bluuebunny/crossref_metadata_2025_split huggingface dataset:
IL-22 is a target for the treatment of anaplastic lymphoma kinase-positive anaplastic large cell lymphoma (P6296)
Authors: Lim Chrissie , McFarland Adelle , Savan Ram 
Published: January 2023

Abstract
We have shown that leukemic T cells from anaplastic lymphoma kinase-positive anaplastic large cell lymphoma (ALK+ALCL) patients aberrantly express IL-22R1, the private receptor for IL-22, on their surface. This causes these cells to resemble a Th17 inflammatory cell type. The expression of IL-22R1 leads to IL-22 production, which then feeds back in an autocrine manner. This signaling also stimulates IL-17 in these cells that results in damaging systemic inflammation, a common complication seen in ALK+ALCL patients. We have shown that IL-22 may be a potential prognostic marker as it is likely a survival factor for the leukemic cells. The objective of the current study is to identify the role of the IL-22/IL-22R1 signaling pathway in ALK+ALCL development and during treatment. Since forced expression of IL-22R1 is sufficient to skew T cells to a Th17 phenotype and induce neutrophilia in vivo, targeting IL-22 may reduce inflammatory disease in ALK+ALCL patients. As IL-22 has proliferative effects, we hypothesized that IL-22 contributes to persistence of ALK+ALCL. We show that neutralizing anti-IL-22 antibody has a cytotoxic effect against an ALK+ALCL cell line when combined with a monoclonal anti-CD30 antibody. If neutralization of IL-22 shows therapeutic promise in our ALK+ALCL model, it could have rapid translational turnover.
E. coli fimbriae serve as an IL-35 agonist stimulating indoleamine 2,3 dioxygenase for protection against collagen-induced arthritis (P5154)
Authors: Kochetkova Irina , Maddaloni Massimo , Thornburg Theresa , Hoyt Teri , Pascual David 
Published: January 2023

Abstract
Orally delivered colonization factor antigen I (CFA/I) fimbriae stimulate IL-35 producing Foxp3+CD39+CD4+ regulatory T cells (Tregs) essential against CIA in C57BL/6 mice. To begin to understand how CFA/I-induced IL-35 mediates protection, recent studies have begun to examine immunosuppressive mediators by dendritic cells (DCs). It was found that IDO was induced in splenic, mesenteric lymph nodes (MLNs), and inguinal, axillary, and iliac lymph nodes (LNs) DCs in protected mice. We hypothesize that CFA/I-induced IL-35 could directly regulate development of IDO+ DCs to sustain anti-inflammatory responses. By flow cytometry, recombinant IL-35 was found capable of binding DCs in vitro suggesting direct action of IL-35 upon DCs. Mice deficient in IL-35 (EBI3-/-) were unresponsive to CFA/I fimbriae and unable to induce IDO. Similar results were observed in CD39-/- mice correlating with the lack of clinical interventions. To inhibit IL-35's impact, CFA/I- or PBS-dosed C57BL/6 mice were treated in vivo with rabbit IgG anti-mouse IL-35 during arthritis manifestation resulting in 50% suppression of intracellular IDO expression along with significantly enhanced CIA severity when compared to nonimmune IgG-treated mice. Adoptive transfer of CFA/I-induced CD39+CD4+ Tregs to mice induced with CIA elevated IDO expression by DCs. Thus, our findings demonstrate the effect of induced IL-35 by Tregs upon IDO+ DCs in conferring protection against CIA. Supported by NIH P01 AT-04986.
Dissecting the molecular mechanisms for the increased risk of T1D in HLA-DQ8/DQ2 heterozygous individuals (P5182)
Authors: Chow I-Ting , James Eddie , Gates Theresa , Yang Junbao , Torres-Chinn Nadia , Kwok William 
Published: January 2023

