bluuebunny/crossref_metadata_2025_split huggingface dataset:
Intra-epithelial T lymphocytes from the cervical-vaginal mucosa of healthy women contain a majority population of CD4+ cells and express high levels of the HIV-1 co-receptor CCR5. (P3341)
Authors: Swaims Alison , Haaland Richard , Lupo Davis , Evans-Strickfaden Tammy , Kohlmeier Jacob , Haddad Lisa , Hart Clyde 
Published: January 2023

Abstract
Current HIV sexual acquisition models for women posit that cell-free virus is carried into genital tissues by antigen presenting cells in the mucosa and that virus also penetrates the mucosa through microabrasions. The potential for genital mucosal intra-epithelial lymphocytes (IELs) to act as primary targets for HIV acquisition in women has not been well-characterized due to the difficulty of isolating and characterizing these cells free of blood contamination. We developed a method to enhance recovery of IELs from the lower genital tract mucosa of women using an atraumatic cervical-vaginal lavage procedure. This procedure was effective for collecting IELs with a T cell resident memory (T¬RM) phenotype from 10 healthy women without the blood contamination that often occurs using cytobrush or vaginal biopsy methods. We detected a high frequency of mucosal CD103+ expression on the CD8+ TRM cells, confirming their identity as IEL cells. Conversely, little to no CD103 expression was detected on CD4+ TRM cells. Moreover, CD4+ cells comprised up to 80% of the T cell population collected from CVL compared to CD8+ T cells (10-20%), with greater CCR5 expression compared to T cells in the peripheral blood. These results show that our atraumatic isolation method is effective for characterizing IELs in the female genital mucosa and that the abundance of CD4+ CCR5hi TRM cells in this population may likely be a highly vulnerable target for HIV-1 sexual transmission.
TLR4-based IL-8 reporter cell line as a tool for screening of the LPS-mediated immunomodulators to control TLR4 signaling pathway. (P3305)
Authors: Lee Hyun-Ku , Luo Yi Feng , Chitta Sriram , Singh Gita , Rosenberg Jonathan , Singh Sujay 
Published: January 2023

Abstract
The lipopolysaccharide (LPS) receptor complex of TLR4, MD-2 and CD14 plays an important role in sensing LPS-mediated immune signals, which provide immediate host protection and stimulate the host immune responses. Despite the fact that TLR4-mediated innate immune signals are required to combat invading pathogens, when signaling is not well regulated, uncontrolled activation can result in the disruption of host homeostasis leading to chronic inflammation and septic shock. Hence it is crucial to develop an assay system that can identify immunomodulators to control TLR4 signaling pathway. Since IL-8 is a major cytokine induced by TLR4, we developed the reporter cell line which stably expresses TLR4, MD-2 and CD14 proteins as well as an optimized Renilla luciferase reporter gene under the transcriptional control of IL-8 promoter. Functional activity of the cell line was evaluated by LPS dose response, TLR4 antagonist test and IL-8 induction suppressor test. The LPS EC50 of 4.3 ng/ml, TLR4 inhibitory peptide IC50 of 14.85 uM and an IL-8 induction suppressor, curcumin IC50 of 15.29 uM against the cell line were assessed. Using this cell line, a series of natural compounds was successfully screened for TLR4 signaling modulators, among which several compounds were identified as potential TLR4 signaling modulators. Taken together, it is proposed that this reporter cell line can be used as a tool for analyzing TLR4 signaling pathway as well as screening of TLR4 signaling modulators.
Development of a conditional NIK knockout mouse (P3293)
Authors: Brightbill Hans , Jackman Janet , Wu Lawren 
Published: January 2023

Abstract
NF-κB Inducing Kinase (NIK) is a kinase that is critical for the induction of non-canonical NF-κB signaling downstream of BAFF, CD40L, lymphotoxin, and RANK. Mice that are deficient in NIK or that have the ALY mutation that disrupts NIK function display defects in secondary lymphoid organ development, B cell populations and responses, and other immune cell phenotypes. We have developed a conditional allele of NIK and have utilized it to study the effects of NIK deletion in adult mice upon tamoxifen-induced deletion. Our conditional NIK KO mice may have broad utility for the study of the effects of NIK deficiency in adult mice and in specific cell types and tissues.
Preservation of allogeneic aortic grafts in anhydric sodium chloride attenuates host immune reaction (P2227)
Authors: Gewartowska Magdalena , Fronczak-Baniewicz Malgorzata , Buyanovskaya Olga , Moscicka-Wesolowska Maria , Maksymowicz Michal , Olszewski Waldemar 
Published: January 2023

