bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-12 Drives IFN-γ-Dependent Autoimmune Kidney Disease in MRL-FaslprMice
Authors: Schwarting A. , Tesch G. , Kinoshita K. , Maron R. , Weiner H. L. , Kelley V. Rubin 
Published: December 2022

AbstractIL-12 is secreted by kidney tubular epithelial cells in autoimmune MRL-Faslpr mice before renal injury and increases with advancing disease. Because IL-12 is a potent inducer of IFN-γ, the purpose of this study was to determine whether local provision of IL-12 elicits IFN-γ-secreting T cells within the kidney, which, in turn, incites injury in MRL-Faslpr mice. We used an ex vivo retroviral gene transfer strategy to construct IL-12-secreting MRL-Faslpr tubular epithelial cells (IL-12 “carrier cells”), which were implanted under the kidney capsule of MRL-Faslpr mice before renal disease for a sustained period (28 days). IL-12 “carrier cells” generated intrarenal and systemic IL-12. IL-12 fostered a marked, well-demarcated accumulation of CD4, CD8, and double negative (CD4−CD8− B220+) T cells adjacent to the implant site. We detected more IFN-γ-producing T cells (CD4 &gt; CD8 &gt; CD4−CD8− B220+) at 28 days (73 ± 14%) as compared with 7 days (20 ± 8%) after implanting the IL-12 “carrier cells;” the majority of these cells were proliferating (60–70%). By comparison, an increase in systemic IL-12 resulted in a diffuse acceleration of pathology in the contralateral (unimplanted) kidney. IFN-γ was required for IL-12-incited renal injury, because IL-12 “carrier cells” failed to elicit injury in MRL-Faslpr kidneys genetically deficient in IFN-γ receptors. Furthermore, IFN-γ “carrier cells” elicited kidney injury in wild-type MRL-Faslpr mice. Taken together, IL-12 elicits autoimmune injury by fostering the accumulation of IFN-γ-secreting CD4, CD8, and CD4−CD8− B220+ T cells within the kidney, which, in turn, promote a cascade of events culminating in autoimmune kidney disease in MRL-Faslpr mice.
The MHC class II-associated invariant chain regulates TLR7 trafficking and signaling in B cells (APP3P.100)
Authors: Manoury Benedicte , Maschalidi Sophia , Achour Karim , Lennon-Duménil Ana-Maria , Tohmé Mira 
Published: December 2022

Abstract
Intracellular Toll-like receptors localise in endosomes and sense nucleotides from viruses and bacteria. This recognition induces their conformational changes resulting in the production of proinflammatory cytokines and MHC class II (MHCII) antigenic presentation. TLR9 trafficking and signalling has been well described, however, regulation of Toll-like receptor 7 which recognizes single-stranded RNA from viruses in endosomes is still largely unknown. Increasing evidences describe a cross-talk between proteins that regulate both innate and adaptive responses. For example, UNC93B1, a chaperone essential for intracellular TLRs trafficking also interferes with the MHCII antigen presentation pathway and MHCII molecules were recently described to regulate TLR9 signaling. Here, we show, that in B cells, in contrast to dendritic cells, at the steady state and after stimulation, TLR7 and TLR9 reside in the lysosomes together with the MHCII molecule, the invariant chain (Ii) and H-2 DM. Following stimulation by TLR7 ligand, Ii specifically interacts with TLR7 but not with TLR9 and its adaptor molecule MyD88. Surprisingly, in the absence of Ii chain or when its expression is downregulated upon viral infection, TLR7 but not TLR9 is localized in the endoplasmic reticulum leading to an exacerbation of TLR7 innate but not adaptive function. This suggests a new role for Ii chain by acting as a negative regulator of TLR7 signaling in B cells
Targeting antigen to CD40 reinforces dendritic cells to cross-prime antigen-specific CD8+ T cells (APP3P.110)
Authors: Yin Wenjie , Gorvel Laurent , Li Dapeng , Joo HyeMee , Duluc Dorothée , Upchurch Katherine , Gu Chao , Ouedraogo Richard , Xue Yaming , Wang Zhiqing , Zurawski Sandra , Gorvel Jean-Pierre , Zurawski Gerard , Oh SangKon 
Published: December 2022

