bluuebunny/crossref_metadata_2025_split huggingface dataset:
ABfinity™ Antibodies - A novel approach to antibody development (144.20)
Authors: Allen Michael , Kim Yun , Dubbels Bradley , Low Quentin , Leong Louis , Huuskonen Jarkko 
Published: January 2023

Abstract
Antibody development falls into two major categories. Polyclonal antibodies are simple and inexpensive to produce but suffer from high variability and low reproducibility. Monoclonal antibodies developed through traditional hybridoma technologies are more expensive and time consuming to produce but have less variability. We have developed ABfinity™ technology which allows us to combine the advantages of these technologies to develop species-independent recombinant monoclonal antibodies from peripheral blood of immunized animals in less than 3 months. ABfinity™ technology allows us to culture and select antigen-specific B cells, clone the corresponding IgG heavy and light chain genes and express them in mammalian cells in a rapid and high-throughput manner. ABfinity™ antibodies are tested in a variety of applications including western blot, imaging, flow cytometry, and immunoassay. Lot-to-lot variability is virtually zero and many antibodies are extremely potent compared to currently available antibodies. In addition to de novo antibody development, ABfinity™ technology can be used to recover IgG genes from existing hybridomas to rapidly develop a recombinant antibody. Finally, because of the recombinant nature of ABfinity™ antibodies we can add affinity tags and other modifications that greatly enhance their use in downstream applications. Through ABfinity™ technology we can mix-and-match specificities, isotypes, species, and modifications to create truly modular antibodies.
Tumor cells engineered to codisplay on their surface 4-1BBL and LIGHT costimulatory proteins as a novel approach for cancer immunotherapy (144.24)
Authors: Sharma Rajesh , Yolcu Esma , Elpek Kutlu , Shirwan Haval 
Published: January 2023

Abstract
Tumor cells genetically modified to express immunomodulatory molecules as vaccine have shown efficacy against cancer. However, such vaccines are not only cost/labor and time intensive, but also have safety repercussions. As an alternative, we developed the ProtEx™ technology that involves modification of the cell membrane with biotin and engineering with proteins of interest chimeric with core streptavidin. We herein demonstrate that tumor cells can be engineered simultaneously with two SA-chimeric costimulatory ligands, SA-4-1BBL and SA-LIGHT. The choice of 4-1BBL and LIGHT were due to their immune activating pleiotropic functions and stimulatory effects on tumor stroma, respectively. Using TC-1 tumor cells as a transplantable cervical cancer mouse model, we demonstrated that these cells can be engineered simultaneously with both molecules. Vaccination with irradiated TC-1 cells engineered with both costimulatory proteins resulted in 100% efficacy in prophylactic and &gt;57% efficacy in therapeutic TC-1 settings. In marked contrast, TC-1 cells displaying these molecules singly showed limited efficacy. Vaccine efficacy was associated with the generation of primary and memory T cell, IFN-γ, and Ab responses, and enhanced CD8+ T cell infiltration into the tumor. Engineering tumor cells in a rapid and effective manner to simultaneously display on their surface a collection of immunostimulatory molecules with distinct functions presents a novel approach for cancer immunotherapy.
A novel M cell-based strategy to boost immune responses to oral vaccines: induction of intestinal villous M cells by RANKL pretreatment leads to enhanced vaccine uptake and improved antibody responses (144.26)
Authors: Knoop Kathryn , Butler Betsy , Kumar Nachiket , Williams Ifor 
Published: January 2023

Abstract
An established strategy for improving immune responses to mucosal vaccines relies on targeting ligands to promote vaccine uptake by microfold (M) cells. M cells are antigen-sampling epithelial cells mostly found in the epithelium covering Peyer’s patches and other organized mucosal lymphoid tissues, but also present on a subset (~7%) of small intestinal villi as villous M cells (vM). Recently we showed that 4 days of systemic treatment (50 μg s.c. per day) with RANKL, a TNF superfamily cytokine, induces differentiation of vM on 100% of small intestinal villi, dramatically increasing the total number of available vM and leading to large increases in uptake of fluorescently labeled bacteria (30-fold) and 200 nm microspheres (100-fold). The present study was done to test a novel strategy for enhancing mucosal vaccine responses to oral vaccines based on RANKL pretreatment to induce supraphysiologic numbers of vM. Mice treated daily with RANKL beginning 4 days prior to a single oral immunization with 2 x 108 fixed OVA-expressing E. coli had an average 8-fold increase in the serum IgM anti-OVA response measured 7 to 11 days later. This effect of RANKL on the antibody response also synergized with the use of LPS as a systemic adjuvant given i.p. at the time of oral immunization. These results provide a proof of principle demonstration that RANKL-mediated induction of vM prior to oral immunization is a novel and effective strategy capable of enhancing the potency of mucosal vaccines.
Distinct role of ICOS in the generation of regulatory T cells induced by CD40-activated B cells (145.12)
Authors: Zheng Jian , Liu Yinping , Lau Yu-Lung , Tu Wenwei 
Published: January 2023

