bluuebunny/crossref_metadata_2025_split huggingface dataset:
Live imaging of peripheral T cell-DC interactions in the islets: An emerging axis of autoimmune infiltration (48.15)
Authors: Friedman Rachel S. , Melli Kristin , Sorensen Caitlin M. , Tang Qizhi , Krummel Matthew F. 
Published: January 2023

Abstract
Defining processes of islet infiltration and destruction in Type 1 diabetes (T1D) is important for its prevention and preventing destruction of transplanted islets in diabetic individuals. We hypothesize that T1D onset and persistence is maintained by T cell-dendritic cell (DC) interactions in both the draining pancreatic lymph node (PLN) and the islets. In the steady-state, we found that intra-islet DCs are a uniform semi-mature population that constitutively take up β cell antigens in preparation for antigen presentation. Islet-antigen specific CD8 T cells activated in the PLN trafficked to the islets where they induced maturation of islet DCs. Leukocytic infiltration of the islets and islet DC maturation were dependent on IFN-γ production by T cells. Blockade of antigen processing following T cell activation in the PLN also blocked islet DC maturation but not islet infiltration. This suggests that the DCs require productive antigen-presenting interactions with T cells in the islets for maturation to occur. Through 2-photon imaging of the islets, we visualized sustained interactions of islet-antigen specific CD8 T cells with islet DCs and characterized features of cytotoxicity and dwell-time in this setting. Taken together, these data suggest that directed and synaptic delivery of IFN-γ to intra-islet DCs induces their maturation.
This work was funded by the Larry L. Hillblom Foundation and the Juvenile Diabetes Research Foundation.
Natural killer cells suppress autoimmune myocarditis in mice (48.23)
Authors: Ligons Davinna L. , Talor Monica V. , Rose Noel R. , Cihakova Daniela 
Published: January 2023

Abstract
Autoimmune myocarditis is induced by the adoptive immune response; however, there is evidence that the innate immune system plays a major role in modulating disease. Natural killer (NK) cells can function in early innate responses, as well affecting the later adaptive immune response. Depletion of NK cells has been shown to exacerbate the development of virus-induced myocarditis. We hypothesized that NK cells also modulate the virus-free model of myosin-induced experimental autoimmune myocarditis (EAM). To study the action of NK cells, we immunized BALB/c mice on days 0 and 7 with myocarditogenic peptide MyHC (614-629) in complete Freund adjuvant. The mice were treated with PBS, rabbit IgG or anti-asialo GM1 antibody every day for 6 days prior to the first immunization and every 2 days following day 0 until day 21. We found that depleting NK cells significantly increased the severity of disease, and serum antibodies specific to the myocarditogenic peptide. Splenic CD4+ T cells were significantly more activated in NK depleted mice compared to controls. Cytokine analysis of heart homogenates revealed a significant increase in eotaxin and IL-5 in the hearts of mice that were depleted of NK cells. Our data suggest that NK cells suppress the adoptive immune response and modulate eosinophil migration and T cell activation during EAM in mice.
Lupus immunotherapy using CD4 targeted nanoparticles (48.29)
Authors: Look Michael W. , Stern Eric , Wang Qin , DiPlacido Leah , Craft Joseph E. , Fahmy Tarek M. 
Published: January 2023

