bluuebunny/crossref_metadata_2025_split huggingface dataset:
Isolation of A Human GM-CSF Antibody with Potent Neutralizing Activity from a Subject with Idiopathic Pulmonary Alveolar Proteinosis (96.24)
Authors: Chan-hui Po-Ying , Cox Tom , Grondin Roxanne , Grandea Glen , Frey Steve , Branum Mark , Ward Courtney , Fox Ray , Olsen Ole , Hammond Phil , Cieplak Witold , Moyle Matthew 
Published: January 2023

Abstract
Background. GM-CSF has been implicated in the development of autoimmune disorders. The efficacy of neutralizing anti-GM-CSF has been demonstrated in several animal disease models rheumatoid arthritis, asthma and lung inflammation. While humanization of murine antibodies (Abs) and phage-display technology have generated potential therapeutic leads currently in development, these engineered leads face potential challenge of serum stability and immunoreactivity. We have isolated a fully human anti-GM-CSF from a patient of idiopathic pulmonary alveolar proteinosis (iPAP) who produces anti-GM-CSF as autoantibody. Method. To isolate natural human Abs, we used a short term memory B cell culture system to generate Ab-containing supernatants and screened for anti-GM-CSF that inhibited the proliferation of TF-1 cells in response to human GM-CSF. The specificity for GM-CSF was then confirmed via secondary binding screening in Alphascreen assays. Recombinant human Abs were isolated from the corresponding memory B cells using RT-PCR and subcloning. Results. By analyzing the serum from several iPAP patients for the neutralizing activity against TF-1 proliferation, we selected one peripheral blood sample for Ab isolation. TCN-350 was isolated with IC50 of 27 pM for TF-1 inhibition. It binds human GM-CSF with an affinity of 39 pM and cross-reacts with rabbit GM-CSF with an affinity of 1 nM. The high neutralization potency of TCN-350 warrants further evaluation as a therapeutic lead.
Revertant muscle fibers expressing dystrophin do not tolerize the immune system in Duchenne muscular dystrophy: lessons learned from a Phase I clinical trial (96.9)
Authors: Campbell Katherine , Rodino-Klapac Louise , Sahenk Zarife , Shilling Chris , Lewis Sarah , Bowles Dawn , Gray Steven , Li Chengwen , Galloway Gloria , Malik Vinod , Coley Brian , Clark Reed , Li Juan , Xiao Xiao , Samulski Jade , McPhee Scott , Samulski R.Jude , Mendell Jerry , Walker Christopher 
Published: January 2023

Abstract
Duchenne muscular dystrophy (DMD) is characterized by progressive muscle weakness due to mutation of the dystrophin gene. Muscle inflammation is an unexplained feature of DMD. T lymphocytes dominate the inflammatory infiltrate but antigen(s) they target have not been identified. Here we evaluated T cell immunity to dystrophin in 6 humans with DMD before and after delivery of a gene therapy vector to skeletal muscle. The vector transgene encoded a miniaturized version of human dystrophin that restores muscle function in animal models of DMD. Sustained transgene expression was not observed in any of the subjects. Non-self epitopes encoded by the vector transgene, but deleted from the defective dystrophin gene, provoked CD4+ and CD8+ T cell responses in a subset of the subjects. Dystrophin-specific T cell immunity was unexpectedly detected in two subjects before gene therapy. Epitopes were mapped to revertant dystrophin protein spontaneously expressed in rare muscle fibers when alternative splicing restores the correct open reading frame. Delivery of the minidystrophin gene to skeletal muscle of one subject provoked a very rapid increase in the frequency of these self-reactive T cells. T cell priming by idiosyncratic expression of revertant dystrophin could explain muscle inflammation and contribute to the pathogenesis of DMD. Recall of auto-reactive T cells also has important clinical significance in the design and monitoring of experimental therapies for this disease.
Exercise increases regulatory T cell function and decreases Th2 and Th17 cytokine production in healthy and asthmatic mice (97.15)
Authors: Lowder Thomas , Dugger Kari , Estell Kim , Deshane Jessy , Schwiebert Lisa 
Published: January 2023