Abstract
Inducible co-stimulator (ICOS) belongs to CD28 and CTLA-4 cell-surface receptor family and is broadly expressed in activated T cells. ICOS has been shown to regulate the interactions between T and B cells. However, the role of ICOS in the induction of regulatory T cells (Tregs) by allogeneic CD40-activated B cells remains to be clarified. In this study, we assayed the expressions of ICOS and ICOSL in CD4+ and CD8+ Tregs induced by CD40-activated B cells and examined the roles of them in the generation and function of these Tregs. With the blockade of ICOS-ICOSL interaction by ICOS-Ig, we found that the generation of both CD4+ and CD8+ Tregs was significantly decreased as compared to control. Interestingly, this blockade during the induction of Tregs only abolished the suppressive capacity of CD4+ Tregs but did not affect the suppressive function of CD8+ Tregs. The loss of suppression in CD4+ Tregs was related to the decrease of the surface expression of CTLA-4 in CD4+ Tregs, which resulted from the impaired transfer of CTLA-4 from cytoplasma to cell surface. In conclusion, ICOS-ICOSL interaction plays an important role in the generation of CD4+ and CD8+ Tregs, and mediates the suppressive functions of CD4+ Tregs by regulating CTLA-4 expression on cell surface induced by allogeneic CD40-activated B cells.
Tumor-sensitized, Vβ spectratype-selected CD8+ T cells promote graft-versus-leukemia responses in the absence of severe lethal graft-versus-host disease in a murine model of allogeneic bone marrow transplantation. (145.20)
Authors: Fanning Stacey , Berger Stephanie , Korngold Robert , Friedman Thea 
Published: January 2023

Abstract
The successful use of allogeneic blood and marrow transplantation (BMT) in the treatment of hematological malignancies is hampered by the fact that many donor T cells responsible for mediating graft-versus-leukemia (GVL) effects are also involved in development of graft-versus-host disease (GVHD). Optimizing outcomes for allo-BMT depends upon successful separation of GVHD and GVL responses. Our work focuses on identifying host reactive and tumor reactive T cells based on TCR Vβ CDR3-size spectratype analysis. In this study, we used a MHC-matched, miHA-mismatched murine model of BMT (B10.BR→CBA) and a CBA-derived myeloid leukemia, MMC6. Previous studies have shown this GVHD model to be mediated by CD4+-independent CD8+ T cells. Here, we show that the Vβ13 family is uniquely skewed in the B10.BR anti-MMC6 CD8+ T cell response. Transplantation of CD8+Vβ13+ T cells at the dose equivalent of 10x106 CD8+ T cells, the dose at which recipient mice develop severe lethal GVHD, did not mediate GVHD or GVL. Increased doses of Vβ13+ T cells led to increased GVL responses, however, recipient mice began to exhibit symptoms of GVHD. Of most interest, when recipients were given MMC6 presensitized Vβ13+ donor T cells, they displayed a significant GVL response in the absence of lethal GVHD. These results indicate that Vβ spectratyping can be used to identify T cells uniquely skewed in GVL responses and suggest that tumor-presensitized T cells can promote GVL in the absence of GVHD.
De novo induction of antigen-specific regulatory T cells by islet transplantation (145.34)
Authors: Kim Yong-Hee , Yoon Il-Hee , Hong So-Hee , Cho Hyoung-Su , Nam Hye-Young , Park Chung-Gyu 
Published: January 2023