Abstract
In case of infection of artificial arterial grafts, replacement by allografts becomes a method of choice. Unfortunately, allografts evoke strong immune reaction. For this reason, a method for successful long-term preservation of arterial allografts with unchanged morphology and low antigenicity is needed. We preserved the fragments of rat or pig aorta in anhydric NaCl and stored them in 4oC for up to 24 months, and subsequently transplanted them into rat or pig recipients for up to 24 or 6 months respectively. Routine hematoxyline/eosine and trichrome azan stainings, as well as electron microscopic studies showed the preserved anatomical structure of transplanted aortae and revealed normal structure of elastin fibers, the presence of fibroblasts between elastin bundles and single endothelial-like cells. No significant differences between preserved transplanted and non-preserved aortae stained for CD31, CD54, RECA-1 were observed. We also observed only slight infiltration of ED1 (CD 68), OX6 (MHC clas II) and W3/13 (CD43) positive cells around the allografts and this immunological reaction was only weakly expressed. No regional lymph nodes reaction was noticed. These observations suggest that preservation in anhydric NaCl may serve as a novel method to obtain low immunogenicity arterial allografts.
Correlations between cytokines, depression and anxiety in patients with colorectal cancer (P2000)
Authors: Miranda Diego , Azevedo Lucas , de LIma Tais , Veronez Luciana , Feres Omar , da Rocha José , Bisson Gabriela 
Published: January 2023

Abstract
Colorectal cancer (CRC) is a leading cause of cancer mortality in the world. Depression and anxiety are the most prevalent psychological disorders in patients with CRC. This study aims to investigate whether there is a correlation between anxiety, depression and serum cytokines in CRC patients. The study group consisted of 19 patients hospitalized for surgical resection of CRC and 20 healthy volunteers comprised the control group. Depression and anxiety were analyzed using the Hospital Anxiety and Depression Scale and serum levels of IL-1, IL-6, IL-12, TNF-α, TGF-β and IL-10 were measured by ELISA. We found that among CRC patients, 47.4% and 52.6% manifested clinically significant levels of anxiety and depression, respectively and 57.9% manifested a combination of severe anxiety and depression. Furthermore, these patients had increased serum levels of IL-1, IL-6 and TNF-α, but lower concentrations of IL-12. Analysis of correlation between HADS scores and serum levels of cytokines in CRC patients revealed a positive association of anxiety and/or depression with IL-1, IL-6 and TNF-α and a negative correlation with IL-12. These initial results indicate a correlation between serum levels of pro-inflammatory cytokines, anxiety and depression in CRC patients and suggest that cytokines are involved in the pathophysiology of these comorbidities in this sample.
Tumor-expressing CCR6 plays a critical role in human colorectal cancer metastasis (P2018)
Authors: Wang Honglin 
Published: January 2023

Abstract
The chemokine receptor CCR6 has been recently shown to be closely associated with liver metastases of colorectal cancer (CRC). However, the direct evidence for whether tumor-expressing CCR6 is a prognostic marker for the survival of CRC patients, and whether it plays a critical role in CRC progression in vivo is lacking. Here we found that levels of CCR6 were markedly increased in CRC cell lines, particularly un-regulated in metastatic cell lines and primary CRC clinical samples. Up-regulated CCR6 expression by CRC cells predicted poor survival in patients. Overexpression of CCR6 in HCT116 cells increased their proliferation, migration, and colony formation activity in vitro and promoted metastatic potential in vivo. We further demonstrated that CCR6 activated Akt signaling, up-regulated metastasis genes and down-regulated metastasis suppressor genes. Finally, we showed that selective targeting tumor-expressing CCR6 dramatically inhibited the growth of CRC in mice. Thus, our data indicate that CCR6 could be used as a novel prognostic marker for the survival of CRC patients, and targeting tumor-expressing CCR6 may represent a promising therapeutic strategy for CRC.
High-mobility group box 1 promotes NK and DC function as well as crosstalk: studies of selective knockout in immune cells (P2008)
Authors: Lotze Michael , Vernon Philip , Li Guanqiao , Liang Xiaoyan 
Published: January 2023

