bluuebunny/crossref_metadata_2025_split huggingface dataset:
The role of DAMPs in the host immune and remodeling response to xenogeneic biologic scaffold materials (141.27)
Authors: Brown Bryan N. , Daly Kerry A. , Lotze Michael T. , Badylak Stephen F. 
Published: January 2023

Abstract
Biologic scaffolds composed of extracellular matrix (ECM) have been used in pre-clinical and clinical applications with few reports of adverse immunologic reactions. Following implantation, ECM scaffolds are readily remodeled by the host; however, the mechanisms underlying the host immune and remodeling responses are not fully understood. A variety of molecules within ECM scaffolds, as well as their degradation products, have been shown to influence the host response. Damage associated molecular pattern molecules (DAMPs) are released during non-apoptotic cell death, and may act as either pro- or anti-inflammatory mediators. Consequently, the presence of DAMPs within a biologic scaffold material may direct the host immune and remodeling responses following implantation. In the present study, the presence of several well-established DAMPs within ECM scaffold materials was evaluated. In addition, the presence of DNA and the redox state of the scaffold microenvironment, which may influence the inflammatory potential of DAMPs, were assessed to determine their effects upon host immune and remodeling responses. The results of the present study demonstrate the complex nature of the interactions between the host immune response, local microenvironment, and tissue remodeling, following the implantation of biologic scaffold materials.
WIP peptide stabilizes WASP in cells from Wiskott-Aldrich syndrome (WAS) patients with mutations in the WIP-binding domain (33.16)
Authors: Massaad Michel , Giliani Silvia , Notarangelo Luigi , Geha Raif S. 
Published: January 2023

Abstract
WAS is an X-linked immunodeficiency characterized by eczema, thrombocytopenia and recurrent infections. WAS-protein (WASP) links cell surface signals to the actin cytoskeleton. WASP is important in T cells for IL-2 production, proliferation, and migration. The WAS-interacting protein (WIP) stabilizes WASP. WASP level is severely reduced in T cells of WIP KO mice. The majority of missense mutations in WAS patients are in the WIP binding domain (WBD) of WASP and interfere with WIP binding, resulting in low WASP levels.
We identified a 35 a.a. fragment of WIP (termed nWIP), which when fused to the C-terminal of EGFP (EGFP-nWIP), interacts with WASP in Jurkat cells. When introduced into WIP KO T cells, EGFP-nWIP increased the level of WASP to normal, as determined by FACS analysis. EBV-transformed B cell lines from WAS patients with mutations in the WBD of WASP expressed low levels of WASP, but had normal mRNA levels. Introduction of EGFP-nWIP into these lines resulted in increased levels of WASP, up to normal. Two cell lines from patients with WAS mutations outside the WIP-binding domain did not show an increase of WASP after introduction of EGFP-nWIP.
Our results demonstrate that nWIP stabilizes WASP and ameliorates WASP levels in B cells from WAS patients with missesense mutations in the WBD of WASP. We will determine if nWIP will correct the functional defects in cells from these patients. (Supported by USPHS grant HL059561 and the PERKIN fund.)
CXCR6/CXCL16 is required for optimal NKT cell and dendritic cell interactions (33.2)
Authors: Veinotte Linnea Lora , Johnston Brent 
Published: January 2023

Abstract
Natural killer T (NKT) cells are a conserved T cell sublineage which rapidly produces cytokines upon activation by glycolipid antigens (i.e. α-GalCer) presented by dendritic cells (DC). Cross-talk between NKT cells and DCs is important in the initiation of anti-tumour responses. The objective of this study was to determine the role of CXCR6/CXCL16 in NKT cell-DC interactions. The chemokine receptor, CXCR6, is highly expressed on NKT cells, while its ligand, CXCL16, is expressed on DCs. When liver NKT cells were cultured with recombinant CXCL16 in vitro they were not activated directly. However, CXCL16 did enhance IFN-γ production in vitro when NKT cells were stimulated with suboptimal doses of plate bound anti-CD3. The presence of CXCL16 on the surface of DC is also important for NKT cell production of IFN-γ. NKT cells cultured with α-GalCer loaded CXCL16+ DC produced more IFN-γ than those cultured with CXCL16- DC. CXCL16+ DC stimulated more NKT cell IFN-γ production than CXCL16- DC in vivo as well. CD8+ DC constitutively express CXCL16 while all other DC subsets upregulate CXCL16 expression following α-GalCer stimulation. This is dependent on NKT cells since CXCL16 upregulation did not occur in NKT cell deficient (Jα18-/-) mice. DC maturation is also attenuated in the absence of CXCL16 signaling. These results suggest that CXCR6/CXCL16 is required for full NKT cell and dendritic cell activation/maturation.
Orchestration of B cell function by the ERM protein ezrin (34.2)
Authors: Gupta Neetu , Bagheri Nayer , Parameswaran Neetha 
Published: January 2023

