bluuebunny/crossref_metadata_2025_split huggingface dataset:
IgG immune complexes promote systemic lupus erythematosus by elevating BAFF and inducing T cell infiltration into the glomeruli (P4036)
Authors: Kang SunAh , Fedoriw George , Jiang Chuancang , Diaz Marilyn , Vilen Barbara J. 
Published: January 2023

Abstract
In systemic lupus erythematosus, the presence of autoantibodies, and the formation of immune complexes are closely related to disease pathogenesis. Although the deposition of immune complexes in the kidneys is a characteristic of SLE, the impact of immune complexes during the onset of disease remains unclear. Here, we show that FcγR/IgG-bound immune complexes containing nuclear self-antigens are displayed on DCs and MFs from C57BL/6 mice at low levels (3-fold), while increased in MRL/lpr mice (10-15 fold). To investigate the biological function of accumulated IgG-immune complexes, we developed a passive antibody transfer system using AID-/-MRL/lpr mice. When treated with anti-nucleosome antibody (IgG), AID-/-MRL/lpr mice accumulated IgG-immune complexes on DCs and MFs, coincident with a dramatic increase in the numbers of splenic B cells, BAFF-secreting DCs, and serum autoantibody titers. The mice also developed lupus nephritis reminiscent of MRL/lpr mice, and showed infiltration of T cells into the glomeruli. These effects were not observed in MRL/lpr mice lacking FcγRI. Our data suggest a mechanistic link between IgG-immune complexes, BAFF secretion, and glomerular infiltration of immune cells. To investigate the role of BAFF in autoimmunity and T cell infiltration into the glomeruli, we will partially neutralize BAFF to restore serum BAFF levels to that of non-diseased mice. This will allow us to define the role of BAFF in autoimmunity, and lupus nephritis.
Cre-based deletion of COX-2 within B cells in vitro: steps toward specific nanoparticle-driven COX-2 inactivation in vivo. (P4044)
Authors: Haque Shabirul , Li Jianhua , de Marco Ario , Ishikawa Tomo-o , Herschman Harvey , Spretz Ruben , Noriega Sandra , Nunez Luis , Mongini Patricia 
Published: January 2023

Abstract
Several lines of evidence indicate that COX-2 and downstream prostaglandin (PG) positively influence germinal center development and Ig heavy chain class switching. Although activated B cells upregulate COX-2 and downstream effector molecules, it is unclear whether autocrine production of PG is a major driver of B cell autoimmunity. One approach to investigate this is through Cre-mediated, B cell lineage-specific COX-2flox recombination in mice prone to autoimmunity. Because Cre insertion into B lineage genes, CD19 and mb-1, compromises expression of molecules needed for BCR-triggered autoimmunity, we are seeking to achieve B cell-specific Cre-Lox recombination through an alternative route: B cell-specific NP bearing Cre as a payload. We here report that purified recombinant Cre protein rapidly achieves specific in vitro recombination within PCR amplicons of COX-2 with inserted pLox sites. Furthermore, one hour exposure of viable splenocytes from NOD.B10 COX-2flox mice to Cre protein at 37oC results in ~ 90% recombination. Cells treated with active Cre and subsequently stimulated with BCR-L, IL-4 and BAFF, display significantly less COX-2 protein upon reculture than cells similarly treated with heat-inactivated Cre. We additionally report the preparation of CD22-specific nanoparticles composed of PEG and PLA. In future studies, similar NP will be employed to target Cre specifically to B cells in Sjogren’s Syndrome-susceptible NOD.B10 COX-2flox mice.
The development of genetically modified K562 cells as artificial antigen presenting cells for NKT cells (P4310)
Authors: Tian Gengwen , Jena Bipulendu , Liu Daofeng , Heczey Andras , Torikai Hiroki , Lee Dean , Cooper Laurence , Metelitsa Leonid 
Published: January 2023