Abstract
High-mobility group box 1 (HMGB1) is an evolutionarily ancient, highly abundant nuclear protein present in all eukaryotes that has distinct compartmental roles in the nucleus, in the cytosol, and within the extracellular space. Within the nucleus it regulates VDJ recombination in B and T cells as well as promotes access to nuclear hormone/nuclear hormone receptors and p53 and p73 transcriptional complexes. Within the cytosol it promotes and sustains autophagy by displacing Beclin 1 from BCL2 and is critical for mitochondrial quality control. When secreted or released by stressed or necrotic cells, HMGB1 serves as the prototypic Damage Associated Molecular Pattern Molecule (DAMP). We created homozygous floxed HMGB1 mice which deleted exons 2 and 3 in back crossed NKp46-cre or CD11c-cre mice to allow specific deletion in NK or cDC respectively. Loss of HMGB1 in NK cells (NKH-) was associated with diminished number in the spleen, reduced proliferative capacity in response to IL-2 and enhanced tumor growth in a liver metastases model. Conversely loss of HMGB1 in cDC was associated with marked decrease in maturation in response to LPS or CD40 ligation but diminished tumor growth in the same liver model. Markedly decreased maturation of DC by NKH- with decreased CD80, CD86, Class II and CD83 was noted. These studies convincingly demonstrate the critical and central role of HMGB1 in immune regulation.
Differential migration of myeloid-derived suppressor cell subsets to tumor-draining lymph nodes in a murine model of breast cancer (P2014)
Authors: Matson Vyara , Loppnow Jessica , Steeber Douglas 
Published: January 2023

Abstract
Myeloid-derived suppressor cells (MDSC) expand in cancer and suppress the anti-tumor immune response. MDSC are morphologically and functionally divided into monocytic (MO-MDSC) and polymorphonuclear (PMN-MDSC) subsets, identified as CD11b+Ly-6G-Ly-6Chigh and CD11b+Ly-6G+Ly-6Clow, respectively. MDSC accumulate primarily in the blood and spleen, but their suppressive function is limited to the tumor and tumor-draining lymph nodes (TDLN). We sought to determine the involvement of adhesion molecules in MDSC migration to TDLN using the 4T1 murine breast cancer model. Both MDSC subsets expressed L-selectin and LFA-1, although L-selectin levels were higher on MO-MDSC. To determine whether this differential L-selectin expression affected MDSC migration to TDLN, we analyzed tissue distribution and in vivo migration of wild type and L-selectin-deficient MDSC. In the blood and spleen of wild type mice, the PMN-MDSC subset predominated (MO-MDSC/PMN-MDSC ratio of 0.1), while the MO-MDSC subset predominated in the TDLN (ratio of 1.5). By contrast, in L-selectin-deficient mice, the MO-MDSC/PMN-MDSC ratios in the TDLN (0.3) and in the blood and spleen (0.1) were similar. This differential tissue distribution was consistent with increased TDLN entry of wild type, but not L-selectin-deficient, MO-MDSC relative to PMN-MDSC in adoptive transfer experiments. Taken together, these results suggest a migratory preference of MO-MDSC to the TDLN dependent on L-selectin.
Cytokine response and morphologic characterization of chicken bone-marrow derived dendritic cells during infection with high and low pathogenic avian influenza viruses. (P6355)
Authors: Kapczynski Darrell , Jiang Hai Jun 
Published: January 2023

Abstract
Dendritic cells are professional antigen-presenting cells of the immune system that function to initiate primary immune responses. They are specialized at capturing and processing antigen for immune presentation. Infection of chickens with avian influenza can result in a wide range of clinical disease, depending if the virus is of low or high pathogenicity. Low pathogenic avian influenza viruses typically induce mild clinical signs, whereas highly pathogenic avian influenza can result in death within days. In these studies, chicken dendritic cells were co-cultured with high and low pathogenic avian influenza viruses of H5 and H7 subtypes to determine the innate immune response, effect on cell morphology, and examine viral growth. A strong proinflammatory response, including interleukin-1β, and stimulation of the interferon response pathway was observed at 8 hours post infection. Microscopically, the dendritic cells altered their appearance after influenza infection to a rounded shape, and eventually died, most likely through apoptosis. Differences in onset of morphologic changes were observed between H5 and H7 influenza subtypes. Increases in viral titers demonstrated that both pathotypes of influenza could replicate in dendritic cells. The increased activation following infection may be indicative of dysregulation of the immune response typically seen with high pathogenic avian influenza infection.
Co-infection with Streptococcus pneumoniae and influenza A virus leads to increased disease and a decrease in the size and functional capacity of the influenza specific CD8+ T cell response (P6363)
Authors: Blevins Lance , Wren Jack , Holbrook Beth , Johnson John , Pang Bing , Swords Edward , Parks Griffith , Alexander-Miller Martha 
Published: January 2023

