bluuebunny/crossref_metadata_2025_split huggingface dataset:
Galectin-3 deficiency reduces the severity of experimental autoimmune encephalomyelitis (99.5)
Authors: Lukic Miodrag L , Jiang Hui-Rong , Al Rasebi Zakeya , Mensah-Brown Eric , Shahin Allen , Xu Damo , Fukada Sandra , Liu Fu-Tong , Liew Foo 
Published: January 2023

Abstract
Galectin-3 (Gal-3) is a member of β-galactoside-binding lectin family and plays an important role in inflammatory. However, the precise role of Gal-3 in autoimmune diseases remains obscure. We have investigated the functional role of Gal-3 in experimental autoimmune encephalomyelititis (EAE) following immunization with myelin oligodendrocyte glycoprotein (MOG)35-55 peptide. Gal-3 deficient (Gal-3-/-) mice developed significantly milder EAE and markedly reduced leukocyte infiltration in the CNS compared with similarly treated wild-type (WT) mice. Gal-3-/- mice also contained fewer monocytes and macrophages but more apoptotic cells in the CNS than did WT mice. Following Ag stimulation in vitro, lymph node cells from the immunized Gal-3-/- mice produced less IL-17 and IFN-γ than did those of the WT mice. In contrast, Gal-3-/- mice produced more serum IL-10, IL-5 and IL-13 and contained higher frequency of Foxp3+ regulatory T cells in the CNS than did the WT mice. Furthermore, bone marrow-derived dendritic cells from Gal-3-/- mice produced more IL-10 in response to LPS or bacterial lipoprotein than did WT marrow-derived dendritic cells. Moreover, Gal-3-/- dendritic cells induced AG-specific T cells to produce more IL-10, IL-5 and IL-12, but less IL-17 than did WT dendritic cells. Taken together, our data demonstrate that Gal-3 plays an important disease-exacerbating role in EAE through its multifunctions roles in preventing cell apoptosis and increasing IL-17 and IFN-γ synthesis, but decreasing IL-10 production.
Kinetic differences in the activation of CD4+ T cells during the initiation of tolerance and autoimmunity (99.50)
Authors: Katzman Shoshana , Villarino Alejandro , O'Gorman William , Nolan Gary , Abbas Abul 
Published: January 2023

Abstract
The initial encounter of a T cell with its antigen-presenting cell (APC) defines a major regulation point, relaying information to the responding T cell. We have assessed the interaction of T cells with APCs under conditions of tolerance and activation, by evaluating the response of auto-reactive CD4+ T cells upon recognition of self-antigen. In this model DO11.10 CD4+ T cells specific for OVA peptide (323-339) are transferred into transgenic mice, which express OVA as a systemically secreted soluble protein (sOVA-Tg). Previous studies in the lab have shown that in a "full" sOVA host, the auto-reactive T cells become tolerant, while in an "empty" or lymphopenic sOVA Rag-/- host, T cells respond with uncontrolled activation and host mice develop autoimmunity. Using 2-photon microscopy we have evaluated the velocity and movement during the initial encounter of T cells with APCs under conitions of tolerance or activation. Our data suggests that under both conditions the antigen specific cells initially stop on APCs, this interaction is retained in sOVARag-/- (autoimmune) mice but lost in sOVA (tolerant) hosts. In addition, the initial activation through the T cell receptor, as determined by pS6 staining, is comparable during early activation. This signal is retained in sOVARag-/- hosts but in contrast rapidly declines in sOVA hosts. These data show that the loss of early activation may define the tolerant phenotype. Understanding the interactions of self-reactive T cells with APCs presenting self-antigen is key to understanding the induction of tolerance vs. immunity and how this balance is maintained is integral for the design of therapies in the future. Research Support-NIH (F32)
Skin Tumor Responsiveness to IL-2 Treatment Correlates to CD8 Treg Expansion in an Immunocompetent Mouse Model. (100.15)
Authors: Foureau David , McKillop Iain , Jones Chase , Amin Asim , White Richard , Salo Jonathan 
Published: January 2023

