bluuebunny/crossref_metadata_2025_split huggingface dataset:
Modeling Immune Effector Responses in Alveolar Epithelial Cell Driven Lung Injury and Fibrosis
Authors: Venosa Alessandro , Tomer Yaniv , Jamil Sarita , Beers Michael Francis 
Published: January 2023

Abstract
Mutations in the Surfactant Protein-C gene [SP-C] have been linked to pulmonary fibrosis (PF), with the missense isoleucine to threonine substitution at position 73 [I73T] as the most common. Clinically, PF is characterized by unrelenting tissue scarring and respiratory failure, intermitted by episodic exacerbations, bouts of inflammatory cell influx of unknown cause, which drastically reduce patient prognosis. Current murine models of PF rely on exposure to toxic agents or overexpression of pro-fibrotic signaling molecules in order to generate fibrotic endpoints. This reduces the overall relevance to human disease. To address these shortcomings, we developed a murine model of fibrosis designed to expressed mutant SP-CI73T. The resulting phenotype is accompanied by elevations in bronchoalveolar lavage cell counts, diffuse alveolar damage, and mortality. RNA and protein analysis demonstrate SP-CI73T epithelial cells initiate immune cell recruitment via CCL-2, IL-5, CCL-11, and CCL-17. Flow cytometric analysis highlighted temporally definite waves of inflammatory cells: an initial Ly6C+ inflammatory monocytes influx (3d), transient neutrophilic influx, and subsequent alveolar eosinophilia (2wk). RNA-seq analysis of sorted Ly6C+ monocytes revealed both inflammatory (inos, Il-6) and pro-fibrotic (col1a1, col1a2) phenotype. Pharmacologic and genetic monocyte ablation reduced inflammatory burden and improved survival of SP-CI73T mice. Taken together, we show that our mutant SP-CI73T model recapitulates the clinical features of PF. These findings support the concept that epithelial dysfunction is a key component of fibrogenesis and that inflammatory monocytes is responsible for disease exacerbation.
Transient immunosuppression blocks iBALT formation in the lung induced by helper-dependent adenoviral vector during gene delivery
Authors: Cao Huibi , Zhou Peter , Duan Cathleen , Hu Jim 
Published: January 2023

Abstract
Lung is an immune sensitive organ with presence of all immune cells at the frontline of immunity. Under chronic infection conditions, inducible Bronchus-associated lymphoid tissue (iBALT) can be formed to defend foreign infectious organisms. We wonder whether viral vectors for gene therapy could induce iBALT formation and whether this process can be controlled through immune modulation. To test these ideas, we repeatedly administered a helper-dependent adenoviral (HD-Ad) vector to mouse lungs and examined iBALT formation. HD-Ad vectors are based on adenovirus but with all viral coding sequences deleted and is one option for large gene delivery. In our previous studies, HD-Ad vectors have exhibited high in vivo efficiency in transduction of mouse, rabbit and pig airway epithelial cells, including stem/progenitor cells such as basal cells. HD-Ad vectors expressing therapeutic gene encoding cystic fibrosis transmenbrane conductance regulator (CFTR) rescue CFTR function in CF patient primary cells. In our mouse studies, we found that HD-Ad vectors induced iBALT formation and antibody production. We used cyclophosphamide or HD-Ad expressing IL-10 to modulate the host immune responses and to examine the effects on iBALT formation. Comparing with control group (HD-Ad vectors only), the mice treated with cyclophosphamide or IL-10 expressing HD-Ad vector showed a significant reduction of iBALT formation and infiltration of inflammatory cells, including CD4+ and B220+ as well as production of anti-Ad antibodies and neutralizing antibodies. Our results indicate that transient modulation significantly mitigated immunopathology and extended transgene expression in mouse airways following readministration of HD-Ad vectors.
Natural Killer cell immune synapse formation and cytotoxicity is controlled by mechanical tension of the target
Authors: Simmonds P , Friedman Daniel , Hale Alexander , Hodson Nigel W , White Michael RH , Davis Daniel M 
Published: January 2023