Abstract
Most of our understanding of influenza A virus and its interaction with the adaptive immune response has been generated in the context of infection with a single pathogen. It is clear however that infection with influenza virus can lead to increased susceptibility to subsequent infection, often Streptococcus pneumoniae. The potential for a second infection to regulate the ongoing anti-influenza response is not clear. To address this important question we have developed an in vivo co-infection model to test the effects of pneumococcus on the ongoing CD8+ influenza specific T cell response. We show that co-infection with A/PR 8/34 (H1N1) and the non-invasive EF3030 strain of S. pneumoniae leads to increased disease. Within the lungs of the co-infected animals we observe a significant decrease in the size of the flu specific CD8+ T cell response. This decrease is not present in the lung draining lymph nodes, indicating selective regulation at the site of bacterial/viral co-infection. In addition to the decrease in size, the effector function of these CD8+ T cells is qualitatively diminished as measured by their ability to co-produce interferon γ and tissue necrosis factor α. These results demonstrate a dynamic interplay between the two concurrent pathogens and the immune system and emphasize the importance of increasing our understanding of how polymicrobial infections modulate the immune response to individual pathogens.
The expression of PD-1/PDL-1 on CNS cells following Theiler’s virus infection is elevated by IL-6 and inhibited by type I IFN (P6365)
Authors: Jin Young-Hee , Hou Wanqiu , Kang Hyun Seok , Koh Chang-Sung , Kim Byung S. 
Published: January 2023

Abstract
Theiler’s murine encephalomyelitis virus (TMEV) infection in the central nervous system (CNS) of susceptible mice results in an immune-mediated demyelinating disease, which is considered as a relevant viral model for human multiple sclerosis. We previously showed that the expression of positive costimulatory molecules is higher on microglia of TMEV-resistant B6 mice compared to TMEV-susceptible SJL mice. In this study, we analyzed the expression levels of negative costimulatory molecules, PD-1 and PDL-1, in the CNS of TMEV infected SJL and B6 mice. Our results indicated that TMEV infection induces the expression of both PD-1 and PDL-1 on microglia and macrophages in the CNS but not in the periphery. The expression of only PD-1 on CNS-infiltrating macrophages, but not on resident microglia, was considerably higher in TMEV-infected SJL mice compared to TMEV-infected B6 mice. We further showed that the presence of IL-6 is necessary to induce the upregulated expression of PDL-1 after TMEV infection. TMEV infection appears to provide both IL-6 and other undefined signal(s) for the upregulation. The expression of PD-1 and PDL-1 in the CNS of TMEV-infected mice was increased in the absence of signaling by type I IFNs, TLR3 or MDA5-mediated pathway. Taken together, these results strongly suggest that the expression of PD-1 and PDL-1 in the CNS is upregulated by IL-6 plus an unknown signal(s) following TMEV infection, and downregulated type I IFNs induced via TLRs and MDA5 pathways.
Crosspresentation with epitope-tagged antibodies targeted to different molecules on the dendritic cell surface (P5032)
Authors: Stewart Danielle , Prechl Jozsef , Schreiber Heidi , Sandor Matyas 
Published: January 2023

Abstract
Tuberculosis (TB) is one of the most prevalent global health crises in the world today and is caused by the bacterium Mycobacterium Tuberculosis (Mtb). While CD4+ T cells are known to be critical in the protective response against Mtb, the importance of CD8+T cell responses are gaining attention. The current vaccine BCG has limited and variable efficacy. BCG induces a strong CD4, but weak CD8+ T cell response. Targeting epitopes to specific dendritic cell receptors is an efficient way to induce T cell immunity. We created recombinant antibodies that recognize the pro-phagocytic receptor DEC205 (CD205), the co-stimulatory molecule CD40, and the phagocytic/activating Fc receptors 2/3 (FcR II/III) (CD16/32). Fused to the Fc portion of the antibodies is an Mtb immunodominant peptide, as well as a CD8+ T cell epitope. Individually, or in combination, these antibodies induce robust CD4+, antigen specific immune response in vivo. Here, we report that these DC receptor-specific recombinant antibodies cross present and cross prime antigen-specific CD8+ T cells. These responses were measured by activation and proliferation both in vitro and in vivo studies. These recombinant antibodies give us a new tool to understand how DC surface receptors influence the processing and cross-presentation of antigens to T cells. In future studies, we will use these recombinant antibodies in combination with BCG in a murine mycobacterium infection model to induce protective immunity against Mtb.
HLA-F and MHC-I open conformers are ligands for natural killer cell receptor KIR3DL2. (P5004)
Authors: Geraghty Daniel , Goodridge Jodie , Burian Aura , Lee Ni 
Published: January 2023