Abstract
HLA-DQ8trans is the HLA molecule that confers the highest risk for human type 1 diabetes (T1D), yet the reason for this risk remains elusive. Using tetramer-guided epitope mapping, we discovered a high degree of peptide overlap between DQ8 and DQ8trans for antigens from influenza A viruses, with only one peptide exclusively presented by DQ8trans. Among influenza epitopes shared by DQ8, DQ2 and DQ8trans, cross-reactivity was observed between DQ8 and DQ8trans, but not between DQ8 and DQ2 or between DQ8trans and DQ2. In agreement with foreign antigen observations, glutamic acid decarboxylase 65 (GAD65)-, and islet tyrosine phosphatase (IA-2)-specific T cell clones isolated from DQ8 or DQ8/DQ2 patients responded to peptides loaded on DQ8 or DQ8trans molecules. The autoantigenic peptides bound to DQ8 and DQ8trans in the same binding registry, but peptides presented by DQ8trans bound with higher affinity to both T cell clones and in vitro expanded T cells. Proliferation assays and cytokine ELISA revealed that DQ8trans elicited stronger functional responses from cross-reactive autoantigenic T cells and created an environment that favored the expansion of self-reactive T cells by preferentially promoting the secretion of IFN-gamma from cross-reactive T cells. Overall, our data suggest that the presence of higher affinity HLA-DQ8trans molecules enhances self-reactive DQ8 and DQ8trans cross-reactive T cell responses, leading to increased susceptibility to T1D in DQ8/DQ2 individuals.
NADPH oxidase 2 activity modulates antigen processing and epitope repertoires (P5186)
Authors: Lemmon Jason , Balce Dale , Rybicka Joanna , Yates Robin 
Published: January 2023

Abstract
Phagocytes such as macrophages and dendritic cells are essential to both the innate and acquired immune systems of mammals. To produce effective immune responses, material antigens must first be processed and presented by phagocytes. The importance of antigen processing is often overlooked, which is exemplified by current therapies focusing on downstream immune processes in T cells and B cells. However, these therapies often result in modulation of the immune system as a whole. Instead, it would be ideal to modulate antigen processing to prevent the generation of epitopes able to stimulate and re-stimulate specific T cells. Antigen processing within phagosomes is a dynamic process influenced by several environmental factors such as pH and oxidative burst. Of particular importance to our lab is the oxidative burst generated by the protein complex NADPH oxidase 2. Our lab has previously published data showing that regulation of NADPH oxidase 2 activity results in modulation of phagosomal proteolysis through modulation of cysteine cathepsin activity. We further hypothesize that protein antigen processing and epitope repertoire generation can be predicted and more importantly influenced by changing the activity of NADPH oxidase 2. Using a combination of fluorometric data, HPLC, and mass spectrometry we have begun to show, through preliminary data, that epitope repertoires can indeed be influenced without knocking down antigen processing and presentation as a whole.
Modeling the effect of PKCθ on T cell motility in lymph nodes (P5192)
Authors: Cannon Judy , Asperti-Boursin Francois , Oruganti Sreenivasa , Letendre Kenneth , Moses Melanie 
Published: January 2023

Abstract
The adaptive immune response is an integrated process that requires the interaction of T lymphocytes and antigen-presenting cells (dendritic cells) within the context of lymph nodes. Under homeostatic conditions, T lymphocytes actively “scan” the lymph node cortex, interacting with multiple dendritic cells (DCs) in search of cognate antigen. T cell motility is driven by CCR7-CCL21/19 and this chemokine-driven motility dictates the rate of interaction between T cells and DCs in lymph nodes and sets the properties of T cell - dendritic cell encounters. We recently found that CCR7 signaling alone without T cell receptor stimulation can activate PKCθ in naïve T cells. Using 2-photon imaging as well as in vitro migration assays, we show that PKCθ is required for optimal T cell migration. We have now generated a computer model to recapitulate the motility of both wild type and PKCθ-deficient T cells in the lymph node. By comparing the behavior of WT versus PKCθ-deficient T cells in our model, we will be able to determine whether the absence of PKCθ compromises interactions between T cells and dendritic cells in the lymph nodes, and therefore the initiation of an adaptive immune response. Our model will allow us to test not only the role for PKCθ in regulating T cell search for antigen, but also the role for other signaling molecules that may regulate T cell motility in vivo.
Deficiency of the leukocyte integrin inhibitor, Del-1, exacerbates experimental autoimmune encephalomyelitis through increased Th17 cell infiltration. (P5198)
Authors: Lim Jong-Hyung , Choi Eun Young , Samus Maryna , Lee Seung Hwan , Kim Hyesoon , Hajishengallis George , Chavakis Triantafyllos 
Published: January 2023