Abstract
CD8 Tregs play an important role in graft tolerance and autoimmune disease status. Interleukin-2 (IL-2) induces Foxp3 expression by activated human CD8 T cells in vitro, and expands circulating CD8 Foxp3+ T cells in melanoma patients. IL-2 is FDA approved for the treatment of advanced melanoma, yet positive outcomes are limited. The aim of this study was to determine the phenotype and distribution of CD8 Foxp3 expressing T cells in skin tumor-bearing mice receiving IL-2. In normal or skin tumor-bearing mice, Foxp3 expressing CD8 T cells were a rare but naturally occurring cell subset. Primarily located in skin-draining lymph nodes CD8 Foxp3+ T cell expressed both activated CTL (CD28+, CD44+) and Treg (CTLA4+, PD1lo, NKG2Alo) markers. Following high dose IL-2 treatment, a dramatic increase in CD8 Treg prevalence was observed in a subcutaneous-tumor model that is non-responsive to IL-2 treatment (B16-F10 melanoma cell line). Furthermore, distribution analysis in this (B16-F10) model demonstrated increased CD8 Treg expression in the circulation and tumor-draining lymph nodes. Conversely, in the IL-2 responsive MCA-205 fibrosarcoma-bearing mouse model no significant changes in CD8 Treg were measured, either in the bloodstream or within the tumor-draining lymph nodes. In conclusion, this study characterized CD8 Foxp3+ T cells (CD8 Treg) as a new immunosuppressive cell subset, the expansion of which was associated with skin tumor that failed to respond to IL-2 treatment.
Mechanical disruption of the primary tumor using magnetic beads inhibits metastasis of breast cancer. (100.24)
Authors: Bouchlaka Myriam , Wilkins Danice , Fuchs Alan , Evrensel Cahit , Murphy William 
Published: January 2023

Abstract
Successful anti-tumor effects are weakened by removal of the tumor antigen pool (i.e. surgery) or use of cytoreductive and immunosuppressive therapies (i.e. chemotherapy and radiation). We hypothesize that mechanical disruption of the tumor in situ will augment immune responses due to antigen release. Iron particles were injected as magneto-rheological fluid (MRF) into an orthotopic primary breast cancer and followed by application of a magnetic field to exert force and shear stress on the tumor. This will result in disruption of the tumor’s architecture and induction of classical “danger signals”: necrotic death, release of tumor antigen with activation of a localized immune response. To promote systemic immune responses we combined MRF treatments with immunotherapy (IT) at suboptimal dosages. Tumors treated with daily application of magnetic field showed increased necrotic cell death, recruitment of activated DCs (i.e. MHC II+, CD83+) to the draining LNs with production of proinflammatory cytokines/chemokines. We then combined MRF treatment with IT to determine if systemic anti-tumor effects could be achieved. Not only have we observed suppression of tumor growth on the contralateral side treated with MRF and IT but also we saw absence of metastasis to the bone marrow of animals treated with either MRF and IT or MRF and magnetic field alone. These data suggest that MRF and magnetic field application to the primary tumor synergize with immunotherapy in disseminated cancer.
Identification of an inhibitory ganglioside responsible for immune suppressive effects associated with ovarian cancer (100.27)
Authors: Oelke Mathias , Webb Tonya J. , Giuntoli, II Robert L. , Lopez Pablo H.H. , Li Xiangming , Schnaar Ronald , Tsuji Moriya , Schneck Jonathan 
Published: January 2023

Abstract
Tumors often use mechanisms to prevent or suppress immune recognition. One such mechanism is the shedding of immune-suppressive glycolipids into the local microenvironment. Previously we reported that pretreatment of CD1d expressing stimulator cells with ascites fluid from ovarian cancer patients inhibited the activation of NKT cells, one of the earliest lines of defense in an anti-cancer immune response. Here we identify the inhibitory factor responsible, ganglioside GD3, which we isolated from the polar lipid fraction of ovarian cancer-associated ascites. GD3 inhibited CD1d-mediated NKT cell activation directly, without antigen processing. Purified GD3 binds with high affinity to CD1d, even when compared to the well-established experimental CD1d activator, α-galactosylceramide. Mechanistically we show that factors in ascites fluid as well as purified GD3 bind directly to the CD1 antigenic binding site, and this presumably displaces endogenous NKT cell ligands. These data indicate that shedding of GD3 by ovarian cancer tumors is a mechanism of immune evasion that inhibits the activation of an anti-tumor NKT cell response.
Inflammation-Induced Myeloid-Derived Suppressor Cells have enhanced resistance to apoptosis. (100.31)
Authors: Chornoguz Olesya , Grmail Lydia , Sinha Pratima , Artemenko Konstantin , Zubarev Roman , Ostrand-Rosenberg Suzanne 
Published: January 2023

