bluuebunny/crossref_metadata_2025_split huggingface dataset:
Enhanced susceptibility to alcohol-induced bacterial translocation, immune activation, and persistent inflammation in patients with alcoholic hepatitis: a prospective observational study
Authors: Li Wei , Amet Tohti , Xing Yanyan , Lan Jie , Liangpunsakul Suthat , Puri Puneet , Kamath Patrick , Sanyal Arun , Shah Vijay , Radaeva Svetlana , Crabb David , Chalasani Naga , Yu Andy 
Published: January 2023

Abstract
Up to 30% chronic heavy drinkers develop a spectrum of severe alcoholic liver diseases, including alcoholic hepatitis (AH). Alcohol-induced impairment of intestinal integrity and microbial translocation into the portal vein lead to immune activation and play a major role in the pathogenesis of AH. To better understand why AH develops only in some heavy drinkers and the effect of alcohol abstinence on immunity, we conducted a prospective study to compare plasma LPS levels and peripheral blood immunoprofiles between 68 AH patients and 65 heavy drinkers without liver disease (HDC) who were followed up to 12 months. Plasma samples were also obtained from 31 health donors (HD). Plasma LPS was detected in a higher portion of AH patients than HDC (80% vs 49%) at baseline, and in only 8% HD. However, LPS levels were not different between AH an HDC subjects with detectable LPS. Compared with HDC, AH patients had higher baseline plasma levels of sCD14, sCD163, sICAM-1, sVCAM-1, IL-22, TNF-α, IL-8, IP10, IL-4, IL-6, IL-9, IL-10, FGF2, IL-7, IL-15, and TGF-α, but lower levels of the anti-inflammatory chemokine MDC. AH patients also had more activated, yet dysfunctional monocytes, T cells, and B cells as they expressed higher levels of CD38 and CD69, but lower levels of HLA-DR, CD80, and CD86. In spite of having significant bacterial translocation, HDC showed little evidence of immune activation. With alcohol abstinence, levels of many dysregulated immune markers normalized in AH patients. However, TNF-α, IL-8, IL-10, sCD163, and sVCAM-1 levels remained higher.

Conclusion
AH patients were more susceptible to alcohol-induced bacterial translocation and immune activation. Alcohol abstinence reduced but did not fully reverse immunological abnormalities.
Lack of central memory cell generation by PD-1-deficient T cells under helper-deficient conditions
Authors: Kim Woojin , Ju Ji-Min , Choi Eun Young 
Published: January 2023

Abstract
An immune check point molecule PD-1 is associated with CD8 T cell exhaustion in tumor-infiltrating and chronic infection situations. Previously, we have demonstrated that CD8 T cells specific for a transplantation antigen, minor histocompatibility antigen H60, are exhausted when primed under CD4 helper-deficient conditions, showing PD-1hi phenotype, and the exhaustion is ascribed to inefficient antigen-clearance. We examined whether the helpless CD8 T cells could respond to re-challenge with H60 in absence of PD-1-mediated signaling using PD-1-deficient mice. Upon re-challenge with H60 of the PD-1-deficient mice, which had been primed against H60 under helper-deficient conditions, the H60-specific CD8 T cells expanded vigorously, exhibiting a memory-like response. Despite the restored proliferative potential to the antigen-re-encounter, the H60-specific PD-1-deficient CD8 T cells did not generate CD44hiCD6Lhicentral memory T cells under helper-deficient priming condition. They exhibited an exhausted status, with an increased CXCR3+KLRG1+ Tint cell fraction. When transferred to naïve hosts, PD-1-deficient helpless CD8 T cells showed reduced generation of central memory cells compared to their wild type counterparts. These results suggest that PD-1 signaling influences fate of effector CD8 T cells and provide information regarding CD8 T cell memory generation.
CD46 is a potent co-stimulatory receptor on CD8+ T cells
Authors: Hansen Aida Solhoej , Slater Josefine , Biltoft Mette , Höllsberg Per 
Published: January 2023

