bluuebunny/crossref_metadata_2025_split huggingface dataset:
Role of the innate and adaptive immune system in omental anti-tumor immunity (165.31)
Authors: Sedlacek Abigail , Gerber Scott , Lord Edith 
Published: January 2023

Abstract
Tumor metastasis to the peritoneal cavity is often observed in patients with primary ovarian cancer as well as resected colon cancers. Metastatic cells frequently attach to the omentum, a peritoneal adipose tissue that contains many immune aggregates (IA). Interestingly, it is to these IA that the tumor cells initially bind and proliferate. Our lab has been investigating the capability of harnessing these IA to generate productive anti-tumor immune responses using the tumor cell line Colon38, a colon adenocarcinoma syngeneic to C57BL/6 mice. Immunization of mice with lethally irradiated Colon38 cells grown in serum-free media results in a marked increase in the number of CD8 T cells as well as their expression of activation markers. Challenge of the mice with live Colon38 14 days after immunization results in no observable tumor growth on the omentum and rechallenge experiments also yield no tumor growth. Unexpectedly, when we challenge mice with an alternate tumor cell line, B16.F0, we also observe no tumor growth on the omentum. These results have been repeated in the BALB/c strain of mice using syngeneic tumor cell lines. Our data not only highlight the uniqueness of the peritoneal cavity, but also suggest that this immunization strategy generates an initial non-specific anti-tumor response that likely involves components of the innate immune system as well as a long-term tumor-specific response involving CD8 T cells.
Aminophylline inhibits breast and bladder tumor growth in mice due to CXCR3-dependent adenosine A2B receptor blockade (165.32)
Authors: Cekic Caglar , Sag Duygu , Linden Joel 
Published: January 2023

Abstract
The activation of adenosine A2A or A2B receptors on immune cells inhibits their ability to suppress tumor growth. In this study, we evaluated the anti-tumor effects of aminophyllilne (AMO) a nonselective antagonist of A2A and A2B receptors that is used clinically to treat asthma. Intra-tumor injection of AMO inhibited the growth of syngeneic MB49 bladder cells subcutaneously injected into C57BL/6 mice and the growth and lung metastasis of luminescent syngeneic 4T1 breast tumor cells inoculated into the mammary fat pats of Balb/c mice. Although AMO inhibited the growth of MB49 in A2AR-/- mice it failed to do so in Rag1-/-, A2BR-/- or CXCR3-/- mice. The selective deletion of the A2BR or CXCR3 from bone marrow cells also blocked tumor suppression by AMO. Transcript levels of interferon-γ and CXCL10, and activation of CD4+ and CD8+ T cells, were increased in tumors treated with AMO, while numbers of CD34+/CD45- endothelial cell precursors were decreased. We conclude that: 1) the anti-tumor effects of non-selective adenosine receptor blockade by AMO are primarily mediated by the A2BR; 2) CXCR3 receptors are required for anti-tumor effects of AMO and influence both T cell trafficking and chemokine-mediated angiogenesis; and 3) AMO may be useful for proof-of-principle studies to determine the efficacy of adenosine receptor antagonists as tumor suppressors alone or in conjunction with tumor vaccines.
Molecular regulation of BCL2 to IGH translocation in follicular lymphoma (165.38)
Authors: Bai Jirong , Lange Miles , Xie Wanqin , Hong Song Yang , Huang Lin , Jones Dallas , Yu Zhihong , Ti He , Su Kaihong , Swanson Patrick , McKeithan Timothy , Chan John , Zhang Zhixin , Kai Fu 
Published: January 2023

Abstract
The hallmark of Follicular lymphoma (FL) is the aberrant translocation t(14; 18)(q32; q21) involving the IgH locus on chromosome 14 and BCL2 gene on chromosome 18. Previous analysis showed that approximately 70% of the BCL2/IgH translocations involve the BCL2 major break point region, known as the BCL2-MBR. Accumulating studies suggested that the BCL2-MBR to IgH translocation may be mechanistically similar to IgH DH to JH or VH to DJH recombination. Our analysis of 42 BCL2/IgH break points showed that 60% of them contain clearly identifiable DH genes, suggesting their origin from VH to DJH recombination. Our recent studies showed that the B lineage specific transcriptional factor Pax5 activates IgH VH to DJH recombination through binding to VH gene coding regions and recruiting RAG complexes. Based on this finding, we hypothesize that Pax5 also controls BCL2 to IgH chromosomal translocations. Our initial sequence analyses identified multiple potential Pax5 binding sites within the 150 bp BCL2-MBR. Electrophoresis Mobility Shift Assay (EMSA) demonstrated that Pax5 binds to the individual Pax5 binding sites as well as the full length BCL2-MBR. The BCL2-MBR displays Pax5-dependent enhancer activity when artificially placed in front of a minimal CMV promoter driven luciferase reporter gene. We also found that purified GST-RAG1 and RAG2 core proteins bind to and cleave at the BCL2-MBR. We speculate that Pax5 binding to the BCL2 MBR may enhance BCL2 to IgH recombination.
Epigenetic memory: tolerant T cells cannot escape their fate (165.40)
Authors: Schietinger Andrea , Delrow Jeffrey , Greenberg Philip 
Published: January 2023

