bluuebunny/crossref_metadata_2025_split huggingface dataset:
Anti-DR5 antibody eliminates inflammatory macrophages rebalancing IL-23/Th17 axis and regulatory T cells in arthritis (147.26)
Authors: Li Jun , Hsu Hui-Chen , Yang PingAr , Wu Qi , Li Hao , Mountz John 
Published: January 2023

Abstract
Dysregulated macrophages (MΦ), IL-23/Th17 axis, and regulatory T cells (Tregs) have been implicated in the pathogenesis of rheumatoid arthritis (RA). Therapeutic strategies that can restore the homeostasis of these factors in RA are unknown. Death receptor 5 (DR5) can mediate apoptosis upon binding to an anti-human DR5 antibody, TRA-8. To investigate the immunomodulatory effects of TRA-8, a 3kb mouse promoter/Floxed-STOP/humanized mouse DR5 transgenic (Tg+) mouse was generated and crossed with lysozyme M.Cre mice to achieve MΦ specific expression. Collagen II (CII)-induced arthritis was induced by chicken CII. TRA-8 (0.2 mg/mouse) was I.V. weekly initiated on day 30 until mice were sacrificed 3-mo post CII. TRA-8 treatment resulted in a 60% reduction of CD11b+ MΦ in the draining lymph nodes (LN) of Tg+ mice. Within the CD11b+ cells, ~60% of the Gr1+Ly6G+ activated MΦ and ~90% of the IL-23+ MΦ were eliminated by TRA-8. The depletion also leads to elevation of CD4+Foxp3+ Tregs (3.6% to 5.9%, P&lt;0.05) and reduction of the CD4+IL-17+ Th17 cells (1.4% to 0.6%, P&lt;0.05). After TRA-8, joint histopathology showed a significantly decreased MAC3+ MΦ infiltration, synovial hyperplasia, and destruction (arthritis score: 9.8 versus 1.5, P&lt;0.01). Neither systematic toxicities nor increased infection were found. Our study suggested that TRA-8 can be developed into a novel biologic which depletes the IL-23 secreting inflammatory macrophages (MΦ) to restore the balance between Th17 and Tregs.
Role of Th1, Th2 and Th17 cytokines on the expression of various inflammatory cytokines, defensive proteins and chemokines by primary human epithelial keratinocytes (148.10)
Authors: Rahman Saifur , Collins Mary , Williams Cara , Ma Hak 
Published: January 2023

Abstract
Psoriasis and Atopic Dermatitis are common autoimmune/inflammatory diseases of the skin. Although the exact etiology is not clear, recent data suggest a dysregulated interplay between skin infiltrates and keratinocytes may play a role in disease progression. To better understand this relationship primary human fetal epithelial keratinocytes were cultured in the absence or presence of various Th1 (IFNg), Th17 (IL-17, IL-21 and IL-22) and Th2 cytokines (IL-4, IL-13) that are known to play a role in either psoriasis or AD. Our results demonstrate that individual treatment with either Th1 or Th17 cytokines (IFN-γ, IL-17 or IL-21) induced the proinflammatory cytokine gene expression of IL-1a and IL-6. However, the Th2 cytokines IL-4 and IL-13 had no effect on expression of the proinflammatory molecules. Additionally, IFN-γ or IL-22 alone or in combination increased the expression of ICAM-1, highlighting the potential for these cells to facilitate leukocyte recruitment to the skin epidermis. Interestingly, only treatment with IL-22 individually or in combination was found to markedly increase Defensin b, Filaggrin, S100A7 and S100A8, genes important in maintaining and protecting the integrity of the skin whereas treatment with only IL-21 or in combination markedly increased the expression of Twist-1, a signature gene for epithelial mesenchymal transition. Taken together, these results suggest various cytokines have unique and redundant roles in regulating keratinocyte activity.
The role of T cells in the development of arthritis in Inherited Inflamed Joints mice. (148.12)
Authors: Shaum Katherine , Combs Matt , McMahon Eileen 
Published: January 2023