Abstract
Appropriate B cell responses hinge on finely balanced signal transduction pathways as well as regulated membrane remodeling. Membrane remodeling requires its uncoupling from the underlying cortical cytoskeleton but its regulation and impact on B cell function is poorly understood. We have identified ezrin, a member of the ezrin-radixin-moesin (ERM) family, as a regulator of membrane-cytoskeletal uncoupling in response to both B cell antigen receptor (BCR) and chemokine stimulation. Both stimuli induce dephosphorylation of ezrin on amino acid T567 in the actin-binding domain of ezrin, resulting in conformational inactivation of ezrin. Expression of a mutant of ezrin (T567D) that constitutively links the membrane to the actin cytoskeleton interferes with BCR-induced lipid raft coalescence, and SDF-1-dependent B cell migration. Our data support a critical role for T567 dephosphorylation in the early membrane remodeling events associated with antigen and chemokine signaling. Mice with a conditional deletion of ezrin in the B cell lineage exhibit defects in peripheral B cell homeostasis. Ezrin-deficient B cells from these mice show impaired in vitro migration and in vivo homing. Our data suggest that ezrin has the potential to shape activation and migration associated with B cell homeostasis and immune response.
Supported by grants from NIH (DK068292), Cancer Research Institute, Department of Defense.
Dual function for a vision-related molecule: retinoic acid in the eye may contribute to ocular immune privilege (39.18)
Authors: Caspi Rachel R , Zhou Ru 
Published: January 2023

Abstract
Ocular immune privilege is a complex phenomenon that encompasses both local and systemic mechanisms. Ocular fluids contain TGFβ as well as several neuropeptides that are thought to contribute to ocular immune privilege. Retinoic acid (RA) and its metabolites are abundant in the eye, as they constitute an integral part of the chemistry of vision. RA was recently shown to be involved in immune regulation at mucosal surfaces but its role, if any, in immune responses involving the eye has not been studied. We therefore examined the ocular fluids for immunoregulatory activity that could be attributed to RA. Confocal and real-time RT-PCR analyses showed expression of RA-synthesizing enzymes RALDH 1, 2 and 3 in the cornea and retina. Aqueous humor (AH) added to T cells undergoing Th1 or Th17 polarization inhibited acquisition of effector function as determined by intracellular staining for IFNγ and IL17, and instead caused their massive conversion into FoxP3+ Tregs. This involved TGFβ-driven upregulation of RARα and could be reversed by antagonizing either RA or TGFβ using LE540 or anti-TGFβ antibodies. In addition, AH-conditioned dendritic cells had reduced ability to support acquisition of effector function by T cells that had not themselves been exposed to AH. Our data support the notion that the role of RA in the eye is not limited to the visual process but also contributes to ocular immune privilege.
NIH intramural funding
PI3Kgamma regulates mucosal immune responses to epicutaneous immunization with cholera toxin as adjuvant (39.2)
Authors: Papenfuss Tracey  L , Satoskar Abhay , Lu Bao , Boyaka Prosper N 
Published: January 2023

Abstract
PI3Kgamma is a phosphoinositide3-kinase predominantly expressed in leucocytes and activated by G-protein-coupled receptors. Cholera toxin (CT) is an AB5 enterotoxin that binds to GM1 ganglioside and catalyzes ADP-ribosylation of Gsalpha. CT is also an effective adjuvant that induces systemic and mucosal immunity. We examined the role played by PI3Kgamma in the mucosal adjuvant activity of CT for epicutaneous vaccines. PI3Kgamma KO and control C57BL/6 mice were immunized three times by transcutaneous application of 50 micrograms of CT with 1 mg ovalbumin (OVA). Control mice developed significant levels of OVA-specific IgA Abs in the serum and in fecal extracts. The adjuvant activity of CT in control mice also resulted in high OVA-specific IgG responses in the serum with IgG1&gt;IgG2b&gt;&gt;IgG2a. Interestingly, the magnitude of OVA-specific serum IgG responses and fecal IgA Abs, were significantly enhanced in PI3Kgamma KO mice. The lack of PI3Kgamma signaling also enhanced Ab responses to CT itself, but did not affect the Th1/Th2 profile of responses. These results suggest that PI3Kgamma plays a regulatory role on immune responses to epicutaneous vaccines with ADP ribosyl-transferase adjuvants.
Characterization of antibody responses to soy proteins following oral vaccination with adjuvanted soymilk formulations (39.39)
Authors: Oakes Judy L. , Piller Kenneth J. , Bost Kenneth L. 
Published: January 2023