Abstract
Killer Immunoglobulin-like receptors (KIR) are innate immune receptors expressed by NK and T cells classically associated with the detection of missing-self through loss of their respective MHC ligand. Some KIR specificities for allelic classical class I MHC (MHC-I) have been described, while other KIR receptor-ligand relationships, including those associated with non-classical MHC-I, have yet to be clearly defined. We report here that KIR3DL2 and the non-classical antigen HLA-F, expressed as a free form devoid of peptide, physically and functionally interact. We further extend those results to include classical MHC-I open conformers as ligands, defining new allelic relationships with KIR3DL2 and MHC-I. The data collectively suggest a broader, previously unrecognized interaction between MHC-I open conformers - with HLA-F as a prototypical example - and KIR receptors, acting in an immunoregulatory capacity centered on the inflammatory response.
How does HLA-DM catalyze peptide exchange? Insights into the molecular mechanism of HLA-DM action from structural and biochemical studies (P5012)
Authors: Stern Lawrence 
Published: January 2023

Abstract
Efficient antigen presentation in the class II MHC pathway requires the non-classical MHC-II protein HLA-DM to catalyze peptide exchange on classical MHC-II molecules. Recent crystal structures of HLA-DM bound to the peptide-loading inhibitor HLA-DO and to a tethered MHC-peptide complex have clarified the nature of the HLA-DM / MHC-II interaction and revealed substantial MHC-II conformational lability around the N-terminal side of the peptide binding site. However, the molecular mechanism by which HLA-DM binding to a MHC-peptide complex induces peptide release still is not clear, and several conflicting models have been proposed. We evaluated the roles of various MHC-peptide and MHC-DM interactions in equilibrium binding and peptide exchange kinetics studies. The results suggest a mechanism in which DM binding induces MHC conformational changes that result in loss of key peptide main-chain hydrogen bonding interactions in the vicinity of the P1 residue, and alteration of peptide side-chain interactions in the P1 pocket, leading to release of peptide and formation of a meta-stable DM-MHC complex. Peptide binding driven by interactions all along the binding site can reverse these conformational changes. Thus resistance to HLA-DM mediated peptide exchange and ultimately peptide immunogenicity is related to the strength of MHC-peptide interactions throughout binding site and not simply occupancy of the P1 pocket.
Monoclonal antibodies to transmembrane proteins (P4022)
Authors: Brown Michael , Chambers Ross , Onisk Dale , Joaquim Tony , Stafford Joe , Lindpaintner Klaus , Keter Daniel , Stave James 
Published: January 2023

Abstract
Transmembrane proteins, including multipass transmembrane proteins like GPCRs and ion channels, are important targets for therapeutic monoclonal antibody (mab) discovery. Therapeutic antibodies to this class of proteins are generally targeted to extracellular domains displayed on the surfaces of living cells. Challenges associated with developing antibodies to this class of targets are small numbers of extracellular amino acids, membrane-dependent protein conformation, difficulty in expression at high levels, high amino acid sequence homology of human and mouse proteins, and post-translational modifications. DNA immunization strategies with full-length constructs and high throughput flow cytometry screening of mab binding to transfected and control cells was used to generate and identify large numbers of mabs to CXCR4 and ADORA2A (GPCRs) and CD20. Panels of mabs were generated for all 3 targets with low numbers of hybridoma fusions. For each target the mab gene sequences were shown to be unique and contain levels of somatic hypermutation comparable to existing benchmark therapeutic antibodies. Functional assays including apoptosis and receptor modulation (calcium flux and cAMP modulation) further demonstrated that the technical approach generated diverse panels of antibodies that exhibit functional activity as good or better than existing benchmark therapeutic antibodies.