Abstract
Natural Killer (NK) cells can kill infected or transformed cells directly via the formation of a lytic synapse at the cell-cell interface. Diseased cells often present with altered mechanical properties, and cell stiffness is a feature of tumour cell migratory potential. However, little is known about how target cell stiffness impacts the assembly of the NK cell synapse or the subsequent functional response.
To study this, we set out to create surrogate targets with specific ligands presented on cell-sized beads of different stiffness. Sodium alginate cell-sized microbeads were synthesised where chemical composition was altered to produce beads of low (9kPa), medium (34kPa), or high (254kPa) stiffness. These were coated with antibodies against the NK cell activating receptor NKp30 and the integrin LFA-1 to model the 3D interaction between NK cells and target cells.
Imaging revealed that later steps in NK-cell activation differed significantly, including the degree of cytoskeletal arrangement, F-actin accumulation, microtubule organising centre polarisation and perforin accumulation. Stiffer beads induced polarisation of the MTOC in 59% of conjugates analysed compared to only 7% with low stiffness targets. Functionally, NK cell degranulation was higher when forming conjugates with stiffer targets. The metabolic profiles were similar between NK cells stimulated overnight by antibody-coated beads, regardless of their stiffness. Altogether, these data imply that stiffer targets are more potent in triggering the initial activating signals in NK cells. Thus, immune synapse mechanosensitivity represents a novel form of NK cell regulation.
Phenotypically Friendly Allospecific NK Cells Differentially Suppress Allospecific T cell Responses in Prenatal Hematopoietic Chimeras
Authors: Kang Hee Kap , Ott Katherine , Turner Lucas , Redden Catherine , Alhajjat Amir , Shaaban Aimen 
Published: January 2023

Abstract
Prenatal allospecific tolerance hinges on the developmental selection of NK cells expressing a friendly phenotype that is not cytotoxic to allogeneic donor cells. We wondered if prenatally educated friendly NK cells influenced allospecific T cell responses as an additional mechanism supporting prenatal tolerance. Therefore, we hypothesized that prenatally educated friendly NK cells exert suppressive effects on the induction of allospecific T cell responses.
To challenge this hypothesis, we examined the suppression potential of prenatally educated friendly NK cells through gain and loss of function experiments in stable prenatal Balb/c into B6.Thy1.2 chimeras (engrafter mice). We first depleted allospecific NK cells in host engrafter mice and then adoptively transferred Thy1.1+ naïve responder T cells along with CFSE-labeled allogenic or syngeneic target cells into the engrafter hosts. We found that the depletion of allospecific Ly49D+ NK cells in engrafter mice increased Thy1.1+ T cell alloimmunity in vivo. Conversely, adoptive transfer of friendly NK cells suppressed the induction of allospecific T cell responses in host mice. Furthermore, cultured friendly NK cells from engrafter mice showed stronger suppression on the induction of T cell alloimmunity in vitro than that of naïve mice. Lasltly, NK cell-mediated suppression of allospecific T cells required cell contact.
From these results, we conclude that prenatally educated friendly NK cell exert enhanced suppressive effects on T cell alloimmunity in vivo and in vitro. Depending on the context of their prenatal education, NK cells can differentially influence T cell responses in prenatal chimeras.
Changes in the gut microbiota caused by ASBTi suppress intrahepatic steatosis and inflammation in a mouse model of non-alcoholic fatty liver disease (NAFLD)
Authors: Matsui Masahiro , Asai Akira , Okamoto Norio , Nishikawa Tomohiro , Ohama Hideko , Tsuchimoto Yusuke , Fukunishi Shinya , Higuchi Kazuhide 
Published: January 2023

Abstract
Although the estimated prevalence of NAFLD is approximately 25%, an effective treatment has not been established. Recently, gut microbiota has been shown to affect the pathogenesis of NAFLD. In this study, the protective effect of Apical Sodium-dependent Bile acid Transporter inhibitor (ASBTi) against NAFLD was investigated in mice fed a high-fat diet for 12 weeks (HFD mice) via changing their gut microbiota. ASBTi mice were mice treated with ASBTi (2.5 μmol/kg/day) during HFD feeding. For fecal microbiota transplantation (FMT), stools were collected from HFD mice and ASBTi mice. Fecal materials obtained from HFD mice and ASBTi mice were transplanted to normal mice, fed HFD for 12 weeks, and designated as HFD-FMT mice and ASBTi-FMT mice, respectively. Microbiota in these groups of mice were analyzed by 16S rRNA gene sequencing. The degrees of hepatic steatosis and inflammation in these mice were determined pathologically. The shannon index of the microbiota from ASBTi mice (3.31±0.19) and ASBTi-FMT mice (3.29±0.18) was significantly high as compared to that from HFD mice (2.90±0.19) and HFD-FMT mice (2.89±0.16). Steatosis and inflammation in the liver of HFD mice and HFD-FMT mice were shown to be pathologically severe and NAFLD Activity Score (NAS) was 7.67±0.58 in HFD mice and 5.67±2.08 in HFD-FMT mice. However, those in the liver of ASBTi mice and ASBTi-FMT mice were improved and NAS was reduced in ASBTi mice (1.67±0.58) and in ASBTi-FMT mice (2.67±0.58). These results suggest that changes in the gut microbiota caused by ASBTi suppress intrahepatic steatosis and inflammation in NAFLD mice. Elobixibat can be expected to prevent fatty liver.
Malaria-driven atypical memory B cells are blinded to soluble but not membrane-associated antigens
Authors: Sohn Haewon , Ambegaonkar Abhijit A. , Kwak Kihyuck , Manzella-Lapeira Javier , Brzostowski Joseph , Pierce Susan K. 
Published: January 2023