Abstract
Exercise training has been shown to significantly reduce Th2 cytokines, airway hyper-responsiveness (AHR), and lung pathology in asthmatic mice. Some of these changes have been postulated to be due to regulatory T cells (Tregs). In these experiments, mice were induced into an asthmatic state (OVA) and exercised (45min, 3d/wk, at 60-70% of VO2max) for a total of 10 bouts. Tregs isolated from the spleens, lungs, and mediastinal lymph nodes of exercise-trained mice and stimulated in vitro were able to suppress effector cells at a significantly greater rate than Tregs from sedentary mice irrespective of whether or not they were induced to an asthmatic state (p &lt; 0.05). Blocking TGF-β did not reduce Treg-mediated suppression in the exercise-trained group, but suppression was decreased in control mice following anti-TGF-β administration. In vitro Treg:effector co-cultures demonstrated significant reductions in Th2 and Th17 cytokine production in exercise-trained mice compared to their sedentary counterparts. Surface expression of Tregs, as assessed by Foxp3EGFP, was significantly higher in exercise-trained asthmatic mice compared to sedentary controls. We show for the first time that exercise training increases Treg expression and function in asthmatic and non-asthmatic mice.
iNKT cells regulate plasmacytoid dendritic cell population in bone marrow (98.16)
Authors: Wen Xiangshu , Eriksson Anna , Kim Peter , Singh Ram 
Published: January 2023

Abstract
Plasmacytoid dendritic cells (pDC) develop and proliferate in bone marrow and migrate to peripheral organs. pDCs produce IFNα and play a role in viral infections and autoimmune diseases. Mechanisms that regulate the development of pDC in the bone marrow are not fully understood. Invariant natural killer T (iNKT) cells comprise of a small population of lymphocytes in the bone marrow. Here, we examined the role of iNKT cells on pDC population in the bone marrow in vivo and in vitro. Bone marrow cells from wild-type, CD1d-deficient, and iNKT cell transgenic (Vα14-Tg) BALB/c mice were cultured with Flt3 ligand (FL) and αGalCer. We found that although all DCs were activated, as determined by expression of MHCII and CD86, in the presence of activated iNKT cells, the proportion of pDCs (CD11c+PDCA1+B220+) was consistently reduced in the presence of activated iNKT cells. Similar results were found in vivo, where an injection of FL and αGalCer reduced the proportion of pDCs, whereas the proportion of myeloid DCs (CD11c+CD11b+) was unchanged or slightly increased, in the bone marrow and spleen. Interestingly, FL+αGalCer increased CCR7 expression on bone marrow pDCs, but not on myeloid DCs, suggesting that iNKT cells may contribute to the egress of pDCs from the bone marrow. Such effect of FL+αGalCer injection was not seen in CD1d-deficient mice. In summary, these data suggest a role of iNKT cells in the development and homeostasis of pDCs.
Regulation of CD8 T cell responses by follicular dendritic cell-bound antigen (98.17)
Authors: Erlebacher Adrian , Curotto de Lafaille Maria , Lafaille Juan , Carroll Michael , McCloskey Megan 
Published: January 2023

Abstract
Follicular dendritic cells (FDCs) are well known to retain the immune complexes that drive antibody affinity maturation. Here, we present evidence that FDC-bound antigens are also persistently sampled by bone marrow-derived DCs for presentation to CD8 T cells. These results are based upon our previous observation that ovalbumin (OVA) expressed in transmembrane form by the placenta is hematogenously released during pregnancy to accumulate on maternal FDCs throughout all secondary lymphoid organs. We now show that OVA loading onto FDCs is complement-dependent and that bound OVA is retained for several weeks after delivery. Strikingly, antigen retention correlated with a prolonged postpartum phase of OVA presentation to CD8 T cells that could be abrogated by the pharmacological ablation of FDC networks. Furthermore, bone marrow chimera experiments demonstrated that FDC-bound OVA could be acquired de novo by DCs arising in the postpartum period. The presentation of placental OVA acquired from FDC-associated depots was highly tolerogenic as it induced T cell deletion even in the presence of exogenous adjuvants. These results suggest an unanticipated level of control over peripheral tolerance induction potentially relevant to the pathogenesis of systemic lupus erythematosus, an autoimmune disease characterized by impaired tolerance to apoptosis-associated antigens opsonized by the complement system. Supported by NIH grant RO1-AI062980.
Regulatory function of myeloid-derived suppressor cells is restricted to inflammatory site. (98.25)
Authors: Haverkamp. Jessica , Ratliff Timothy 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSC) are identified in mice as Gr-1+CD11b+ cells able to suppress T cell proliferation. Suppressive function of MDSC is linked to expression of arginase I and inducible nitric oxide synthase (iNOS) and can be augmented by inflammation. While inflammation is linked to MDSC function, it is unknown if MDSC isolated from the inflammatory site possess equal regulatory function as those in distal sites such as the spleen. Using the POET-3 model of prostate inflammation, we show Gr-1+CD11b+ cells isolated from inflamed prostates express elevated levels of ARG I and iNOS. In contrast, cells from the spleens of mice with prostate inflammation do not express detectable levels of ARG I or iNOS. However, when function of Gr-1+CD11b+ cells from spleens or the inflammatory site were tested in conventional suppression assays, all cells demonstrated suppressive function. RT-PCR analysis revealed that arginase I and iNOS mRNA were induced in MDSC during suppression assays, in part through IFN-γ. Thus conventional suppression assays induce functional suppressor cells and do not test their actual activation state in vivo. To better evaluate the function of Gr-1+CD11b+ cells we developed a short term suppression assay designed to limit exposure to IFN-γ during culture. Only cells from the inflammatory site suppressed T cell proliferation. These data demonstrate the novel finding that only Gr-1+CD11b+ cells at the inflammatory site are functional MDSC.
Novel chalcone JSH 4-4 inhibit NF-kB-regulated inflammatory gene expression via targeting LPS-binding site of MD-2 (98.4)
Authors: Roh Eunmiri , Lee Heun-Sik , Kwak Jeong-Ah , Jung Sang Hun , Han Sang-Bae , Kim Youngsoo 
Published: January 2023