Abstract
T cell receptor transgenic mouse on RAG knock out background does not have regulatory T cells (Tregs). We could induce adaptive Tregs in one of such mouse strain Marilyn by HY-antigen-mismatched islet transplantation. We transplanted male C57BL/6 islet to streptozotocin-induced diabetic female Marilyn mouse. Marilyn mouse is HY(minor histocompatibility antigen in Y chromosome)-specific TCR transgenic mouse on RAG2 knock out C57BL/6 background. Naïve Marilyn mouse rejects male skin. But, male islets were accepted by Marilyn mouse and induced adaptive Tregs. After acceptance of islet grafts, we grafted male C57BL/6 skin to confirm the tolerance to male antigen. In that skin graft, induced Tregs could prevent rejection to male’s skin. We could also detect Tregs after female islet transplantation. But, they were donor-origin Tregs, and those Tregs could not protect the male skin from rejection. In conclusion, we can make de novo induction of Tregs by islet transplantation, and those Tregs are antigen-specific Tregs.
Drak2 is required for T cell mediated allogeneic transplant rejection (145.9)
Authors: Weist Brian , Walsh Craig 
Published: January 2023

Abstract
Drak2 is a serine threonine kinase that has previously been shown to negatively regulate activation of T cells through the T cell receptor. Additionally, T cells lacking Drak2 are hypersensitive to low levels of stimulation, but are surprisingly resistant to autoimmunity in an EAE model. Since it has been hypothesized that Drak2 deficient T cells die after activation, we tested the ability of these T cells to respond to allogeneic stimulation, which would mimic transplantation from an allogeneic donor. Using in vitro mixed lymphocyte reactions, Drak2-/- CD8+ T cells responded less to allogeneic stimulation compared to wildtype T cells when analyzed by CFSE dilution. Additionally, allogeneic tumors were used to determine the role of Drak2 during an in vivo allogeneic transplant. Rag2-/- gamma C-/-mice adoptively transferred with purified Drak2-/- T cells were not able to clear allogeneic tumors contrary to wildtype T cell recipients. Additionally, transgenic Bcl-xL expression in Drak2-/- T cells rescued the alloresponse, illustrated by restored ability to clear allogeneic tumors. Next, we aim to determine if Drak2 blockade is sufficient to prevent rejection in a pancreatic islet transplant model.
TNF promoter region allele frequencies in the White population and their expression levels (34.11)
Authors: Selby Sandra , Posch Phillip 
Published: January 2023

Abstract
Tumor necrosis factor (TNF) is a pro-inflammatory cytokine that plays a variety of roles in immune regulation and disease. The TNF regulatory region contains numerous SNPs some of which affect expression. We previously identified nine alleles of TNF promoter region from random samples, but allele frequencies in different populations and diversity of the gene, as well as expression levels driven by the promoter alleles are not known. The TNF promoter and gene were sequenced from 82 random healthy White individuals. Three novel promoter alleles and two novel SNPs were identified. Novel SNPs also were found in intron 1 and the 3’ UTR. Promoter allele p*001 was the most common allele (55.49%). Allele p*002 (15.85%), p*006 (14.02%) and p*003 (7.32%) also were frequent, while all other alleles were found at less than 5% frequency. Differences were noted in an initial analysis of expression levels in T cells driven by the TNF promoter alleles. Allele p*004 was significantly increased in unstimulated and PMA/ionomycin conditions when compared to allele p*001. Expression was decreased for alleles p*005, p*006, and p*008 in comparison to allele p*001 in stimulated and unstimulated conditions. Allele p*007 showed decreased expression only in unstimulated conditions, while allele p*002 showed reduced expression only under stimulated conditions. These data confirm some of the previously reported impacts of SNPs on TNF expression and suggest an impact of additional SNPs on TNF levels.
IL-17A plays a pro-atherogenic role in atherosclerosis. (34.8)
Authors: Butcher Matthew , Smith Emily , Phillips Tracy , Prasad Konkall-Matt , Kolls Jay , Ley Klaus , Galkina Elena 
Published: January 2023