Abstract
A common feature of many chronic human infectious diseases, including malaria, includes the large expansion of a population termed atypical memory B cells (MBCs) in peripheral blood, which are phenotypically and transcriptionally distinct from naïve B cells and classical MBCs. One shared distinguished phenotype among these is their high expression of a variety of inhibitory receptors including FcγRIIB. Despite the potential importance of atypical MBCs, if or how they function in chronic infectious diseases has yet to be clearly known. We reported earlier that in malaria atypical MBCs, in contrast to naïve B cells and classical MBCs, are refractory to activation by soluble antigen. Thus, atypical MBCs have been described as hypo-responsive to antigen-driven B cell activation and believed to be ‘exhausted’ and to contribute to the poor acquisition of immunity in chronic infectious diseases. However, we show the evidence here that atypical MBCs respond robustly to membrane-associated antigen by signaling through the B cell antigen receptor, gathering and internalizing antigen for presentation and initiating differentiation toward plasma cells. We discovered that on antigen-containing membranes the immune synapses of atypical MBCs actively exclude the inhibitory receptor FcγRIIB thus allowing BCR signaling. The high expression of inhibitory receptors by atypical MBCs may serve to restrict B cell responses to antigens presented on cell surfaces, avoiding the potential deleterious effects of chronic soluble antigen exposure.
IL12RB1 allelic expression imbalance in polarized human TH cells
Authors: Mejia Oscar Rosas , Williams Amanda , Robinson Richard 
Published: January 2023

Abstract
Allelic Expression Imbalance (AEI) occurs when two alleles of a given gene are expressed at different levels, either due to epigenetic inactivation and/or genetic variation in regulatory regions. Human autosomal genes are widely presumed to be equally expressed from both alleles (biallelic expression); this is not always the case, however, as AEI affects expression of odorant receptor and adhesin genes in neurons, as well as immunoglobulin (Ig) and T-cell receptor (TCR) genes in lymphocytes. Ig and TCR gene AEI is an important regulator of immune development and disease resistance; therefore, it is reasonable to believe AEI of other immune receptors may similarly impact immune development and disease resistance. Here we demonstrate that the human cytokine receptor gene Interleukin 12 Receptor Beta 1 (IL12RB1) exhibits AEI in differentiated T cells, as measured by a significant deviation of the allelic mRNA expression ratios from that of the allelic gDNA ratios. Specifically, naïve human CD4+ T cells were purified from healthy adult donors and incubated with polarizing cytokines to drive their differentiation into specific T helper subsets (e.g. TH0 cocktail: IL-2. TH1 cocktail: IL-2, anti-IL-4, IL-12. TH17 cocktail: IL-2, IL-1b, TGF-b1, IL-23, anti-IFNg, anti-IL-4). The extent of IL12RB1 AEI was then quantified and related to T cell expression of IFNg (TH1) and IL17 (TH17). Collectively, our data suggest that IL12RB1 AEI associates with the expression of anti-mycobacterial effector genes in T cells. Future experiments will determine the epigenetic or molecular mechanism behind IL12RB1 AEI.
REV-ERB-dependent heme signaling regulates Th17 cell activity
Authors: Mosure Sarah A , Solt Laura A 
Published: January 2023

