bluuebunny/crossref_metadata_2025_split huggingface dataset:
Functional impairment of regulatory T cell in multiple low dose streptozotocin induced murine diabetes (P3099)
Authors: Singh Kailash , Kadesjö Erik , Lindroos Julia , Sandler Stellan , Thorvaldson Lina 
Published: January 2023

Abstract
Despite intense research on the role of the regulatory T cells (Treg) , the kinetics of regulatory T cells in the early development of T1D is still not clear. We have examined the proportions of CD4+CD25+Foxp3+ regulatory T (Treg) cells and CD4+ T helper (Th) cells at close intervals in the multiple low dose streptozotocin (MLDSTZ) murine diabetes model using flow cytometry, PCR and immunostaining. The Foxp3 mRNA levels were elevated in the spleen, pancreas and PDLNs of MLDSTZ treated mice. Foxp3 immunostaining in pancreas and spleen suggested that the infiltration of Treg cells increased in pancreas and spleen of MLDSTZ treated mice. The proportions of CD4+CD25+FoxP3+ Treg cells increased in MLDSTZ treated mice from day 7 in pancreatic draining lymph nodes (PDLNs) and day 21 in spleen. The proportions of CD4+ Th cells in MLDSTZ treated mice decreased on day 7 but increase from day 10, at which time the numbers of Tregs increased suggesting that the increase in Tregs fail to control the ongoing immune insult. Impaired mRNA expression levels of IL-35 and IL-10 in PDLNs and spleen of MLDSTZ suggest a functional impairment of the Tregs. Elevated IL-17 mRNA level in spleen of MLDSTZ mice on day 21 further provide evidence of a functional defect in the Tregs. In conclusion Tregs are upregulated in the development of MLDSTZ. However, the upregulation does not protect from hyperglycemia possibly because of a functional deficiency in the Treg population in MLDSTZ treated mice.
Disease exacerbation in multiple sclerosis patients is characterized by loss of terminally differentiated neuroantigen-specific CD8+ Tregs (P3115)
Authors: Cunnusamy Khrishen , Baughman Ethan , Franco Jorge , Ortega Sterling , Greenberg Benjamin , Frohman Elliot , Karandikar Nitin 
Published: January 2023

Abstract
Multiple sclerosis (MS) is an inflammatory, demyelinating disease of the central nervous system (CNS) that afflicts more than 400,000 people in the US. Although the etiology of the disease is unknown, pathogenic T cells are thought to underlie MS immune pathology. In contrast to the current paradigm, we recently showed that MS patients harbor CNS-specific CD8+ T regulatory cells (CD8 Tregs) that are deficient during disease relapse. In the current study, we demonstrate that the neuroantigen-specific CD8 Tregs were cytolytic and eliminated pathogenic CD4+ T cells. Sorting of CD8+ T cells using an array of surface cellular markers revealed that the CD8 Tregs were terminally differentiated (CD27-, CD45RO-). The CD8 Treg-mediated suppression was perforin, granzyme B, and interferon-γ-dependent. Interestingly, we found that MS patients with acute disease exacerbation displayed a significant loss (averaging 25%) in the terminally differentiated CD8+ T cells, with a concurrent loss in perforin and granzyme B expression. In order to restore the regulatory potential of impaired CD8 Tregs during exacerbation, we pre-treated exacerbation-derived bulk CD8+ T cells with the cytokine IL-12 and significantly increased the suppressive capability of the cells by ~48% through upregulation of granzyme B and perforin. Our studies uncover the immune suppressive mechanism of neuroantigen-specific CD8 Tregs, and may contribute to the design of clinically relevant immune therapies for MS patients.
Prognostic immunological markers in patients with relapsing-remitting multiple sclerosis: a prospective magnetization transfer brain imaging study (P3117)
Authors: Markovic-Plese Silva , Tao Yazhong , Zhang Xin , Weinstock-Guttman Bianca , Hayward Brooke , Dangond Fernando , Zivadinov Robert 
Published: January 2023