Abstract
Dendritic cells are major antigen-presenting cells that can efficiently cross-prime antigen-specific CD8+ T cells. Thus, targeting antigen to dendritic cells via surface receptors is an appealing strategy to mount CD8+ T cell-mediated immunity against intracellular pathogens and cancers. Nonetheless, which targeted receptor is the most efficient at priming CD8+ T cells remains elusive. Herein, we report the superior function of CD40 over nine different lectins and scavenger receptors at priming antigen-specific CD8+ T cells. A quantitative analysis of intracellular trafficking of antibody-bound receptors revealed that αCD40 monoclonal antibody localized mainly at the plasma membrane and subsequently accumulated at early endocytic compartments whereas αLOX-1 and αDectin-1 monoclonal antibodies localized at both early and late endocytic compartments in dendritic cells. Regardless of the differences in their subcellular localizations, targeting antigen to CD40 and lectins resulted in the same pattern of peptide epitope-specific IFNg+CD8+ T cell responses. We also report that poly(I:C) can significantly enhance both CD8+ and CD4+ T cell responses elicited by targeting antigen to CD40 in both human in vitro and in human CD40 transgenic mouse in vivo settings. This study provides key information for the rational design of DC-targeting vaccines against cancers and intracellular pathogens.
Implications of T cell receptor cross-reactivity on the CD4+ T cell repertoire (APP3P.111)
Authors: Nelson Ryan , Jenkins Marc 
Published: December 2022

Abstract
Naïve CD4+ T cell populations with T cell receptors (TCR) specific for different major histocompatibility complex II (MHCII)-bound foreign peptides vary in size for unknown reasons. Negative selection on self peptides could play a role. We found that MHCII-binding nonamer peptides only had to have the same residues at five positions to be recognized by the same TCR. This degree of TCR cross-reactivity between self and foreign peptides reduced the sizes of foreign peptide-specific T cell populations via clonal deletion. Small foreign p:MHCII-specific populations that underwent a greater degree of cross-reactive negative selection were on average lower affinity and produced more variable effector cell responses to immunization. In contrast, large foreign p:MHCII-specific T cell populations with minimal evidence of negative selection were higher affinity and produced larger responses to immunization. Reciprocally, an incompletely deleted naïve T cell population specific for a tissue-restricted self peptide could be triggered by similar microbial peptides to cause autoimmunity. Thus, TCR cross-reactivity between similar self and foreign peptides influences the size of certain foreign peptide-specific T cell populations, and may allow tissue-restricted self peptide-specific populations to cause autoimmunity after infection.
T cell-dependent and -independent regulatory roles of a new inhibitory molecule VISTA (APP3P.115)
Authors: Ohno Tatsukuni , Kondo Yuta , Azuma Miyuki 
Published: December 2022

Abstract
[Objective] VISTA is a new member of the CD28-B7 family co-inhibitory molecules. VISTA is expressed on T cells and myeloid cells, but not on B cells. It seems that VISTA functions as a co-inhibitory molecule, but the actual contribution as a receptor or ligand has not been clear. Here, we generated a new mAb against VISTA and examined the effects on T-dependent and -independent immune responses. [Results and Discussion] We generated a new anti-VISTA mAb (MIH63). In a T-cell-mediated DNFB-induced contact hypersensitivity model, treatment with MIH63 at sensitization, but not at challenge, enhanced ear swelling. In the MIH63-treated mice, the ratio of CD8+ T cells was markedly reduced, whereas the ratio of IFN-g+ effector CD8+ T cells was enhanced. Blockade of FcgR by 2.4G2 cancelled the reduction of CD8+ T cells by MIH63 without affecting ear swelling and CD8+ T cell activation. These results suggest that MIH63 possesses both antagonistic and ADCC abilities. Adaptive transfer of MIH63-treated sensitized T cells enhanced ear swelling after ear challenge. In a T cell-independent LPS-induced shock model, treatment with MIH63 markedly exacerbated the mortality and increased plasma TNFα and IL-6 levels and liver transaminases (AST/ALT). Our results demonstrated that treatment with anti-VISTA mAb enhanced both T cell-dependent and -independent immune responses, suggesting regulatory roles of VISTA. We will further report the actual contributing cells that express VISTA.
B cell directed tolerogenic fusion proteins (APP3P.117)
Authors: Zhang Ai-Hong , Wang Hong , Kim Yongchan , Scott David 
Published: December 2022