Abstract
Previous studies have shown that chalcone has beneficial effects with antioxidant and anti-inflammatory activities. However, molecular basis for these effects not yet fully understood. In this study, a chalcone derivative, 2’, 4’-dihydroxy-6’-isopentyloxy chalcone (JSH 4-4) was discovered as an inhibitor of NF-kB activation. To demonstrate molecular target affected by JSH 4-4, we examined a homotypic interaction of TLR4, using Ba/F3 cells stably expressing TLR4-Flag, TLR4-GFP CD14 and MD-2, seperately. As a result, JSH 4-4 inhibited the LPS-induced TLR4 oligomerization. Moreover, we demonstrated that JSH 4-4 inhibit LPS-induced NF-kB activation via targeting LPS-binding site of MD-2. This mechanism of action could contribute to suppressive effect of JSH 4-4 on NF-kB-regulated production of inflammatory mediators such as nitric oxide, COX-2 and cytokines. TLR4-mediated NF-kB signalling occurs following sequential activation of IRAK family and TAK1. JSH 4-4 inhibited LPS-induced phosphorylation of IRAK-1 and TAK1. In addition, JSH 4-4 significantly inhibited IKK activity and phosphorylation of IkBa, resulting in sequential prevention of downstream events, including degradation of IkBs, nuclear translocation of NF-kB p65. In conclusion, JSH 4-4 inhibit NF-kB-regulated inflammatory mediators via targeting the receptor complex MD2-TLR4 in macrophages, thus JSH 4-4 might be considered as a potential agent for the treatment of NF-kB-associated inflammatory diseases.
Abrogation of tolerance with Flt3L requires CD154 expression (98.9)
Authors: Gurung Prajwal , Griffith Thomas 
Published: January 2023

Abstract
Soluble proteins are poorly immunogenic, and will often lead to Ag-specific tolerance. Similarly, Ag associated with apoptotic cells will frequently induce tolerance. These types of Ag are not very “dangerous”, so that recruitment and stimulation of professional APC is minimal. Treatment with the hematopoietic growth factor Fms-like tyrosine kinase 3 ligand (Flt3L) results in tremendous DC expansion, and previous studies have shown that Flt3L treatment can prevent the establishment of tolerance. Yet, the exact mechanism by which Flt3L treatment enhances immune responses remains undefined. I.V. injection of apoptotic cells or soluble Ag into wt mice induces tolerance, but this tolerance is abrogated by treatment with Flt3L. Interestingly, similar abrogation of tolerance was not observed in CD154-/- mice, suggesting a role for CD40-CD40L interaction. Further analysis of Flt3L-treated mice found reduced expansion of CD11c+ cells in CD154-/- mice versus B6 mice. This reduction in CD11c+ cells correlated with reduced T proliferation, as determined by CFSE dilution, in Flt3L-treated CD154-/- mice compared to B6 mice. Phenotypic analysis of the CD11c+ DC from Flt3L-treated B6 and CD154-/- mice showed reduced costimulatory molecule expression, especially CD40, in the CD154-/- CD11c+ DC. Together, these data suggest that maximal DC mobilization by Flt3L requires CD40-CD154 binding, which primes the DC for stimulating immunity even against Ag that normally induce tolerance.
Roles of Bcl6 and Blimp1 in CD4 T follicular helper (Tfh) lineage commitment (99.11)
Authors: DiToro Daniel , Johnston Robert , Crotty Shane 
Published: January 2023

