bluuebunny/crossref_metadata_2025_split huggingface dataset:
Silicon-based peptide microarrays for epitope mapping of serum autoantibodies from patients with systemic lupus erythematosus with single-amino acid resolution (P4030)
Authors: Haddon David , Jarrell Justin , Liu Chih-Long , Price Jordan , Tangsombatvisit Stephanie , Credo Grace , Xu Guangyu , Varma Madoo , Baechler Emily , Utz Paul 
Published: January 2023

Abstract
We have developed a silicon-based peptide microarray with &gt;5,700 features, corresponding to 891 unique peptides derived from the U1-70 protein, a well-known target of autoantibodies in systemic lupus erythematosus (SLE). U1-70 is a component of the U1-small nuclear ribonucleoprotein (U1-snRNP) complex, which is one of five snRNPs that comprise the spliceosome. The peptides on the microarray represent every possible overlapping peptide, up to 21 amino acids in length, spanning amino acids 110 to 170 within the U1-70 RNA binding domain. The microarray includes regions of U1-70 targeted by SLE patient autoantibodies, although the precise epitopes are not known. Additionally, the region includes a peptide (131-150) that our lab recently identified as a target of autoreactive CD4+ T cells. We have validated the microarray using commercial anti-U1-70 antibodies, and have shown that this array is capable of sensitive and specific determination of their minimal epitopes. We went on to evaluate a cohort of SLE (n=24) patients and identified two minimal reactive epitopes, peptides 116-121 (n=3) and 143-148 (n=1), of their U1-70 autoantibodies. Further analysis of a larger SLE cohort (n=80) by peptide ELISA revealed additional patients with reactivity to peptides 116-121 (1/80) and 143-148 (5/80). Currently we are correlating reactivity to these peptides with patient clinical information in an effort to better understand the development and function of anti-U1-70 autoantibodies in SLE.
Higher cytotoxic capacity in peripheral blood of sarcoidosis patients (P3135)
Authors: Wahlström Jan , Enger Tobias , Lorenz Daniel , Rao Ramana , Forsslund Helena , Rahman Jubayer , Skold Magnus , Kullberg Susanna , Eklund Anders , Brighenti Susanna , Grunewald Johan 
Published: January 2023

Abstract
BACKGROUND. Sarcoidosis is a granulomatous disease of unknown etiology, mainly affecting the lungs, where greatly increased numbers of CD4+ T cells typically are seen. However, CD8+ T cells can sometimes make up the majority of cells recovered by bronchoalveolar lavage (BAL). Previous studies have extensively studied Th1 cytokine-producing T cells and established their role in the pathological process. Little is however known about cytotoxic cells in sarcoidosis. METHODS: Peripheral blood (from 17 sarcoidosis patients and 11 healthy controls) and BAL fluid (from 19 patients and 4 controls) was obtained. Flow cytometric analysis of cell subsets was performed after staining with antibodies against the surface molecules CD3, CD4 and CD8 as well as antibodies against the cytotoxic intracellular mediators perforin, granzyme B and granulysine. The PBMC cytotoxic activity was quantified with the 51Cr release assay. RESULTS: Sarcoidosis patients had a significantly higher percentage of both perforin+ (p &lt; 0.001) and granzyme B+ (p &lt; 0.05) CD8+ T cells in blood. In BAL, however, a tendency in the opposite direction was seen. Higher cytotoxic activity was observed in PBMC from patients with sarcoidosis compared to controls. CONCLUSION: Cytotoxic cells, including CD8+ T cells, may be involved in the propagation or regulation of the inflammatory process in sarcoidosis, but further studies are needed to delineate their role in this disease.
γδ T cells and Th17 collaborate in CD18hypo PL/J psoriasiform dermatitis (P3139)
Authors: Gatzka Martina , Hainzl Heidi , Peters Thorsten , Singh Kamayani , Tasdogan Alpaslan , Wlaschek Meinhard , Scharffetter-Kochanek Karin 
Published: January 2023