Abstract
CD1d-restricted type-I Natural Killer T cells (NKTs) have been shown to mediate anti-tumor immune responses in mouse models and are associated with improved outcome in several types of cancer. However, the therapeutic application of NKTs has been limited by low numbers and functional defects of these cells in patients with cancer. To provide a means for effective ex-vivo NKT-cell expansion for cell and gene therapy applications we explored native and engineered properties of K562 cells to function as artificial antigen presenting cells (aAPC). K562 cells express an activating allele of HLA-C and propagate NK cells which compete with NKTs in the same culture. Therefore, we used zinc finger nucleases to delete the endogenous HLA-C gene in K562 cells and transduced parental and HLA-Cnull K562 cells with CD1d cDNA followed by single cell sorting and clonal expansion. The clones exhibited a range of CD1d expression. They were pulsed with a NKT ligand, αGalactosylceramide and tested as aAPC for NKTs using CFSE proliferation assay. We found that aAPC clones with an intermediate level of CD1d expression induced the highest rate of NKT-cell proliferation. No difference was observed between K562/CD1d and HLA-CnullK562/CD1d clones when purified NKTs were used as responders. In contrast to K562/CD1d, HLA-CnullK562/CD1d clones did not expand NK cells when added to primary PBMC. Therefore, these HLAnullCD1d+ aAPC can be used to selectively expand primary NKTs for adoptive cell therapy.
Development of a fluorescent dual-cytokine cellular immune response assay: preliminary results from sipuleucel-T treated subjects (P4312)
Authors: Wei Jessica , Perdue Nikole , Sheikh Nadeem , Dela Rosa Corazon , Trager James 
Published: January 2023

Abstract
In the Phase III IMPACT study evaluating sipuleucel-T, an immunotherapy for asymptomatic or minimally symptomatic metastatic castrate resistant prostate cancer, post-treatment antigen-specific cellular responses were detected using an IFNγ ELISPOT assay. While this assay is a robust readout of memory responses, it only detects one cytokine, providing minimal information of the responding cells. As clinical trial samples are limited, simultaneous detection of multiple cytokines is a pragmatic alternative that provides additional information without using more cells. The UV-fluorescent FluoroSpot assay detects multiple cytokines, providing an understanding of the phenotype of the immune response. FluoroSpot results were concordant to the ELISPOT data; antigen-specific IFNγ responses were seen after treatment in sipuleucel-T subjects, indicating treatment produced immunological memory. However, we saw that IL-2 secreting cells were maintained throughout treatment, whereas IL-5 secreting (Th2) cells decreased after the first infusion. Most importantly, the FluoroSpot assay showed the generation of polyfunctional T cells (IFNγ/IL-2 and IFNγ/IL-5) after treatment. The presence of the distinct dual cytokine functional T cell signatures supports the supposition that sipuleucel-T induces proliferative antigen-specific T cell immunological responses. The multi-analyte FluoroSpot assay may provide insight into the roles of other different cell populations after sipuleucel-T treatment.
IL-33 precursor regulates inflammation in vivo in a T1/ST2-independent, Th2-independent fashion (P6270)
Authors: Atamas Sergei , Lockatell Virginia , Kopach Pavel , Todd Nevins , Luzina Irina 
Published: January 2023

Abstract
IL-33 precursor (IL33PR) released from damaged cells is known to bind to the receptor T1/ST2 and activate cells in culture. However, we found that in patients with interstitial lung diseases (ILD) and in the bleomycin mouse model of lung injury, pulmonary expression of IL33PR is elevated intracellularly and intranuclearly. Recombinant adenoviral gene delivery of IL33PR to mouse lungs in vivo did not lead to IL-33 maturation and produced a phenotype that was strikingly different from that induced by mature IL33 (MIL33). IL33PR induced pulmonary lymphocytosis and neutrophilia, whereas MIL33 induced eosinophilia, goblet cell hyperplasia, and increases in Th2 cytokines. The effects of MIL33 were, but the effects of IL33PR were not, abrogated by gene deficiency of T1/ST2. When combined with bleomycin injury, IL33PR potentiated lymphocytosis, collagen accumulation, and the expression of proinflammatory and profibrotic cytokines TGF-β, IL-6, MCP-1, and MIP-1α in the lungs. Gene expression profiling in cultured fibroblasts and epithelial cells overexpressing IL33PR revealed 10 - 100 fold increases in HSP70 mRNA, further confirmed by RT-Q-PCR. Previous reports have associated HSP70 with ILD. Immunohistochemical analyses showed strong HSP70 protein increases in mice challenged with bleomycin, IL33PR gene delivery, and particularly in mice challenged with these factors combined. Thus, IL33PR promotes ILD by inducing proinflammatory and profibrotic cytokines and HSP70.
Lymphopenia modulates levels of STAT1 expression leading to enhanced CD4 T cell responsiveness to Type-I IFN (P6272)
Authors: Le Saout Cecile , Hasley Rebecca , Imamichi Hiromi , Tcheung Lueng , Luckey Megan , Smith Mindy , Hu Zonghui , Sneller Michael , Rehm Catherine , Lane H. , Catalfamo Marta 
Published: January 2023