Abstract
We are exploring a novel platform technology using soybean seed-based formulations for the development of subunit vaccines that can be delivered via an oral route. One requirement for an effective subunit vaccine formulation is the inclusion of an adjuvant in order to elicit a robust immune response within the intestinal mucosa. When considering a soybean seed based formulation it is critical that there is no immune response to native soy proteins when administered orally with an adjuvant. One of the many advantages of a soybean seed-based platform for vaccine development is the fact that virtually every animal and human has been exposed to soy proteins given that soy is a major component of almost every diet. We hypothesize that routine consumption of soy proteins in a diet should effectively tolerize against soybean proteins and therefore abrogate the adjuvant effect of soy proteins. To test this hypothesis, we orally vaccinated groups of mice with formulations containing high (15mg) and low (1mg) doses of soybean seed extracts, along with cholera toxin (CT) as an adjuvant. As expected, there were no detectable sera IgE or IgG response in either group of vaccinated mice. Furthermore, there was no detectable IgA response against soy proteins in fecal extracts of immunized animals. Immune responses to CT in all groups were as predicted with a robust IgG response reflecting a Th2 subtype and an IgA response in fecal extract. Collectively, these data demonstrate that adjuvanted soybean-based formulations do not induce immune responses to soybean seed proteins, and underscore the vast potential for a novel soybean-based platform technology that can provide safe, stable and cost-effective vaccines worldwide.
Differential effects of mucosal adjuvant cholera toxin (CT) on macrophage cyclooxygenase-2 (COX-2) and endoplasmic reticulum (ER) stress (39.44)
Authors: Kogiso Mari , Pantuso Traci , Henriksen Ruth Ann , Dorey C. Kathleen , Shibata Yoshimi 
Published: January 2023

Abstract
CT is known to induce enhanced COX-2 activity and increased release of prostaglandin E2 (PGE2) in macrophages (MØ) and mucosal cells; CT also induces ER stress and unfolded protein responses. Since CT alone does not induce COX-2 transcription in MØ, post-transcriptional mechanisms up-regulating COX-2 appear essential, but remain elusive. To determine whether CT-induced ER stress is associated with regulation of MØ COX-2, RAW 264.7 MØ were treated overnight with CT (0.2 and 1 µg/ml) followed by 6 h treatment with heat-killed Mycobacterium bovis bacillus Calmette-Guérin (BCG, 5, 50, and 100 µg/ml). We found that pre-treatment with CT caused dose-dependent inhibition of BCG-induced COX-2 expression and marked elevation of growth arrest- and DNA damage-inducible gene 153 (GADD153), suggesting increased ER stress. Expression of glucose-regulated protein 78 (GRP78), an ER chaperone, was unchanged. In sharp contrast, if MØ had been pre-treated with BCG overnight before 6 h CT treatment, COX-2 expression was enhanced, GADD153 was not induced, and GRP78 expression was unchanged. Either pre- or post-treatment with CT resulted in inhibition of BCG-induced TNFα production. Our results indicate that the differential COX-2 regulation by CT is dependent on an ER stress/GADD153 pathway. Further studies will be conducted to assess whether the ER stress-dependent biphasic COX-2 expression is also observed in mucosal MØ and epithelial cells in CT-treated animals. (Supported by NIH HL71711, DOD DAMD 17-03-1-0004, BCCRP 06BS-04-9615)
The role of FasL and IFNγ in a model of AAV-mediated chronic liver injury in the mouse (39.52)
Authors: Spahn Jessica , Azadniv Mitra , Clementi Alicia , Crispe I. Nicholas 
Published: January 2023