Abstract
Upon activation CD8+ T cells may proliferate extensively and become specialized effector cells with an important role in clearing viral infections. Similar to CD4+ T cells, precursor CD8+ T cells are thought to be dependent on a co-stimulatory signal through CD28. CD46 is another co-stimulatory receptor on CD4+ T cells that promotes proliferation and secretion of IFNγ followed by secretion of high amounts of IL-10, thereby inducing a regulatory phenotype. To our knowledge, the co-stimulatory function of CD46 on CD8+ T cells is yet unknown.
CD4+ and CD8+ T cells were isolated from peripheral blood mononuclear cells from healthy donors and activated in vitro with plate-bound αCD3/αCD28 or αCD3/αCD46 for 72 hrs. The proliferation was examined by CFSE staining and the secretion of IFNγ and IL-10 by ELISA. As expected, we observed increased proliferation as well as secretion of IFNγ and high amounts of IL-10 in the CD4+ T cells upon CD46 co-stimulation compared to CD28 co-stimulation. Intriguingly, CD46 co-stimulation also promoted proliferation of CD8+ T cells and secretion of high amounts of IFNγ compared to CD28 co-stimulation. In contrast to the regulatory phenotype of CD4+ T cells, it appears that CD46-costimulation promotes the effector response of CD8+ T cells, as we observed elevated IFNγ and only limited induction of IL-10 secretion in CD8+ T cells when compared with CD4+ T cells.
In conclusion, these results suggest that CD46 co-stimulation has a functional significance on both proliferation and effector functions of CD8+ T cells.
TET2 and TET3 regulate stability and function of regulatory T cells.
Authors: Nakatsukasa Hiroko , Yoshimura Akihiko 
Published: January 2023

Abstract
Regulatory T cells (Tregs) play a pivotal role in regulating immune response and maintaining immunological tolerance, and are characterized by the expression of Foxp3. Thymus-derived Tregs have been shown to be relatively stable compared with iTregs generated from naive T cells with TGFβ in vitro. Treg stability is maintained by the epigenetic modification, such as DNA demethylation of Treg-specific demethylation regions (TSDRs) during Treg development in the thymus. In fact, the TSDRs of iTreg are hypermethylated. Although DNA demethylase, ten-eleven translocation (TET) family proteins have been proposed to be involved in the DNA demethylation of TSDRs, actual contribution of TET family proteins in Treg stability and function remains to be elucidated. To investigate the role of TET proteins in Tregs, we generated Treg-specific Tet-deficient (Foxp3YFPCre+Tet2f/fTet3f/f; FDKO) mice. FDKO mice died at an average of 20 weeks of age with splenomegaly and lymph adenopathy accompanied by uncontrolled activation of T cells and B cells. These FDKO mice developed inflammation in the gut, and both Foxp3+ and Foxp3− T cells were expanded and produced high levels of IL-17 especially in gut mucosa and mesenteric lymph nodes. Transfer experiment of FDKO Tregs into lymphopenic mice revealed that Foxp3 expression in FDKO Tregs was unstable and Treg cells which have lost their Foxp3 expression were accumulated. TSDRs of FDKO Tregs were hyper-, but not completely, methylated compared with WT Tregs, indicating that there are TET2/3 dependent and independent mechanisms for TSDR demethylation. These data indicate that TET2 and TET3 are redundantly involved in TSDR demethylation and are important for the stability and homeostasis of Tregs.
Regulation of CD11c+ T-bet+ IgM+ Memory B-cells by TNFα
Authors: Popescu Maria , Cabrera-Martinez Berenice , Winslow Gary M. 
Published: January 2023

Abstract
Our studies of ehrlichial infection have identified a population of CD11c+ T-bet+ IgM memory B-cells that arise within 30 days post-infection. We and others have shown that CD11c+ T-bet+B cells require for their development or function a number of factors, including T-bet, IFNγ, CD4 T-cells, IL-21, and TLRs. Here, we addressed a possible role for TNFα in the generation of CD11c+ T-bet+IgM memory B-cells, because aside from being a major inflammatory cytokine, it can regulate germinal center development, and mediate apoptosis. We observed an 8-fold increase in the number of CD11c+IgM memory B-cells (also CD73+) in TNFα-deficient mice, relative to wild-type mice, on day 16 post-infection. The B cell population persisted at much higher frequency and number on day 30 post-infection. The changes in the IgM memory cell population were associated with a concomitant 10-fold relative increase in the frequency of GL7+ CD38lo CD95+germinal center B cells in TNFα-deficient mice. These latter data contrast with published studies that have reported that TNFα is required for GC development, suggesting that the requirement we have observed for TNFα is context-dependent. These data suggest that there may be a positive relationship between GC development and generation of CD11c+ IgM memory cells, and that TNFα may act to mediate IgM memory cell homeostasis by regulating apoptosis. Ongoing studies will address mechanisms whereby TNFα contributes to the genesis and maintenance of CD11c+ T-bet+IgM memory B-cells.
Ubiquitin Signaling: Systematic Profiling in the Immune Response
Authors: Song Changcheng , Lu Xiaolong , Akimov Vyacheslav , Singh Rajesh , Mason Mark , Blagoev Blagoy 
Published: January 2023