Abstract
The contribution of IL-17 to the pathogenesis of psoriasis is substantiated by the therapeutic efficacy of antibodies against the common p40 subunit of IL-12 and IL-23, like Ustekinumab, suppressing Th17 and Th1 signature cytokine production. To dissect the roles of distinct IL-17 producing T cell subsets in psoriasis, we systematically analyzed the occurrence of γδ and CD4+ T cells in blood, lymph nodes and skin of CD18hypo PL/J mice, developing psoriasiform dermatitis at 12-14 weeks of age due to reduced expression of CD18/β2 integrin to 2-16% of wild-type levels, and assessed the effects of blocking different targets within the IL-23/IL-17 axis. Severity of CD18hypo PL/J psoriasiform dermatitis correlated with a loss of skin-resident Vγ5+ T cells and concurrent skin infiltration with inflammatory γδTCRlow cells preceded by increases in Vγ4+ T cells in local lymph nodes. Moreover, inflammatory CD18hypo PL/J γδ T cells enhanced allogeneic CD4+ T cell responses in vitro. Injection of diseased CD18hypo PL/J mice with anti-γδTCR, -IL-17, and -IL-23/p19 antibodies resulted in resolution of skin inflammation and eliminated pathological γδ T cells. These results for the first time demonstrate a critical function of pathological skin-infiltrating γδ T cells in a complex psoriasis model and of wild-type CD18 levels in their suppression.
Increased expression of PRRs and NOS in patients with endometriosis (P3143)
Authors: Park Dong Choon , Yeo Joon Hyung , Yeo Seung Geun 
Published: January 2023

Abstract
Objective: To assess these immune responses, we evaluated the levels of expression of Toll-like receptors (TLR)-1, -2, -4, -5, and -9; nucleotide-binding oligomerization domains (NOD)-1 and -2; interleukins-1β, -6, -8, -10, and -12; interferon-γ; tumor necrosis factor-α; inducible nitric oxide synthase (iNOS) and endothelial NOS (eNOS); and immunoglobulins (Igs) in patients with endometriosis. Methods: The levels of TLRs, NODs, cytokines, and NOS mRNAs in peritoneal effusion were assessed by real time polymerase chain reaction, and IgG, IgA and IgM concentrations were measured by enzyme-linked immunosorbent assays (ELISA) in 40 patients with and 40 without endometriosis. Findings from the two groups were compared. Results: We observed expression of all Pattern Recognition Receptors (PRRs), cytokines, and NOS mRNAs and immunoglobulins in the effusion fluid of patients with and without endometriosis. The levels of TLR-2 and -9; NOD-1 and -2; iNOS and eNOS mRNA and CA 125 were significantly higher in the endometriosis than in the non-endometriosis group (p&lt;0.05 each). Moreover, PRRs, cytokine, and NOS showed significant correlations (p&lt;0.05). Conclusions: PRRs, cytokines, and NOS, which act cooperatively in the innate immune response, are closely associated with endometriosis. Increased expression of PRRs and NOS in peritoneal fluid may be associated with endometriosis.
Pathophysiology of cytopenias in an autoimmune lymphoproliferative disease linked to bone marrow fibrosis (P3165)
Authors: Mailloux Adam , Zhang Ling , Moscinski Lynn , Bennett John , Yang Lili , Yoder Sean , Sokol Lubomir , Loughran Thomas , Epling-Burnette Pearlie 
Published: January 2023

Abstract
Large Granular Lymphocyte (LGL) leukemia is associated with unexplained cytopenias, autoimmune features, and CD57+CD8+perforin+ T-cell lympoproliferation. Like many autoimmune disorders, the pathophysiology of cytopenias in LGL leukemia is unknown. Bone marrow (BM) pathology from 24 LGL leukemia patients was retrospectively studied using H&E, reticulin, and trichrome-stained sections, and revealed that BM fibrosis was present in 21 out of 24 patients (88%). Fibrosis, a previously undefined complication, was significantly associated with splenomegaly, the presence of cytopenias, and coexistance of autoimmune diseases. LGL cell numbers in the BM, but not peripheral blood, was associated with fibrosis severity. Colony forming assays in the presence of exogenous bioartificial collagen matrix using healthy bone marrow mononuclear cells (BM-MNCs) revealed a direct inhibitory effect of collagen on progenitor growth. Consistent with microarray analysis, primary mesenchymal stromal cultures (MSCs) from LGL leukemia BM displayed heightened collagen deposition. In co-culture assays, collagen matrix produced by LGL MSCs directly inhibited healthy hematopoietic progenitor growth support. This study demonstrates that excessive collagen matrix deposition by deregulated MSCs may result from the local BM accumulation of activated CD8+ LGL leukemia T-cells. Therefore, BM fibrosis should be explored as a mechanism for secondary cytopenias associated with autoimmune diseases.
Design of less immunogenic therapeutic factor VIII proteins (P3169)
Authors: Pratt Kathleen , James Eddie , Puranik Komal , Kwok WIlliam , Ettinger Ruth 
Published: January 2023