Abstract
The purpose of this study is to deduce the influence of inflammation on apoptosis of Myeloid-Derived Suppressor cells. Myeloid-derived suppressor cells (MDSC) accumulate in patients and animals with cancer where they mediate systemic immune suppression. MDSC induced in heightened inflammatory environments with high levels of IL-1β and/or IL-6 accumulate more rapidly, are more potent suppressors of T cell activation, produce more IL-10, and have a greater ability to down-regulate macrophage production of IL-12, as compared to MDSC that develop in environments with lower levels of these cytokines. To determine how inflammation enhances MDSC levels and activity we used mass spectrometry to identify proteins produced by MDSC induced in highly inflammatory vs. less inflammatory settings. Proteomic pathway analysis identified Fas pathway and caspase network proteins, leading us to hypothesize that inflammation enhances MDSC accumulation by down-regulating Fas-mediated MDSC apoptosis. Using cytochrome C and activated caspase 3 and caspase 8 as indicators of apoptosis, confocal microscopy and flow cytometry demonstrated that inflammation-induced MDSC have enhanced resistance to Fas-mediated apoptosis. These results suggest that an inflammatory environment protects MDSC against extrinsic-induced apoptosis resulting in MDSC with a longer in vivo half-life and may explain why MDSC accumulate more rapidly and to higher levels in inflammatory settings.
Coordination of hepatic and myeloid signal transducer and activator of transcription 3 in governing liver carcinogenesis and regeneration (100.36)
Authors: Wang Hua , Park Ogyi , Lafdil Fouad , Yin Shi , Horiguchi Norio , Gao Bin 
Published: January 2023

Abstract
Activation of signal transducer and activator of transcription 3 (STAT3) occurs virtually in most liver diseases, but how STAT3 links liver regeneration and cancer remains elusive. Using cell-specific STAT3 deletions mouse strains, we have investigated the interplay of myeloid and hepatic STAT3 in controlling liver regeneration induced by partial hepatectomy (PHx) and hepatocarcinogenesis induced by diethylinitrosamine (DEN). We show that in DEN model hepatic STAT3 plays an important role in mediating a cytoprotective action and enhancing tumor growth, while myeloid STAT3 also significantly affects tumor growth via its anti-inflammatory effect. Furthermore, STAT3 is activated in myeloid cells after PHx and its conditional deletion leads to enhanced innate inflammatory response and liver regeneration. Conditional deletion of STAT3 in both hepatocytes and myeloid cells results in strong elevated inflammatory response, activation of STAT1, apoptosis of hepatocytes and a dramatic reduction in survival after PHx, whereas additional global deletion of STAT1 protects against these effects. These data show that the interplay of myeloid and hepatic STAT3 signaling is essential to prevent acute liver failure during liver regeneration through tempering a strong innate inflammatory response mediated by STAT1 signaling. Secondly, orchestral crosstalk between hepatic and myeloid STAT3 enables survival of mutagenized cells and subsequent tumor promotion in DEN-induced hepatocarcinogenesis.
Role of human lymphocyte activation gene 3 in tumor infiltrating lymphocytes (101.12)
Authors: Bruno Tullia , Noonan Kimberly , Grosso Joseph , Durham Nicholas , Goldberg Monica , Getnet Derese , Yen Hung Rong , Pyle Kristin , Borrello Ivan , Pardoll Drew , Drake Charles 
Published: January 2023

Abstract
Among the various molecules that regulate T cell function, lymphocyte activation gene 3 (LAG- 3) has garnered significant interest. LAG-3 is expressed on activated T cells, B cells, NK cells, tumor infiltrating lymphocytes (TILs), and plasmacytoid dendritic cells. We previously showed that LAG-3 was relatively over-expressed on HA-specific transgenic T cells rendered anergic in vivo by encounter with cognate self antigen. In this system, regulatory activity could be functionally blocked with a LAG-3 specific monoclonal antibody (Huang et al). Observations in our lab using LAG-3 knockout mice demonstrate that CD8 T cells undergo enhanced homeostatic proliferation in vivo if LAG-3 is absent. Currently, we are conducting studies to understand the role of LAG-3 in human cancer. Via microarray, we compared CD4+25+GITR+ (Treg) T cells from the prostate to CD4+25-45RA+ (naïve) T cells from the peripheral blood. We found LAG-3 and CTLA-4 to be relatively upregulated in prostate infiltrating T regulatory cells. Also, at the expression level, we observed variable levels of LAG-3 expression on patient CD8 tumor infiltrating T lymphocytes (TILs). At a functional level, we have shown that a human anti-human LAG-3 antibody enhances T cell proliferation and cytokine function in a mixed lymphocyte reaction. Further studies that are currently underway suggest that LAG-3 could be a promising candidate for enhancing anti-tumor immunotherapy in a clinical setting.
Beta7 integrin deficiency suppress cutaneous tumor formation (101.15)
Authors: Kai Hiromichi , Kadono Takafumi , Ohmatsu Hanako , Tomita Manabu , Asano Yoshihide , Sugaya Makoto , Sato Shinichi 
Published: January 2023