Abstract
The pathogenesis of systemic lupus erythematosus is mediated by collaborations between CD4 T cells and B cells with autoantibody production. We propose that the use of biodegradable, nanoparticulate materials that are targeted against CD4 T cells and carry immunosuppressive drugs and proteins can be used to treat lupus. Biodegradable nanoparticles exhibit excellent therapeutic features such as biocompatibility, targeting potential, and controlled release rates of its encapsulated cargo. Here, we report results demonstrating the therapeutic efficacy of using nanoparticles to suppress lupus in NZB/W F1 mice, a lupus prone animal model. NZB/W F1 mice were administered weekly treatments of nanoparticle therapy, and disease progression and survival were monitored. Results show that nanoparticle therapy can delay disease progression and improve survival, using an immunosuppressive drug dosage in nanoparticles that is at least 16-fold less than the amount of immunosuppressive drug administered in buffer. These results suggest that nanoparticle immunotherapy can be used to effectively treat lupus at a lower dosage amount and frequency of dosage than conventional therapeutic regimens.
This research is funded by the Wallace H. Coulter Foundation. ML is funded by a National Defense Science and Engineering Graduate (NDSEG) fellowship from the U.S. Department of Defense.
B cells play a critical role for renal injury in an experimental model of systemic sclerosis (48.3)
Authors: Chen Cindy Y , Delaney Tracy A , Jane Connor , Kiener Peter , Coyle Anthony J , Herbst Ronald 
Published: January 2023

Abstract
Systemic sclerosis (SSc) is a complex autoimmune disease involving autoreactive B and T cells and excessive extracellular matrix deposition in target organs such as kidneys. We have developed a novel human collagen V (hCollV) induced autoimmune model which shares some common pathological and immunological features with Ssc. Mice immunized with hCollV have increased proteinuria, renal fibrosis, serum autoantibodies (anti- RNP, -SSA, -collagen I, II) (p&lt;0.01), and renal IgG depositions by day 42. Levels of proinflammatory cytokines and chemokines associated with tissue fibrosis, such as IL-6 and MCP-1, were elevated in the sera of immunized mice when compared to controls, as were gene expression levels in the kidneys by qPCR analysis. Depletion of B cells by afucosylated anti-hCD19 antibody resulted in reduced proteinuria, tissue fibrosis, and IgG depositions in the kidneys of immunized mice, decreased cytokines and chemokines levels, and greatly declined serum autoantibodies. Therefore, our data indicated that the B cells play crucial roles in causing renal damage in the hCollV-induced systemic sclerosis model, suggesting a potential therapeutic target to benefit Ssc patients.
Mapping Susceptibility Loci for Immune Mediated Nephritis in 129/svJ Mice (49.19)
Authors: Xie Shangkui , Li Li(Lisa) , Xie Chun , Li Li , Han Jie , Wang Tao , Zhou Joseph , Mohan Chandra 
Published: January 2023

Abstract
Laboratory strains of mice differ in their susceptibility to immune-mediated nephritis. Whereas NZW, DBA1 and 129/svJ strains of mice are highly susceptible to anti-GBM induced nephritis, most other laboratory strains, including C57BL/6 and BALB/c have significantly less disease. In order to uncover susceptibility genes for immune-mediated nephritis, a panel of 156 (129 x BALB/c)F2 mice were bred, challenged with anti-GBM antibodies and phenotyped for nephritis. About 40% of these F2 progeny had evidence of severe nephritis. Genotyping and linkage analysis revealed 2 strongly recessive 129/svJ loci for glomerulonephritis, one on chromosome 2 (linked to rs6295520, P=0.0008) and the second on chromosome 14 (linked to rs3701623, P=0.0004). Proteinuria was strongly linked to 3 recessive 129/svJ loci, including rs3677657 on chr. 7 (P=0.0026), rs3711424 on chr. 13 (P=0.0001), and the chromosome 14 locus for GN. Among these, the loci on Chr 7 and 14 were also strongly linked to glomerular crescent formation, operating additively.
To summarize, recessive loci on chromosomes 2, 7, 13 and 14 account for the enhanced susceptibility to nephritis in 129/svJ mice. Congenic dissection studies and candidate gene testing are in progress.
Class II MHC-Binding Peptides are Active Over a Broad Range of Affinities for MHC (49.29)
Authors: Mage Michael , Levin Ditza , Revilleza Maria J , Boyd Lisa , Natarajan Kannan , Shevach Ethan , Margulies David 
Published: January 2023