Abstract
Objective: To identify predictive immunological biomarkers in patients with relapsing-remitting multiple sclerosis (RRMS) correlating with brain magnetization transfer imaging measures. Background: Baseline immunological biomarkers may help predict response to treatment. Methods: Intracellular cytokine staining and gene expression studies were performed on CD4+ and CD8+ T cells derived from 23 patients with RRMS and 15 healthy controls (HCs) at baseline (BL) and 6 months after subcutaneous interferon (IFN) β-1a treatment. The correlation between the changes in volume of normal-appearing brain tissue (NABT) and T2 lesion volume (LV) measured by voxel-wise magnetization transfer ratio (VW-MTR) with BL immunological parameters was investigated. Results: The percentage of BL IL-22-positive T cells was higher in patients than in HCs (CD8+, p=0.008; CD4+, p=0.039). A higher percentage of IL-22-positive CD8+ cells at BL correlated with a greater amount of decreasing VW-MTR T2 LV (p=0.031) in treated patients at 6 months, consistent with more prominent demyelination in T2 lesions, and a higher percentage of IFN-γ-positive CD8+ cells at BL correlated with a greater amount of decreasing VW-MTR NABT volume (p=0.007), consistent with more prominent demyelination in NABT. Conclusions: Production of IL-22 and IFN-γ by CD8+ cells is predictive of more prominent demyelinating changes.
Abnormalities in regulatory T cells and natural killer cells in major depressive disorder (P3127)
Authors: Savitz Jonathan , Tan Chibing , Taylor Ashlee , Drevets Wayne , Teague Kent 
Published: January 2023

Abstract
Major depressive disorder (MDD) is associated with increased levels of acute phase proteins, pro-inflammatory cytokines, and monocytosis, indicative of immune activation. However, in apparent contrast to evidence of depression-associated inflammation, a reduction in natural killer cell (NKC) activity is also a well-replicated finding, leading to the hypothesis that MDD is an “immunosuppressive” disorder. Here used FACS analysis to identify key subtypes of T cells and NKC in 36 healthy controls (HC) and 34 physically healthy, unmedicated patients with MDD matched for age and body mass index. After controlling for group differences in gender, we found a greater percentage of monocytes and Treg cells (relative to total live cells) in the MDD group. In addition, there was a smaller percentage of putative regulatory NKC (CD56+CD16-) but not cytotoxic NKC (CD56+CD16+) or NK T-cells (NKT) in the MDD patients. In wild-type mice, a microglia-driven, T cell-dependent protective response has been shown to be restricted by Treg cells, raising the possibility that the elevation of Treg cells in the MDD group may impair the resolution of acute pathophysiological changes leading to chronic inflammation. The depression-associated reduction in putative regulatory NKC but not effector NKC favors the hypothesis that MDD is associated with inflammation rather than immunosuppression.
The role of MMP-9 and serine proteases in the cleavage of surfactant protein D by stimulated PMNs and in sputum from CF patients (P3131)
Authors: Bratcher Preston , Gaggar Amit 
Published: January 2023

Abstract
Although the ability of serine proteases to cleave surfactant protein D (SP-D) has been well characterized, it has recently been reported by our group that matrix metalloprotease 9 (MMP-9) is able to cleave SP-D as well. However, our initial study examined MMP-9-mediated cleavage of SP-D in vitro using purified proteins. In order to expand upon this study in a more physiological context, we collected supernatants from primary polymorphonuclear leukocytes stimulated with LPS and fMLP and incubated them with purified SP-D in the presence of a general serine protease inhibitor (PMSF) and/or specific inhibitors against MMP-9 and neutrophil elastase (NE). All inhibitors were able to decrease the amount of detectable cleavage of SP-D, and none of them alone were able to completely inhibit cleavage. We were only unable to detect cleavage when a combination of PMSF and the MMP-9 inhibitor were used, suggesting that MMP-9 is the only non-serine protease released from PMNs with the ability to cleave SP-D. In addition, we examined the cleavage of endogenous SP-D in sputum samples from patients with cystic fibrosis (CF), and found that the MMP-9 inhibitor also significantly attenuated SP-D degradation seen when CF sputum was incubated at 37°C over time. These experiments suggest that MMP-9 is able to cleave SP-D in vivo, and that MMP-9 degrades SP-D in the airways of CF patients.
Increased circulating Th17 cells and elevated serum levels of TGF-beta and IL-21 involved in human non-segmental vitiligo development (P3141)
Authors: Zhou Li , Shi Yuling , Li Kai , Hamzav Iltefat , Lim Henry , Mi Qing-Sheng 
Published: January 2023