Abstract
Hemophilia A patients lack functional factor VIII (FVIII), which is essential for normal blood clotting. Approximately 1 in 4 patients receiving replacement therapy by infusions of FVIII develop neutralizing antibodies that can cause serious bleeds. Tetramer-guided epitope mapping (TGEM) experiments identified an HLA-DRB1*01:01-restricted T-cell epitope in 2 subjects with missense substitution A2201P. The same epitope was identified using samples from a subject with severe hemophilia. T-cell clones were isolated by single-cell sorting and expansion of tetramer-positive CD4 T cells. Peptide-MHC binding assays using recombinant DR0101 and FVIII peptides with systematic arginine substitutions identified anchor residues F2196, M2199, A2201 and S2204. Recombinant FVIII-C2 domain with substitution F2196A abrogated proliferation of 4 clones. Six FVIII proteins with rational substitutions designed to interfere with epitope presentation (F2196A, F2196K, F2196L, M2199A, M2199W and M2199R) were produced in a BHK expression system. All had pro-coagulant activities, measured by both clotting and chromogenic assays, similar to that of wild-type FVIII. Their immunogenicity is now being evaluated. These experiments provide proof of principle for the design of less immunogenic, functional FVIII proteins targeted to specific subsets of patients.
Diminished B cell response and peripheral B cell abnormalities in patients with SLE are linked to disturbances of the spleen tyrosine kinase (P4056)
Authors: Fleischer Sarah , Daridon Capucine , Dörner Thomas 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by a breakdown of self-tolerance, autoantibody production, B cell hyperreactivity and an impaired B cell homeostasis. Recent data suggested that an impaired B cell response could also lead to autoimmunity. In this study, the BCR downstream signaling spleen tyrosine kinase Syk of healthy donors (HD), SLE, rheumatoid arthritis (RA) and primary Sjögren’s syndrome (pSS) patients was analyzed in detail and revealed new insights into peripheral B cell abnormalities based on intracellular characteristics. Interestingly, SLE B cells showed a significantly lower but long-lasting Syk (Y352) phosphorylation after BCR engagement compared to HD (p&lt;0.05). Furthermore, a significant enlarged frequency of Syk(bright) B cell population has been identified within the CD27(-) B cell subset in the blood of SLE patients (p&lt;0.001), but no correlation with the disease activity was observed. Notably, the frequency of Syk(bright) B cells was not increased in RA or pSS patients. Further characterization showed that Syk(bright) cells exhibit an increased basal level of phosphorylated Syk, expressed higher levels of CD19 and CD95 and mainly do not express CD38 as compared to Syk(dim) cells. Thus, SLE patients show a diminished but long-lasting BCR response and exhibit an enlarged unique Syk(bright) B cell subset that could be responsible for the hyperactivity of SLE B cells.
Germinal center B cells but not TFH require T cell-derived IL21 in a model of autoimmune arthritis (P4068)
Authors: Block Katharine , Huang Haochu 
Published: January 2023

Abstract
IL21 is a pluripotent cytokine known to promote germinal center B cell differentiation and is thought be an autocrine factor for TFH differentiation and maintenance. The effects of autocrine IL21 on TFH have been addressed in several mouse models, but its requirement is still disputed. We investigated the importance of IL21 as a T cell autocrine factor in a model of autoimmune arthritis. Adoptive transfer of KRN transgenic CD4+ T cells into appropriate hosts drives germinal center formation and autoantibody production against glucose-6-phosphate isomerase, leading to inflammation and joint destruction. By comparing transfer of T cells deficient in IL21 to transfer of T cells lacking IL21R we were able to distinguish whether T cells themselves require IL21 to induce disease or whether it is required exclusively by other cell types. We found that T cells deficient in IL21 did not induce germinal center formation or autoantibody production, but had normal TFH differentiation. Transfer of T cells lacking IL21R was similar to wild-type transfer in antibody titer, GC B cell frequency, and TFH frequency both early and late in disease. To test whether IL21R signaling was required by autoreactive B cells to induce disease, KRN T cells and IL21R sufficient or deficient B cells were transferred into T and B cell deficient hosts. Mice that received IL21R deficient B cells did not develop arthritis, demonstrating that B cells require IL21 from TFH for a proper germinal center response.
Combination of agonist anti-OX40 therapy with CTLA-4 blockade augments anti-tumor effector CD4 and CD8 T cells (P4318)
Authors: Redmond William , Linch Stefanie , Kasiewicz Melissa 
Published: January 2023