Abstract
Cell signaling induced by cytokine receptors is regulated by ubiquitin (Ub) modification. Ub forms polymerized chains to encrypt the coding for the alteration of signaling molecules in immune response. Here we report a systematic approach for profiling Ub coding. Ubiquitylation of specific target proteins in stimulated and unstimulated human PBMCs and T Leukemia (Jurkat) cells are profiled by ELISA and Immunoblotting UBiTest and Ubi-Omics methods. The cells are treated with the de-ubiquitylase USP7 inhibitor P50378, proteasome inhibitor bortezomib, or a ubiquitin ligase inhibitor. Poly-ubiquitylated cellular proteins is affinity isolated using Tetra Ub Binding Entities (TUBEs), which bind Ub with high affinity (nmol). TUBEs that are selective for isolated K48, K63 and M1 (linear) poly-ubiquitylated proteins are employed. Then, these polyubiquitylated proteins were digested with Lys-C and immunoprecipitated with UbiSite antibody. Ub-rich peptides were analyzed by LC-MS/MS. This UbiSite-Omics approach affords the advantages of both identifying N-terminal ubiquitylation and distinguishing ubiquitination from modification by Ub-like proteins. These data provide proteome-wide profiling of Ub-omics not achievable by traditional proteomics methods and suggest that the TUBE and UbiSite based technology will reveal new Ub landscapes in immune cells. This novel approach will facilitate the discovery of pharmacological markers for drugs, and enable cell biologists to identify patterns of polyUb proteins.
Effects of temporary BLyS treatment of rhesus macaques
Authors: Scholz Jean , Pahar Bapi , Madej Joanna B , Soldemo Martina , Trunch Wendy Lala , Guenaga Javier , Myles Arpita , Tran Karen , Wyatt Richard T , Hedestam Gunilla B Karlsson , Lackner Andrew , Cancro Michael 
Published: January 2023

Abstract
BLyS is a survival cytokine that regulates peripheral B cell numbers and transitional B cell throughput. In mice, exogenous BLyS administration yields elevated pre-immune B cell numbers, an increased proportional representation of transitional B cells, and shifts in repertoire composition (1–3). Consistent with these observations, BLyS pretreatment alters the quality of antibody responses to HIV gp140 in mice, yielding enhanced neutralizing responses (4). While it is assumed that BLyS plays analogous roles in other species, similar studies in nonhuman primates are lacking. Accordingly, we have assessed the effects of exogenous BLyS treatment in rhesus macaques. Here we treated juvenile rhesus macaques with 0.05 mg/kg recombinant human BLyS over ten days. Physical exams and blood cell counts indicate no adverse effects of treatment except a decrease in lymphocyte counts during the treatment periods. We find that the numbers and proportional representation of transitional B cells (CD20+ IgM+ CD10+) increase during and immediately following BLyS treatment, returning to pre-treatment levels within 20–30 days. There is a small increase in plasma anti-dsDNA antibody, and a significant increase in anti-recombinant human BLyS antibody indicating immunogenicity in this species. Together, these results indicate that BLyS plays similar roles in rodents and primates. They further suggest that deliberate manipulation of BLyS may be an effective approach in rhesus macaques for expanding B cell repertoire diversity and altering the quality of antibody responses.
Estimate of within clones and between clone selection suggests a multi-stage selection process in the production of high affinity B cell receptors.
Authors: Benichou Jennifer Israel Cohen , Louzoun Yoram 
Published: January 2023

Abstract
During an immune response, B cells undergo a process of clonal expansion, somatic hypermutation of the immunoglobulin genes and affinity-dependent selection. Over a lifetime, each B cell may participate in multiple rounds of affinity maturation as part of different immune responses. We characterized these two time-scales for selection which are apparent in the structure of B cell lineage trees. The “trunk” of the tree consists of mutations that are shared across all members of a clone, and the branches form a “canopy” consisting of mutations that are shared by a subset of clone members.
We show that the framework regions are subjected to a negative selection at all stages. However, CDR regions are affected to purifying and antigen-driven positive selection on the short term, which leads to a net positive selection in the long term. Overall, we found that long-term selection is strongly dependent on the heavy chain variable gene family.
To the selection in the tree canopy, we propose a novel mixed lineage tree/sequence based method to detect within population selection as defined by the effect of mutations on the average number of offspring. Specifically, we propose to measure the log of the ratio between the number of leaves in lineage trees branches following synonymous and non-synonymous mutations (LONR), and show that selection in CDR region occur at a different stage.
Adenosine regulation of danger signaling
Authors: Cekic Caglar , Akdemir Imran , Koyas Altay , Kayhan Merve , Savas Ali Can 
Published: January 2023

