bluuebunny/crossref_metadata_2025_split huggingface dataset:
Increased ERK1/2 activation leads to sustained T cell receptor signaling in rheumatoid arthritis (49.8)
Authors: Singh Karnail , Deshpande Pratima , Pryshchep Sergey , Weyand Cornelia M , Goronzy Jorg J 
Published: January 2023

Abstract
Immune responses to citrullinated neoantigens indicate a tolerance defect in rheumatoid arthritis (RA). We examined the hypothesis that abnormal TCR signaling in RA T cells lowers the activation threshold. T cells from 33 seropositive RA patients and 33 controls were stimulated by CD3/CD28 crosslinking or superantigen-coated dendritic cells. Activation-induced signaling was quantified by PhosFlow, Western blotting, and confocal microscopy. Basal as well as post-stimulation p-ERK was higher in naïve, memory and effector CD4 and CD8 T cells from RA patients compared to controls (p&lt;0.01 to p&lt;0.001). Increased ERK phosphorylation was also observed upon PMA/ionomycin stimulation suggesting a defect distal of early TCR signals. Phosphorylation of ZAP70 and NFk-B was only elevated at late time points, and this increase was completely amenable to MEK1 and ERK inhibitors. Increased ERK responsiveness led to delayed recruitment of SHP-1 to the immunological synapse of RA T cells and lowered the TCR threshold to suboptimal stimulation. Increased ERK responsiveness was also a characteristic finding in the SKG mouse model of RA and preceded clinical symptoms. MEK-1 inhibition with subtherapeutic doses significantly delayed the onset of less severe disease. We propose that increased ERK activity tunes TCR thresholds to impair tolerance mechanism and induce autoimmunity in RA. Supported by RO1-AR41974 & AI 44142.
IC87114, a Selective Inhibitor of PI3Kδ Suppresses Joint Inflammation and Bone Erosion in Collagen-Induced Arthritis in Rat (50.14)
Authors: Puri Kamal D , Steiner Bart H. , Kashishian Adam S , Chen Hao , Gallatin W. Michael , Giese Neill A 
Published: January 2023

Abstract
PI3Kδ plays an essential role in antigen-receptor signaling, proliferation, activation and function of lymphocytes. Mice deficient in PI3Kδ activity show substantial reduction in immunoglobulin levels, partial impairment in chemoattractant-induced neutrophil migration as well as defects in signaling and function of macrophages. Collagen-induced arthritis (CIA), a commonly used model for studying antirheumatic drugs, requires participation of both B and T cells to initiate disease and reproduces many of the pathogenic mechanisms detected in human rheumatoid arthritis. In this study, we have investigated the ability of IC87114, a selective PI3Kδ inhibitor, to reverse the rheumatoid arthritic like state in the CIA model of arthritis. Clinical arthritis was initiated by immunizing animals with collagen followed by a booster dose on day 7. IC87114 or control treatment was initiated when at least one hind paw was significantly inflamed and continued for an 18-day treatment course. IC87114-treatment of arthritic rats reduced the progression and severity of clinical arthritis that was evident within 6 days after initiation of therapy. IC87114-treatment significantly reduced the level of anti-collagen antibodies. Arthritic rats treated with IC87114 had significantly lower radiographic scores compared with control, indicating that treatment with IC87114 was effective in protecting bone integrity. Histopathological evaluation demonstrated that IC87114 was effective in reducing the histological changes induced by rheumatoid arthritis. The effect of IC87114-treatment on these parameters together suggests a therapeutic potential for PI3Kδ-selective compounds to ameliorate rheumatoid arthritis.
NADPH quinone oxidoreductase induction by sulforaphane inhibits synovial hyperplasia and rheumatoid T cell proliferation. (50.15)
Authors: Kim Wan-Uk , Kong Jin-Sun , Yoo Seung-Ah , Yoon Hyung-Ju , Park Yoon-Jung , Cho Chul-Soo 
Published: January 2023