Abstract
T helper 17 (Th17) cells are important for protective immunity at mucosal surfaces, but their dysregulation is implicated in the pathogenesis of autoimmune and chronic inflammatory diseases. A better understanding of the signaling pathways governing Th17 cell-mediated immunity and pathogenicity would aid in the development of novel therapeutics to treat Th17 cell-mediated diseases. We recently showed that the REV-ERB nuclear receptors (REV-ERBα and REV-ERBβ) are critical regulators of Th17 cell development and pathogenicity. As ligand-regulated transcription factors, the REV-ERBs can bind their natural ligand, heme, which modulates REV-ERB function. To investigate heme’s role as a REV-ERB-dependent signaling molecule in Th17 cells, we differentiated naive CD4+ T cells under Th17-polarizing conditions and found heme treatment repressed differentiation. Repression was partially ablated in cells lacking the REV-ERBs, indicating heme can specifically activate the REV-ERBs in Th17 cells. We further found that enzymes involved in heme biosynthesis and transport were upregulated during Th17 cell differentiation, indicating heme is actively produced in Th17 cells. To investigate the role of endogenous heme, we overexpressed REV-ERB mutants incapable of binding heme, which relieved repression of Th17 cell differentiation relative to over expression of wild type (WT) REV-ERBs. RNA-seq of mutant- vs WT-overexpressed cells revealed that heme modulates a specific subset of REV-ERB target genes. Together, our findings suggest that conditions affecting heme levels in Th17 cells can alter the transcriptional phenotype of the cell via heme modulation of REV-ERB activity.
Identifying key transcriptional regulators that establish T cell memory during viral infection
Authors: Tsuda Shanel M , Diao Huitian , Getzler Adam J , Milner Justin J , Goldrath Ananda , Crotty Shane , Pipkin Matthew E. 
Published: January 2023

Abstract
During infections, naive CD8 T cells differentiate into terminal effector cells (TE) that are relatively short-lived, and memory precursor (MP) cells that give rise to long-lived memory CD8 T cells, but the transcriptional control of this process is still unclear. In naïve CD8 T cells, cis-regulatory regions that become accessible in chromatin during the first 24 hours of TCR stimulation and that remain accessible in mature memory T cell subsets are highly enriched with motifs encoding binding sites for the ETS- and bZIP-families of transcription factors (TFs), many of which also overlap Runx-TF binding sites. The ETS and bZIP TF families are encoded by 88 genes, and are differentially expressed between naïve, early effector, and memory CD8 T cell subsets. To interrogate their requirements functionally, we applied an in vivo pooled RNA interference screen using short hairpin RNAs in microRNA contexts (shRNAmirs) to suppress individually all TFs from both families in CD8 T cells responding to lymphocytic choriomeningitis virus (LCMV) infection. In addition, we focused on Ets1, the most highly expressed ETS TF in naïve, effector and memory cell subsets. Ets1 suppression with shRNAmirs impaired overall effector CD8 T cell numbers in vivo, and increased the frequency of TE-like cells at early times after LCMV infection, which resulted in reduced long-lived effector cells and an increased fraction of altered central memory-like cells at later time points. Enforced Runx3 expression in Ets1-suppressed cells partially restored normal MP cell formation at early times. These results suggest that Ets1 and Runx3 are each necessary for the normal differentiation of MP cells, and could function cooperatively to program their differentiation.
Establishment And Application of An in Vitro Cd8+ T Cell Exhaustion System
Authors: Zhao Manzhi , Kiernan Caoimhe H. , Leon Leticia G. , van Meurs Marjan , Brouwers-Haspels Inge , Mueller Yvonne M , Katsikis Peter D 
Published: January 2023

Abstract
Exhaustion is a dysfunctional state of cytotoxic CD8+ T cells (CTL). In vivo models of CTL exhaustion yield very few exhausted CTL, limiting the analysis can be done on these cells. Furthermore, the milieu in these models can obscure the phenotype of exhausted CTL. An in vitro system could therefore greatly facilitate the study of this cell state.
Purified OT-I CTL were cultured and repeatedly stimulated with OVA(257–264) SIINFEKL peptide to induce exhaustion. On day 5, inhibitory receptors (IRs) and cytokine production were assayed. Sorted live CTL were transferred into WSN-OVA influenza infected mice and on day10 post infection the number of the donor CTL were measured. Gene expression changes were determined by RNAseq and DNA methylation sequence.
In vitro exhausted CTL exhibited upregulation of multiple IRs as well as impaired polyfunctionality. They expanded less than controls in vivo. In vitro exhausted CTL exhibited all the transcriptomic characteristics of in vivo exhausted CTL.
Repeated stimulation of CTL with their specific peptide, induced all the functional, phenotypic and transcriptomic characteristics of exhausted CTL. This in vitro system can be used to identify genes and signalling pathways involved in exhaustion and facilitate the screening of reagents that prevent/reverse CTL exhaustion.
C C Chemokine Receptor 7 Differentially Regulates Trafficking of T Cells to the Bone Marrow During the Adaptive Immune Response
Authors: Vines Charlotte M , Cervantes Jaisel , Vazquez Aaron , Bill Colin A 
Published: January 2023