Abstract
To maintain the size of T cell pools throughout life, homeostatic mechanisms regulate both survival and proliferation through IL-7 and TCR signaling. IL-7 (through lymphopenia-induced proliferation/LIP) can also mediate expansion of autoreactive clones leading to the onset of autoimmune disease or enhance effector function against tumors and viruses. In the present study, we hypothesized that in a setting of chronic inflammatory environment, such as HIV infection, LIP could contribute to the immune activation leading to CD4 T cell depletion and CD8 T cell expansion. Type-I IFN is an essential cytokine in host defense during viral infection. In HIV infected patients, the genes associated with γc cytokines and Type-I IFN signaling were differentially expressed in CD4 and CD8 T cells. Using a lymphopenic murine model, we present evidence that, in vivo, IL-7 differentially regulated the expression of the total-Signal Transducer and Activator of Transcription 1 (t-STAT1) in CD4 and CD8 T cells undergoing LIP and in so doing enhanced CD4 T cell responsiveness to Type-I IFN. In this setting, chronic treatment with IFN-α led to decreased CD4 T cell counts and CD8 T cell expansion. This interplay between IL-7 and Type-I IFN might be advantageous in immunity against pathogens, however chronic stimulation of this pathway could be deleterious for CD4 T cell homeostasis and may contribute to the aberrant immune activation and eventual CD4 T cell depletion observed during HIV infection.
Blocking of PD-L1 on therapeutic dendritic cells modulate in vitro T cells proliferation and differentiation to Th1 and Th17 subpopulations (P4230)
Authors: Lopez Mercedes , Behrens Carolina , Falcon-Beas Cristian , Tempio Fabian , Salazar-Onfray Flavio 
Published: January 2023

Abstract
DCs constitute a promising treatment against melanoma. Recently, we showed the effectiveness of an allogeneic melanoma lysate DC-based immunotherapy for improving long-term survival in melanoma patients. Moreover, an elevated proportion of regulatory T lymphocytes (Tregs) was observed in non-responder patients. PD-L1 is an important co-inhibitory molecule expressed on DCs binding to PD-1 on T cells. This interaction promotes the induction, conversion and maintenance of Tregs, suggesting a role in immunosuppression. Here, we studied if the blockade of PD-1-PD-L1 interaction reverses Tregs differentiation and increases T helper populations. Monocyte-DCs were generated in the presence of pro and anti-inflammatory factors, and the PD-L1/PD-1 signaling was blocked by anti PD-L1 mAbs. Allo-stimulatory capacity of DCs was impaired, and inversely correlated with PD-L1/CD86 ratio. However, blocking PD-L1 on DCs increased of Th1 and Th17 proliferation, concomitant with a Tregs reduction, independently of the DCs phenotype and functional characteristics. We demonstrated that the PD-L1/CD86 ratio play a key role in the functional capability of DCs. Therefore, blocking this molecule on DCs becomes an attractive strategy to improve the efficacy of DCs-based vaccines in melanoma patients.
Recognition and neutralization of bovine herpesvirus-1 by bovine antibody variable heavy- and light-chain domains (P4182)
Authors: Pasman Yfke , Nagy Eva , Kasuhik Azad 
Published: January 2023

Abstract
Mono- and multimeric single chain variable fragments (scFvs), expressed in Pichia pastoris, are capable of neutralizing Bovine Herpesvirus type-1 (BoHV-1) in vitro [Koti et al. 2011. Vaccine 29; Pasman et al., 2012. Clin Vaccine Immunol 19]. The cDNA encoding these scFvs originated from a mouse x cattle hybridoma that secreted bovine IgG1 capable of neutralizing BoHV-1. The molecular modelling of heavy (VH) and light (VL) chain domains predicted that the VL might only have a supportive role in antigen recognition. VH and VL were expressed as single domains (Fd), in P. pastoris with an objective to assess their role in viral recognition and neutralization. The ability to recognize and neutralize BoHV-1 of the monomeric scFv3-18L, VH and VL linked via 18 amino acid linker, was compared to the ability of the individual FdVH and FdVL domains, in an ELISA and in vitro plaque reduction assays. These experiments demonstrated that while FdVH, but not FdVL, recognized BoHV-1 in an ELISA, FdVH did not neutralize BoHV-1. The complexities associated with such antibody functions will be discussed. [Supported by NSERC Canada]
Leptin antagonist reduces STAT1/3 activation, lessens Smad7 expression, restores TGF-β1 signaling and induces regulatory T cells (Tregs) to ameliorate chronic colitis (P4208)
Authors: Singh Udai , Singh Narendra , Busbee Brandon , Guan Hongbing , Price Robert , Taub Dennis , Mishra Manoj , Nagarkatti Mitzi , Nagarkatti Prakash 
Published: January 2023