Abstract
Beta7 integrin, a cell adhesion molecule, associates either with the alpha4 or with the alphaE integrin subunit. Alpha4beta7 integrin is expressed on most leucocytes and is essential for their migration to gut-associated lymphoid tissues by interacting with its primary ligand, MAdCAM-1. Although involvement of beta7 integrin in intestinal immunity is well-established, little has been known about the engagements of beta7 integrin in cutaneous immune responses, especially cutaneous anti-tumor immunity. In this study, we injected B16 murine melanoma cells into the skin superficial to the ear cartilage of beta7 integrin-deficient mice. The tumor formation in the ears from beta7 integrin-deficient mice were reduced compared to that from C57BL/6 mice. Local cytokine mRNA expression levels were measured by real-time PCR in the ear tumors, and IFN-gamma mRNA levels tended to increase in beta7 integrin-deficient mice compared with wild-type mice. To evaluate leukocyte subsets in the tumors, immunohistochemical analysis of the ear tumor was performed. Numbers of infiltrating CD4+Tcells per unit area were decreased in beta7 integrin-deficient mice compared with wild-type mice. Overall, these results indicate an important role of beta7 integrin in tumor immunity.
Comparative screening of human antibody libraries derived from autoimmune vs. cancer patients for affinity reagents directed against tumor-associated antigens (101.20)
Authors: Zhao Aizhi , Cheng Yi , Coukos George , Siegel Don , Scholler Nathalie 
Published: January 2023

Abstract
In our previous study we showed that scFv against tumor vasculature marker could be isolated from a yeast-display scFv library derived from an autoimmune patient (TTP). However, scFv library derived from cancer patients may yield more diverse and higher affinity reagents against tumor-associated markers. To address this question, we constructed a yeast display scFv library from PBMC and ascites B cells of 5 ovarian cancer (OC) patients. To clone the antibody repertoires into the yeast display vector, we designed a new set of primers capable of amplifying all antibody sub-families and we optimized a three-fragment gap repair method using VH, VL and linearized vector. While scFv assemblage by overlap PCR was not performing well, the recombination of VH, VL and linearized vector in yeast yielded 1x108 transformant per electroporation. Sequencing of 50 clones demonstrated that all of them contained different full-length scFv inserts. We are currently screening the OC scFv library with human TEM1 protein, to compare the frequency and affinity of the scFv against tumor markers when retrieved from OC scFv library vs. TTP scFv library. In conclusion, our findings so far indicate that three-fragment gap repair is a valid and cost-effective strategy to construct large yeast-display libraries. We anticipate that the comparative screening of the TTP and OC libraries using tumor markers may give indications about the repertoire and anti-tumoral specificity of cancer patient antibodies.
In vivo impact of proteasome- and TAP-dependent MHC class II processing. (130.12)
Authors: Robinson Tara , Eisenlohr Laurence 
Published: January 2023

Abstract
In order to be recognized by CD4+ T cells, viral protein antigens must be degraded intracellularly and resultant peptide fragments loaded onto MHC class II molecules. Classically, class II antigen processing is thought to occur via endosomal proteases and loaded onto MHC molecules in a late endosomal compartment with the assistance of the chaperone molecule H2-M. Previously, our lab has demonstrated that the H2-IEd-restricted epitope NA79-93 of the influenza A/PR/8/34 neuraminidase protein must be degraded by the proteasome and requires the presence of TAP- both molecules previously associated exclusively with the MHC class I pathway. Furthermore, this pathway is largely inaccessible when the source of viral antigen is an inactivated, non-replicating virus. A substantial fraction of the CD4 T cell response is directed towards epitopes processed via this endogenous pathway. In the current study we extend this work to examine how endogenously processed epitopes 1) affect T cell priming, and 2) contribute to providing protection from viral challenge. Differences in CD4 T cell function and the restricted repertoire elicited in the absence of endogenously processed class II epitopes contribute to differences noted after adoptive transfer and challenge with influenza. These results may provide important insights into rational vaccine design.
T cell epitopes derived from autoantigens are selected differently than those from exogenous antigens (130.15)
Authors: Kim Ae , Hartman Isamu , Boronina Tatiana , Cole Robert , Sadegh-Nasseri Scheherazade 
Published: January 2023