Abstract
Although it is widely believed that non-segmental vitiligo (NSV) results from the autoimmune destruction of melanocytes, the detailed immune mechanisms remain poorly understood. Th17 cells, a subset of CD4+ helper T cells, have been identified to be implicated in a variety of human autoimmune diseases. To comprehensively evaluate the role of Th17 cells and the pathways driving these T cell differentiations in NSV, we had compared the frequencies of peripheral blood Th17 cells and other IL-17 producing T cells by flow cytometry between 42 active NSV patients with 45 race-, gender- and age-matched healthy control subjects. Serum levels of IL-17A and Th17 cell differentiation related cytokines including TGF-beta1, IL-6, IL-23, IL-22 and IL-21 were detected by ELISA. Our results showed the increased circulating Th17 cell frequencies and elevated serum IL-17A, TGF-beta1 and IL-21 levels in NSV patients. Further more, the increased Th17 cell frequencies are positively correlated with serum TGF-beta1 levels and the body surface area (BSA) in NSV patients. These results indicate the involvement of Th17 cells in the development and progression of NSV, and more importantly, our data highly suggest that elevated serum TGF-beta and IL-21 could contribute to enhanced Th17 cell differentiation in NSV.
Calpastatin prevents NF-κB mediated hyperactivation of macrophages and attenuates colitis (P3151)
Authors: Rose Aaron , Huang Zhi , Hoffmann Fukun , Denk Tobias , Hashimoto Ann , Hoffmann Peter 
Published: January 2023

Abstract
Calpain enzymes proteolytically modulate cellular function and have been implicated in inflammatory diseases. In this study, we found that calpain levels did not differ between intestinal tissues from inflammatory bowel disease (IBD) patients and healthy controls, but IBD tissues showed increased levels of the endogenous calpain inhibitor, calpastatin (CAST). To investigate the role of CAST in the immune system during IBD, mice were x-ray irradiated and reconstituted with either CAST knockout (KO) or wild-type (WT) bone marrow and subjected to dextran sulfate sodium (DSS)-induced colitis. CAST KO recipients with induced colitis exhibited more severe weight loss, bloody diarrhea, and anemia compared to WT controls. Histological evaluation of colons from KO recipients with colitis revealed increased inflammatory pathology. Macrophages purified from the colons of KO recipients had higher IL-6, TNFα, and IFNγ mRNA levels compared to WT controls. Mechanistic investigations using siRNA and KO bone marrow to generate CAST deficient macrophages showed that CAST deficiency during activation with bacterial PAMPs including heat-killed E. faecalis or CpG DNA led to increased IκB cleavage, NF-κB nuclear localization, and IL-6 and TNFα secretion. Thus, CAST plays a central role in regulating macrophage activation and limiting pathology during inflammatory disorders like IBD.
CXCL9 and γδ T cells are increased in patients with allergic conjunctivitis (P3159)
Authors: Jimenez-Martinez Maria C , Alonso Miguel , Estrada-Garcia I Iris , Estrada-Parra Sergio , Perez-Tapia Mayra , Garfias Yonathan 
Published: January 2023

Abstract
Allergic conjunctivitis (AC) is a frequent disease that affects the ocular surface, The study of immune physiopathology of allergic conjunctivitis is focused on T helper cells. Recently it has been suggested that γδ T lymphocytes are necessary for the maintenance of the ocular allergic response, however there are not studies in humans about the role of γδ T lymphocytes in human AC. The aim of this study was to evaluate the frequency of γδ T cells and chemokines in tears of AC patients. Patients: 31 patients were included (16 men and 15 women), AC diagnosis was based on clinical history, physical and eye examination. PBMC were obtained from patients and healthy controls and frequency of γδ T cells was evaluated by flow cytometry; tear samples were also collected to determine chemokine concentrations by CBA. Results: γδ T lymphocytes were 11.9 times increased in AC-patients (22.63 ± 2.9%) compared to healthy donors (1.9 ± 1.9%) (p=0.002), chemokine determination in tears showed increased MIG/CXCL9 in AC patients (1604 ± 1404 pg/ml) when compared with healthy subjects (568 ±841pg/ml) p&lt; 0.02. Conclusion: γδ T cells could be also involved in human allergic conjunctivitis as is has been demonstrated in animal models, it is possible that γδ T cells could migrate to conjunctiva due to several chemokines such as CXCL9, significant increased in AC patients, sustaining the local inflammation in conjunctiva
Interferon and TNF family soluble inflammatory mediators are detected prior to and increase at Systemic Lupus Erythematosus classification in serum samples from military personnel acquired from the Department of Defense Serum Repository (P3087)
Authors: Munroe Melissa , Anderson Jourdan , Niewold Timothy , Tsokos George , Keith Michael , Harley John , James Judith 
Published: January 2023