Abstract
We previously demonstrated that retroviral delivery of antigen-IgG fusion proteins in B cells ex vivo rendered these cells tolerogenic. Such B cells, upon adoptive transfer, could block or reverse the adverse immune responses in rodent models of multiple sclerosis, type 1 diabetes and hemophilia. In order to create a non-viral­ delivery system taking advantage of the tolerogenicity of IgG fusion proteins directed into B cells, we have prepared a fusion comprising anti-human CD20 single chain variable fragment and with target T-cell epitope on a human IgG4 scaffold. We term this fusion a B-cell-specific antigen IgG tolerogen (BAIT). As an example, the BAIT fusion containing a DR1 restricted T cell epitope FVIII2191-2210 (BAIT-FVIII2191) was prepared in a CHO cell expression system via stable transfection. Ex vivo analyses demonstrated that biotinylated BAIT-FVIII2191 could be specifically taken up by CD20+ve B cells from human PBMC. In this report, we will evaluate the presentation of B-cell directed BAIT-FVIII2191 to FVIII2191-2210 specific human T cells, and then test in vivo using DR1-transgenic FVIII KO mice. The results obtained from these studies will facilitate the translation of our successful B-cell gene therapy approach into a novel protein therapeutic for adverse immune responses, such as anti-FVIII inhibitor development in hemophilia A.
NLRC5 regulates MHC-I expression and immunity during viral infection (APP3P.101)
Authors: Lupfer Christopher , Stokes Kate , Kanneganti Thirumala-Devi 
Published: December 2022

Abstract
NLRC5 is a member of the NOD-Like receptor family and reportedly functions in the regulation of type I interferons and inflammasome activation. In addition, NLRC5 functions as a transcriptional transactivator of MHC-I gene expression. We have observed that Nlrc5-deficient mice infected with influenza A virus have normal type I interferon responses and inflammasome activation but have dramatically reduced H-2Db and H-2Kb major histocompatibility complex (MHC)-I allele expression on virtually all myeloid and lymphoid cells. Furthermore, Nlrc5-/- mice have diminished capacity for CD8+ T cell responses directed toward influenza A virus following infection. These data suggest that during influenza A virus infection, NLRC5 plays an important function in the generation of adaptive immunity but has no detectable role in the innate immune response.
Retina-specific T cells provide neuroprotection in a mouse model of glaucoma (BA6P.140)
Authors: Kielczewski Jennifer , Horai Reiko , Caspi Rachel 
Published: December 2022

Abstract
Glaucoma is a disease that leads to degeneration of retinal ganglion cells (RGC) and is potentially blinding if left untreated. Based on the concept of “beneficial autoimmunity”, we examined whether naïve, retina-specific T lymphocytes can provide neuroprotection to retinal ganglion cells in an optic nerve crush injury glaucoma model. Purified T cells from mice that express the transgenic R161H TCR for the retinal antigen IRBP, as well as eGFP (R161H-eGFP), were adoptively transferred into wild type (WT) mice. Also, irradiated bone marrow (BM) chimeras were stably engrafted with &lt;10% circulating R161H-eGFP cells. Recipient mice were subjected to retrograde fluorogold labeling for quantification of RGC prior to an optic nerve crush injury. Retinas were imaged 4 or 7 days later for quantification of RGCs and donor R161H-eGFP T cells were detected by confocal microscopy. In both types of recipients, T cell influx into the eye and neuroprotection were apparent, but differed in kinetics. In adoptively transferred mice, R161H T cells were detected in the eye 4 days post injury and were associated with 27% less RGC loss compared to controls infused with normal polyclonal eGFP+ T cells (p&lt;0.05). In the BM chimeric mice, major influx of R161H-eGFP cells occurred at 7 days post injury and was associated with a 16% statistically significant reduction in RGC loss. We conclude that naïve retina-specific T cells can mediate neuroprotection of retinal ganglion cells.
A novel rodent model for autoimmune pulmonary alveolar proteinosis (BA6P.123)
Authors: Ferretti Andrew , Fortwendel Jarrod , Gebb Sarah , Barrington Robert 
Published: December 2022

Abstract
Pulmonary alveolar proteinosis (PAP) is a rare lung syndrome caused by the accumulation of surfactants in the alveoli. The most prevalent clinical form of PAP is autoimmune (aPAP) whereby IgG autoantibodies neutralize the cytokine colony stimulating factor 2 (Csf2, formerly GM-CSF). Csf2 is a pleiotropic cytokine that promotes the differentiation, survival, and activation of alveolar macrophages (AM), the cells responsible for surfactant catabolism. IgG-mediated neutralization of Csf2 thereby inhibits AM homeostasis and function, leading to an innate immunodeficiency with increased susceptibility to opportunistic infections. Importantly, there are no rodent models for this disease, and therefore underlying immune mechanisms regulating Csf2-specific IgG in aPAP are not well understood. Herein, we identify autoimmune-prone Rasgrp1-deficient (KO) mice as the first rodent model of aPAP: 1) analysis of lung sections revealed an eosinophilic, periodic acid Schiff positive proteinaceous accumulation in the alveoli; 2) KO mice exhibit reduced pulmonary compliance; 3) AM from KO mice are enlarged, exhibit impaired protein levels and have reduced surfactant degradation; and 4) serum Csf2-specific IgG are increased (ELISA) in KO mice. Finally, while control mice infected intranasally with Aspergillus fumigatus clear conidia readily, KO mice failed to clear conidia from the lungs. Collectively these data suggest that KO mice represent the first rodent model for aPAP.
Ex vivo killing of primary melanocytes by syngeneic mononuclear cells from active, autoimmune vitiligo lesions in Smyth chickens (BA7P.152)
Authors: Falcon Daniel , Dienglewicz Robert , Erf Gisela 
Published: December 2022

