bluuebunny/crossref_metadata_2025_split huggingface dataset:
Co-delivery of adjuvant and subunit antigens via a nanoparticle platform induces tissue-associated and systemic adaptive immune responses (P4409)
Authors: Dunkle Alexis , Blanchette Craig , Boone Tyler , Corzett Michele , Fischer Nicholas , Hoeprich Paul , Driks Adam , Rasley Amy 
Published: January 2023

Abstract
Vaccination is a critical countermeasure against infection for both biodefense and public health. As many pathogens are encountered at mucosal interfaces, a delivery system that induces robust mucosal immunity is paramount. Here, we investigate a nanolipoprotein particle (NLP) containing the Toll-like receptor 4 agonist monophosphoryl lipid A (MPLA) as a novel vaccine platform using disparate model antigens. Co-administration of MPLA-NLPs with ovalbumin significantly enhances the expansion of OVA-specific T cells in mice in vivo after a single intranasal inoculation, indicating that MPLA-NLPs function as efficient adjuvants. To combine the adjuvant effects with antigen delivery, we engineered MPLA-NLPs to accommodate attachment of His-tagged proteins. Intranasal administration of MPLA-NLPs conjugated to Bacillus anthracis antigens induces specific T cell responses in lung-draining lymph nodes and significant antibody responses in serum and bronchoalveolar lavage. Experiments to assess the immunogenicity of multiple antigens in combination on a single particle and to compare between co-administered formulations are ongoing. These data further support a role for NLPs as a flexible vaccine platform that allows co-delivery of antigen and adjuvant to lung-associated immune tissues and promotes robust adaptive immune responses.
Culture conditions promoting innate immunity and homeostatic proliferation generate highly enriched, MUC1- or HER2/neu-specific CD4+ and CD8+ T cells (P4419)
Authors: Cohen Peter , Pathangey Latha , McCurry Dustin , Gorman Jessica , Viso Carole , Gendler Sandra , Disis Mary 
Published: January 2023

Abstract
Although tumors from melanoma patients can often provide a source of therapeutically active T cells for adoptive therapy, it has historically proved challenging to employ peripheral blood as the T cell source for any type of cancer. We have observed nonetheless that HER2-specific T cells can be Ag-driven and enriched in culture when PBMCs from HER2-vaccinated breast cancer patients are treated with exogenous IL12. We hypothesized that stimulating innate immunity could further improve PBMC cultures by activating the DC subpopulation more effectively than adding exogenous rIL12. Bulk PBMC exposure to optimized TLR agonists produced vast quantities of IL12 and IL23, and upregulated HLA-DR, B7.1 and CD40 in the DC subpopulation, effects not produced by exogenous IL12. Subsequent IL7 exposure emulated homeostatic proliferation, selectively causing the Ag-driven T cell subset both to proliferate faster and to strongly resist apoptosis. Ag-specificity within two weeks approached 80-100% of both CD4+ and CD8+ T cells for recall Ags and 1-15% for a wide array of MUC1- and HER2-derived peptides, including the ability to distinguish glycoforms. Culture-expanded T cells retained a young CD28+/CD56- phenotype, uniformly expressed ROR-γ during culture, variably produced IL-2 and/or IL-17, and uniformly expressed T-bet and secreted IFN-γ when reexposed to Ag. Such T1/T17 bipotency may be ideal for anti-tumor adoptive therapy and highly efficient for use in Ag discovery.
Molecular mechanism of the Vγ9Vδ2 T lymphocytes anergy. (P4385)
Authors: Poupot Mary , Fruchon Severine , Boissard Frederic , Aruebo Marie-Eve , Cendron Delphine , El Faqihi-Olive Fatimah , Pont Frederic , Bardouillet Laure , Mekaouche Mourad , Ingoure Sophie , Sicard Helene , Dubreuilh Guy , Fournie Jean-Jacques 
Published: January 2023