Abstract
Sulforaphane (SFN), derived from cruciferous vegetables, has been previously shown to induce phase II enzyme and to have anti-carcinogenic activity. In this study, we investigated whether SFN regulates synoviocyte hyperplasia and T cell proliferation in rheumatoid arthritis (RA). SFN induced synoviocyte apoptosis by regulating Bcl-2/Bax expression and pAkt activity. In addition, SFN dose-dependently inhibited T cell proliferation and the production of IL-17 and TNF-alpha by RA mononuclear cells stimulated with anti-CD3 antibody, but enhanced the enzyme activities of NADPH quinone oxidoreductase-1 (NQO1), a representative phase II enzyme. SFN inhibition of synoviocyte survival was abrogated by co-treatment with curcumin, an inhibitor of NQO1 or by downregulation of NQO1 transcripts. The intra-peritoneal administration of SFN to mice with type II collagen (CII)-induced arthritis suppressed arthritis severity, anti-CII antibody levels, and T cell proliferative response to CII. The productions of IL-17, TNF-alpha, IL-6, and IFN-gamma from lymph node and spleen cells of these mice also were markedly reduced by SFN treatment. Moreover, in mice with anti-CII antibody-induced arthritis, a passive model of arthritis that is independent of adaptive immunity, SFN treatment reduced arthritis severity and synovial hyperplasia. In summary, NQO1 induction by SFN was found to inhibit synoviocyte proliferation and TH17 cell activation in vitro. Moreover, the anti-arthritic and immune regulatory effects of SFN were confirmed in vivo, which suggests that SFN may offer a possible treatment option for RA.
Targeted ablation of human CD30 positive cells with an anti-CD30-MMAE antibody-drug conjugate (SGN-35) in vivo inhibits antigen-specific secondary immune responses (50.17)
Authors: Oflazoglu Ezogelin , Zeng Weiping , Klussman Kerry , McEarchern Julie A , Law Che-Leung , Grewal Iqbal S , Sievers Eric L , Gerber Hans-Peter 
Published: January 2023

Abstract
CD30 is a member of the tumor necrosis factor receptor superfamily whose normal physiologic expression is restricted primarily to relatively small subpopulations of activated T- and B-cells. In abnormal physiologic states, CD30 has been found on inflammatory cells in patients with a variety of chronic inflammatory diseases such as atopic dermatitis and asthma. In addition, malignant Reed-Sternberg and anaplastic large cell lymphoma cells express particularly high levels of CD30. Cells expressing CD30 have been shown to be activated and produce cytokines during inflammatory responses. SGN-35 is an antibody-drug conjugate that binds to human CD30, ablates CD30+ tumor cells in vitro, and strongly inhibits growth of HL and ALCL tumors in vivo. In the present study, the effects of SGN-35 on antigen specific secondary immune responses were evaluated. Compared to untreated cells, SGN-35 induced a dose-dependent inhibition of T cell proliferation, with maximum inhibition of 70% following antigen stimulation in vitro. In the hu-SCID mouse tetanus toxoid model, antigen-specific immunoglobulin responses in SGN-35 treated mice were much lower compared to untreated controls (p=0.0019). Taken together, these data demonstrate specific targeting of subsets of activated lymphocytes and suggest that SGN-35 may be employed clinically to decrease antigen specific cells and immunoglobulin that damage tissue in autoimmune and inflammatory diseases.
Differential effects of anti-TNF-a mAb and the TNF-R-Fc fusion molecule on fibroblast-like synoviocyte apoptosis in arthritis. (50.19)
Authors: Pattacini Laura , Salvarani Carlo 
Published: January 2023