Abstract
To stimulate T cells, antigenic peptides must not only bind the presenting MHC molecule, but must form a complex of sufficient stability to interact with the T cell receptor (TCR). In a murine autoimmune gastritis model based on the transgenic expression of MHC-II restricted T cell receptors (TCR) directed against peptides from the alpha chain of the gastric parietal cell proton pump, the H/K ATPase, previous studies mapped residues 630-641 (PITAKAIAASVG), and residues 889-900 (PLLCVGLRPQWE) as the epitopes recognized by TXA23 and TXA51 transgenic T cells respectively. We studied the dissociation kinetics of peptides with various amino acid substitutions (including alanine-substitution at each non-alanine position). Binding to IAd was compared to T cell stimulatory activity of the same peptides in MHC-peptide complexes. Dissociation half lives at pH 5.3 (similar to endosomes) varied from ~20 hours to over 600 hrs. Peptide/MHC stability is greatest at neutral pH, where a nonstimulatory peptide (CLIP) and a highly stimulatory peptide (GHNLAVGLRPQWHE) had half lives approximately tenfold longer than at pH5.3. We find that a half-life of &gt;20 hours at pH 5.3 is sufficient for a peptide with the necessary epitopic residues to retain immunogenicity. These results can be explained in the context of the X-ray structure of the IAd/PLLCVGLRPQWE complex. Supported by the NIAID intramural research program.
A GMCSF-neuroantigen (NAg) fusion protein targets NAg to dendritic cells (DC) and is an effective therapy for experimental autoimmune encephalomyelitis (EAE) (50.10)
Authors: Blanchfield J. Lori , Mannie Mark D. 
Published: January 2023

Abstract
The GM-CSF and M-CSF cytokines were tested as domains in cytokine-NAg fusion proteins to assess targeting of NAg to different APC subsets. Fusion proteins were designed with a cytokine N-terminal domain and the encephalitogenic peptide 69-88 of guinea pig myelin basic protein (MBP) as the C-terminal domain. Studies measuring T-cell activation in vitro, prevention of EAE, and treatment of EAE showed that GMCSF-NAg was the most potent fusion protein, represented by the following rank order of activity (GMCSF-NAg &gt; MCSF-NAg &gt; GP69-88). GMCSF-NAg was 1000-fold more potent than GP69-88 in stimulating MBP-specific T-cell proliferation. The mechanism by which GMCSF-NAg promoted T-cell activation involved cytokine receptor-mediated uptake of NAg by antigen presenting cells (APC), because free GM-CSF inhibited the GMCSF-NAg potentiated response. GMCSF-NAg potently targeted NAg to dendritic cells and macrophages in vitro, but not to B or T cell APC. Covalent linkage between GM-CSF and NAg was required for enhanced potency of GMCSF-NAg in vitro and for the prevention and treatment of EAE in vivo. In conclusion, GMCSF-NAg potently targeted self-antigen to myeloid APC subsets and caused profound antigen-specific tolerance in EAE.
This study was supported by research grants from the National Multiple Sclerosis Society and the Brody Brothers Endowment Fund.
Gene therapy promoting inflammation-triggered IL-10 production ameliorates collagen induced arthritis (50.21)
Authors: Henningsson Louise , Eneljung Tove , Lidberg Ulf , van de Loo Fons , van den Berg Wim , Gjertsson Inger 
Published: January 2023