Abstract
Upon activation with phosphoantigens (PAgs), human and non-human primates circulating TCRVγ9Vδ2 T lymphocytes release pro-inflammatory cytokines, mediate tumor cell cytotoxicity and proliferate through an IL2-dependent clonal expansion. Together, these features make it versatile and attractive candidates for new cancer immunotherapies (Kabelitz, D et al., 2007, Canc.Res., 67(1):5-8). Despite a potent and rapid response of the TCRVγ9Vδ2 T lymphocytes in vitro and in vivo, repeated stimulations with PAgs desensitize selectively this T cell lineage in humans and non-human primates (Cendron, D. et al., 2007, Eur.J.Immunol., 37(2):549-65; Sicard, H. et al., 2005, J.Immunol., 175(8):5471-80). Thus, we investigated the gene expression pattern associated with this PAg-selective anergy of these lymphocytes. TCRVγ9Vδ2 T cells were isolated from blood samples from PAgs-injected macaques (3 repeated injections) for transcriptome studies. After the third stimulation, in addition to some genes corresponding to the inhibition of cell cycle, a significant gene signature emerged from the desensitized γδ T cells transcriptome involving the PPARα and CARM1 pathways, based on up-regulation of both EP300, HSD17B4, PRKAR2B and RB1 genes while RARA and PRKCA (PKCα) were down-regulated. Moreover, inhibition of proliferation and cytokine production by PPARα agonists-treated human TCRVγ9Vδ2 T lymphocytes confirmed the implication of PPARα pathway in the γδ T cells desensitization by PAgs.
Tolerance induction using Ag-ECDI-PLGA prevents autoimmunity in the BDC2.5 AT model for T1D (P5208)
Authors: Prasad Suchitra , Xu Dan , Miller Stephen 
Published: January 2023

Abstract
Type 1 Diabetes (T1D) is an autoimmune disease characterized by the infiltration and destruction of insulin producing β-cells of the pancreatic islets by autoreactive T-cells. Treatment with autoantigenic peptide(s) chemically crosslinked with ECDI to splenocytes or other cellular carriers (Ag-ECDI-SP) can prevent and/or reverse murine autoimmune diabetes. Due to the inherit limitations of cellular therapy in a clinical setting including the sourcing and production of Ag-ECDI-SP, here we explore the use of biodegradable poly(lactic-co-glycolic acid) (PLGA) micro-particles as substitutes for splenic donor cells. Treatment with p31-ECDI-PLGA protected against disease in the BDC2.5 adoptive transfer model of T1D. Mice tolerized with p31-ECDI-PLGA mice had reduced levels of total infiltrating BDC2.5 CD4 T cells in the pancreas when compared to mice treated with the non-diabetogenic MOG35-55-ECDI-PLGA control. Additionally, BDC2.5 T cells isolated from the pancreas of p31-ECDI-PLGA treated mice had reduced levels of inflammatory cytokines, such as IFN-gamma, TNF-alpha and IL-17. These data demonstrate the efficacy of an antigen specific therapy to modulate a highly pathogenic autoimmune response using an FDA approved biodegradable PLGA microspeheres. Thus, this carrier system provides a more viable alternative to the cell based approach that can be readily translated in clinical trials.
Pharmacological repression of RORγ is therapeutic in the collagen-induced arthritis experimental model (P5214)
Authors: Chang Mi Ra , Lyda Brent , Kamenecka Theodore , Griffin Patrick 
Published: January 2023

Abstract
The nuclear receptor RORγ has been shown to play an essential role in the development of Th17 cells. Previously, it was reported that the potent and selective RORγ inverse agonist SR2211 was effective at modulating RORγ activity in cell culture. More recently we demonstrated that the RORγ inverse agonist SR2211 can repress expression and production inflammatory cytokines in Th17 cells and LPS stimulated RAW264.7 cells. Here we present in vivo efficacy of SR2211 in an experimental model of arthritis, the murine collagen induced arthritis or CIA model. Significantly reduced Joint CIA scores were observed in SR2211 treated mice as compared with vehicle in the acute phase of the model. CIA represents an autoimmune environment characterized by the accumulation of inflammatory cells into the synovium leading to the destruction of joints. In this model cytokines have been shown to be important regulators of synovial inflammation. Administration of SR2211 in CIA mice led to an increase in IFNγ in ex vivo stimulated draining lymph nodes. As IFNγ has been shown to be beneficial for the treatment of rheumatoid arthritis. In previous work, patients treated with IFNγ showed significant therapeutic potential of RA the data presented suggest the potential of targeting RORγ to repress inflammatory T cell function and macrophage activation in arthritis. Thus, SR2211 has potential utility in the treatment of inflammatory disease.
CRAC channel inhibitors block cytokine production with a pattern distinguishable from cyclosporine (P5216)
Authors: Ramos Stephanie , Butler Lori , Kimbler Trevor , Cao Jianguo , Grey Jonathan , Rogers Evan , Wang Zhijun , Whitten Jeffrey , Roos Jack , Stauderman Kenneth , Velicelebi Gonul 
Published: January 2023