Abstract
The IIJ (Inherited-Inflamed-Joints) mouse strain spontaneously develops an inflammatory, possibly autoimmune, arthritis at an early age. While only a third develop disease, incidence displays a distinct female-bias and mice have elevated Ig, autoantibodies, and an increased percent of double negative (DN; CD4-CD8-CD3+) T cells in their secondary lymphoid organs. The goal of our research was to further examine the T cell compartment. The thymi of AR IIJ mice were significantly decreased in size compared to non-arthritic (NAR) IIJ and SJL controls. Flow cytometry revealed significant disregulation in thymocyte subpopulations and immunohistochemistry indicated disrupted cortical/medullary organization. AR IIJ mice also had increased thymic apoptosis based on Annexin V/PI staining and TUNEL assays were performed to investigate the spatial localization of apoptosis. While it remains unclear whether the DN T cells in AR IIJ mice contributes directly to arthritis development, we used flow cytometry to show that they were present in the blood before clinical symptoms began. Finally, since the IIJ strain was originally derived from a transgene-negative mouse from a breeding colony of 5B6 TCR transgenics, we used flow cytometry and PCR to verify the absence of the transgenes. Overall, our studies confirm that IIJ mice have significant disruption in the T cell compartment and that this disruption occurs before onset of clinical disease suggesting a possible causal role in pathogenesis.
Enhanced T cell responses to modified and unmodified myelin epitopes in subjects with multiple sclerosis (148.19)
Authors: James Eddie , Kwok William 
Published: January 2023

Abstract
MS is an autoimmune disease characterized by infiltration of CD4+ T cells and other immune cells into the central neural system, leading to demyelination and neuronal damage. Myelin antigens are known to undergo post-translational modifications such as citrullnation and phosphorylation. However, responses to the modified forms of these antigens have been minimally studied. We used a systematic approach to investigate CD4+ T-cell responses to modified and unmodified myelin epitopes. We first characterized the ability of DR1501 to accommodate citrulline or arginine containing sequences, demonstrating that citrulline is accepted within three of its binding pockets. Based on previously published methods, we developed an algorithm to predict DR1501 restricted epitopes within modified and unmodified peptides from MBP, PLP, and MOG. Predicted epitopes that were able to bind to DR1501 protein were used to stimulate PBMCs from MS and healthy control subjects with DR1501 haplotypes. A subset of these sequences elicited tetramer positive T cell responses in the majority of subjects with MS, but not healthy controls. In some cases citrullination or phosphorylation altered the antigenicity of these peptides. These findings highlight the potential importance of post-translationally modified sequences in multiple sclerosis and suggest specificities that could be used for immune monitoring or antigen specific therapies.
Chemokine-directed B cell trafficking in rabbit GALT during diversification of the primary antibody repertoire (149.4)
Authors: Lanning Dennis , Knight Katherine 
Published: January 2023

Abstract
Rabbits preferentially rearrange the 3’-most VH gene segment during VDJ gene rearrangement. Early in life, rabbit B cells diversify these VDJ genes in gut-associated lymphoid tissue (GALT) in response to signals acquired from intestinal commensal bacteria, and thus generate a diversified primary antibody repertoire. To gain insight into the cell-cell and cell-microbe interactions required for VDJ gene diversification, we examined B cell surface expression levels of four chemokine receptors that mediate trafficking to important sites within GALT during repertoire diversification. We stained B cells from two- through seven-day-old appendix with fluorescence-labeled Ig-fusion proteins of the chemokines CXCL12, CCL20, CXCL13 and CCL21 and analyzed them by flow cytometry. The changes we observed in relative chemokine receptor expression levels during the first week of life suggest a model in which B cells, upon entering appendix follicles, first traffic to the follicle-associated epithelium, where they likely encounter commensal bacteria sampled from the intestinal microbiota. B cells subsequently migrate to, and likely receive additional signals from, the follicular dendritic cell network. Upon becoming activated, B cells then traffic to the basal region of the follicle, where they proliferate and diversify their VDJ genes, thereby generating the primary antibody repertoire.
Characterization of monoclonal antibodies to human CXCR7 (149.9)
Authors: Bonnevier Jody , Loges Naomi , Bostrom Kerri , Stump Julie , Brumbaugh Kathy , Munshi Cyrus , Ni Jessie 
Published: January 2023