Abstract
T-cell tolerance to self antigens is required to prevent autoimmunity. Since most tumor antigens are self antigens a critical challenge is to develop strategies that break T-cell tolerance to such antigens without causing autoimmune injury. A hallmark of tolerant T-cells is the failure to respond to antigen, but the precise regulatory mechanisms remain largely unknown. Previously we showed that strategies which “force” tolerant T-cells to proliferate abrogate tolerance. To understand the mechanisms by which repetitive cell division rescues tolerant T-cells, we studied the effect of homeostasis-driven proliferation (HP) on tolerant T cells. Following adoptive transfer into lymphopenic hosts, tolerant T cells became functional, even in a tolerogenic environment. However, T-cell rescue was only transient and tolerance was re-imposed after the host became lymphoreplete, even if no self antigen was present. The finding that rescued T-cells reacquired tolerance in non-tolerogenic hosts challenges the prevailing paradigm that tolerance maintenance requires continuous exposure of T-cells to self antigen. Instead, mRNA and microRNA profiling revealed that tolerant T-cells harbor a tolerance-specific molecular signature; while the programming can be temporarily overridden, it is eventually reasserted, dictated by an “epigenetic memory”. These studies on the regulation of T-cell tolerance have revealed novel targets for cancer immunotherapy and the treatment of autoimmune disease.
Arginase-I is expressed by ckit+IL-7Ra+ populations in cryptopatches (166.11)
Authors: Bando Jennifer , Liang Hong-Erh , Locksley Richard 
Published: January 2023

Abstract
Arginase-I is expressed by alternatively activated macrophages during type-2 inflammation and is strongly induced by the cytokines IL-4 and IL-13. In order to track arginase-I expression, our lab developed a YFP-arginase-I (Yarg) reporter mouse. In the naive Yarg mouse, we detected arginase-I-expressing cells that were CD45+ckit+IL-7Ra+, but lacked the common myeloid and lymphoid cell surface markers CD11b, CD11c, Gr-1, B220, CD3, and NK1.1. In the small intestine, these cells were found in cryptopatches and expressed RORγt and LTβ RNA, consistent with a lymphoid tissue inducer (LTI) phenotype. Indeed, we found Yarg+RORγt+ cells that were present in adult and one-day-old neonatal mouse small intestine. Interestingly, we also identified Yarg+RORγt- cells that were located in cryptopatches and were still present in the lamina propria of RORγt-deficient mice. These ckit+IL-7Ra+ subsets were the primary Yarg+CD45+ cells in several organs. Thus, ckit+IL-7Ra+ cells are a previously unappreciated arginase-I-expressing population in naive mice.
Differential activation of cytokine signaling pathways in the colon of Crohn’s disease and ulcerative colitis patients. (166.12)
Authors: Christophi George , Rong Rong , Holtzapple Philip , Massa Paul , Landas Steve 
Published: January 2023

Abstract
Cytokine signaling pathways play a central role in the pathogenesis of inflammatory bowel disease (IBD). Numerous clinical studies have shown that several inflammatory cytokines are elevated in active IBD and recently genome-wide association studies further confirmed and advanced our knowledge of intricate immune pathways that confer susceptibility to IBD. This study aims to delineate the cytokine signaling pathways in colonic mucosa of CD and UC patients, explore possible molecular markers for IBD diagnosis, and identify novel immune-mediators of IBD pathogenesis. Based on established clinical diagnosis and strict histopathological criteria, 93 colon biopsy samples were from severe/active UC or CD, quiescent UC or CD, and normal control subjects. Gene expression is analyzed by quantitative real-time RT-PCR. This study analyzes the expression of more than 30 genes, including cytokines, signaling molecules, and downstream inflammatory mediators. Cytokines like IL-17, IL-6 and IL-33 and transcription factors like T-bet, GATA-3, Foxp3are differentially regulated between UC and CD and among disease severity. These data provide insights into the pathogenesis of IBD though identifying novel immune-mediators and elucidating distinct signaling networks in CD and UC. Finally, several biologic agents are novel promising IBD treatments and this study provides an easy approach that can be used to identify molecular markers that can predict the response to those individual treatments.
HSP70-mediated antigen-dependent nonclassical MHC class Ib-restricted anti-tumor responses in the frog Xenopus laevis (170.15)
Authors: Nedelkovska Hristina , Robert Jacques 
Published: January 2023