Abstract
Engagement of the T cell receptor (TCR) triggers complex signaling cascades that result in various effector T cell responses. A critical component of TCR signaling is store-operated calcium entry (SOCE) that occurs through the activation of calcium release-activated calcium (CRAC) channels. Activated CRAC channels cause an increase in cytoplasmic calcium, which results in the transcription of calcineurin/NFAT-dependent genes involved in T cell proliferation and cytokine production. Both CRAC channel inhibitors and calcineurin inhibitors are known to inhibit T cell function. However, the relative impact of CRAC channel inhibitors and calcineurin inhibitors on Th1, Th2, Th17, and Treg functions are not known. Using anti-CD3/anti-CD28 to stimulate cytokine production by peripheral blood mononuclear cells (PBMCs), we demonstrate that novel CalciMedica CRAC channel inhibitors have a pattern of cytokine inhibition that is different from the calcineurin inhibitor cyclosporine (CsA). CRAC channel inhibitors inhibit Th1 and Th17 cytokines (IFNγ and IL-17) more potently than Treg and Th2 cytokines (IL-10 and IL-4), while CsA inhibits IL-10 more potently than IFNγ, IL-4, and IL-17. These results suggest that CRAC channel inhibitors and CsA have different potencies against different effector T cell subsets, and that CRAC channel inhibitors may achieve a more favorable balance of effects on pro- versus anti-inflammatory pathways in immune disorders.
Identification and characterization of T cell epitopes expressed on the surface of cancer cells. (P2002)
Authors: Rozanov Dmitri , Reddy Ashok , David Larry , Burrows Gregory , Rantala Juha , Levin Trevor , Wilmarth Philip , Anur Pavana , Chiotti Kami , Spellman Paul 
Published: January 2023

Abstract
In our search for better ways to stimulate anti-tumor T cell-mediated immune response we optimized powerful technique known as “reverse” immunogenetics, in which MHC I-bound peptides are recovered and sequenced using highly sensitive mass spectrometry. Using this technique we have identified more than 3,500 peptides eluted from the surface of 20 breast carcinoma cell lines of 4 different subtypes. By using sequence data for each breast carcinoma cell line we predicted bound peptide consensus sequence for a particular allele of HLA-A, -B, and C molecules. Next, we determined the frequency of the identification of both peptide and protein that is associated with this peptide in subtype-specific and all analyzed cell lines. This allowed us to associate some MHC I-presented peptides/proteins with a specific subtype of breast cancer as well as shared peptides/proteins. Some of the identified proteins are tumor specific and immunogenic. For example, Aldolase A, which is MHC I-presented in many cell lines, has been identified by Serex analysis in lung cancer and breast cancer patients. Lastly, using gene expression data for 60 breast carcinoma and normal cell lines as well as for 708 tumors and 329 normal tissues we have sorted all identified proteins in accordance with the level of their expression in non-transformed and transformed cells. This arrangement allowed us to identify, for the first time, several tumor specific genes that could be good targets for T cell-based therapy.
Memory to self-antigens in Systemic Lupus Erythematosus (P4020)
Authors: Chen Yunying , Wermeling Fredrik , Petersen Sven , Kaiser Ylva , Karlsson Mikael 
Published: January 2023

Abstract
Systemic Lupus Erythematosus (SLE) is a systemic autoimmune disease associated with auto-IgG antibodies, which are produced due to defective clearance of apoptotic cells (AC) and loss of tolerance to self-antigens. SLE follows flare and remission courses; disease symptoms can come rapidly and unpredictably, suggesting the presence of immune memory to self-antigens in patients. However, experimental evidence and molecular mechanisms behind this are not known. We set up a mouse model by injecting normal mice with syngenic AC weekly for 4 weeks that breaks tolerance and induces auto-IgG production. One month later the serum auto-IgG drops to pre-immune level, these mice can rapidly respond to a single AC injection and produce increased auto-IgGs. This memory-like response is transferable into naïve mice. Serum level of auto-IgG subclass IgG2, but not IgG1, is dramatically increased in the memory response compared to that in the primary response, which contribute to pathogenic effects. The memory auto-IgGs are produced via both germinal center and plasmablast pathways. Spectratyping shows that B-cell repertoire is more skewed in memory response than that in pre-immune and primary response. These observations demonstrate an inducible immune memory to self-antigen in normal mice and establish a model for study of SLE flare-like response in mice. In addition, this study suggests that the formation of immune memory to self-antigens contributes to the disease relapse in SLE patients.
Detection and characterization of autoreactive B cells in patients with Rheumatoid Arthritis (P4024)
Authors: Titcombe Philip , Barsness Laura , Giacobbe Lauren , Gillespie Emily , Peterson Erik , Mueller Daniel 
Published: January 2023