Abstract
Processes that lead to clinical illness in SLE years before diagnosis are not well characterized. This study evaluates the temporal relationship between autoantibody production, cytokine levels and the onset of SLE. Serial sera from 60 SLE cases before diagnosis with no ACR criteria to SLE classification (average timespan=4.4 years) and matched healthy controls (HC) were obtained from the DODSR. Sera samples were tested for autoantibodies (BioPlex2200), C-reactive protein (hs-CRP), Von Willebrand’s Factor (VWF) and 32 soluble inflammatory mediators, including cytokines, chemokines, and soluble receptors. Patient samples before ACR classification had significant (p &lt; 0.01) alterations in 16 soluble mediators of inflammation, as well as VWF and hs-CRP. TNF Receptor levels, TNFRI, TNFRII, BLyS, and APRIL, dramatically increased as lupus classification approached (p &lt; 0.001). The increase in TNFR shedding parallels the accumulation of autoantibodies in SLE cases. Interferon (IFN)-associated mediators of inflammation, IFN-γ, IP-10, MIG, and MIP-1α (p &lt; 0.01), were also increased leading up to classification. That these alterations are present prior to the transition to active SLE suggests that multiple perturbations in immune-mediated inflammatory processes occur long before clinical classification and suggest that high-risk, pre-clinical individuals, can be identified before their illness is clinically manifested and damage occurs.
Expansion of a marginal zone B cell-like population in lymph nodes of mice developing collagen-induced arthritis (P4042)
Authors: Palm Anna-Karin , Kleinau Sandra 
Published: January 2023

Abstract
The importance of B cells in collagen-induced arthritis led us to investigate whether particular B cell subsets participate in the autoimmune response to collagen type II. We show here that MZB cells in the spleen are naturally self-reactive to collagen type II and are the first cells to expand after collagen type II immunization. In contrast, the autoimmune collagen type II response in lymph nodes develops one week later. Due to this time lapse we hypothesized that the MZB cells may be involved in initiating the autoimmune response in the lymph nodes. We did indeed find a small MZB-like B cell population in the lymph nodes that expressed B220+CD23loCD1dhi, thereby being distinguishable from FOB cells. The MZB-like cells expanded almost two-fold after immunization with collagen type II but not the control antigen ovalbumin, making this phenomenon specific to the autoimmune response. The MZB-like cells displayed an antigen activated phenotype with increased cell surface expression of MHCII, FcγRIIb and CD70. Moreover, after immunization the MZB-like cells increased their expression of CXCR5, implying that they are migrating cells. Further investigation of the specific function of the MZB-like cells is warranted. In conclusion, MZB cells in the spleen trigger the autoimmune collagen type II response in collagen-induced arthritis, and a MZB-like population in the lymph nodes is specific for the autoimmune reaction and may be of importance for disease pathogenesis in this model.
Delivery of a genetic cancer vaccine containing a small molecule inducible immune adjuvant by in vivo electroporation (eVac) (P4332)
Authors: Collinson-Pautz Matthew , Seethammagari Mamatha , Decker William , Spencer David , Levitt Jonathan 
Published: January 2023

Abstract
The use of dendritic cell vaccines to treat cancer has significant potential, however that potential has yet to be achieved in treating patients. Fortunately, pioneering biotechnologies are paving the way for more effective cancer vaccines. Our lab has developed a drug inducible MyD88/CD40 (iMC) composite adjuvant that promotes robust cytotoxic T cell priming in iMC activated DCs. Despite the iMC innovation, the use of patient-tailored ex vivo DC vaccines is impractical for widespread use, due to issues of scalability and cost. We have therefore begun to address the delivery of DC vaccines as “off-the-shelf” therapies using in vivo electroporation (EP) of plasmid DNA (pDNA) encoding both the iMC adjuvant and tumor antigen (eVac). Intradermal EP of pDNA encoding the model antigen LacZ in mice suggests that LacZ-specific T cell responses were induced, as mice receiving LacZ + EP prophylactically were either protected from establishment of B16/LacZ tumors or demonstrated significantly slower tumor growth compared to controls. In mice bearing pre-established B16/LacZ tumors, LacZ-eVac, but not EP with LacZ antigen alone significantly reduced tumor burden and increased survival. Taken together, these data indicate that eVac with iMC-antigen is an effective “off-the-shelf” cancer vaccine. These findings warrant elucidation of the underlying immunological mechanisms responsible for the anti-tumor responses observed with eVac treatment to guide further refinement of the vaccine.
Altered adaptive and TNF-superfamily soluble inflammatory mediators mark impending Systemic Lupus Erythematosus disease flare in European-American lupus patients after influenza vaccination (P6282)
Authors: Munroe Melissa , Anderson Jourdan , Merrill Joan , Guthridge Joel , Roberts Virginia , Air Gillian , Thompson Linda , James Judith 
Published: January 2023