Abstract
Del-1 is an endogenous endothelial-derived inhibitor of LFA-1-mediated leukocyte adhesion (Choi et al. Science 2008). As Del-1 has high expression in brain, we assessed here its role in neuroinflammation in experimental autoimmune encephalomyelitis (EAE), the rodent model of multiple sclerosis. We found decreased Del-1 expression in the inflamed spinal cord of mice subjected to the EAE protocol. Moreover, Del-1-deficiency in mice resulted in increased EAE severity, associated with increased inflammatory cell recruitment to the inflamed spinal cord. Immunohistochemistry showed that demyelination and axonal damage of the spinal cord was higher due to Del-1-deficiency. Recently, we proved that Del-1 suppressed IL-17-triggered inflammation (Eskan et al., Nat Immunol, 2012). Consistent with this finding, we found that the infiltration of IL-17 producing CD4+ T cells (Th17 cells) was significantly increased at the onset of the disease. This resulted in enhanced infiltration of CD45highCD11b+ monocytes/macrophages that was up to the effector phase of EAE. Subsequent studies revealed that IL17R-/- deficiency reversed the aggravated EAE phenotype of Del-1-deficiency, accompanied by a decrease of leukocyte infiltration into the spinal cord. Our results suggest that Del-1 deficiency exacerbates EAE disease severity by elevating Th17 cell infiltration to the inflamed spinal cord, which implies that Del-1 could be used as a therapeutic approach for neuro-inflammatory disorders.
Identification of novel cell surface markers on mouse and human TH17 cells (P6354)
Authors: Bonnevier Jody , Guerra Alberto , Goetz Christine 
Published: January 2023

Abstract
TH17 cells are a subset of CD4+ T helper cells that protect against extracellular bacteria, but may also be involved in autoimmune disease progression. Consequently, basic and clinical research on TH17 cells has become a prominent and active area of research in immunology. The TH17 subset is defined by the ability to produce cytokines, such as IL-17 and IL-22, and also by the expression of cell surface markers, such as the IL-23 receptor. Much research effort in TH17 cell biology is the search for surface molecules that can be used to consistently characterize TH17 cells in different immune environments, and ultimately lead to potential therapies for autoimmunity. To date, only a few surface molecules have been found that are exclusively expressed by TH17 cells, making the use of surface markers for this cell type a challenge. Our objective was to identify new surface markers to distinguish TH17 cells from other subsets of helper CD4+ T cells. In order to accomplish this goal, we used a high-throughput flow cytometer to screen over 700 surface antibodies on mouse and human TH17 cells. We identified 26 novel surface markers in human TH17 cells that also coexpress IL-22, IL-17, and RORgT, confirming these markers were on functional TH17 cells. Interestingly, 4 of these markers (BTLA, CD200, CD99, and IL-18Ra) were also identified in mouse TH17 cells. Further study of these markers will enable the development of therapies for autoimmunity and cancer research.
Simultaneous multicolor flow cytometric detection and analysis of IL-17A and IFN-γ mRNA and protein levels expressed in human T lymphocytes (P6364)
Authors: Wu Xiaowei , Martin Jody , Park Emily , Ernst David , Berry Jody , Sepulveda Homero , Rohrer Jurg 
Published: January 2023

Abstract
The ongoing development of multiparameter flow cytometers, fluorescent dyes and specific antibodies for surface and intracellular markers provides valuable tools to analyze and define cellular and molecular mechanisms that underlie complex biological systems. These tools have contributed significantly to both basic and clinical research applications including immune system monitoring, drug screening, and diagnostic tests. Recently-developed techniques and reagents allow for the specific, amplified fluorescent detection of different RNA transcript levels expressed in individual cells. In this study, we evaluated the use of multicolor flow cytometry for the simultaneous analysis of differentially coexpressed mRNA and protein levels in primary cells. Using a human Th17-like cell model system, we demonstrate that in vitro-differentiated Th17-like cells can be simultaneously probed for their differentially expressed levels of IL-17A and IFN-γ proteins as well as IL-17A and IFN-γ mRNAs and IL-23 receptor mRNA. These results suggest that numerous potential new mRNA biomarkers can now be defined and used along with conventional immunofluorescent staining of protein biomarkers, to more fully characterize the nature of cells that function in complex biological systems. Simultaneous multicolor flow cytometric analysis of mRNA and protein in individual cells constitutes an important new toolset that can provide novel information from basic and clinical research investigations.
Natural killer cell impairment is associated with liver fibrotic progression in chronic HBV infection (P3361)
Authors: Zhang Zheng , Zhao Juanjuan , Li Yonggang , Wang Fu-Sheng 
Published: January 2023