Abstract
B cell tolerance to self antigen is critical for preventing several autoimmune diseases, including Rheumatoid Arthritis (RA), but how tolerance is maintained or broken in physiologic settings is not well understood. Our lab recently developed a novel protein-tetramer enrichment strategy for detecting and analyzing rare self antigen-specific B cells within the endogenous polyclonal repertoire of mice. We have now adapted this strategy to study human peripheral blood B cells specific for two different cyclic citrullinated peptides (CCP) that mimic the self-protein epitopes targeted by autoantibodies in a majority of RA patients. RA patients with epitope-specific anti-CCP IgG serum antibodies (Ab+ RA) display an increased frequency of CCP-specific B cells compared to Ab- RA patients and healthy controls. Ab+ RA patients also demonstrate an increased frequency of CCP-specific B cells that have undergone isotype-switching and memory cell differentiation (IgD- CD27+), and their numbers correlate with anti-CCP antibody levels. Future work tracking self antigen-specific B cells in humans will focus on identifying factors important for preserving or restoring B cell tolerance.
Extracellular Granzyme B: a key player in aneurysm pathogenesis (P3072)
Authors: Granville David , Ang Lisa , Bleackley Chris , Zhao Hongyan 
Published: January 2023

Abstract
Granzyme B (GZMB) is a pro-apoptotic protease that is released by cytotoxic immune cells and internalized by target cells in a perforin-dependent manner. However, GZMB contributes to aortic aneurysm pathogenesis through a perforin-independent, extracellular process involving extracellular matrix proteolysis. In vitro and in vivo studies have identified decorin, a key regulator of collagen organization, fibrillogenesis and collagen tensile strength, as a GZMB substrate. We hypothesized that GZMB contributes to aortic aneurysm through the degradation of decorin leading to reduced collagen organization and tensile strength resulting in aortic rupture. To induce aneurysm, apoE-/- mice were implanted with osmotic minipumps that constitutively released angiotensin II. Mice were injected with the GZMB inhibitors, serpin A3N or anti-GZMB neutralizing antibody. A dose-dependent reduction in the frequency of aneurysm rupture was observed in mice that received inhibitor. Reduced GZMB and decorin proteolysis were observed in inhibitor-treated mice. Adventitial collagen from serpin-treated mice exhibited higher fibre density and reduced fibril size irregularity. GZMB was also elevated in human aortic aneurysm. GzmB was expressed in lymphocytes in the intraluminal thrombus and in mast cells in the aneurismal aortic adventitia. In summary, extracellular GZMB contributes to decorin degradation resulting in reduced collagen organization and loss of aortic wall integrity leading to aneurysm.
Th40 T cells are pathogenic in multiple sclerosis and EAE the mouse model of MS (P3121)
Authors: Wagner David , Waid Dan , Corboy John , Schreiner Teri 
Published: January 2023

Abstract
Multiple sclerosis (MS) is a neurodegenerative disease with established autoimmune components. We described a unique T cell subset expressing CD40, Th40 cells that achieve effector status in autoimmune diseases. Furthermore we discovered that Th40 cells are detected in human subjects and that during autoimmunity these T cells are significantly (p &lt; 10(-7) expanded in number compared to normal controls. Th40 cells produce both IFNg and IL-17, suggesting an interesting option for pathogenicity development. The majority of patients carried the DR15 (DR2) haplotype, but a significant number do not. Th40 cells were predominantly CD45RO (memory phenotype) and unlike Th40 cells in T1D express both CCR5 and CXCR3. In many patients, an antigen response profile was determined; i.e. some patients respond to MOG while others respond to PLP etc. In the EAE mouse model of MS we discovered that prior to EAE onset levels of Th40 cells are systemically very low, disease induction increased Th40 cell percentages and numbers drastically. When Th40 cells are isolated from EAE induced mice , disease can be adoptively transferred to scid recipients. Histology reveals that Th40 cells infiltrate brain and spinal cord. Th40 cells isolated from the brains of stage 3 EAE mice are pro-inflammatory.
Impaired lymphocyte response in patients with chronic kidney disease (P3075)
Authors: Mansouri Ladan , Lundahl Joachim , Jacobson Stefan , Paulsson Josefin 
Published: January 2023