Abstract
Metabolic and immune related activities converge as main triggers of adenosine accumulation in extracellular space. Adenosine by engaging adenosine A2A and A2B receptors strongly suppresses innate and adaptive immune responses. Although adenosine receptors are being targeted in preclinical and clinical studies how different danger signals are regulated by adenosine is poorly understood. Here we showed that adenosine receptor stimulation strongly inhibited inflammatory responses while sparing type-I interferon responses downstream of different danger signals in dendritic cells and macrophages. Mechanistically, danger signals associated with MyD88-dependent inflammatory pathways such as LPS and CpG but not the danger signals associated with IRF3/Type-I interferon pathways such as pA:U and cGAMP increase the expression adenosine A2A and A2B receptors. Adenosine was shown to increase the expression of NR4A nuclear hormone receptors to inhibit NF-κB activation. Although adenosine did not influence NF-κB phosphoactivation expression of anti-inflammatory NR4A1 was increased after adenosine receptor stimulation in the presence of TLR ligands known to activate MyD88 pathway but not in the presence of cGAMP and pA:U. Overall these results indicate that there is a differential modulation of danger signaling by adenosine rather than overall supression. Our results have important implications for developing combinatorial approaches to target adenosine and danger signaling pathways to cure immune-related diseases.
The role of SHP-2 on NK cell development and functions
Authors: Miah S.M. Shahjahan , Sexl Veronica , Yang Wentian , Brossay Laurent 
Published: January 2023

Abstract
The SH2 domain-containing tyrosine phosphatase SHP-2 (PTPN11) has been shown to play essential roles in immune cell signaling. Several inhibitory receptors expressed on natural killer (NK) cells associate with SHP-2 to exert downstream functions. Here we examined the influence of SHP-2 on NK cell development and effector activity. Because SHP-2 is required for embryonic development in mice, we generated mice conditionally deficient for SHP-2 in NK cells. SHP-2fl/fl mice were crossed to NKp46-Cre mice in order to inactivate Ptpn11 in NKp46+ cells. These mice were also crossed to R26R-EYFP reporter mice to track cells with active Cre recombinase. We found an increase in NK cell frequency and number in these mice compared to littermate controls. The frequency of mature NK cells (CD11b+CD27−) was also increased in the periphery while decreased in the bone marrow. In addition, the homeostatic proliferation of SHP-2-deficient NK cells was impaired. However, effector functions were not affected significantly. These included IFN-γ production at the peak of the NK cell response during MCMV infection, rejection of β2-microglobulin deficient splenocytes, and degranulation via Ly49H using M157-transfected cells as targets. Taken together, our data suggest that SHP-2 is dispensable for NK cell effector functions and education, but is important for NK cell maturation.
Reactive oxygen species modulate immune cell effector function
Authors: Lee Seung-Hwan , Almutairi Saeedah , Ali Alaa Kassim 
Published: January 2023

Abstract
During an infection or carcinogenesis, the stimulation of immune cells may lead to elevated intracellular levels of reactive oxygen species (ROS), and this oxidative stress can cause damage to the cell. Interestingly, previous reports have shown that ROS produced through cell activation play a critical role as signaling messengers in immune responses. Yet, little is known in regard to the role of ROS in natural killer (NK) cell effector functions. Our data suggest that the levels of ROS increase in NK cells upon stimulation. Therefore, we hypothesize that, upon stimulation of NK cells, ROS accumulate and provide positive signaling that leads to the upregulation of NK cell effector functions. To test this hypothesis, we examined NK cell function and proliferation in Glrx2-deficient mice in which the ROS levels are elevated in the mitochondrial matrix due to a defect in the redox system. We found that lack of Glrx2 in NK and T cells does not improve their cytotoxic potential or their ability to produce IFNg. However, Glrx2-deficient NK cells showed enhanced proliferation when stimulated with IL-2, indicating that during prolonged stress, the elevated levels of ROS drive NK cells to undergo robust cell division. This finding will elucidate a link between the levels of ROS and immune cell proliferation during viral infection and carcinogenesis.
Siglec-E negatively regulates the activation of Toll-like Receptor 4 by controlling its endocytosis
Authors: Chen Guoyun , wu Yin , Ren Dongren 
Published: January 2023