Abstract
Current treatments of rheumatoid arthritis are focused on suppression of the immune response irrespective of the inflammatory status. Interleukin (IL) -10 is a cytokine with anti-inflammatory properties. Lentiviral vectors has been used to insert an additional IL-10 gene, driven by an inflammation-inducible promoter, into the mouse genome. This enhanced endogenous production of IL-10 would allow an arthritic host to inhibit episodes of flares whilst protecting form joint destruction. Bone marrow transplants were performed to introduce the virus into DBA/1 mice. The mice were immunized with collagen type II and arthritis was evaluated throughout the experiment. The mice treated with the IL-10 construct showed a delayed onset of disease and a significantly less severe arthritis compared to controls. The frequency of arthritis in the control group increased to 70% while only 30% of the IL-10 treated mice were affected. The use of viral vectors expressing IL-10 significantly ameliorates the severity of collagen induced arthritis. Further experiments are ongoing to elucidate the mechanism of protection.
This study was supported by grants from the Swedish Medical Research Council, Göteborgs Läkaresällskap, King Gustav V's 80 Year Foundation, Reumatikerförbundet i Göteborg, Professor Nanna Svartz Foundation, Thölen and Kristler Foundation, the Swedish Foundation for Strategic Research, the 6th Framework Program of the European Union, NeuroproMiSe, LSHM-CT-2005-01863, AUTOCURE, LSHM-CT-2005-018661
Role of TLRs in the induction of tolerogenic CD8a- dendritic cells by GM-CSF (50.34)
Authors: Prabhakar Bellur S , Ganesh Balaji B , Pilli Tania , Bhattacharya Palash , Gopisetty Anupama , Vasu Chenthamarakshan 
Published: January 2023

Abstract
GM-CSF is a potent dendritic cell (DC) growth factor that plays an essential role in DC differentiation. We examined the potential of GM-CSF to impart tolerogenic potential to CD8a+ and CD8a- sub-populations of DCs. We show that GM-CSF selectively modulates myeloid CD8a-, and not plasmacytoid or CD8a+, DCs in vivo. GM-CSF treatment rendered CD8a- DCs tolerogenic which was characterized by the diminished production of inflammatory cytokines IL-12 and IL1β by these DCs. Self antigen, mouse thyroglobulin (mTg), presentation by GM-CSF exposed CD8a- DCs, relative to control CD8a- or other DCs, caused a significant increase in the frequency of Foxp3+ T cells among CD4+ T cells from mTg-primed mice. The ability of GM-CD8a- DCs to induce Foxp3+ T cells was severely abrogated upon addition of IL-12, and not IL1β, into the culture and indicated that reduced IL-12 production rendered them tolerogenic. Further, we studied the effects of treatment with various TLR ligands to either augment or abolish the effects of GM-CSF on CD8a- DCs. Interestingly, addition of TLR ligands such as LPS and CpG further augmented the ability of GM-CSF exposed CD8a- DCs to induce Foxp3+Tregs. These results indicated that exposure of GM-CD8a- DCs to TLR4 and TLR9 ligands can enhance their tolerogenic potential and strongly suggested that pro-inflammatory immune stimuli may confer an enhanced antigen specific tolerogenic effect on GM-CSF exposed CD8a- DCs. NIH 1RO1AI058190
Protection from diabetes after subcutaneous insulin B:9-23/IFA immunization is mediated by Tregs that require IL-10, IFNg and NO but not IL-4 (50.39)
Authors: Fousteri Georgia , Dave Amy , Juntti Therese , Bot Adrian , von Herrath Matthias 
Published: January 2023

Abstract
Induction of antigen-specific tolerance in type 1 diabetes (T1D) to auto-antigens such as the insulin B chain peptide 9-23 (B:9-23) has been proven beneficial when conducted in conjunction with incomplete Freud's adjuvant (IFA) in the NOD (non-obese diabetic) mice. Due to the B:9-23/IFA ongoing clinical trials, the ill-defined mechanisms involved in achieving tolerance after B:9-23/IFA immunization could become a setback for this otherwise attractive T1D prevention strategy. Here we report that a single subcutaneous (s.c.) B:9-23/IFA immunization is as effective at later stages of the disease as at a younger stage, and loses its protective effect after IFNg or IL-10 neutralization but not in the absence of IL-4. Interestingly, when CD4+CD25+ or IFNg-producing cells from B:9-23/IFA protected mice were transferred into pre-diabetic NOD recipients, protection from T1D was conveyed, indicating that a dominant regulatory Treg-mediated effect was operational that depended on IFNg as well as NO production. In agreement, a significant increase in Treg numbers in the spleen and pancreatic draining lymph nodes (PLN) after s.c. B:9-23/IFA immunization was observed. Thus, protection from T1D after s.c. B:9-23/IFA immunization occurs through IFNg/NO and IL-10-mediated suppressive mechanisms.
Characterization of Critical Residues of the Granzyme B Inhibitor, Serpina3n (50.43)
Authors: Marcet Palacios Marcelo , Bleackley Chris 
Published: January 2023