Abstract
Ligation of the TNF receptor family co-stimulatory molecule OX40 (CD134) with an agonist anti-OX40 mAb enhances anti-tumor immunity by augmenting CD4 and CD8 T cell clonal expansion, effector differentiation, as well as turning off the suppressive activity of FoxP3+CD4+ regulatory T cells (Treg). Moreover, the targeted blockade of the checkpoint inhibitor CTLA-4 on CD4 effector and Treg cells enhances anti-tumor immunity by boosting effector T cell responses and alleviating the suppressive capacity of Treg. However, monotherapy with anti-OX40 or anti-CTLA-4 mAb can have limited efficacy, particularly against poorly immunogenic tumors. Therefore, we investigated whether combination immunotherapy could overcome this defect to generate optimal therapeutic responses. We demonstrate that combined anti-OX40/anti-CTLA-4 therapy significantly enhanced tumor regression and survival of TRAMP-C1 prostate or MCA-205 sarcoma tumor-bearing hosts. Mechanistic studies revealed that combined anti-OX40/anti-CTLA-4 therapy elicited CD8 T cell proliferation and differentiation as well as the expansion of effector CD4 T cells producing Th1 and Th2 cytokines, but did not inhibit the expansion or suppressive function of Treg cells. Together, these data demonstrate that combined anti-OX40/anti-CTLA-4 immunotherapy elicits potent anti-tumor immunity and indicate that clinical evaluation of this combination regimen to boost tumor immunotherapy is warranted.
Inhibition of T cell protein tyrosine phosphatase enhances IL-18-dependent hematopoietic stem cell expansion (P6268)
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 augment 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 increase in the number of hematopoietic progenitors in murine bone marrow. This effect could be reproduced using a short (48 h) treatment with a pharmacological inhibitor of TC-PTP in murine bone marrow, as well as in human bone marrow peripheral blood and cord blood. We also demonstrate that the ex-vivo use of TC-PTP inhibitor only provides a temporary effect on stem cells and did not alter their capacity to reconstitute all hematopoietic components in vivo. We establish that one of the mechanisms whereby inhibition of TC-PTP mediates its effects involves the IL-18 signaling pathway, leading to increased production of IL-12 and IFN-gamma by progenitor cells. Together, our results reveal a previously unrecognized role for IL-18 in contributing to the augmentation of the stem cell pool, and provide a novel and simple method to rapidly expand progenitor cells from a variety of sources using a pharmacological compound.
Expression and clinical significance of CCR9 in prostate cancer (P6276)
Authors: Singh Shailesh , Singh Rajesh , Gupta Pranav , Lillard, Jr. James , Grizzle William 
Published: January 2023

Abstract
Despite recent advances in diagnosis and treatment, prostate cancer is the second leading cause of cancer-related deaths in US. Precise etiology and molecular target to treat prostate cancer is yet to be defined. In recent years, involvement of chemokines and their corresponding receptors in pathogenesis of prostate cancer has gained tremendous attention. We have previously shown that CC chemokines receptor-9 (CCR9) is significantly expressed in prostate cancer cell lines compared to normal epithelial cells derived from prostate. Furthermore, we have shown higher invasive and migratory potential of prostate cancer cells under the chemotactic gradient of CCL25, the only known natural ligand of CCR9. The aim of this study was to establish the clinical relevance of CCR9 in prostate cancer using tissue micro array (TMA). The TMAs were stained for CCR9 and virtual slides were created using an Aperio ScanScope scanning system to numerically analyze the Immuno-intensity of CCR9. The ScanScope generated true color digital images of each stained sample, which were viewed and analyzed using Aperio ImageScope (version 6.25 software). Our data show higher expression of CCR9 in prostate cancer tissues compared to match benign tissues. Interestingly, CCR9 immuno-intensity was stronger in tissues with Gleason grade 8 and above. This study suggests the potential involvement of CCR9 in pathogenesis of prostate cancer.
Autophagosome-enriched cancer vaccine strategy provides a novel non-genetic method to engineer leukocytes to express molecules that may augment cancer immunotherapy. (P4252)
Authors: Paustian Christopher , Hilton Traci , Van De Ven Rieneke , Haley Daniel , Twitty Chris , Hu Hong-Ming , Fox Bernard , Aung Sandra 
Published: January 2023