Abstract
In humans autoimmune vitiligo is characterized by the loss of pigment-producing melanocytes in the skin. The vitiligo-prone Smyth line (SL) chicken together with the MHC-matched parental Brown line (BL) and vitiligo-resistant Light Brown Leghorn line of chickens constitute a spontaneous, non-transgenic animal model for this disorder. Cell-mediated immunity (CMI) is known to contribute to the etiopathology of vitiligo. Furthermore, CMI was demonstrated in vivo in vitiliginous SL chickens based on the delayed wattle response (DWR) to melanocyte lysates. Immunohistochemical analysis of DWR-positive tissue revealed extensive T cell infiltration indicative of a CMI response. In order to examine CMI in SL vitiligo (SLV) ex vivo we developed a co-culture assay using primary effector and target cells from growing feathers (the target tissue). Mononuclear cells (MNCs) were harvested from feathers of chickens with active SLV and BL controls. Target cells (melanocytes) were isolated from pigmented feathers prior to development of SLV. To detect melanocyte-specific death a modified flow cytometric method of Lee-MacAry et al (2001) was used. Regardless of whether melanocytes were isolated from chickens that did or did not eventually develop SLV, co-culture with MNCs from SLV chickens resulted in greater melanocyte death compared to controls. Use of this ex vivo system with sorted MNCs from active, autoimmune SLV lesions will help identify roles of individual subsets in melanocyte death.
Differential effects of IFNβ on IL-12, IL-23, and IL-10 expression in TLR-stimulated dendritic cells (BA7P.157)
Authors: Yen Jui-Hung , Kong Weimin , Hooper Kirsten , Emig Fran , Rahbari Kate , Kuo Ping-Chang , Scofield Barbara , Ganea Doina 
Published: December 2022

Abstract
Autoimmune disease multiple sclerosis (MS) is characterized by immune cell infiltration in the CNS leading to cumulative disability. IFNβ, used clinically in remitting-relapsing MS, reduces lesion formation and rate of relapses. Although the molecular mechanisms are not entirely elucidated, myeloid cells appear to be a major target for the therapeutic effects of IFNβ. Dendritic cells (DC) play a critical role in experimental models of MS, through their effect on Th1/Th17 cell differentiation and expansion. Here we studied the effects of IFNβ on DC expression of cytokines involved in Th1/Th17 differentiation. Administration of IFNβ to EAE mice inhibited IL-12 and IL-23 expression in splenic DC and reduced in vivo differentiation of Th1/Th17 cells. IFNβ affected cytokine expression in TLR-stimulated DC in a similar manner in vitro, inhibiting IL-12 and IL-23 and stimulating IL-10 at both mRNA and protein levels, by signaling through IFNAR. We investigated the role of the signaling molecules STAT1/STAT2, IRF1 and IRF7, and of the PI3KàGSK3 pathway. Inhibition of IL-12 p40 and p35 by IFNβ was mediated through STAT1/STAT2, whereas inhibition of IL-23 was STAT1-depedent, and the stimulatory effect on IL-10 expression was mediated through STAT2. Although IFNβ induces IRF-7 and IRF-1, neither IRF mediated the effects of IFNβ on IL-12, IL-23, or IL-10. We found that the PI3K pathway mediated IL-12 inhibition but did not interfere with the inhibition of IL-23 or stimulation of IL-10.
Natural killer cells protect against cardiac inflammation and fibrosis by directly limiting eosinophilic accumulation (BA7P.144)
Authors: Ong SuFey , Ligons Davinna , Barin Jobert , Wu Lei , Talor Monica , Diny Nicola , Fontes Jillian , Gebremariam Elizabeth , Rose Noel , Cihakova Daniela 
Published: December 2022