Abstract
Atherosclerosis is a disease of large arteries that is characterized by the formation of atherosclerotic plaques within the arterial wall and the immune response accompanies this process. To date, the contribution of IL-17A+ T cells in atherogenesis remains unclear. To test whether atherosclerotic conditions induce IL-17+ cell generation, we analyzed atherosclerosis-prone apolipoprotein E-deficient (Apoe-/-) and control C57BL/6 mice for IL-17A. CD4+ and γσ+ IL-17A+ T cells were significantly increased in the aortas, spleen and lamina propria of Apoe-/- vs. C57BL/6 mice. To analyze the role of IL-17A in vivo, Apoe-/- mice were transfected with adenovirus constructs expressing soluble IL-17 receptor (Ad-IL-17RA:Fc) or adenovirus-Luciferase controls. After a 15 week western diet, Ad-IL17RA:Fc-treated mice had reduced plaque burden, macrophage infiltration, and CXCL1 expression compared to controls. Atherosclerosis was attenuated in newly generated IL-17A-deficient Apoe-/- (Il-17a-/-Apoe-/-) mice vs. Apoe-/- mice. The expression of CX3CL1, CXCL1, CXCL2, CXCL5, IFNγ, IL-6 and TNFα was also reduced within the aortas of Il-17a-/-Apoe-/- mice. The adhesion of Monocytes to Il-17a-/-Apoe-/- aortas were diminished vs. Apoe-/- controls in ex vivo assays. Together, these results suggest that atherosclerotic-prone conditions induce Th17 and γσ+ IL-17A+ T cells, and accelerate atherosclerosis through the induction of aortic chemokines and recruitment of monocytes to atherosclerotic lesions.
Plasma membrane-derived vesicles (PMVs) carry TGF-β and promote adhesion and differentiation of THP-1 monocytes to macrophages (35.15)
Authors: Ansa-Addo Ephraim , Inal Jameel 
Published: January 2023

Abstract
Microvesiculation is a ubiquitous cellular mechanism, which occurs as a result of exocytosis, to release exosomes (between 50-100nm) or by direct release of vesicles from the cell surface membrane, known as plasma membrane-derived vesicles (PMVs) (0.1-1μm). Plasma membrane-derived vesicles are small, intact membrane vesicles, which are released by most cells upon activation with various extracellular stimuli and contain numerous proteins and lipids characteristic of the cell of origin. Among these are cytokines, for example transforming growth factor-β (TGF-β), a multifunctional protein, well documented to function in the regulation of cell proliferation, differentiation, morphogenesis and phenotype expression. Since TGF-β plays an important role in cell regulation, we tested whether TGF-β-bearing PMVs derived from THP-1 cells could themselves regulate the growth and differentiation of monocytes and their leukaemic counterparts. We show that addition of PMVs to monocytes or THP-1 cells did indeed cause significant reductions in the growth rate and induced their terminal differentiation into macrophages. Further investigations also showed that PMVs isolated from cells with high TGF-β content, caused significantly higher reductions in the growth rate of THP-1 cells as compared to PMVs from cells, which express less TGF-β. These results present a possible autocrine-like mechanism involving monocyte-derived PMVs by which monocytes terminally differentiate into macrophages.
Didox (3,4-Dihydroxybenzohydroxamic acid) is a potent anti-oxidant and anti-inflammatory compound (35.17)
Authors: Rice Charles , Gallicchio Vincent , Matsebatlela Thabe 
Published: January 2023

Abstract
Didox (3,4-Dihydroxybenzohydroxamic acid) is a synthetic antioxidant and one of the most potent ribonucleotide reductase inhibitors developed to date. Little is known about the potential for Didox as an anti-inflammatory agent. After an analysis of more than 200 oxidative stress- and inflammatory-associated genes in mouse RAW264.7 macrophages we show that Didox significantly lowers LPS-induced expression of inflammatory genes (iNOS, IL-1α, IL-1β, IL-6, TNF-α, NF-KB, Cox-2, p38α, TLR4, GM-CSF, G-CSF, CCL2, CCL4, others), while upregulating expression levels of anti-oxidant and anti-inflammatory genes (Catalase, glutathione peroxidase, IL-10, superoxide dismutase, and peroxiredoxins, others). In addition, Didox dose-dependently inhibits LPS-induced IL-6 secretion and nitric oxide production. Buthionine Sulfoximine- and LPS-induced ROS production are also suppressed by Didox. We also show that phagocytosis of yeast particles is decreased as well. In terms of cellular signaling, Didox prevents NF-KB activation and translocation of NF-kappa beta p65 subunits in LPS-treated cells. These results suggest that the mechanism behind anti-oxidant and anti-inflammatory properties of Didox may be mediated, at least in part, via modulation of NF-kappa beta signaling. Didox may, therefore, may hold promise in managing chronic diseases exacerbated by LPS-induced oxidative stress and inflammation in macrophage-mediated inflammation.
Macrophage migration inhibitory factor promotes central nervous system pathology in a model of neuroinflammation (35.8)
Authors: Cox Gina , Alexander J. , Kithcart A. , Williams J. , Smith K. , Shawler T. , Satoskar A. , Whitacre C. 
Published: January 2023