Abstract
CD80 expression by APC provides signal 2, a critical element in priming T cell immune responses to antigen. As limited CD80 expression on APC can result in reduced immune responses, strategies that increase CD80 may have benefit. We report here that the autophagosome-enriched vaccine named DRibbles (DRiPs and SLiPs in Blebs), derived from human tumor cells transfected with a plasmid encoding huCD80, are highly efficient at transferring membrane proteins onto human PBMCs. When PBMCs were incubated with a DRibble vaccine derived from a CD80 expressing breast-tumor cell line, a significant increase in surface expression of CD80 was observed on PBMCs within 5 minutes (p&lt;0.001). This effect was time and concentration-dependent and repeatable across multiple donors (N=5) in multiple experiments (N≥16). DRibble vaccine could transfer CD80 onto as high as 60-86.4% of monocytes and 41-86.1% of CD4+ T cells. Transfer of CD80 to CD8 T cells was substantially less effective. Transfer of membrane-associated proteins between leukocyte populations is a well-documented phenomenon, however, this is the first report of transfer of membrane-associated proteins from cell-free autophagosomes. Current efforts are investigating the functional properties associated with increased expression of CD80. Given the critical role of CD80 in augmenting immune responses this provides a novel strategy to further augment the response to DRibble vaccines, which are being explored in clinical trials.
Age related pharmacokinetic and pharmacodynamic response of intranasal rGM-CSF (P4190)
Authors: Resetar Erin , Eichinger Katherine , Anderson Kacey , Empey Kerry 
Published: January 2023

Abstract
Respiratory syncytial virus (RSV) is one of the leading causes of respiratory tract infections in children. Nearly every child will be infected by the age of two, yet there is currently no vaccine or effective treatment for RSV infection. Previous studies have shown that immunotherapy, such as recombinant GM-CSF (rGM-CSF), expedites viral clearance, likely due to increased activation and accumulation of pulmonary antigen-presenting cells (APCs). This study examines the pharmacokinetics and pharmacodynamics of a weight-based, inhaled rGM-CSF dose in infant compared to adult mice. Using a Luminex assay, the concentration of rGM-CSF in digested lung tissue and blood was analyzed over 24 hours. In lung digest, rGM-CSF was rapidly detected in both ages with levels peaking at 1 hour and decreasing to endogenous levels by 8 hours. rGM-CSF levels in the blood peaked at 0.5 hours and decreased by 4 hours in pups, while adult levels peaked at 1 hour and decreased by 8 hours. Whole lung was analyzed by flow cytometry for macrophage activation at 4, 8, 24, and 48 hours post dose. In pups and adults, CD11c+/CD11b+ expression increased, suggesting activation or infiltration of APCs. CD86 and MHC class II expression increased significantly in adults, with a more marked MHCII increase at 48 hours. Using a non-compartmental pharmacokinetic modeling approach, we predict that increasing the dose from 50 to 170ng/g will achieve similar biological outcomes in pup compared to adult mice.
Proliferation promotes functional changes in NK cells during viral infection. (P6054)
Authors: Tarrio Margarite , Biron Christine 
Published: January 2023

Abstract
Following mouse cytomegalovirus (MCMV) challenge, NK cells control viral replication by killing infected cells and producing IFNγ in response to IL-12 induced at day 1.5 after infection. Previous work in our laboratory has demonstrated that under conditions of high viral replication, NK cells make significant amounts of IL-10 and deliver immunoregulatory functions at 3 to 4 days of infection (Lee et al., J. Exp. Med. 206:2235, 2009). The condition is accompanied by extensive NK cell proliferation. To test the hypothesis that proliferation is required for the changing function, culture systems were developed examining NK cell responses to IL-12 with and without division supported by high dose IL-2. Proliferating NK cells were induced to express the IL-10-reporter GFP in response to IL-12. One cell division was sufficient as was entrance into cell cycle measured by incorporation of BrdU. In contrast to intracellular IFNγ, proliferation was required for IL-10-GFP expression. Cell transfer studies into MCMV-infected mice demonstrated that IL-10-GFP levels were elevated with increasing divisions. When evaluated either in vitro or in vivo, decreasing intracellular IFNγ expression correlated with increasing IL-10-GFP expression as divisions progressed. Taken together, the studies reveal a previously unappreciated role for proliferation in shaping NK cell responses to infection. (Supported in part by the National Institutes of Health, USA.)