Abstract
In our study, we tested the hypothesis that different anti-TNF-a drugs had different effects on apoptosis of fibroblast-like synoviocytes (FLS) obtained from rheumatoid arthritis (RA) patients. FLS and peripheral blood mononuclear cells (PBMC) from RA patients were co-cultured and incubated with one of the 3 biologics for 6 days and cell death and signal transduction pathways examined. Whereas all 3 biologics induced an increase in FLS apoptosis, etanercept was the most efficacious, leading to a 3-fold increase in apoptosis induction (p&lt;0.01). All of the biologics activated PTEN-FAK pathway and up-regulated Bax. However, whereas infliximab and adalimumab caused tmTNF dephosphorylation and Bcl-2 upregulation, etanercept had very little effect. The incubation with the molecule MAK195 - the F(ab)'2 fragment of the anti-TNF-a - as well as the cross-linking of etanercept, resulted in reduction of apoptosis induction and upregulation of Bcl-2. In our study, we have shown that the three drugs have a different efficacy in apoptosis induction. Whereas all of them can activate a pro-apoptotic pathway, only the anti-TNF-a antibodies can also induce a reverse signaling, which, in FLS, results in the upregulation of Bcl-2. This finding might be relevant in the choice of the therapy for patients with RA, since etanercept could be more indicated in patients with a particularly aggressive FLS phenotype.
Potent Therapeutic Effect of Apeaz™ Cream in the Mouse Collagen Induced Arthritis Model (50.20)
Authors: nuhaily Samer , Yei Soonpin , Meza Miguel , Incardona Francesca , Esber Heny J , Damaj Bassam B 
Published: January 2023

Abstract
Rheumatoid arthritis (RA) is a chronic autoimmune disease of unknown etiology, characterized by chronic inflammation in the joints and subsequent destruction of the cartilage and bone. OTC NSAIDs are only approved for temporary pain relief and do not qualify as Disease-modifying antirheumatic drugs (DMARDs). Apeaz™ is an OTC cream approved for temporary relief of pain resulting from arthritis. The main ingredient in Apeaz™ is methyl salicylate as anti-inflammatory with glucosamine sulfate, menthol, camphor and methyl sulfonyl ethane as inactive excipients. Because Collagen-induced arthritis (CIA) is a murine experimental disease model sharing a number of clinical, histological and immunologic features with RA, we used it to study the effect of topically applied Apeaz™, once a day, on the pathogenesis of arthritis. The results show that treatment with Apeaz™ has great benefit at the clinical and pathological levels, as the therapeutic effect of Apeaz™ was associated with the downregulation of both inflammatory and autoimmune components of the disease. Treatment with Apeaz™ completely abrogated joint swelling and destruction of cartilage and bone as demonstrated by histopathology and X-ray analysis of joints. In addition, Apeaz™ reduced the levels of inflammatory cytokines in the joints of arthritic mice. Our data suggest that Apeaz™ could be a viable candidate as a DMARD for RA.
Infection with VSV ameliorates autoimmune disease in FcγR2B deficient mice (50.23)
Authors: Kole Hemanta K , Deane Jonathan A , Bolland Silvia 
Published: January 2023

Abstract
Both genetic and environmental factors, including infection by pathogens, affect the development of autoimmune diseases such as lupus. We are investigating the effect of viral infections on the lupus model of FcγR2B-/- mice. We infected FcγR2B-/- mice a single time at 2 months of age with vesicular stomatitis virus (VSV) and observed the development of autoimmune disease for the following 6 months. Infection with VSV significantly reduced autoantibody titers in serum and improved survival of FcγR2B-/- mice. B cell development in the bone marrow was not affected in VSV infected mice whereas memory B cells, and germinal center, plasma, and IgG producing mature B cell numbers were reduced. VSV infected FcγR2B-/- mice had lower number of follicular helper T cells and CD11c+ cells in the spleen. Increased survival in these mice correlated with a reduction of inflammatory cytokines and chemokines in the serum. The fact that a single round of infection with VSV confers long time protection from lupus in this mouse model indicates that the suppressor mechanism underlying the VSV effect on autoimmunity could be an excellent candidate for therapies seeking long term efficacy.
Histone/Protein Deacetylase Inhibitors (HDACi) and DNA Methyltransferase Inhibitors (DNMTi) Enhance Human Treg Function Through Epigenetic Regulation of Foxp3 (50.27)
Authors: Akimova Tatiana , Wang Liqing , Golovina Tatiana N , Mikheeva Tatiana V , Riley James L , Hancock Wayne W 
Published: January 2023