Abstract
As models of chronic liver injury in the mouse are limited, we have developed such a model in the C57B6 mouse that utilizes Adeno-associated viral vectors (AAV) engineered to express both enhanced Green Fluorescent Protein and the CD8+ epitope SIINFEKL. After injection of this vector directly into the liver, 1x104 SIINFEKL-specific OT-1 cells are administered. Injection of these cells results in an increase in OT-1 cell numbers in the liver until Day 20 correlating with liver damage from Days 10-25. Addition of these cells does not cause decreased vector DNA but significantly decreases vector mRNA expression. To define the mechanisms by which OT-1 cells mediate their effect, we focused on both IFNγ and Fas. In looking at the role of these two molecules, we have found that IFNγ is important in the activation and/or recruitment of the OT-1 cells as IFNγR KO hosts have a significant decrease in the number of OT-1 in the liver at Day 20 compared to their WT controls. The same has been found when comparing WT and Faslpr hosts indicating a role of FasL in the same activation or recruitment processes. FasL has been shown by Suzuki et al to play a role in costimulation in a peptide model, but this phenomenon has not been seen in other viral or vector models1. The role of these two molecules in our AAV model may give clues to the molecular events that occur in other types of hepatitis.
Suzuki I JI 2000 Nov 15; 165(10):5537-43.
Support: NIH RO1 AI064463
ICAM-1 mediated NK cytotoxicity is enhanced by thymosin beta 4 (40.12)
Authors: Lee Ha-reum , Cho Dae-Ho 
Published: January 2023

Abstract
Thymosin beta 4, which is the major G-actin sequestering protein, has been shown to have ubiquitous distribution and multiple biological activities. However, thymosin beta 4's functions in relation to natural killer (NK) cells are still unknown. In this study, we show that synthetic thymosin beta 4 peptide increases NK cell cytotoxicity in a dose-dependent manner on human NK cell line, NK-92MI. Addition of thymosin beta 4 enhances NK cell aggregation and intercellular adhesion molecule-1 (ICAM-1) surface expression that are involved in NK cell cytotoxicity. Neutralizing ICAM-1 activity impairs thymosin beta 4-enhanced NK cell cytotoxicity. Furthermore, thymosin beta 4 stimulates degranulation of cytolytic granules, which is verified by CD107a assay, and up-regulates NK cell cytotocixity on primary human NK cells. Our findings revealed that thymosin beta 4 enhances NK cell cytotoxicity mediated by ICAM-1 through secretion of cytolytic granules to target cells. This suggests that thymosin beta 4 is a key activator of NK cell cytotoxicity.
Increased peripheral natural Tregs in prostate cancer patients is correlated with PSA (40.13)
Authors: Koksoy Sadi , Cicek Nilüfer , Akin Yigit , Erdogru Tibet , Baykara Mehmet , Ciftcioglu Mehmet Akif , Yucel Selcuk 
Published: January 2023

Abstract
The aims of this study were to analyze peripheral natural regulatory T cells (Treg) in Prostate Cancer (PCa) patients, regarding density and correlation with serum Prostate Specific Antigen (PSA). Peripheral Tregs from 75 subjects undergoing diagnostic prostate biopsies were enumerated by two sets of markers, CD4+CD25highFoxp3+ and CD4+CD25+CD127low/-. Association between peripheral Treg and serum PSA values was first determined in the entire population, including people with no prostate pathology (healthy) , PCa and Benign Prostate Hyperplasia (BPH) patients, and second, in 9 PCa patients before and after curative prostatectomy. Peripheral Treg frequencies were significantly increased in PCa patients (n=30) compared with BPH patients (n=30) and healthy subjects (n=15) by both methods (p&lt;0.01). The percentage of Treg in BPH patients was also slightly but significantly higher than that of healthy subjects. Importantly, the increase paralleled the elevation in total serum PSA levels, demonstrating a strong positive correlation (r =0.70 by both methods). Additionally, prostatectomy concomitantly restored both peripheral Treg numbers and serum PSA levels. In conclusion, peripheral Treg densities are not only increased in PCa and BPH patients but are also strongly correlated with PSA, suggesting that PSA may have a role in induction and/or maintenance of increased proportion of Tregs in these people.
Immunologic mechanisms against an SV40 viral oncoprotein require humoral and cell-mediated pathways to achieve tumor immunity (40.25)
Authors: Lowe Devin B , Shearer Michael H , Aldrich Joel , Jumper Cynthia A , Kennedy Ronald C 
Published: January 2023