Abstract
The etiology of Crohn’s disease (CD) and ulcerative colitis (UC), the two major forms of inflammatory bowel disease (IBD) are still unknown. However, there is evidence that chronic intestinal inflammation caused by excessive immune response to mucosal antigens affects millions of people around the world. Studies clearly support the role for TGF-β1 in the initiation and maintenance of tolerance by induction of Tregs to control gut inflammation. Leptin is a satiety hormone produced primarily by adipose tissue, which increases during colitis progression. In the present study, we investigated the mechanism of how pegylated leptin antagonist (PG-MLA) mediates the inhibition of experimental colitis. The PG-MLA significantly attenuates the clinical score and reverses pathogenesis including a decrease in body weight associated with colitis. PG-MLA reduces systemic and mucosal inflammatory cytokines. Further, PG-MLA increases the percentage of systemic and mucosal Tregs and TGF-β1 expressing CD4+ T cells. In addition STAT1/3 activation in immune cells, and expression of Smad7 in colon also lessens after PG-MLA treatment. These findings clearly suggest that PG-MLA diminishes Smad7 expression, reduces STAT1/3 activation and restores TGF-β1 signaling that leads to the induction of Tregs resulting in amelioration of chronic colitis. These results support a link between inflammation and nutritional status in controlling immune self-tolerance by inducing Treg responsiveness and function.
Abnormally short serum half-lives of chimeric and human IgGs in NOD-SCID mice (P4184)
Authors: Pop Laurentiu , Liu Xiao-yun , Pop Iliodora , Vitetta Ellen 
Published: January 2023

Abstract
The immunodeficient mice are the standard animal model in antibody research, which include the inbred NOD-SCID strain. We report here the aberrant pharmacokinetics of chimeric and human IgGs with a human IgG1 Fc domain in NOD-SCID mice. These IgGs displayed very short in vivo half-lives (25.0±0.4 hrs and 27.6±1.3 hrs, respectively) when injected at a dose of 10 μg. At a dose of 500 μg the half-lives were extended (131.1±3.1 hrs and 220.3±14.3 hrs, respectively), but still shorter than those observed in other strains of mice. The half-lives of these IgGs in NOD MrkTac, BALB/c, Swiss Webster and another immunodeficient strain of mice, C.B-17 SC-R, were similar to previous reports (300-400 hrs). In contrast, no differences in half-lives were observed for murine, rat, bovine and goat IgGs in any mouse strain studied here. RT-PCR for FcRn cDNA showed no difference in FcRn gene sequence and the binding affinity of chimeric and human IgGs to the FcRn-positive intestinal brush border of suckling mice was similar for all mice. However, chimeric and human IgGs showed higher in vitro binding affinities for Fc gamma receptors on macrophages from NOD-SCID mice vs. other strains of mice studied, suggesting that at low doses, these antibodies might bind preferentially to Fc gamma receptors-expressing macrophages, impairing their pharmacokinetics. Taken together, the NOD-SCID mice and their derivatives may not be suitable models for studying the in vivo activity of chimeric and human IgGs.
Therapeutic clearance of the virally infected nervous system is mediated by noncytopathic T cell interactions with resident myeloid cells (P4220)
Authors: Herz Jasmin , McGavern Dorian 
Published: January 2023

Abstract
Several viruses can infect the mammalian nervous system, some with devastating consequences, and others with little or no overt pathology. Adoptive immunotherapy is an approach that has shown promise in clinical studies for the treatment of CMV, EBV and adenovirus infections. Our laboratory models immunotherapy by transferring memory T cells into mice persistently infected from birth with lymphocytic choriomeningitis virus. Here, we demonstrate that memory T cells can completely purge the brain of persistently infected mice without causing blood brain barrier breakdown or severe tissue damage. This is accomplished by a tailored release of chemoattractants that recruit adaptive immune cells, but few pathogenic innate immune cells such as neutrophils and inflammatory monocytes. Interestingly, memory T cells enlist the support of nearly all brain resident myeloid cells (microglia) by converting them into CD11c-expressing antigen-presenting cells (APC). Two-photon imaging studies revealed that anti-viral CD8 T cells are more likely to decelerate and form stable interactions with brain-resident APC than CD4 T cells. Importantly, microglia do not undergo cell death following T cell engagement and appear to protect themselves by upregulating serine protease inhibitors like Spi-6. We propose that non-cytopathic viral clearance from the brain by therapeutic memory T cells results from tailored chemoattractant production and interactions with resident myeloid cells protected by Spi-6.
NB4 cells are superior to HL-60 cells as a basis for the Activated Transfected Killer (ATAK) cell platform (P4224)
Authors: Lin Lin , Tan Brandon , Pantapalangkoor Paul , Spellberg Brad 
Published: January 2023