Abstract
The heat shock proteins (hsps) hsp70 are highly conserved molecular chaperones that elicit potent T cell responses against the antigens (Ags) they chaperone in both mammals and Xenopus. We have shown that frogs immunized with hsp70 generate CD8 T cell responses against the Xenopus thymic tumor 15/0 that expresses several nonclassical MHC class Ib genes, but not classical MHC class Ia. Therefore, we hypothesized that hsp70 can prime class Ib-mediated anti-tumor unconventional CD8 T cells in an Ag-dependent manner. To test this, we produced Xenopus recombinant tagged hsp70 proteins (both the cognate hsc73 and the inducible hsp72) from stable 15/0 tumor transfectants. Antigenic peptides can be easily removed from both hsps by ATP treatment. We used a cross-presentation assay to adoptively transfer hsp-pulsed antigen presenting cells and showed that both hsp72 and hsc73-Ag complexes, but not ATP treated Ag-negative hsp70s, elicit class Ib-mediated CD8 T cells responses resulting in protection from 15/0 tumor challenge. From expression studies, we further postulate that these responses involve one or a few class Ib gene products among which XNC10 is the prime candidate due to its lymphoid tissue distribution. To further elucidate the role of class Ib molecules in anti-tumor immunity, we developed a reverse genetic approach combining transgenesis and siRNA technology and obtained F1 Xenopus clones with silenced expression of class Ib in vivo that will be used for further analysis.
Optimization of immunoassays to monitor canine immune system (170.19)
Authors: Delamere Patricia , Nadwodny Kimberly , DeVinney Brooke , Fuller Claudette 
Published: January 2023

Abstract
Drug related changes to lymphocyte populations and immune functions can indicate immunotoxicity in many species, including the dog. Immunoassays are routinely performed in human, monkey, and rodent samples, but few validated immunoassays specific for the dog exist. Here, we present five immunoassays optimized for functional and phenotypic analysis of canine adoptive and innate immunity. These assays were evaluated in canine peripheral blood and/or tissue to characterize immunomodulation effects and pharmacodynamics of stimuli and/or pharmaceuticals.
Effect of larval thymectomy on T-cell development and immunoglobulin class switch in Xenopus laevis (170.5)
Authors: Mashoof Sara , Du Christina , Criscitiello Michael 
Published: January 2023

Abstract
The gut associate lymphoid tissues (GALT) are the digestive tract’s immune system and play a crucial role in the evolution of the vertebrate adaptive immune system. Mammalian immunity depends on antibody production by B cells with T cell help. Our knowledge about adaptive immunity in cold blooded vertebrates is fragmentary. The frog Xenopus laevis is an ideal model for such studies. Amphibians were the first animals to evolve Ig isotype class switching. It is unclear if T cell help is phylogenetically more fundamental or class switch recombination. To determine whether T cell help is required for gut-specific IgX induction (assumed analogue of mammalian IgA) in X.laevis we need a model with no T -cells. We thymectomize 10-12 day tadpoles which provide us with a useful T -cell deficient model. Thymectomized tadpoles were monitored 15-20 days after surgery to check for any regrowth of thymus. TCR reverse transcriptase PCR was performed from peripheral blood of cleanly thymectomized animals. PCR results show there are no TCR α or γ mRNA transcripts in thymectomized frogs indicating that T-cell development is fully thymus dependent. Thymectomized animals validated by PCR will be orally immunized to test whether it will yield gut specific IgX response, and if IgX is T-dependent. Our preliminary experiments show higher induction of IgX in orally immunized animals with competent thymus.
On the trail of regulatory T cells in blood and tissues of Mycobacterium paratuberculosis infected cattle (170.8)
Authors: Coussens Paul , Murphy Brooke , Sipkovsky Sue , Colvin Chris , Roussey Jon 
Published: January 2023