Abstract
Manipulation of CD4+CD25+FOXP3+ Treg cells may be a valuable new approach to therapy for transplantation and autoimmunity. We report the first data involving epigenetic modulation of the suppressive functions of human Tregs. CD4+CD25- T cells and CD4+CD25+ Treg cells from 14 healthy donors were isolated by magnetic beads. CFSE-labeled CD4+CD25- T cells were stimulated with CD3 mAb and irradiated PBMC, co-cultured for 3-4 d with Tregs and varying concentrations of HDACi (sodium butyrate, valproic acid) and/or DNMTi (decitabine, zebularine, RG108), and cell divisions determined by CFSE dilution. Control wells without drugs provided serial suppression data specific for each donor, and wells without Tregs showed maximal cell divisions that were not markedly impaired by drugs. We found that HDACi or DNMTi directly increased expression of Foxp3 and Foxp3-dependent genes and increased the suppressive activity of human Tregs by up to 3-fold (p&lt;0.05). This approach also markedly improved outcomes in parallel studies of murine Tregs in vitro and in vivo, including in murine models of autoimmunity and transplant rejection. We conclude that epigenetic-based therapies using clinically approved drugs may provide the means to usefully upregulate Treg functions in patients with autoimmunity or post-transplantation.
Amelioration of Lupus Nephritis in MRL/lpr mice by Adoptive Transfer of IgM Anti-dsDNA antibodies (50.28)
Authors: Zhao Minglang , Jiang Chuancang , Diaz Marilyn 
Published: January 2023

Abstract
The role of IgM autoantibodies in lupus nephritis (LN) remains controversial. Our previous work with AID-deficient MRL/lpr mice demonstrated that IgM autoantibodies do not contribute to LN. In fact, these mice experienced an abrogation of LN in spite of extremely high levels of IgM autoantibodies, and their survival levels exceeded those of mice with B cells but lacking antibodies. This suggests that IgM autoantibodies may play a protective role in LN. To determine if IgM autoantibodies do in fact protect against LN and to delineate the characteristics of protective autoantibodies, MRL/lpr mice were passively transferred with serum, IgM anti-dsDNA antibodies and IgM non-autoantibodies collected/generated from AID-sufficient and -deficient MRL/lpr mice. MRL/lpr mice treated with the serum from AID-deficient MRL/lpr mice (having higher levels of IgM autoantibodies but no IgG) showed an apparent reduction in the severity of kidney disease, while the mice treated with the serum from AID-sufficient MRL/lpr mice (having both IgM and IgG autoantibodies) seemed to have increased renal impairment. MRL/lpr mice treated with Anti-dsDNA monoclonal IgM either from AID-sufficient or -deficient MRL/lpr mice displayed a reduction in the severity of kidney damage with a delayed onset of proteinuria, significantly less deposition of immune complexes, inflammatory infiltration, and apoptotic cells in the kidneys. In contrast, MRL/lpr mice treated with non-autoreactive monoclonal IgM did not seem to delay the onset of nephritis. Therefore, passive transfer of autoreactive IgM ameliorated renal impairment in MRL/lpr mice, demonstrating a protective role of IgM autoantibodies in the pathogenesis of LN.
* Zhao and Jiang contributed equally to this work
A critical mediator of leukocyte extravasation during inflammatory disease (50.8)
Authors: Kithcart Aaron P , Sielecki Thais , Short A , Mavrikis G , Williams J , Smith K , Shawler T , Gienapp I , Satoskar A , Whitacre Caroline C 
Published: January 2023