Abstract
A distinct CD4 effector T cell subset, called T follicular helper cells (Tfh), is required for proper B cell responses in vivo. The mutually antagonistic transcription factors Bcl6 and Blimp1 play central but opposing roles in Tfh differentiation (Johnston et al., Science 2009). Bcl6 is both necessary and sufficient for in vivo Tfh differentiation, while Blimp1 specifically inhibits it. Expression of both Bcl6 and Blimp1, controlled temporally and spatially by cell-cell contact and multiple cytokines, is necessary for a normal CD4 T cell response. Naïve CD4 T cells express Bcl6 mRNA prior to, and independent of, commitment to the Tfh lineage. Bcl6 contains multiple functional domains and can affect different transcriptional outcomes depending on the local chromatin and co-repressor environment. Bcl6 repression of Blimp1 requires the co-repressor MTA3, which binds the Bcl6 RDII domain. We show that a loss of function mutation in the Bcl6 RDII domain results in a total failure of CD4 T cells to proliferate in vivo following infection with LCMV. We posit that this proliferation defect is a result of the failure to control expression of Blimp1, a strongly anti-proliferative protein, immediately following activation. Conversely, failure to attain adequate Blimp1 expression also results in an abnormal antiviral CD4 T cell response. Taken together, our findings suggest that the roles of Bcl6 and Blimp1 in CD4 T cells extend beyond Tfh lineage commitment.
SLAM-associated protein (SAP)-deficiency impedes the function of T follicular helper cells in germinal center formation (99.3)
Authors: Lu Kristina , Cannons Jennifer , Kanno Yuka , O'Shea John , Schwartzberg Pamela 
Published: January 2023

Abstract
Mutations affecting SAP lead to XLP, an immune disorder with impaired antibody responses. SAP-/- mice show defective formation of germinal centers (GC), due to a defect in T:B cell interactions. T follicular helper cells (Tfh) are important for providing help to B cells and GC formation. To investigate the role of SAP in Tfh cells for GC formation, we have developed a modified protocol to generate “Tfh-like” cells in vitro, resulting in IL-21 producing cells with elevated expression of hallmark Tfh markers CXCR5, ICOS, PD-1, BTLA, and SLAM family receptors CD84 and Ly108, along with enhanced levels of Bcl6 transcripts. Cell transfer of control Tfh-like cells into SAP-/- hosts can rescue GC formation upon immunization significantly better than polarized Th1, Th2 or Th17 cells, suggesting these cells act as functional Tfh cells in vivo. In vitro differentiation of SAP-/- cells is similar to control cells, yet, upon cell transfer, SAP-/- Tfh-like cells are unable to support GC formation. Interestingly, differentiation of Tfh-like cells is not terminal, as they can be repolarized to produce other distinct Th cell cytokines. Similarly, other Th cell populations can be repolarized to Tfh-like cells. Analyses of epigenetic modifications of transcription factors also supports plasticity between these Th cell lineages. These findings provide further insight into Tfh cell biology and suggest that SAP is critical for Tfh cell function in GC formation and immune homeostasis.
High-throughput characterization of MHC I-associated peptides presented by B lymphoblastoid cells (100.11)
Authors: Granados Diana Paola , Yahyaoui Wafaa , Côté Caroline , Muratore-Schroeder Tara , Thibault Pierre , Perreault Claude 
Published: January 2023

Abstract
Sensing of the MHC I-peptide (MHC I-p) repertoire allows elimination of infected or transformed cells. High-throughput strategies used to study MHC I-p include i) the use of secreted forms of MHC I transfected into model cell lines, or ii) chemical or metabolic labeling of MHC I, which is limited to certain MHCs and to cell culture model systems. Here, we used a quantitative proteomics approach to characterize and compare for the first time the MHC I-p repertoire associated to unlabeled HLA-A,B molecules of untransfected B lymphoblastoid cells (B-LCLs). Peptides were obtained from B-LCLs from 3 subjects by mild acid elution, fractionated by liquid chromatography and analyzed by nanoLC-MS/MS on a LTQ-Orbitrap mass spectrometer. We have identified more than 1000 unique 8-11mers associated to 10 HLA-A,B alleles: HLA-A*0101,0201,0205,0301,2902; B*0801,1501,4403,5001,5701. We found MHC-dependent differences in the predicted binding affinity of identified peptides. We used bioinformatic tools to identify the genes source of peptides and to characterize and compare the proteins encoding the MHC I-p repertoire. Peptides were encoded by genes located in all chromosomes and the mitochondria, and were significantly enriched for genes located in chromosomes 17 and 6. Our analysis revealed an enrichment of genes implicated in cell cycle, transcription and protein synthesis, as well as in immune and viral infection-related pathways, reflecting a B cell-specific signature.
The mechanism of resistance to parasite infection. (100.14)
Authors: Grover Harshita , Reese Michael , Blanchard Nicolas , Gonzalez Federico , Chan Shiao , Robey Ellen , Shastri Nilabh 
Published: January 2023