Abstract
Based on previous observations, it is proposed that induced regulatory T cells (Tregs) form an important aspect of immune responses to Mycobacterium avium subspecies paratuberculosis (MAP) in cattle. Tregs are typically CD4+CD25+ and express the FoxP3 transcription factor. Removal of CD25+ T cells from peripheral blood mononuclear cells (PBMCs) isolated from MAP infected cattle prior to stimulation with MAP antigens enhances production of IFNγ mRNA and reduces production of IL-10 mRNA from remaining cells. To more definitively identify antigen-specific Tregs in MAP infected cattle, we have begun to analyze cell populations using multi-color flow cytometry and intracellular staining. Tregs in tissues have been visualized using multi-color confocal microscopy and immunohistochemical staining. As expected, stimulation of PBMCs from infected cattle with MAP enhances the percent CD3+ cells that are also CD25+ (25%, +/-2.8%), relative to nil stimulated cells (6.4%, +/- 3.8%). Expression of the FoxP3 transcription factor is more variable with some animals showing up regulation following stimulation with MAP (20%-50% of CD25+ cells) relative to nil stimulation (~6%). Thus, in some cattle, CD25+FoxP3+ cells do indeed form a significant proportion of cells responding to MAP antigen stimulation. FoxP3 positive cells are also a significant feature in many lesions associated with MAP infection, although it is not yet known if these cells are antigen specific.
T1D-derived osteoclasts display defective LPS-tolerance (44.1)
Authors: Catalfamo Dana , Neiva Kathleen , Calderon Nadia , Wallet Shannon 
Published: January 2023

Abstract
Bone and joint abnormalities are frequent co-morbidities of type 1 diabetes (T1D). The complexity of T1D itself and added complexity of bone and joint co-morbidities necessitates well-controlled and innovative approaches to assess the totality of potential causes for these co-morbidities. Objective: To elucidate osteoclast-specific mechanisms responsible for increased inflammation induced bone resorption in T1D by evaluating the phenotype and activation of T1D osteoclasts (OCs). Methods: Bone marrow-derived OCs from a T1D mouse model (NOD) and controls (NOR and C57BL/6) were evaluated for their ability for form resorption pits with SEM. Luminex was used to evaluate cyto/chemokine secretion to determine activational status and FACS analysis of calcitonin receptor (CR), integrin beta3, RANK and IL-10R was used to determine phenotype. All experiments were performed in the presence or absence of TLR stimulation. Results: NOD-derived OCs resorb more bone than controls in the presence of LPS while control OCs display LPS-induced tolerance. T1D-derived OCs also secrete higher levels of osteoclastogenic mediators and show increased RANK yet decreased CR expression compared to controls. Conclusions: These data suggest that NOD-derived OCs display defective LPS-tolerance when compared to controls which may be due to defects in osteoclastic regulatory mechanisms.
Secondary light chain gene rearrangements in the generation of anti-DNA autoantibodies in the chronic graft-versus-host model of SLE (44.15)
Authors: Eisenberg Robert , Choudhury Arpita , Luning Prak Eline , Schlissel Mark 
Published: January 2023

Abstract
The loss of B-cell tolerance to nuclear autoantigens is central to the pathogenesis of SLE, but it is not known which tolerance checkpoints fail in the production of anti-DNA autoantibodies. We have utilized the chronic GVH model of SLE to target the loss of tolerance to mature, splenic B cells (JI 179, 5564, 2007). We have now extended these studies to show that the production of these autoantibodies involves the switching of κ-chain alleles in mature B cells. We have distinguished the two κ alleles by utilizing a genetic locus in which the human κ constant region has replaced the mouse κ constant region. Either pure human κ+ CD93- splenic B cells or pure mouse κ+ CD93- splenic B cells, isolated from C57BL/6.huκ/moκ heterozygous mice, transferred to rag1-/- recipients and challenged with CD4+ bm12 T cells, produced both moκ+ and huκ+ anti-DNA and anti-chromatin autoantibodies. Even huκ+ cells that have their moκ locus in the germ line configuration produced moκ+ autoantibodies in this cGVH transfer system. These data suggest that the creation of lupus autoantibodies can occur at least in part by the acquisition of a new immunoglobulin receptor in mature B cells through peripheral secondary light chain genetic recombination (receptor revision). We postulate that abnormal T cell help is required at that stage either to initiate receptor revision and/or to permit the survival and activation of the B cells with their newly acquired autoreactivity.
Contribution of VH replacement products to the generation of autoantibodies in systemic lupus erythematosus (44.19)
Authors: Yu Yangsheng , Fan Run , Wang Zhixin , Yue Yinshi , Huang Lin , Li Song , Su Kaihong , Zhang Zhixin 
Published: January 2023