Abstract
Multiple sclerosis (MS) is a demyelinating neurological disease affecting more than 400,000 Americans. MS pathology involves activation of autoreactive lymphocytes and their migration across the blood-brain barrier (BBB) into the central nervous system (CNS). MS patients have a higher level of the ubiquitously expressed, proinflammatory cytokine MIF. Mice lacking MIF have fewer leukocytes in the CNS following induction of experimental autoimmune encephalomyelitis (EAE), an animal model for MS. We found using bone marrow chimeras that mice expressing MIF from myeloid cells had significantly more severe disease than MIF KO mice or mice expressing MIF only in non-myeloid cells (CDI±SD; 28.0±17.9 vs. 10.3±17.9 or 4.7±10.4). Using immunohistochemistry, we found more infiltrates in the CNS of mice expressing MIF from myeloid cells. We utilized a small molecule inhibitor of MIF to evaluate whether disruption of MIF activity could inhibit migration of leukocytes. Mice had reduced severity of EAE following inhibitor administration and reduced migration of leukocytes into the CNS. Other groups have shown that blocking MIF reduces expression of adhesion molecules on the BBB. Our data suggests that during inflammatory disease, MIF from myeloid cells is critical for leukocyte migration and an inhibitor of MIF reduces migration and subsequent clinical disease. (Supported by NIH grant AI 064320 and NMSS grant RG3272)
A proteomics study of MHC-associated peptide profiles displayed by macrophages after stimulation in vitro with human low density lipoproteins (78.12)
Authors: Attiyah Sama M , Bermudez-Fajardo Alexandra , Oviedo-Orta Ernesto 
Published: January 2023

Abstract
Recognition of modified self proteins by macrophages usually induces the generation of peptide fragments derived cytoplasmic pool of cells that can be presented by the major histocompatibility complex (MHC). Despite the potential for presentation of a wide range of peptides only a limited number of peptides are displayed by the complexes. This generates peptide patterns that imprint a signature associated with particular cellular or disease processes. The aim of this study was to investigate whether stimulation of macrophages with human low density lipoprotein (LDL) generates peptide patterns on macrophage MHC class I and II in vitro. The study also assessed the effectiveness of peptide elution methods for peptide extraction. To accomplish this we stimulated J774.2 cells with native or oxidised LDL particles and extracted the MHC-associated peptides using citrate-phosphate buffer at pH 3, 4 & 5. Samples were then analysed by LC/MS/MS. MASCOT analysis was carried out to identify the protein source of peptides and IEDB and NetMHCII 1.0 were used to predict the peptide-binding affinity to MHC class I and II complexes. We identified 54 peptides derived from macrophages stimulated with ox-LDL, 33 peptides derived from cells treated with n-LDL and 10 peptides identified from untreated cells. Peptide derived from proteins involved in macrophage DNA and RNA replication, signal transduction, cytoskeleton and lipid metabolism were identified. We found a differential pattern of MHC-associated peptide expression in treated macrophages. These findings contribute to future identification of disease-associated peptides for their potential use in the development of therapeutic and/or diagnostic tools.
Antigen presentation and T cell activation by Coccidioides-Ag2/PRA-cDNA-transfected primary murine dendritic cells (78.21)
Authors: Awasthi Shanjana , Vilekar Prachi 
Published: January 2023