Abstract
Malignant pleural mesothelioma (MPM) is an aggressive disease with survival prognosis less than one year. Interestingly, MPM samples have been reported to express the viral oncoprotein, Simian virus 40 (SV40) large tumor antigen (Tag). To this end, our laboratory has characterized protective SV40 Tag-specific immunologic mechanisms following prophylactic immunization in a murine experimental pulmonary metastasis model. Recent results indicate that ADCC functions as a primary in vivo anti-tumor mechanism since SV40 Tag antibody and Fcγ receptors are required immunologic-components in the observed protection from lung metastasis. We also find that with an antibody response due to vaccination, CD8+ T lymphocytes are required T cell subsets in the effector phase response of tumor challenge in vivo. The nature of the CD8+ T cell response appears to be CTL-driven based upon a systemic increase in Th1 cytokines by flow cytometry and tumor antigen-specific splenocyte production of IFN-γ when assessed by ELISpot and ELISA assays. In all, our results support the hypothesis that an SV40 Tag antibody-based ADCC pathway drives the activation of a cell-mediated response leading to protection from tumorigenic growth. These data also have implications for the construction of an effective MPM-specific vaccine that elicits broad-based immunity in individuals at high-risk for developing SV40 Tag-expressing tumor cells.
Cancer Immunoediting by the Innate Immune System (40.34)
Authors: Bui Jack D , Vermi William , Ngolab Jennifer , Arthur Cora , White J. Michael , Uppaluri Ravindra , Schreiber Robert D. 
Published: January 2023

Abstract
Although components of innate and adaptive immunity have been shown to work together to protect the host against cancer development and sculpt tumor immunogenicity (i.e., promote cancer immunoediting), it remains unclear whether innate immunity is capable of manifesting tumor-editing functions on its own. To address this question, we compared the immunogenicities of sarcoma cells derived from MCA-treated wild type (WT) mice, RAG2-/- mice lacking adaptive immunity, or RAG2-/- x IL-2Rγc-/- mice which lack natural killer (NK) cells and adaptive immunity. Consistent with our previous reports, all sarcoma cell lines from MCA-treated WT mice were poorly immunogenic and grew progressively when transplanted into syngeneic WT recipients. In addition, 40% of sarcoma cell lines from MCA-treated RAG2-/- mice displayed high immunogenicity and were rejected. Interestingly, MCA-sarcoma cells from RAG2-/- x IL-2Rγc-/- mice were highly likely to be immunogenic since 60% were rejected when transplanted into WT mice. Moreover, these highly immunogenic cell lines displayed delayed growth when transplanted into RAG2-/- mice and became heavily infiltrated with macrophages that expressed high levels of MHC class II and the glycolipid asialoGM1. This infiltration required IL-2Rγc function and resulted in tumor editing. Our results conclusively show that innate immunity can manifest IL-2Rγc-dependent tumor editing function in the absence of adaptive immunity. We propose that macrophages and NK cells collaborate as innate tumor editors.
Regulatory CD4+CD25+FOXP3+ T-cells can be Converted into Cytotoxic Effectors (40.36)
Authors: Peikert Tobias , Coughlin Kathleen , Van Keulen Virginia , Hansen Michael , Pease Larry 
Published: January 2023

Abstract
Background: Binding of the human IgM antibody B7-DC XAb to B7-DC on dendritic cells (DC) has profound immune modulatory effects. It enhances antigen presentation by DC, repolarizes T-helper (Th) 2 to Th1 cell responses, rapidly generates cytotoxic effectors and reprograms regulatory T-cells (Treg) into non-suppressive, cytokine producing autoimmune effectors. We hypothesized that co-culture of B7-DC XAb treated DC (DCXAb) with Treg results in the rapid generation of cytotoxic effectors.
Methods: Bone marrow derived DC from wild type C57BL6 mice (WT-B6) were treated for 24 hours with either B7-DC XAb or a control human IgM antibody in the presence of melanoma cell lysate (B16). Subsequently, DCXAb were co-cultured with Treg (CD4+CD25+FOXP3+), Non-Treg (CD4+, CD25-), cytotoxic CD8+ T-cells isolated from WT-B6 mice and Treg from CD8 knockout mice (CD8 -/-). After re-isolation these T-cells were used in cytotoxic T-lymphocyte (CTL) assays targeting B16 and Thymic lymphoma (EL4) tumor cells.
Results: Treg isolated from either WT-B6 mice or CD8 -/- mice co-cultured with B16 lysate treated DCXAb but not control antibody treated DC transformed into cytotoxic effectors targeting B16 but not EL4 tumor cells. Their level of cytotoxicity was comparable to that of CD8+ cells co-cultured with DCXAb. In contrast Non-Treg co-cultured with DCXAb did not kill in similar CTL assays.
Conclusion: Co-culture of Treg with antigen treated DCXAb rapidly generates antigen specific cytotoxic effectors.