Abstract
Systemic lupus erythematosus (SLE) is characterized by the over production of high affinity autoantibodies that play important roles in SLE pathogenesis. However, it is not clear how these antibodies are generated and selected during the course of lupus. Our sequence analysis show that VH replacement products are highly enriched in IgH genes derived from different autoimmune diseases, including SLE. To gain insight into the mechanisms responsible for autoantibody generation in SLE, we performed single cell PCR analysis to study the functional antibody repertoires in active SLE patients. Analysis of 102 recombinant antibodies derived from the naive B cells and plasmablasts of the SLE patients show that 4.8% of the antibodies derived from naïve B cells and 26.7% of the antibodies derived from plasmablasts are autoreactive. Detailed sequence analysis reveal that in the plasmablasts of two SLE patients, VH replacement products contribute to more than 30% of IgH genes. These VH replacement products have long CDR3 regions enriched with charged amino acids, and more than 60% of them encode anti-nuclear or poly-reactive autoantibodies. These studies provide the first evidence that VH replacement products directly contribute to the generation of autoantibodies in SLE.
Excess B cell CD40/CD40L signaling promotes CD4 T cell-mediated encephalomyelitis in mice (44.22)
Authors: Shimoda Michiko , Bolduc Anna , Takezaki Mayuko , Tsubata Takeshi , Koni Pandelakis 
Published: January 2023

Abstract
To test the pathogenic role of B cells under excess inflammatory CD40/CD40L signaling, experimental autoimmune encephalomyelitis (EAE) was induced in B cell specific CD40L transgenic (CD40LBTg) and C57BL/6 control mice by immunization with MOG35-55 peptide. Clinical symptoms were monitored using a 0-5 scoring system during days 10-30 after induction. Alternatively, MOG-specific CD4 TCR Tg (2D2) mice were crossed onto a CD40LBTg background and the incidence of spontaneous encephalomyelitis monitored at 4-35 weeks of age. Both CD40LBTg and control mice developed EAE with similar kinetics, with disease onset at around day 15-17 and the peak of disease score at around day 21-25. However, CD40LBTg mice showed significantly more severe disease than wild type mice with average peak clinical scores of 3.0 and 1.5, respectively. Higher frequencies of IL-17+ CD4 T cells were also observed in CD40LBTg mice compared to wild type mice at the EAE disease peak. Also, 60% (6/10) of 2D2 CD40LB double Tg mice showed spontaneous encephalomyelitis-like symptoms (severe hind and/or fore limb paralysis) between 4-6 weeks of age, with IL-17+ and IFNγ+ CD4 T cells in spleen and inguinal lymph nodes. Furthermore, even those without apparent EAE symptoms died before 30 weeks of age. None of the 2D2 or CD40LBTg mice showed such symptoms between 4-35 weeks of age. These results support the notion that excess B cell CD40/CD40L signaling promotes MOG-specific pathogenic CD4 T cell response.
Automated analysis of multi-dimensional B-cell flow cytometry data (44.23)
Authors: Qian Yu , Wei Chungwen , Lee Eun-Hyung , Cambell John , Kaminski Denise , Kong Megan , Liu Yue , Dougall David , Sanz Ignacio , Scheuermann Richard 
Published: January 2023

Abstract
Understanding B cell diversity and its functional correlates requires the combined study of large numbers of surface and intracellular molecules. Advances in multi-parameter flow cytometry (FCM) allow for the independent detection of many fluorochromes on individual cells. However, traditional gating strategies based on single or two-color displays are inadequate for separating cell populations defined in multi-dimensional space. We have developed a novel program, FLOCK, which uses density-based clustering to algorithmically identify cell subsets in multi-dimensional space in an unbiased fashion, reducing operator variability. FLOCK has been used to accurately identify and quantify 17 distinct B cell subsets in human peripheral blood, including novel plasmablast subsets after tetanus vaccination. It is being used to monitor B cell subsets in different autoimmune conditions including systemic lupus erythematosus patients treated with BAFF inhibitors. FLOCK has been assessed by the FlowCAP consortium (Critical Assessment of Population Identification Methods), and achieved encouraging results in a recent FlowCAP competition compared with other automated methods. FLOCK has been implemented in the publically available Immunology Database and Analysis Portal - ImmPort (www.immport.org) for open use by the immunology research community. Core modules of FLOCK will also be integrated into GenePattern of the Broad Institute as a component of the FCM analysis suite.