Abstract
We are developing a cellular immunotherapy to enable neutrophil transfusions to treat neutropenic infections. We have found that HL-60 cells (AML M2 cells) can be transfected with a suicide trap and activated to differentiate into neutrophil-like cells. However, we now report that NB4 (AML M3 cells) result in far superior neutrophil differentiation by morphology, superoxide production, and CD11b expression. NB4-based ATAK cells possess superior microbicidal characteristics against Staphylococcus aureus, Acinetobacter baumannii, and Candida albicans than HL-60-based ATAK cells. NB4 cell microbicidal activity and morphology are similar to freshly harvested neutrophils.. Our current results confirm marked improvement in performance of the technology platform based on use of an NB4 cell line.
Dynamics of natural killer cell responses to hepatitis delta virus infection during the course of interferon alpha treatment (P6070)
Authors: Lunemann Sebastian , Malone David , Manns Michael , Sandberg Johan , Cornberg Markus , Ljunggren Hans-Gustaf , Wedemeyer Heiner , Björkström Niklas 
Published: January 2023

Abstract
Infection with the hepatitis delta virus (HDV) causes the most severe form of viral hepatitis. 15-20 million people are chronically infected with HDV. Hepatitis delta is believed to be an immune-mediated disease however adaptive immune responses against HDV are hardly detectable. This suggests a possible role of natural killer (NK) cells in the pathogenesis of HDV infection. To study this, we collected blood samples from 34 patients with hepatitis delta, 16 out of which were sampled during antiviral treatment with pegylated interferon alpha (peg-IFNalpha). Expression levels of activation and inhibitory NK cell receptors and multi-functional NK cell responses were assessed using 15-color flow cytometry. HDV infected patients exhibited higher baseline frequencies of NK cells. Peg-IFNalpha treatment was associated with a redistribution of NK cell subsets with increased numbers of CD56bright NK cells whereas total and CD56dim NK cells decreased. During treatment the most mature CD56dim NK cell subsets disappeared from the periphery and the frequency of immature CD56dim NK cells increased. Furthermore, the treatment resulted in decreased multi-functional NK cell responses against K562 cells, with and without additional IFNα stimulation. Taken together, we show that HDV infection increases the amount of NK cells. Furthermore our data suggest a reshaping of the NK pool by IFNα during treatment resulting in a loss of mature cells and decreased functional responses.
Reduced immune cells infiltration and increased neuroinflammation in type 2 diabetic mice infected with West Nile virus (P6038)
Authors: Kumar Mukesh , Roe Kelsey , Nerurkar Pratibha , Orillo Beverly , Verma Saguna , Nerurkar Vivek 
Published: January 2023

Abstract
Diabetes is a significant risk factor for developing West Nile virus (WNV)-associated encephalitis (WNVE) in humans. Using a diabetic mouse model, db/db, we recently demonstrated that diabetes enhanced WNV replication and susceptibility of mice to WNVE. Herein, we have examined immunological and pathological events in the brain of wild-type (WT) and db/db mice after WNV infection. C57BL/6 WT and db/db mice were infected with WNV and leukocyte infiltration, expression of cell adhesion molecules (CAM), neuroinflammatory responses and neuronal death were analyzed. We demonstrate that infiltration of CD45+ leukocytes and CD8+ T cells was significantly reduced in the brains of db/db mice, which was correlated with attenuated expression of CAM such as E-selectin and ICAM-1. WNV infection in db/db mice was associated with enhanced inflammatory response in the brain characterized by significantly high levels of multiple cytokines and chemokines. Elevated levels of cytokines correlated with increased astrocytes activation and neuronal damage in the brain of db/db mice. These data suggest that reduced leukocytes recruitment in the brain, in part, due to lower levels of CAM, results in failure to clear WNV infection leading to increased neuroinflammation, which mediates increased neuronal death and mortality in db/db mice. This is the first study to elucidate neuropathogenesis of WNV infection in a diabetic mouse model and can be used to develop therapies to manage WNVE in diabetics.