Abstract
Granzyme B (GrB) is crucial for the immune system in eliminating tumor and virus-infected cells by caspase-mediated apoptosis. GrB also has detrimental effects in allograft tissue rejection and autoimmune diseases. Our aim was to design effective GrB inhibitors as unique immunosuppressive drugs in treating these disorders.
Our lab has defined a novel murine inhibitor of GrB named serpina3n (ser3n). Ser3n is a superior candidate for drug design since it is the only known GrB inhibitor that is secreted extracellularly, making it more plasma stable. We performed scanning mutagenesis of the reactive centre loop (RCL) domain, which confers serpins with their protease specificity, to test which residues have inhibitory properties. Using an in vitro transcribed/translated (IVTT) ser3n protein, we found that the putative P1 residue (Met) and the P3' residue (Lys) are important for ser3n binding to GrB.
Human antichymotrypsin (ACT) is highly homologous to ser3n, but is not a GrB inhibitor. Our goal was to generate a human secretory protein that inhibits GrB. We created an ACT-ser3n chimera by replacing the RCL of ACT with that of ser3n. We then created a mutant ACT-ser3n chimera with a Met-to-Asp mutation that resulted in greater GrB binding than the original chimera, to a degree equal to ser3n binding. This inhibitor will work on pathogenic states when secreted GrB is important. This reveals a potential drug for blocking GrB in autoimmune diseases.
Cordyceps militaris modulates presentation of exogenous antigen associated with MHC class I and MHC class II pathway in dendritic cells. (78.28)
Authors: Shin Seulmee , Kwon Jeonghak , Lee Sungwon , Moon Sunhee , Lee Seungjeong , Lee Chongkil , Cho Kyunghea , Kim Kyungjae 
Published: January 2023

Abstract
Cordyceps militaris has been used in traditional medicine to treat numerous diseases and reported to have antitumor and immunomodulatory activities in vivo and in vitro. The precise mechanism of its immune response activities is unclear. The water extract of Cordyceps militaris (CME) and cordycepin (3'-deoxyadenosine) were prepared. Cordycepin, nucleoside analogue, is a metabolite of C. militaris, whereas CME is included the cordycepin 1.3 ng/mg. How CME or cordycepin modulated immune reaction mechanism in antigen presenting cells (APCs) by ability to present exogenous antigens in association with the major histocompatibility complexes (MHC) in vitro and ex-vivo were examined. Microencapsulated ovalbumin (OVA) was efficiently captured, processed and presented on both MHC class I as well as MHC class II molecules. DC 2.4 cells (H-2Kb) or bone marrow- drived DCs (BM-DCs) generated from the BM cells of C57BL/6 mouse (H-2Kb) were cultured in the presence of CME or codycepin with OVA-microspheres, and the amount of OVA peptide - MHC class I complexes was measured by T cell hybridoma, CD8 OVA 1.3 T cell, that recognizes the OVA-H-2Kb complex and IL-2 production. The increment of the phagocytotic activity and the expression of MHC molecules on DCs by CME (12.5 ~ 200 μg/ml) were shown, while suppression of its activity had shown by cordycepin. CME also increased IL-2 production when we examined the effects of CME on CD4+ T cells. On the other hand, cordycepin inhibited CD4+ T cell responses in a concentration of between 5μg to 40μg. The present study demonstrates the modulation of cross priming capability could be a target of therapeutic immunoregulation.
Key words: Cordyceps militaris, cross presentation, immunomodulator
Chlamydial respiratory infection predisposes to neutrophil dominated allergic airways disease (AAD) (79.5)
Authors: Horvat Jay C , Starkey Malcolm R , Beagley Kenneth W , Gibson Peter G , Foster Paul S , Hansbro Philip M 
Published: January 2023