Abstract
Natural killer (NK) cells have been implicated in inducing fibrosis remission in mice model; however, their roles in fibrotic progression remain to be elucidated in liver fibrosis (LC) patients. We comprehensively characterized peripheral and intrahepatic NK cells in 50 chronic hepatitis B (CHB) patients and 68 HBV-associated LC patients as well as 35 healthy subjects (HC). Both peripheral and intrahepatic NK cell frequencies in LC and CHB patients were significantly decreased, and they expressed lower levels of TRAIL, activation marker, activation receptors and granzyme and perforin and higher levels of inhibitory receptors in LC patients than those from CHB patients, thus displaying the decreased CD107a and interferon (IFN)-γ production and reduced killing capacity against hepatic stallete cells (HSCs). In vitro, HSCs-derived TGF-β could suppress NK cytolytic activity, and blockade of TGF-β significantly enhanced NK cell-derived CD107a and IFN-γ production. The immunohistochemical staining showed that NKp46-positive cells were more enriched in the α-SMA-negative area in livers from LC patients. Finally, NK cell cytolytic activity was also correlated negatively with liver fibrosis scores in HBV infected patients, which is further confirmed by the longitudinal follow-up of LC patients. Our findings may facilitate the rational development of immunotherapeutic strategies to enhance NK activity while limiting or abolishing liver fibrosis in chronic HBV infection.
Exploiting RNA aptamers for cancer therapy (P4407)
Authors: Pratico Elizabeth , Urban Johannes , Johnson Laura , Sullenger Bruce , Nair Smita 
Published: January 2023

Abstract
Nucleic acid ligands (aptamers), both antagonists and agonists, can be used to target receptors on cells with high affinity and specificity. We describe the development of RNA aptamers for several cancer therapeutic applications: (1) targeting antigens to dendritic cells (DCs), (2) immune modulation, and (3) tumor-specific delivery of therapeutics. Our laboratory has successfully engineered a nuclease-resistant RNA aptamer that binds to the macrophage mannose receptor (MMR) expressed on DCs with excellent specificity and high affinity. MMR aptamer-mediated targeting of tumor antigen peptide to DCs resulted in a 4-fold increase of IFN-γ expression, which enhanced cross-presentation and T cell activation in vitro. For immune modulation of T cell function, we have now developed a human OX40 aptamer that is used as an agonist. A multimerized version of the human OX40 aptamer stimulates T cell proliferation and IFN-γ expression. Lastly, we have developed an aptamer to deliver toxic agents to a subset of leukemia and lymphoma cells. This aptamer is engineered into a chimeric molecule possessing two levels of specificity: the binding domain (aptamer) and the agent domain (saporin) that specifically targets and kills leukemia cells (K562) in vitro.
Evaluation of ebolavirus glycoprotein Fc fusion protein as a subunit vaccine (P4417)
Authors: Konduru Krishnamurthy , Shurtleff Amy , Bavari Sina , Kaplan Gerardo 
Published: January 2023

Abstract
Ebolavirus is a Filoviridae that induces severe hemorrhagic fever in humans. Several vaccines based on Filovirus glycoprotein (GP) are under development including recombinant adenovirus, parainfluenza virus, Venezuelan equine encephalitis virus, vesicular stomatitis virus, and subunit GP vaccines such as virus-like particles and soluble protein constructs. Here we describe the development of an ebolavirus subunit vaccine based on GP fused to the Fc fragment of human IgG. We expressed the ectodomain of the Zaire ebolavirus (EBOV) GP fused to the Fc fragment of human IgG1 (EBOVgp-Fc) in mammalian cells and showed that GP undergoes the complex furin cleavage and processing as observed in the membrane-bound GP. In vaccinated mice, EBOVgp-Fc induced high levels of total and neutralizing anti-GP antibodies and elicited T-cell immunity characterized by the production of IFNγ (Konduru et al. 2011). Vaccinated guinea pigs developed a strong humoral response as measured by total and neutralizing antibodies to GP, and completely protected against lethal EBOV challenge. We are currently extending our studies to the monkey model. Our preliminary data indicates that EBOVgp-Fc induces neutralizing antibodies and recall CD8+ T-cell immunity, as assessed by the production of IFNγ and TNFα in PBMC. Further experiments, including protection against lethal challenge in monkeys, will be required to determine whether Filovirus GP-Fc fusion proteins could be developed as a candidate vaccine.
Evaluation of the genetically attenuated E. coli heat-labile enterotoxin (LTR192G) as adjuvant for mucosal and intradermal immunization (P4421)
Authors: Maciel Jr Milton , Dinesh Ravi , Poole Steven , Savarino Stephen 
Published: January 2023