Abstract
The G protein-coupled chemokine receptor CXCR7 binds with high-affinity to CXCL12/SDF-1 and CXCL11/I-TAC. It is also a co-receptor for several HIV and SIV strains. The expression of CXCR7 in human cell types has been studied extensively and analyzed at the transcription and protein levels. Depending on the antibody used, the reported expression profiles of CXCR7 on human cells differ significantly. This calls into question the specificity of some of the antibodies used in the studies. In this poster, we described how our monoclonal antibody panel to CXCR7 was developed, and present vigorous testing results using flow cytometry, Western blot and molecular biology approaches on multiple CXCR7 transfectants, primary cells and cell line, to establish the specificity of our antibodies. Using different specific monoclonal antibodies within our panel, we showed that the profiles of human CXCR7 expression on hematopoietic cells differ. Although the basis of these differences is not known, it likely can be attributed to cell context and the specific epitopes recognized by the different antibodies. Our panel of anti-CXCR7 should be useful for future studies to define interaction partners that interact with CXCR7 on different cell types.
Thymic stromal lymphopoietin can restore functional T cell-independent antibody responses in young mice (150.5)
Authors: Dickinson Gregory , Liu Hongqi , Shriner Anne , Leonard Warren , Alugupalli Kishore 
Published: January 2023

Abstract
Unlike adults, young children cannot mount an antibody response to T cell-independent (TI) antigens such as Pneumococcal polysaccharides (PPS) and the basis for this impairment remains elusive. The ontogeny of B cells early in life is IL-7-independent, whereas in adults it is IL-7-dependent. Despite having TI B cell subsets, young wildtype or adult IL-7-/- mice are impaired in anti-PPS response and do not survive Pneumococcal challenge. In contrast, enforced expression of IL-7 restores protective anti-PPS responses in young mice, indicating that IL-7-dependent B cells are critical for TI responses. Thymic stromal lymphopoietin (TSLP) is structurally similar to IL-7 and utilizes IL-7Rα for signaling, therefore it may also contribute to the development of TI B cell responses. Indeed, we found that young TSLP transgenic mice generate an anti-PPS response and protective immunity comparable to that of adult mice. TSLP can be induced upon activation of TLR2 and TLR9. Borrelia hermsii, an agent of relapsing fever, activates TLR2 and TLR9 signaling and induces a protective TI antibody response even in young mice, suggesting that TSLP induced during this infection could play a role in anti-B. hermsii responses. Indeed, IL-7Rα-/- mice (deficient in both IL-7 and TSLP signaling) have a more severe impairment in anti-B.hermsii responses and suffer more prolonged bacteremia than IL-7-/- or TSLPR-/- mice. These data indicate that TSLP can restore functional TI responses in the young.
IL-21R is critical for the development of memory responses (150.9)
Authors: Lowe Leslie , Rankin Andrew , MacLeod Heather , Keegan Sean , Andreyeva Tatyana , Bloom Laird , Collins Mary , Nickerson-Nutter Cheryl , Guay Heath , Young Deborah 
Published: January 2023

Abstract
The development of long-term humoral immunity, characterized by the formation of high affinity memory B cells and long-lived plasma cells in the bone marrow, is a major goal of vaccination, but the mechanisms involved in the generation of long-term humoral immunity are not well understood. IL-21 can induce B cell differentiation into plasma cells and influence isotype switching to some subclasses of IgG. Using IL-21R KO mice, we show for the first time a role for the IL-21 pathway in the development of long-term humoral immunity. Antigen specific IgG serum responses to the T cell-dependent antigen, NP-CGG, were delayed in IL-21R KOs, but reached comparable titers to those measured in WT within 3 weeks. The formation of germinal centers, which give rise to long-lived plasma cells and memory B cells, was also delayed in IL-21R KO, but reached similar magnitudes as in WT within 2 weeks. IL-21R KO generated normal numbers of NP-specific IgG plasma cells in their bone marrow at ~1.5 months after immunization. NP-specific serum IgG antibody responses were of similar relative affinity in IL-21R KO and WT at this time, indicating that NP-specific B cells affinity matured in IL-21R KO. However, IL-21R KO failed to generate NP-specific secondary IgG antibody responses following rechallenge with NP-CGG. These results indicate that the IL-21 pathway is necessary for the formation of memory B cell responses but not high affinity long-lived plasma cells.
Insight into the mechanism of STAT5 tyrosine phosphorylation in IgE sensitized, antigen-stimulated mast cells (151.13)
Authors: Pullen Nicholas , Barnstein Brian , Wilson Bridget , Falanga Yves , Mizuno Kazuya , Ryan John 
Published: January 2023