Abstract
The parasite, Toxoplasma gondii (T. gondii) can cause severe disease in immunocompromised humans. Susceptibility to T. gondii is also linked to the MHC haplotype: B10.D2 (H-2d) mice are relatively resistant to T. gondii infection than C57Bl/6 mice (B6, H-2b). CD8 T cells, activated by MHC molecules, are known to be that primary mediators of protection from T. gondii, and it is possible that differences in their responses might determine susceptibility to infection. Indeed, in H-2d mice, we recently identified the key immunodominant peptide, HF10, contained in the GRA6 protein that elicited protective CD8 T cell responses. In contrast, we show here that the CD8 T cell response to T. gondii in susceptible B6 mice is weaker and specific for a distinct peptide derived from the ROP5 protein. Furthermore, unlike the HF10 peptide that generated protective CD8 T cells in H-2d mice, immunization with the ROP5 peptide failed to diminish the parasite burden in chronically infected H-2b mice. Thus, susceptibility to T. gondii correlates with MHC-linked processing of specific parasite proteins that elicit differential CD8 T cell responses in mice.
The role of ERAAP polymorphisms in the autoimmune disease Ankylosing Spondylitis (100.17)
Authors: Bakker Arne , Shastri Nilabh 
Published: January 2023

Abstract
Ankylosing spondylitis (AS) is a chronic inflammatory joint disease of the axial skeleton, ultimately leading to fusion of the spine. For over three decades it has been known that the MHC class I molecule HLA-B27 is strongly associated with AS. In addition, the ER-resident aminopeptidase ERAAP (or ERAP1) was recently shown to be associated with as much as 26% of the risk for developing AS in HLA-B27 positive individuals. Multiple non-synonymous SNPs have been identified in the ERAAP gene that increase the risk of AS, but the functional significance of these polymorphisms is not known. The ERAAP protease is an essential component of the MHC class I pathway where it trims antigenic peptides in the ER. The significant association of both HLA-B27 and ERAAP with AS suggests that an altered MHC class I antigen processing pathway could play an important role in this partially T cell-mediated disease. To test this hypothesis we have analyzed the antigen processing capacity of several polymorphic forms of human ERAAP in ERAAP-deficient cells. All the polymorphic ERAAP constructs tested were enzymatically active and able to trim model antigenic peptides in different cell lines, with either no or minor differences in the quantity of total epitopes generated. We will discuss the implications of these findings.
Presentation of citrullinated peptides by class II histocompatibility molecules is associated with autophagy. (100.19)
Authors: Rimer Jamie , Unanue Emil 
Published: January 2023

Abstract
Autoimmune responses to citrullinated proteins are associated with rheumatoid arthritis; however, little is known of the mechanism of citrullination by the antigen presenting cells (APC) of the immune system. Using T cell hybridomas as probes, we found constitutive presentation by class II-Major Histocompatibility Complex (MHC) molecules of a dominant citrullinated peptide from hen egg-white lysozyme (HEL) by either macrophages or dendritic cells. Treatment of APC with 3-Methyladenine (3MA) blocked presentation of the citrullinated peptides after processing unmodified HEL, but presentation of unmodified peptides was not at all affected. In contrast presentation of citrullinated peptides by splenic B cells or B lymphoma cells was not detected under normal culture conditions. However, B cell lines presented citrullinated peptides after serum starvation in a process affected by 3MA treatment or by reducing the expression of the autophagy protein Atg5. Higher levels of citrulline in naturally processed peptides from serum starved B cell lines were detected biochemically. Primary B cells presented citrullinated HEL peptides after engagement of their B cell receptor by antigen or by using anti-immunoglobulin antibodies. Our findings demonstrate a role for autophagy in citrullination of antigen during processing by APC and indicate a link in B cells between engagement of their antigen receptor and presentation of citrullinated peptides.