Abstract
It has become increasingly evident that the bone marrow is an important niche for memory B cells and memory T cells that can be activated to become effector T cells during a secondary immune response. However, signaling events that promote bone marrow persistence of memory cells remain largely unknown. We have found that the C-C chemokine receptor 7 (CCR7) signaling plays an important role in regulating the presence of memory cell populations of the bone marrow. In mice homozygously deleted for CCR7 (CCR7−/−) we find that the memory immune cell repertoire is redistributed following the secondary immune response. Specifically, instead of homing to the lymph nodes or spleen, the CCR7−/− effector memory T cells, which respond quickly to antigen, localize to the bone marrow. Additionally, using a CCR7 conditional knockout mouse, we found that when CCR7 is deleted only in T cells, numbers of activated B cells and antibody titers increase, while the regulatory T cells that would normally suppress the immune response, have reduced numbers within the bone marrow. Moreover, we found that in our CCR7f/f-CD4-Cre mice in the absence of CCR7 in T cells, bone marrow activated B cells increased in numbers on day 30. This was not due to differences in the immune response to B-cell cytokine IL-4, which was not significantly increased in any of the mice tested. Instead, we observed a difference in the overall percentage of activated B cells in CCR7f/f-CD4-Cre mice compared to CD4-Cre controls. From these data we infer that CCR7 may promote differential migration of effector memory T cells into the bone marrow that have a direct effect on the composition of the memory B-cell compartment.
The LncRNA Snhg1 is Essential for Memory CD8 T Cell Establishment
Authors: Zhang Yanyan , Bai Qiang , Ni Ting , Yang Liuqing , Jiang Lubin , Ye Lilin , Wu Yuzhang 
Published: January 2023

Abstract
The efficient induction and long-term persistence of pathogen-specific memory CD8 T cells are pivotal to rapidly curb the reinfection. Recent evidence indicates that lncRNAs possess higher lineage and stage specificity than mRNAs in T cell development, differentiation and function. However, the pivotal lncRNAs regulating CD8 T cell differentiation remain largely unclear. Through profiling of CD8 T cell lineages in this study, the lncRNA Snhg1 with the conserved on-off-on expression pattern in naïve-effector-memory CD8 T cells from mouse to human, is found to be required for memory CD8 T cell generation. Further, Snhg1 is found interacting with the highly conserved vesicle trafficking protein Vps13D to facilitate IL-7Rα membrane location specifically. IL-7 has been reported to sustain memory survival through STAT5-BCL2 axis, but whether IL-7 can directly promote memory differentiation remain cloudy. In this research, Snhg1/Vps13D depletion impair memory formation through IL-7-STAT3-TCF1-Blimp1 axis. Whether TCF-1 is induced by canonical Wnt-β-Catenin signaling in memory CD8 generation remain controversial. This work finds gene Tcf7 is regulated by direct binding of p-STAT3 in CD8 stimulated with IL-7. Furthermore, TCF-1 is found promoting memory with blocking effector mainly through impeding Prdm1 expression directly to balance Blimp1-BCL6 and Blimp1-ID3 axis. Thus, this study proposes a model for lncRNA as critical factor that couples membrane trafficking of cytokine receptor with transcriptional induction to promote memory CD8 T cell establishment.
Siglec 5 - a novel checkpoint receptor in T cells
Authors: Vuchkovska Aleksandra , Iwashima Makio 
Published: January 2023

Abstract
Immune checkpoint molecules have become a major target for designing immunotherapies. Across fourteen malignancies, the FDA has approved six drugs targeting three different inhibitory checkpoint receptors. Even though TCR stimulation through MHC-peptide and CD28 co-stimulation are the major requirements for T cell activation, it is the engagement of checkpoint receptors that dictate the functional outcome of a particular T cell. Here, we describe a new candidate checkpoint receptor, Siglec 5, expressed on activated T cells. To date, Siglec 5 is defined as a self-associated molecular pattern receptor that exhibits an inhibitory response upon interaction with sialic acids, subsequently dampening immune system activation against self. Current literature shows that Siglec 5 is expressed mainly by myeloid cell, such as neutrophils and monocytes, and that T cells do not express any Siglecs, including Siglec 5. However, we found that Siglec 5 is a unique, late-activation marker of T cells, upregulated at 48 hours, with peak expression at 72 hours post-TCR stimulation. Similar to PD-1, Siglec 5 has an ITIM and a SWITCH domain, both well described for their inhibitory function. In this study, we sought to determine the function of Siglec 5 on T cell activation. We found that overexpression of Siglec 5 in T cells results in a significant inhibition of T cell activation measured by reporter NFAT and AP-1 activity. Mutagenesis studies show that both of the intracellular domains, ITIM and SWITCH, of Siglec 5 are required for mediating T cell inhibition. Together, these data demonstrate that Siglec 5 is a previously unknown immune checkpoint molecule found on T cells.