Abstract
Among the challenges to developing a subunit vaccine against the diarrhea-causing pathogen enterotoxigenic E. coli (ETEC) is the selection of a proper administration route and adjuvant. We assessed the adjuvant LTR192G, an attenuated form of the heat-labile enterotoxin (LT), co-administered with dscCfaE protein, the prototype ETEC CFA/I fimbrial tip adhesin, by two different routes. BALB/c mice received dscCfaE+LTR192G by intranasal (IN) or intradermal (ID) immunization on days 0 and 21. Serum, spleen and bone marrow (BM) were collected on day 28. Both regimens elicited comparable serum IgG anti-dscCfaE and functional Ab levels, the latter measured by hemagglutination inhibition (HAI). IN immunization, however, resulted in higher serum and fecal IgA anti-dscCfaE Ab levels, and fecal HAI titers. IgG1 was the only adhesin-specific subclass detected in the ID group, while IN immunization also elicited low titers of IgG2a anti-dscCfaE Ab, suggesting a more balanced Th1/Th2 response. The number of IgG and IgA anti-dscCfaE antibody-secreting cells in spleens and BM were two-fold higher in the IN vs ID group. After in vitro stimulation with dscCfaE protein, similar numbers of IFN-γ spot-forming cells (SFC) were observed in both groups. The IN group showed lower numbers of IL-5 SFC compared to the ID group. The composite results suggest that ID immunization with dscCfaE+LTR192G elicits a mucosal immune response, but the magnitude and immune profile differ from the IN-immunized group.
Artificial antigen-presenting cells plus IL-15 and IL-21 efficiently induce melanoma-specific cytotoxic CD8+CD28+ T lymphocyte responses (P4389)
Authors: Lu Xiaoling , Zhao Yongxiang , Zhou Nuo , Yu Xia , He Jian , Mongkhoune Sodaly , Peng Yi , Xie Yu an , Zhou Sufang , Xie Xiaoxun , Luo Guorong , Li Xi , Fang Yuan 
Published: January 2023

Abstract
Abstract Objective: To develop a novel artificial antigen-presenting system for efficiently inducing melanoma-specific CD8+CD28+ cytotoxic T lymphocyte (CTL) responses. Methods: Cell-sized Dynabeads® M-450 Epoxy beads coated with H-2Kb:Ig-TRP2180-188 and anti-CD28 antibody were used as artificial antigen-presenting cells (aAPCs) to induce melanoma-specific CD8+CD28+ CTL responses with the help of IL-21 and IL-15. Dimer staining, proliferation, ELISPOT, and cytotoxicity experiments were conducted to evaluate the frequency and activity of induced CTLs. Results: Dimer staining demonstrated that the new artificial antigen-presenting system efficiently induced melanoma TRP2-specific CD8+CD28+ CTLs. Proliferation and ELISPOT assays indicated that the induced CTLs rapidly proliferate and produce increased IFN-γ under the stimulation of H-2Kb:Ig-TRP2-aAPCs, IL-15, and IL-21. In addition, cytotoxic experiments showed that induced CTLs have specific killing activity of target cells. Conclusions: The new artificial antigen-presenting system including aAPCs plus IL-21 and IL-15 can induce a large number of antigen-specific CD8+CD28+ CTLs against the melanoma. Our study provides evidence for a novel adoptive immunotherapy against tumors.
Ability of different isolates of Staphlococcal aureus to induce reactive metabolites in human polymorphovuclear leucocytes (P4429)
Authors: Javed Numan 
Published: January 2023

Abstract
Staphylococcus aureus is a gram positive bacterium that causes a number of diseases such as abscesses, infective endocarditis, septic arthritis, etc. It is acquiring resistance against many antibiotics like methicillin due to which its control is becoming difficult. Phagocytes interact with bacteria and phagocytose them. The phagocytosis leads to killing of bacteria by activation of respiratory burst during which different bactericidal metabolites like reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI) are produced. For this one hundred different isolates of S. aureus were obtained from various hospitals of Lahore in which fifty isolates were methicillin resistant while rest of the fifty isolates were methicillin sensitive. Peripheral blood from healthy individuals was used to obtain polymorphonuclear leucocytes by dextran density gradient centrifugation. Superoxide concentration was determined by the reduction of ferricytochrome c and nitric oxide concentration was measured by Griess method.Methicillin sensitive S. aureus (MSSA) has showed more phagocytosis by human polymorphonuclear leucocytes as compare to methicillin resistant S. aures (MRSA). Similarly, a significant difference was observed between two isolates of S. aures to induce ROI and RNI by human PMNs. MRSA produced more mean value of superoxide and nitric oxide as compare to MSSA.