Abstract
In Chronic Kidney Disease (CKD), immune cells are affected due to uremic toxins. Given the central role of T lymphocytes in the progress of CKD, we analyzed lymphocyte proliferative response and immune modulators production to selected stimulators. We applied a method called Flow-cytometric Assay for Specific Cell-mediated Immune-response in Activated whole blood (FASCIA) using whole blood and flow-cytometric analysis for detection of proliferation in addition to a multi-array method (Milliplex 26-plex) to analyze the profile of immune modulators. The absolute number of lymphoblasts, CD4+, CD8+ subpopulations in response to mitogens were equal comparing healthy controls, patients with eGFR &lt;20 (ml/min/1.73 m2) and hemodialysis patients. However, levels of Interleukin (IL)-2, 10 and 15 in the Staphylococcus enterotoxin A -stimulated supernatant as well as levels of IL-15 and IL-5 in the Influenza A vaccine-stimulated supernatants of patients were lower compared to healthy controls. TNF-α, IL-10, IL-12, IL-15, IL-8, MCP-1, IP-10, IFN-α2, IL-1α and Eotaxin levels were higher in the patient plasma. Our results suggest that T-cells have no intrinsic defects in terms of proliferative response to mitogens but an inability to produce selected cytokines due to the uremic milieu or dialysis procedure. The reported aberrations may contribute to the increased frequency of infections in CKD patients and the systemic inflammatory state may be causative for atherosclerosis.
HMGB1, a major damage-associated molecule, is an early and critical mediator in an animal model of uveitis induced by uveitogenic T cells (P3161)
Authors: Shao Hui , Jiang Guomin , Sun Deming , Kaplan Henry 
Published: January 2023

Abstract
It is largely unknown how invading autoreactive T cells initiate the pathogenic process inside the diseased organ in organ-specific autoimmune disease. In this study, we used a chronic uveitis disease model in mice, experimental autoimmune uveitis (EAU), induced by adoptive transfer of uveitogenic, interphotoreceptor retinoid binding protein (IRBP)-specific T cells and showed that high mobility group box chromosomal protein 1 (HMGB1), an important endogenous molecule that serves as a danger signal, is rapidly released from retinal tissue cells into the extracellular matrix and intra-ocular fluid in response to IRBP-specific T cell transfer. HMGB1 levels in the intra-ocular fluid correlated with disease severity. The HMGB1-releasing tissue cells remained intact, showing that this molecule was not released by necrotic/apoptotic cells. HMGB1 release required direct cell-cell contact between retinal tissue cells and IRBP-specific T cells. Administration of HMGB1 antagonists markedly suppressed development of EAU via mechanisms including inhibition of migration of granulocytes/monocytes and Th1 cytokine production. Our study demonstrates that HMGB1 is an early and critical mediator of ocular inflammation initiated by autoreactive T cell invasion.
Role of calcium/calmodulin kinase IV on podocyte function in lupus nephritis (P3081)
Authors: Ichinose Kunihiro , Ushigusa Takeshi , Koga Tomohiro , Tsokos George , Kawakami Atsushi 
Published: January 2023

Abstract
[Objectives] Kidney podocytes and their slit diaphragms settle the integrity of renal basement membrane and prevent urinary protein loss. T cell from patients with systemic lupus erythematosus display increased expression of calcium/calmodulin kinase IV (CaMKIV). Here we evaluated the expression of CaMKIV in kidney biopsy specimens from patients with lupus nephritis (LN) and in a human podocyte cell line (AB8/13) after exposure to IgG from sera of patients with LN. [Methods] We examined the expression level of nephrin, a podocyte marker and CaMKIV in normal and LN kidney specimens by immunofluorescence staining. We cultured human podocytes with IgG purified from sera of normal individuals and LN patient for 24 hr and analyzed CaMKIV mRNA levels. We also performed microarray analysis to determine gene expression. [Results] Although the expression levels of nephrin were decreased, CaMKIV expression was found increased in podocytes of LN kidney biopsy specimens.Interestingly, culture of podocytes in the presence of IgG from LN sera led to 1.5-fold increase in the expression of CaMKIV mRNA.Gene array analysis revealed that the expression of genes related to the regulation of neural precursor cell proliferation decreased significantly (P&lt;0.000441) in podocytes treated with IgG from LN patients. [Conclusion] We suspect that increased CaMKIV expression may contribute to the inability of podocytes to maintain the integrity of glomerular basement membrane in LN patients.