Abstract
Chlamydial lung infections are associated with asthma but it is unknown how these Th1-inducing infections influence Th2-mediated asthma. We investigated the effect of current and resolved chlamydial infection on the development of hallmark features of asthma using a murine model of ovalbumin (Ova)-induced AAD.
BALB/c mice were infected with Chlamydia and had either resolved or current respiratory infection at the time of sensitization to Ova. The effect of infection on AAD was compared with un-infected or non-allergic controls.
Current, but not resolved, infection attenuated hallmark features of eosinophilic AAD; pulmonary eosinophil influx, T cell production of IL-5, mucus secreting cell (MSC) hyperplasia and airways hyper-responsiveness (AHR). Importantly, while current infection suppressed Th2-mediated AAD, robust Ova-specific IFN-γ release from T cells and allergen-driven neutrophilic inflammation were induced. This neutrophilic phenotype correlated with increased pulmonary expression of IL-12 and IL-17 and suppression of thymus and activation-related chemokine (TARC). Inhibition of neutrophil and macrophage chemotaxis into the lungs during current infection reversed neutrophilic inflammation and IFN-γ production in AAD and removed the attenuating effect of infection on MSC hyperplasia and AHR. These changes correlated with decreased IL-12 and IL-17 expression, increased TARC and altered antigen presenting cell function.
Ongoing chlamydial respiratory infections modify key allergen-specific immune responses in AAD with the composition of cellular inflammatory responses to infection crucial in determining the outcome of allergic phenotype.
Supported by the NHMRC
Preliminary characterization of chicken hemaglutination-enhancing factor (81.20)
Authors: Koppenheffer Thomas L. , Hampton Stephen 
Published: January 2023

Abstract
Hemagglutination-enhancing factor (HEF) is a chicken serum macroglobulin first described on the basis of the ability of normal chicken serum to cause pronounced aggregation of allogeneic erythrocytes (RBC) sensitized with non-agglutinating levels of alloantisera. Subsequent work showed that HEF was distinct from the first component of complement, but the identity of HEF and its ligand(s) have not been defined. We found that chicken serum agglutinated homologous RBC and sheep RBC lightly sensitized with specific IgY but not sheep RBC sensitized with rabbit IgG. Thus, the sole ligand of HEF appears to be antigen-bound IgY. HEF activity was found to be heat resistant (56o for 30 min.), unaffected by chelation with 10mM EDTA, and was not inhibited by monosaccharide ligands of C-type lectins. An isolate was obtained from chicken serum following salt precipitation, gel filtration and ion exchange chromatography that contained potent HEF activity, and consisted almost exclusively of IgM. These findings suggest that HEF is a naturally occurring IgM antibody that has the capacity to interact with antigen-bound IgY.
Supported in part by the Ralph & Louise Bricker Endowed Fund
The CD8+ T cell memory state of readiness is actively maintained and reversible (83.18)
Authors: Allam Atef , Giardino Torchia Maria Letizia , Conze Dietrich , Yagita Hideo , Munitic Ivana , Sowell Ryan T , Marzo Amanda L , Ashwell Jonathan D 
Published: January 2023

Abstract
The ability of the adaptive immune system to respond rapidly and robustly upon repeated antigen exposure is known as immunologic memory, and it is thought that T cell progression from the naïve to the memory state is an irreversible differentiation event. Here we report that unlike naïve T cells, which are in the G0 phase of the cell cycle, CD8+ memory T cells exist in a state of readiness in G1. This was maintained by occupancy of members the TNF receptor superfamily, CD27 and 4-1BB, which when disrupted caused memory T cells to revert to G0 and respond like naïve T cells to stimulation. Furthermore, maintenance of the G1 state was compromised in antigen-specific memory T cells of VSV-infected CD27-deficient mice. Manipulating the memory T cell state of readiness might benefit disorders involving chronic ongoing immune responses.
This work was supported by the Intramural Research Program of the National Institutes of Health, Center for Cancer Research,
National Cancer Institute