating combinations were used to assess the effect of pregnancy-associated hormones and fetal antigens on different T-cell subsets and their a2V-ATPase (a2V) expression during pregnancy. Our data shows that a2V was significantly (p≤0.05) increased in thymus recovered from normal mating combinations (♀BALB/cX♂BALB/c and ♀CBAX♂BALB/c) when compared with abortion prone mating combinations (♀CBAX♂DBA). The percentage of NK-T cells (CD3+NKp46+ cells) was significantly (p≤0.05) increased in all studied mating combinations on days 12 and 16 of pregnancy when compared to their respective non-pregnant females and pups. The expression of a2V on NK-T cells was also significantly (p≤0.05) increased as their numbers was increased. The numbers of CD3+CD4+CD8- cells were significantly (p≤0.05) increased, while the numbers of CD3-CD4-CD8- and CD3+CD4+CD8+ cells were significantly (p≤0.05) decreased in all the mating combinations when compared to their respective non-pregnant females and pups. This dynamic change in the numbers of T-cells subsets during pregnancy suggests that T-cells development and differentiation is altered in pregnancy. Moreover, a2V may play a crucial role in differentiation of NK-T cells from different T-cells subsets. Future studies to decipher the mechanism of action of a2V in thymic T-cells development and differentiation will lead to valuable knowledge of thymus development as well as its role in maternal fetal allogeneic relationship.
The role of ID3 in iNKT cell development (115.12)
Authors: Verykokakis Mihalis , Kee Barbara 
Published: January 2023

Abstract
Invariant NKT cells comprise an unconventional T cell population with innate characteristics. Like the conventional αβ T cell lineage, they develop in the thymus from DP precursors after receiving TCR-dependent positive selection signals, although utilizing different pathways. iNKT cells further mature through a series of differentiation stages and acquire their effector properties intrathymically. The ID3 protein is required to reduce E protein activity for proper selection of DP progenitors towards conventional thymocyte lineages. Here, we investigate the requirement for ID3 in the thymic development of iNKT lineage, in vivo. We show that the frequency of iNKT cells in the Id3-/- thymus is higher than in WT littermates, in a cell-intrinsic manner. Id3-deficient post-selected DP cells express higher levels of the canonical Vα14-Jα18 TCRα chain transcripts. Additionally, the transcription factor PLZF, a molecular determinant of the NKT lineage, is expressed in a subset of post-selected DP cells in the Id3-/- mice. However, further thymic iNKT differentiation is blocked at an immature stage. Taken together these results suggest that ID3 restricts the innate T cell potential of DP precursors thus contributing to the lineage fate decisions at the DP stage. However, ID3 is necessary for the completion of the innate T cell program during iNKT differentiation. Ongoing gene expression analysis will identify potent E protein target genes essential for NKT development
Tcf-1 in early lymphoid development (115.14)
Authors: Germar Kristine , Dose Marei , Konstantinou Tassos , Zhang Jiangwen , Wang Hongfang , Lobry Camille , Arnett Kelly , Blacklow Stephen , Aifantis Iannis , Aster Jon , Gounari Fotini 
Published: January 2023

Abstract
In hematopoiesis, the transcriptional networks governing lineage choice from pluripotent progenitors to mature lymphoid cells remain poorly understood. Although some key factors associated with the specification and commitment of T cells have been described, the functional roles of key regulators in orchestrating these developmental programs remain unclear. Using a loss-of-function model, we established the importance of T-cell factor 1 (Tcf-1), a T-cell specific mediator of Wnt signaling, as a gatekeeper of T-cell fate. We show that Tcf-1 is directly activated by Notch signals. Activation of Notch signaling in uncommitted precursors by the thymic stroma is known to initiate the T-cell differentiation program, and we show that Tcf-1 is one regulator first induced in these precursors. Tcf-1 is required at the earliest phase of T-cell determination for progression beyond the early thymic progenitor (ETP) stage, and this requirement is cell intrinsic and Wnt independent. The global expression profile of Tcf-1 deficient progenitors indicates that basic processes of DNA metabolism are downregulated in its absence, and the blocked T-cell progenitors die by apoptosis. While Tcf-1 is dispensable for the development of bone marrow (BM) progenitors and their migration through the blood, we present evidence for a novel role for Tcf-1 in early BM natural killer (NK) development. Our data thus demonstrate multiple roles for Tcf-1 in the roadmap of lymphoid development.