Abstract
TLR4 signaling is critical for providing effective immune protection but must be tightly controlled to avoid inflammation-induced pathology. Previously, we reported extensive and direct interactions between TLR and Siglec families of Pattern Recognition Receptors. Here, we examined the biological significance of this interaction during infection. We found that Siglec-E is required for E. coli-induced endocytosis of TLR4. Siglec-E-deficient dendritic cells infected with E. coli fail to internalize TLR4. This leads to sustained TLR4 on cell surface and activation of NF-κB and MAP kinase p38, resulting in high levels of TNF-α and IL-6 compared with wild-type dendritic cells. In contrast to the signaling events occurring at the plasma membrane, as a result of the inability to internalize of TLR4, Siglec-E-deficient dendritic cells were also defective for TRIF-mediated IFN-β production in response to E. coli infection. Furthermore, we found that accumulation of ubiquitinated-TLR4 and binding of E3 ubiquitin ligase Triad3A to TLR4 was increased significantly in bone marrow-derived dendritic cells from wild-type mice, but not from Siglec-E-deficient mice, after E. coli infection. This represents a newly discovered mechanism that regulates the signaling of TLR4 during E. coli infection.
C-reactive protein modulates myeloid derived suppressor cell function and signaling.
Authors: Jimenez Rachel V , Szalai Alexander J 
Published: January 2023

Abstract
Recently, we provided evidence that C-reactive protein (CRP) plays a causal role in acute kidney injury (AKI) i.e., human CRP exacerbates AKI in CRP transgenic mice (CRPtg) and AKI severity is lessened in CRP knockout mice. The CRP effect in CRPtg was associated with significantly increased renal infiltration by myeloid derived suppressor cells (MDSC). Subsequently, we showed that the AKI exacerbating influence of CRP in CRPtg could be blocked by depletion of MDSCs. In the present work we sought to define the impact of CRP on MDSC ontogeny and function. Using wild type mouse bone marrow (BM) we derived MDSCs and established that CRP enhances their ability to suppress CD3ɛ/CD28-stimulated T cell proliferation in vitro. By flow cytometry we verified that mouse BM-MDSCs express variable amounts of three CRP receptors: LOX-1, FcγRI, and FcγRIIB. Expression of the latter on BM-MDSCs was upregulated upon CRP exposure, as demonstrated by flow cytometry. Furthermore, preliminary data obtained from studies using BM-MDSCs derived from FcγRIIB deficient mice suggest that FcγRIIB participates in CRP-mediated signaling. In their sum, these data suggest that CRP-mediated augmentation of MDSC suppressive function is FcγRIIB-dependent. Ongoing work seeks to define the consequences of CRP-mediated modulation of MDSC functions in vivo and to explore whether FcγRIIB and other CRP receptors are involved.
ROCK2, but not ROCK1 interacts with STAT3 to regulate TH17/TFH gene transcription
Authors: Chen Wei , Nyuydzefe Melanie S. , Weiss Jonathan M. , Zhang Jingya , Waksal Sam D. , Zanin-Zhorov Alexandra 
Published: January 2023

Abstract
Rho-associated kinase 2 (ROCK2) was recently shown to be implicated in regulation of IL-17 and IL-21 secretion in both mice and humans via down-regulation of STAT3 phosphorylation and subsequent transcription activity. Here we report that the ROCK2, but not ROCK1 protein binds phosphorylated-STAT3 (pSTAT3) in both cytoplasmic and nuclear compartments in human CD4+ T cells during T helper 17 (TH17)-skewing activation. The interaction of ROCK2 with pSTAT3 was partially abrogated by treatment of cells with selective ROCK2 inhibitor suggesting that ROCK2 kinase activity is required for formation of JAK-STAT complex and STAT3 phosphorylation. Further analysis by chromatin-immunoprecipitation sequencing (ChIP-seq) revealed that ROCK2 binding is significantly enriched toward promoter regions and peaks at transcription start sites (TSS) relative to genomic DNA. Moreover, 60–70% of ROCK2 and STAT3 peaks are overlapped genome-wide and co-localized to several key genes that control TH17 and T follicular helper (TFH) cell functions. Specifically, the binding of ROCK2 and STAT3 to the IRF4 and Bcl6 genes was validated by ChIP-qPCR analysis performed in human CD4+ T cells activated by TH17-skewing conditions. Finally, siRNA-mediated inhibition of STAT3 attenuated ROCK2 binding to the IRF4 and Bcl6 promoters indicating the critical role of STAT3 in the recruitment of ROCK2 to chromatin and ROCK2-mediated regulation of TH17/TFH gene transcription. Together, the present work provides previously unidentified insights into the molecular mechanism of specific involvement of ROCK2 isoform in regulating STAT3 phosphorylation and transcriptional activity in TH17-driven autoimmune responses.