Abstract
Multiple sclerosis (MS) is a chronic inflammatory demyelinating disease of the central nervous system (CNS). Cerebrospinal fluid collected from MS patients during relapse was found to contain greater levels of macrophage migration inhibitory factor (MIF) than samples collected during remission. We have shown that induction of experimental autoimmune encephalomyelitis (EAE) in MIF-deficient mice results in reduced clinical signs and CNS inflammatory infiltrates relative to wild type controls. However, there was no difference in T cell function in the periphery between the two groups. These findings suggest that MIF acts within the CNS to potentiate disease progression. The potent pro-inflammatory properties of MIF are well established; however, the precise contribution of MIF to neuroinflammation is not well defined. MIF is expressed by many cell types in the CNS including infiltrating macrophages and resident microglia, suggesting that MIF may enhance the inflammatory environment of MS lesions. To assess the contribution of MIF to CNS inflammation, primary microglial cultures were treated with increasing doses of rMIF, which upregulated inflammatory mediators associated with MS and EAE: IL-1β, iNOS, and CCL2. We are currently exploring the role of MIF in vivo using an intraspinal model of neuroinflammation. Taken together, these data suggest that inhibition of MIF may serve as a therapeutic strategy for resolving CNS inflammation.
Paneth cell activation after acute kidney injury causes liver and intestine injury and systemic inflammation in mice (35.9)
Authors: Park Sang Won , Kim Mihwa , Chen Sean , Brown Kevin , Kolls Jay , Lee H. Thomas 
Published: January 2023

Abstract
Patients who develop acute kidney injury (AKI) frequently suffer from extra-renal organ dysfunction and systemic inflammation syndrome. We aimed to determine the mechanisms of ischemic and non-ischemic AKI induced systemic complications. C57BL/6 mice were subjected to sham surgery, 20 min renal ischemia or unilateral nephrectomy. In addition to plasma Cr, plasma ALT markedly increased 5 hrs after both ischemic and non-ischemic AKI. These rises were correlated with peri-portal hepatocyte necrosis, vacuolization, neutrophil infiltration and pro-inflammatory mRNA upregulation. Plasma levels of TNF-α and IL-17 were also elevated after AKI. Small intestine histology after AKI showed massive epithelial necrosis and apoptosis as well as profound endothelial apoptosis. Neutralization or genetic deletion of TNF-α or IL-17 prevented AKI induced hepatic and intestine injury. Remarkable hyperplasia and degranulation of Paneth cells were observed. Laser capture microscopy of Paneth cells showed significant TNF-α and IL-17 mRNA induction after ischemic or non-ischemic AKI. Finally, Paneth cell depletion with dithizone prevented both hepatic and intestinal injury after AKI. Taken together, we propose that Paneth cell activation/degranulation after AKI produces systemic inflammation and hepatic damage via cytokine induction and intestinal barrier dysfunction. Modulating Paneth cell dysregulation may have important therapeutic implications in reducing systemic complications arising from AKI.
Downregulation of intracellular apolipoprotein D in DNA-PKcs-independent V(D)J recombination (36.16)
Authors: Franco Daniel , Reardon Christopher 
Published: January 2023

Abstract
Recombination-inducible mouse pre-B cell lines are models for intracellular events that occur during lymphocyte V(D)J recombination. Using the ts-abl transformed pre-B cell lines with wild type DNA-PKcs or lines from SCID mice that have a DNA-PKcs deficiency, they can be induced to recombine their V(D)J genes segments. One of the events coincident with recombination is G1 cell cycle arrest that is much more dramatic in the SCID pre-B cells. We have recently become interested in intracellular apolipoprotein D (ApoD), an ancient intracellular lipocalin. ApoD has been described to have a role in extending lifespan by modulating cellular stress responses. ApoD would be expected to be upregulated in cells undergoing cell cycle arrest as this is a potentially stressful event for dividing cells. However, when assessing the effects of cell cycle arrest in the pre-B cell lines, we found that the intracellular Apo D actually decreases. The SCID pre-B cells, having a more severe G1 cycle arrest from recombination induction, also show a more dramatic reduction in ApoD levels. It is unclear whether the down-regulation of ApoD is associated with the recombination induction or the cell cycle arrest events. Since the SCID pre-B cells are deficient in DNA-PKcs function, it is tempting to speculate that ApoD gene expression may be influenced by this protein kinase. Nevertheless, these data suggest a previously undescribed role for intracellular ApoD in B-cell V(D)J recombination events.