Abstract
We have established a novel minimalist cell-free MHC class II antigen-processing system for identifying immunodominant epitopes from protein antigens. The system constitutes HLA-DR1, HLA-DM (DM) and three cathepsins. This system not only identifies the immunogenic epitopes from complex protein antigens, but also provides insight to the mechanism of immunodominance. Understanding of how these immunogenic epitopes are selected certainly provides better strategies for developing effective peptide vaccine designs and therapies against infectious diseases, cancer, autoimmune diseases and allergy. Using our cell-free system, and based on the source of the antigens categorized as exogenous antigens versus autoantigens, we find that both DM and cathepsins are crucial in regulating the selection of immunodominant epitopes. Three types of determinants are generated: first, non-dominant epitopes that are sensitive to both DM mediated dissociation, and cathepsins digestion. These epitopes were eliminated immediately from peptide repertoire. Second, T cell epitopes derived from the exogenous antigen are DM resistant and cathepsin sensitive. Third, T cells epitopes derived from an antoantigen that are sensitive to DM mediated dissociation, but resistant to cathepsin digestion. This shows that generation of immunodominant epitopes is regulated differently depending on the source of proteins antigens.
CD23-bound IgE augments and dominates recall responses through human naïve B cells (130.3)
Authors: Griffith Qyana , Liang YanMei , Onguru Daniel , Shah Taslima , Haas Helmut , Mwinzi Pauline , Ganley-Leal Lisa 
Published: January 2023

Abstract
Peripheral blood human naïve B cells express high levels of CD23 and circulate pre-loaded with IgE. The antigen specificity of CD23-bound IgE presumably differs from the B cell receptor (BCR) and may reflect the antigen specific mix of free serum IgE. CD23-bound IgE is thought to enhance B cell antigen presentation raising the question of how a B cell might respond when presented with a variety of antigens and CD23-bound IgE specificities. We recently reported that an increase in CD23+ B cells is associated with resistance to schistosomiasis highlighting the potential importance of CD23-bound IgE in immunity. We sought to determine the relationship between BCR and CD23-bound IgE mediated B cell activation in schistosomiasis. Crude schistosome antigens downregulated basal B cell activation levels in individuals hyper-exposed to infectious worms. However, schistosome-specific IgE from resistant, occupationally exposed Kenyans recovered responses of naïve B cells to schistosome antigen. Furthermore, cross-linking of CD23 overrode intracellular signals mediated via the BCR illustrating its dominating role in B cell activation. Notably, the nature of the cognate antigen appeared to dictate the threshold of antigen-specific CD23-bound IgE required for B cell activation suggesting that CD23-bound IgE functions as a rheostat for B cell responses to antigenic stimuli. These results suggest that CD23-bound IgE augments and dominates host recall responses through naïve B cells.
Cutaneous delivery of Poly(dG) conjugated protein antigens targets and activates DCs in vivo. (130.38)
Authors: Toussaint Helene , Erdos Geza , Williams Melissa , Donahue Cara , Falo Louis 
Published: January 2023

Abstract
Efficient and specific targeting of exogenous antigen to antigen presenting cells (APCs) in vivo is a critical challenge for vaccine design. APCs acquire antigens from a variety of sources and endocytic mechanisms can predetermine intracellular antigen routing, processing and presentation to CD4+ or CD8+ T cells. We have designed an APC targeting strategy based on simple and direct coupling of Poly(dG) oligo deoxynucleotides to protein antigens. In comparison to soluble OVA, we find that conjugates of Poly(dG) and OVA (OVA-Poly(dG)) are rapidly internalized by dendritic cells (DCs) and follow distinct intracellular processing pathways. Functionally, OVA-Poly(dG) is both presented to OVA-specific CD4+ T cells (OT-II cells) and efficiently cross-presented to OVA-specific CD8+ T cells (OT-I cells) by DCs in vitro. Importantly, immunization of naïve animals by direct injection of OVA-Poly(dG) results in efficient priming of potent antigen specific CTL responses. Extending our findings to human skin, we show that I.D. injection of antigen-Poly(dG) conjugates results in efficient uptake by human skin migratory DCs including Langerin+ DCs that also acquire potent T cell allostimulatory function. Our data suggest that Poly(dG) can both target antigens to human skin immune APCs and function as an adjuvant to induce potent CD4 and CD8 mediated T cell immunity.