Abstract
SLE is a waxing and waning disease characterized by major organ involvement, disease flares and immune dysregulation. Understanding mechanisms and identifying biomarkers of flare would help prevent damage and improve management. Plasma samples were obtained from SLE patients on the day of influenza vaccination (BL) and at 6 and 12 weeks post (FU) in 28 European American SLE patients who exhibited SELENA-SLEDAI defined disease flare at 6 (n=13) or 12 (n=15) weeks post-vaccination. Each SLE patient was matched to a unique SLE patient who did not flare (NF), and to a healthy control (HC). Samples from 13 SLE patients with flare were compared to samples from a non-flare year (self nonflare, SNF). Plasma samples were tested for 52 soluble inflammatory mediators using xMAP technology. SLE patients who exhibited disease flare had significantly (p &lt; 0.01) higher levels of BL pro-inflammatory mediators, including Th1, Th2, and Th17-type cytokines compared to NF/SNF SLE patients and HC. Regulatory cytokines, including IL-10 and TGF-β were increased (p &lt; 0.01) at BL in SLE patients not exhibiting disease flare (NF/SNF) compared to SLE flare and HC. TNFR family members, TNFRI, TNFRII, Fas, FasL, CD40L, were increased at BL in SLE flare patients. Select serum cytokines and chemokines are elevated in patients before SLE disease flare, providing important pathways to dissect for mechanism of flare, therapeutic selection or development.
The central role of dendritic cell-produced IL-6 signals in reversing age-related CD4 T cell defects (P4226)
Authors: Brahmakshatriya Vinayak , Rymeski Barbara , Kuang Yi , Swain Susan 
Published: January 2023

Abstract
The response of aged CD4 T cells to vaccines is less than that of young T cells resulting in ineffectiveness of many current vaccines and contributing to the greater susceptibility of the aged to new infections. We find that activation of DC by TLR ligands (*DC), used as APC for naive CD4 T cells, induces their production of high levels of IL-6 that improve aged CD4 T expansion, cytokine production and survival. Importantly, this enhances their ability to differentiate into T follicular helper cells that promote long-term B cell responses and sustained antibody production, a key to vaccine-induced immunity. The enhanced helper function of aged CD4 T cells is lost if the *DC cannot make IL-6, while effects of *DC on young CD4 T cells are not IL-6 dependent. This suggests that loss of responsiveness to IL-6 is central to the aged defects of CD4 T cells. We find that in vitro IL-6 signaling of the aged is blunted such that the dose of IL-6 required to achieve a particular level of IL-6R internalization and phosphorylation of STAT3 is higher in the aged vs. young naïve CD4 T cells. Clearly, the ability of aged CD4 T cells to respond to the strong *DC-mediated IL-6 signal during their interaction is still intact and can restore their function to that comparable to young cells. This *DC-based vaccine strategy to localize pro-inflammatory cytokines to the Ag-specific CD4 T cells, also avoids counterproductive and often-dangerous bystander effects of adjuvants given systemically.
Host DNA released in response to aluminum adjuvant enhances MHCII mediated antigen presentation and prolongs CD4 T cell interactions with dendritic cells. (P4244)
Authors: McKee Amy , Burchill Matthew , Michael Munks , Jin Lei , Kappler John , Friedman Rachel , Jacobelli Jordan , Marrack Philippa 
Published: January 2023

Abstract
Many vaccines include aluminum salts (alum) as adjuvants in spite of little knowledge of alum’s functions. Host DNA rapidly coats injected alum. Here we further investigated the mechanism of alum and DNA’s adjuvant function. Our data show that DNAse co-injection reduces CD4 T cell priming by intramuscularly injected antigen plus alum. This effect is partially replicated in mice lacking STING, a mediator of cellular responses to cytoplasmic DNA. Others have shown that DNAse treatment impairs alum induced DC migration from the peritoneal cavity to the draining LN. However our data shows that DNAse does not affect accumulation of, nor expression of costimulatory proteins on, antigen loaded dendritic cells in lymph nodes draining injected muscles, the site by which most human vaccines are administered. DNAse does inhibit prolonged T cell-DC conjugate formation and antigen presentation between antigen+ DCs and antigen specific CD4 T cells following intramuscular injection. Thus, from the muscle, an immunization site that does not require host DNA to promote migration of inflammatory DC, alum acts as an adjuvant by introducing host DNA into the cytoplasm of antigen-bearing DCs, where it engages receptors that promote MHC II presentation and better DC-T cell interactions.