Abstract
The dendritic cells (DCs) have an important role in antigen presentation and regulation of host defense. Immunization with in vitro modified DCs has been shown to activate protective immune responses. Among various approaches used for modifying DCs, the nonviral genetic transfection of DCs is easy and safe as compared to viral-mediated transfection methods. In this study, we investigated antigen presentation and T cell stimulatory ability of nonvirally, gene-transfected primary murine DCs. We isolated primary murine bone marrow derived DCs and transfected isolated DCs with Ag2/PRA-cDNA (protective epitope of Coccidioides, a fungal pathogen) using TransIT-TKO (Mirus Bio, WI)- a nonviral transfection reagent. We found that upto 30-40% of primary murine DCs were transfected with the plasmid DNA carrying Ag2/PRA-cDNA. The transfected DCs remained viable and expressed the transgene (Ag2/PRA-cDNA)-encoded protein. Only moderate changes were observed in the immunophenotype of genetically-transfected DCs. The activation of T cells was studied by flow-cytometry after co-culture of naive splenic T cells with DCs. The T cells co-incubated with Ag2/PRA-cDNA transfected DCs showed upregulation of cell-surface CD25 and CD69 (activation markers). These results suggest that the primary DCs can be efficiently transfected in vitro using a non viral method; the transfected primary DCs can process and present the antigens and stimulate T cells.
Genetic manipulation of BCG leads to altered presentation of CD4 and CD8 epitiopes from Antigen 85B by mouse dendritic cells. (78.22)
Authors: JAGANNATH CHINNASWAMY , Dhandayuthapani Subramanian 
Published: January 2023

Abstract
The partial efficacy of BCG vaccine has led to a search for more effective vaccines. The immuno-dominant antigen-85B (Ag85B) from BCG vaccine is processed in macrophages and dendritic cells (DCs) dependent upon the phagosomal pH and lysosomal delivery. However, BCG avoids lysosomal localization and its phagosomes are near neutral. We found that induction of autophagy enhanced the lysosomal delivery of BCG vaccine in DCs. This in turn increased the MHC-II dependent presentation of Ag85B which was detected using an Ag85B-specific T cell hybridoma. DCs also showed an enhanced presentation of Ag85B when infected with BCG that was engineered to over-express Ag85B. We found that over expressed (OE)-Ag85B triggered autophagy. Furthermore, Ag85B is secreted from BCG vaccine into the cytosol of DCs although BCG is inefficient in inducing CD8 T cells against Ag85B in mice. Since OE-Ag85B enhanced the MHC-II dependent presentation, we examined whether OE-Ag85B could also be routed through the proteasomal pathway. DCs were infected with BCG-OEAg85B and Ag85B presentation monitored with or without the blockade of proteasomes through lactacystin. Lactacystin enhanced the MHC-II dependent presentation of Ag85B suggesting that OE-Ag85B goes through the proteasome as well as lysosome during autophagy. Mice were then immunized with BCG-OEAg85B and the spleens were found to contain an increased number of CD8 T cells specific for an epitope of Ag85B detected using a pentamer-conjugate. We propose therefore that genetic manipulation of BCG may be used to alter the levels of CD4 and CD8 epitope presentation by DCs and alter vaccine efficacy.
Antigen processing and MHC-II presentation by dermal and tumor infiltrating Dendritic Cells (78.26)
Authors: Gerner Michael Y , Mescher Matthew F 
Published: January 2023

Abstract
MHC-II presentation by DC is necessary for both initial CD4 T cell priming and for peripheral effector function. We have previously found that naïve tumor-specific CD4 T cells were ignorant to poorly immunogenic murine tumors, suggesting a possible insufficiency in DC function. Here, we investigated the phenotype, antigen uptake and MHC-II presentation capacity of normal dermal DC (DDC) and tumor infiltrating DC (TIDC) in lymphoid and peripheral sites. We found that tumors were extensively infiltrated by partially mature TIDC that were phenotypically similar to DDC. However, TIDC were inefficient at MHC-II presentation due to poor intrinsic protein uptake capability. This resulted in both inferior initiation of T cell responses in the draining LN and poor peripheral effector cell accumulation. In addition, toll-like receptor stimulation selectively enhanced MHC-II antigen presentation by DDC but not TIDC in the draining LN, and did not affect overall peripheral antigen uptake of either. These results show that TIDC are functionally distinct from normal interstitial DC, indicating that neoplastic tissues can evade effector CD4 T cells through modification of DC competence. NIH AI34824(M.F.M.), CA82596(M.F.M.), NCI 2 T32(M.Y.G.).