Abstract
Previously we established that STAT5 expression is required for IgE-mediated mast cell function. STAT5 tyrosine phosphorylation is swiftly and transiently enhanced by FcϵRI aggregation in an IL-3- and MAPK-independent manner. However, a more detailed understanding of the signaling apparatus responsible in this situation is wanting. Recently, we have observed that STAT5 co-localizes with FcϵRI during mast cell activation and that lipid raft dissipation interrupts STAT5 activation. Downstream of FcϵRI we find that this mode of STAT5 activity is also independent of BTK, PI3K, Akt, Syk, and Lyn, whereas removal of Fyn kinase ablates the effect. Immunoprecipitation demonstrates that STAT5 is associated with Fyn, and this co-IP decreases concomitant with the induction of IgE-mediated STAT5 tyrosine phosphorylation. Examination of other signaling-related factors reveals that SHP-1 phosphatase deficiency enhances STAT5 tyrosine phosphorylation, even prior to stimulation. Loss of GAB2, an adaptor protein that associates with Lyn and SHP-1, also augments STAT5 activation. Targeting of STAT5A or B with knock-outs and specific siRNA pools shows that IgE-mediated mast cell cytokine production is selectively dependent upon the STAT5B isoform. Altogether these data implicate Fyn as the major positive mediator of STAT5 after FcϵRI engagement, and demonstrate importantly distinct roles for STAT5A and STAT5B in mast cell function.
Adipose eosinophils promote alternatively activated macrophages that maintain metabolic homeostasis (151.21)
Authors: Molofsky Ari , Wu Davina , Liang Hong-Erh , Bando Jennifer , Jouihan Hani , Ricardo-Gonzalez Robert , Chawla Ajay , Locksley Richard 
Published: January 2023

Abstract
Eosinophils are innate Th2-related granulocytes that are elevated in asthma, allergy, and helminth infection, and are frequency associated with alternatively activated macrophages (AAMs). Whereas adipose classically activated macrophages are associated with obesity and type-2 diabetes, AAM help maintain ‘lean physiology’. Adipose AAM can be induced by the cytokine interleukin-4 (IL-4), although a definitive cellular source of IL-4 within adipose tissue is not known. Surprisingly, we find eosinophils are resident within mouse white adipose tissues, where they are the major interleukin-4 (IL-4) expressing cells. Elevated adipose eosinophils promote Arginase-1+ AAM in an IL-4/13 dependent manner, and, in the absence of eosinophils or IL-4/IL-13, adipose AAM are diminished. In mouse models of obesity, adipose eosinophils inversely correlate with weight, and a genetic absence of eosinophils results in increased body fat and insulin resistance. Conversely, infection with the helminth Nippostrongylus brasiliensis leads to prolonged adipose eosinophilia and decreased insulin resistance. Eosinophils are evolutionarily conserved cells whose function has remained elusive, present in high levels in many naturally parasitized human populations. We speculate that eosinophils participate in integrating metabolic and immune signals to maintain homeostasis during chronic parasitism.
Increased activities of cysteine cathepsins in mouse models of asthma (151.23)
Authors: Yin Fangfang , Yu Mang , Liu Chen , Paulick Margot , Verdoes Martijn , Stein Paul , Galli Stephen , D'Andrea Annalisa , Bogyo Matthew 
Published: January 2023

Abstract
Asthma is a severe disorder characterized by chronic inflammation of the airways and increased responsiveness to nonspecific stimuli. During an asthma attack, a number of immune cells are recruited to the lung. In addition to cytokines, these cells secrete a number of proteolytic enzymes or proteases to mediate the inflammation and airway remodeling associated with asthma. Cysteine cathepsins (CCs), a family of lysosomal proteases, have been found to be present in asthma patients, but little is known about their role in asthma pathology. In our study, we have used small molecule activity-based probes that specifically target CCs to investigate the in vivo profiles of CCs in two mouse models of asthma. The two models differ in their mechanism of initiating allergic inflammation by sensitization either in presence of the adjuvant alum (mast cell independent, Alum+) or in its absence (mast cell dependent, Alum-). In the Alum- model, we have found that the activities of CCs, especially Cathepsin S, were elevated in asthmatic lungs. Using immunostaining and flow cytometry, we have discovered that macrophage is a major cell type that produces elevated CCs in asthmatic lungs, while in the fluids of bronchoalveolar lavage, both macrophage and eosinophils are the main cell types that are associated with CC activities. We are now profiling CC activities in Alum+ model. These studies will allow us to gain a complete understanding of CC activities during asthmatic responses.
Low level stimulation of FcϵRI on human skin mast cells modulates expression of adenosine receptors A2aAR and A3AR. (151.5)
Authors: Gomez Gregorio , Schwartz Lawrence 
Published: January 2023