Abstract
Myocarditis is the inflammation of the cardiac muscle and is among the leading causes of sudden cardiac failure in young adults. We demonstrate here that natural killer (NK) cells, a subset of the Type I innate lymphoid cell (ILC) group, suppress experimental autoimmune myocarditis (EAM). EAM was initiated in BALB/c mice by immunization with myocarditogenic peptide emulsified in complete Freund's adjuvant. During disease, activated cardiac NK cells accumulated in the heart, secreted interferonγ, perforin, and granzyme-B, and expressed activation markers CD69, TRAIL, and CD27. The depletion of NK cells during EAM with anti-asialo GM1 antibody resulted in a 10-fold increase from 0.12 - 12.0% in the percentage of eosinophils found in the cardiac tissue at the peak of disease. This was accompanied by significantly increased myocarditis severity and elevated fibrosis. Eosinophils were required for the change in disease severity in the absence of NK cells, as treatment with anti-asialo GM1 did not increase myocarditis severity in eosinophil-deficient ΔdoubleGATA1 mice. We show that NK cells limited eosinophilic infiltration during EAM indirectly through altering eosinophil-related chemokine production by resident cardiac fibroblasts both in vitro and in vivo. Additionally, the co-culture of NK cells and eosinophils directly induced eosinophil apoptosis in vitro. Altogether, we define a new pathway of eosinophilic regulation through interactions with NK cells.
Identification of genes/pathways responsible for resistance vs susceptibility to EAE utilizing HLA class-II transgenic mice (BA8P.163)
Authors: Mangalam Ashutosh , Luckey David , Smart Michele , Sarangi Vivekananda , Giri Shailendra , Rodriguez Moses , David Chella 
Published: December 2022

Abstract
MS is an autoimmune disease of the CNS and among all the genetic factors; MHC class-II molecules show the strongest association. To better understand the role of MHC class-II (HLA-DR/DQ) genes in susceptibility and resistance to MS, we utilized transgenic (Tg) mice expressing HLA class-II gene lacking endogenous mouse class II genes. HLA-DR3 Tg mice are susceptible to PLP91-110 induced experimental autoimmune encephalomyelitis (EAE), whereas DQ8 (DQB1*0302) transgenic mice are resistant. To identify the gene expression changes and pathways required for disease susceptibility vs resistance to EAE, we performed mRNA expression profiling of CD4+ T cells from HLA-DR3 (susceptible) and HLA-DQ8 (resistant) Tg mice. Whole genome expression analysis identified 2945 genes in HLA-DR3 Tg mice (PLP91-110 immunized vs. naïve) and 1053 genes in HLA-DQ8 Tg mice (PLP91-110 immunized vs. naïve). Comparison of modulated genes between DR3 and DQ8 identified 772 genes common to both strains, 2173 genes unique to DR3 whereas 281 genes were unique to DQ8 mice. Ingenuity pathway analysis showed up regulation of pathways involved in T cell signaling in inflammation, NF-kB signaling and those responsible for communication between innate and adoptive immunity. Majority of these pathways have IL1β as a major upstream regulator in disease susceptible HLA-DR3 Tg mice. Thus microarray data indicates an important role of IL1β induced encephalitogenic T helper cell differentiation in disease susceptibility.
Basophils’ IL-4 stifles the function of neonatal DCs and promotes Th2 immunity in newborn mice (CCR4P.212)
Authors: Miller Mindy , Dhakal Mermagya , Barik Subhasis , Chen Weirong , Ukah Tobechukwu , Ellis Jason , Zaghouani Habib 
Published: December 2022

Abstract
The immune system of newborns displays a deficiency in Th1 lymphocytes but excess Th2 cells leading to poor protective immunity against microbes and prevalence of pediatric allergies. The mechanisms underlying these discrepancies are not fully understood. Herein, we show that upon immunization, neonatal basophils readily produce IL-4 which contributes to the prevalence of Th2 cells in two ways. First, IL-4 is known to directly promote naïve T lymphocytes to differentiate into Th2 cells, but surprisingly, it also binds to the type II IL-4 heteroreceptor (composed of IL-4Rα and IL-13Rα1) on dendritic cells and diminishes their function and IL-12 production. Interestingly, the differentiation program of neonatal T cells proceeds under limited IL-12 which leads to an unusual appearance of the very same type II IL-4 heteroreceptor on Th1 cells which will serve as a death marker. Indeed, during secondary antigen challenge, Th2 cells produce copious IL-4 which triggers apoptosis of the Th1 cells via signaling through the heteroreceptor. While this Th1/Th2 imbalance serves anti-inflammatory roles that allow for embryo engraftment during gestation and maintenance of pregnancy, it severely impacts the capacity of neonates to respond to microbial infections and pediatric vaccination. Potential therapeutics that silence basophils or block IL-4 signaling may be able to restore Th1 responses and allow neonates to combat infections and ultimately reduce childhood mortality.