Abstract
Adenosine is a purine nucleoside that can potentiate and inhibit IgE-induced degranulation of mast cells. Extracellular adenosine acts through four G protein-coupled receptors (A1AR, A2aAR, A2bAR and A3AR). This study shows that low level stimulation of IgE receptor (FcϵRI), but not complement factor C5a receptor (C5aR), on human skin mast cells (SMCs) differentially modulates the expression of A2aAR and A3AR. Stimulation of SMCs with the anti-FcϵRIα mAb 22E7 at a concentration that does not induce degranulation temporally induced a 5-fold increase in A2aAR and a 5-fold decrease in A3AR. A2bAR was not affected and A1AR, which is not expressed in resting SMCs, was not induced. Mechanistically, the FcϵRI-induced increase in A2aAR was dependent on the NF-κB transcription factor since it was mostly blocked by the NF-κB inhibitor BAY-11. In contrast, A3AR levels were further reduced when NF-κB was inhibited. Since A2aAR couple predominantly with Gs proteins (adenylate cyclase activation) and A3AR with Gi proteins (adenylate cyclase inhibition), cAMP levels were measured. Exposure to adenosine of SMCs incubated ±22E7 resulted in a significant increase in cAMP levels in 22E7-stimulated SMCs relative to those cultured without 22E7. These data suggest that altered expression of A2aAR and A3AR by low-level stimulation of FcϵRI which promotes increased cAMP production upon adenosine exposure might act to raise the threshold for mast cell degranulation under certain conditions.
MKK3 regulates mast cell IL-4 production through Egr1. (151.9)
Authors: Lin Tong-Jun , MacNeil Adam 
Published: January 2023

Abstract
Mast cells play a central role in allergic inflammation and are activated through crosslinking of IgE receptors, initiating a signaling cascade resulting in production of biologically potent mediators. Signaling pathways in the regulation of specific mediators remain incompletely defined. Here, we examined the role of mitogen-activated protein kinase (MAPK) kinase 3 (MKK3) in IgE-dependent mast cell activation. In an in vivo model of passive cutaneous anaphylaxis, MKK3-deficient mice showed a deficit in late phase IgE-dependent inflammation. To characterize the mechanism of this deficiency, we cultured bone marrow-derived mast cells (BMMCs) from wild-type and MKK3 knockout (KO) mice. We found that IgE-mediated mast cell activation induced rapid MKK3 phosphorylation by 5 minutes, diminishing slowly after 6 hours. In MKK3-deficient BMMCs, phosphorylation of p38 was reduced at early and later time points. Among 40 cytokines tested using a protein array, IL-4 was the only cytokine specifically down-regulated in MKK3-deficient BMMCs. Furthermore, early growth response-1 (Egr1) bound to the promoter of IL-4 in FcϵRI-activated mast cells and Egr1 transcription factor activity was diminished from 1-6 hours in MKK3-deficient BMMCs. Thus, MKK3 is a specific regulator of IgE-induced IL-4 production likely through regulating Egr1 activity in mast cells.
Activation of Nrf2 skews CD4+ cells toward Th2 differentiation (152.15)
Authors: Rockwell Cheryl , Zhang Mingcai , Fields Patrick , Klaassen Curtis 
Published: January 2023

Abstract
Nuclear erythroid 2 related factor 2 (Nrf2) is a transcription factor activated by cell stress, including oxidative and electrophilic stimuli, resulting in the upregulation of a battery of cytoprotective genes. Although best known for its role in detoxication, Nrf2 has also been shown to have a number of anti-inflammatory effects. The role of Nrf2 in T cells is largely unknown, however. Nrf2 can be activated experimentally by a number of different pharmacological agents, including tert-butylhydroquinone (tBHQ), which is also used commercially as a food preservative. The present studies found that the Nrf2 activator, tBHQ, suppresses production of IFNγ in activated CD4+ cells, while concurrently inducing production of IL-4, IL-5, and IL-13. These effects were also observed in CD4+ cells that were stimulated with anti-CD3/anti-CD28 in the presence tBHQ for 5 days and then restimulated without further addition of Nrf2 activators, suggesting that the effects are due to CD4+ cell differentiation. Furthermore, the current studies suggest that activation of Nrf2 diminishes T-bet binding activity, while concurrently enhancing GATA-3 binding activity. Collectively, the present studies suggest that activation of Nrf2 has a suppressive effect on Th1 differentiation, while promoting Th2 differentiation, which likely occurs through inhibition of T-bet activity and potentiation of GATA-3 activity.