bluuebunny/crossref_metadata_2025_split huggingface dataset:
The DH/PH domain of BCR-ABL mediates INKT cell immunosubversion in chronic myeloid leukemia by downregulating cell-surface CD1D expression in myeloid dendritic cells (TUM10P.1038)
Authors: Herbelin Andre , Basbous Sara , Levescot Anais , piccirilli nathalie , GOMBERT JEAN MARC 
Published: January 2023

Abstract
Chronic myeloid leukemia (CML) is associated with a rearrangement between the Abelson (ABL) and the BCR genes, generating a chimeric fusion gene. The constitutive tyrosine-kinase (TK) activity linked to ABL is sufficient to induce malignant transformation while BCR contains a DH/PH domain ensuring activation of the Rho/ROCK pathway that is implicated in the control of mobility of leukemia cells. iNKT cells constitute a distinct population of T cells that recognize glycolipids as α-GC presented by CD1d, and are believed to play a key role in cancer immunosurveillance. CML in chronic phase (CP) is associated with functional deficiencies of iNKT cells. Here, we hypothesized that the anergic status of iNKT cells during CML-CP results from BCR-ABL-dependent dysfunctions in APC. Accordingly, CML mDC downregulated their cell surface expression of CD1d, as compared to mDC from healthy individuals. Since Rho/ROCK pathway is a negative regulator of CD1d expression, we surmised that its enhancement by the DH/PH domain of BCR-ABL is responsible for the impaired cell-surface CD1d expression in CML mDC. In favor of this hypothesis, we showed that treatment of PBMC by Rho/ROCK inhibitors restores in vitro cell-surface expression levels of CD1d on CML-CP mDC as well as expansion of CM-CP iNKT cells in response to α-GC. Altogether, these results indicate that the DH/PH domain of BCR-ABL mediates iNKT-cell immunosubversion in CML patients by downregulating cell-surface CD1d expression in mDC.
Lymphocyte depletion after neoadjuvant chemoradiotherapy for operable gastric cancer. (TUM10P.1039)
Authors: Wydmanski Jerzy , Chwieduk Agata , Kotyla Przemyslaw , Kaletka Agnieszka , Tukiendorf Andrzej 
Published: January 2023

Abstract
The purpose of this study was to assess the effect of preoperative chemoradiotherapy (CRT) for operable gastric cancer on lymphocyte subpopulations counts. A total of 35 patients with adenocarcinoma of the stomach were investigated. The preoperative CRT regime consisted of radiotherapy and chemotherapy. The total dose of 45 Gy was administered in 25 fractions, with 5 fractions per week for 5 weeks. Analysis of lymphocyte subpopulations in peripheral blood was carried out before and during treatment (5 analyses for each patients) using a BD Multitest IMK Kit (Becton Dickinson). The data were analyzed with ANOVA. Before and after CRT, the mean values (MVs) of the subtype counts (SD) were as follow: lymphocytes, 1,788 cells/µl (±694) and 239 cells/µl (±131); CD3+CD4+ cells, 738 cells/µl (353) and 135 cells/µl (55); CD3+CD8+ cells, 562 cells/µl (±398) and 56 cells/µl (±41); CD3-CD19+ cells, 150 cells/µl (±112) and 2 cells/µl (±2); and CD16+CD56+ cells, 329 cells/µl (±155) and 28 cells/µl (±36). Every week, we have observed the gradual depletion of all subset of lymphocytes and those differences were statistically significant (p&lt;0.0000). Before CRT, the MV of the CD4+/CD8+ ratio was 1.58 (SD±0.97 ). This ratio has gradually increased during CRT. At the end of treatment, the MV of the CD4+/CD8+ ratio was 2,96 (SD±2,04), p&lt;0.0000. Preoperative chemoradiotherapy induces depletion of all lymphocyte subpopulations.
Development of high cytotoxic NK cell model for leukemia treatment (TUM10P.1041)
Authors: Twu Yuh-Ching 
Published: January 2023

Abstract
Natural killer (NK) cells play an important role in innate immune system to against virus-infected cells, transformed cells and tumor cells without further stimulation. The killing functions of NK cells regulated by the signals from a large set of activating receptors and inhibitor receptors. Moreover, either the directly interaction with other immune cells, including dendritic cells, macrophages and T cells, or the cytokines secreted from other cells also enhance the NK cell killing activity. The NK cell produces cytokines, such as IFN-γ and TNF-α, recruit immune cells to enhance the inflammatory response as well. Acute myeloid leukemia is characterized by the accumulation of immature cells in bone marrow and affects the hematopoiesis. By standard chemo- and radio-therapy, leukemia could be controlled on the remission status. However, the leukemia stem cells which are resistant to conventional treatments, cause the relapse leading to the death of AML patients. So far it is believed that hematopoietic stem cell transplantation is a curable treatment for AML, but it is limited by the patient age and the availability of HLA-matched donor. According to previous study, NK cells seem to have the potential to kill LSCs and lower the disease relapse. In this study, we try to develop a NK cell model with the ability to eliminate both leukemia cells and LSCs. If success, the highly cytotoxic NK model may apply to clinical AML treatment to prevent the relapse of AML.
HLA-G expressing NK cell subset with suppressive properties is expanded in breast cancer (TUM10P.1043)
Authors: Ostapchuk Yekaterina , Aktas Cetin Esin , Perfilyeva Yuliya , Yilmaz Abdullah , Belyaev Nikolai , Deniz Gunnur 
Published: January 2023

Abstract
NK cells were considered traditionally as cytotoxic effector cells rapidly killing infected and transformed cells. Recently, NK cells have been shown to act as potential regulatory cells interacting with various components of the immune system. HLA-G is an important immune-regulatory molecule.There is evidence that HLA-G expression contributes to escape of malignant cells from immune recognition and destruction by down-regulation of the immune response. Immunosuppressive T cells expressing HLA-G have been recently reported. In healthy donors, we have found a subset of circulating NK cells that constitutively express HLA-G (4.5±1.3, n=11), which occurs more frequently among CD57- than CD57+ NK cells. They are enhanced in response to PHA stimulation and enriched with IL-10- and TGF-β-producing NK cells. HLA-G+ NK do not show cytotoxic activity and strongly decrease both cytotoxicity and expression of perforin, granzyme B and CD107a of HLA-G- NK cells, but do not affect IFN-γ production. HLA-G neutralization antagonized the suppressive effects, leading to a 88% restoration of cytotoxic capacity of HLA-G- NK cells. In breast cancer patients, an increased number of circulating HLA-G+NK cells compared to healthy women (6.8±1.8, n=21; 4.0±1.1, n=6; p=0.001) has been detected. Obtained data suggest that increased number of suppressive HLA-G+ NK cells may be included into cohort of immunosuppressive cells mediating escape of NK-sensitive tumor cells from cytolysis.
SAHA prevents osteoblast-mediated protection of AML cells via upregulation of Nherf-1 (TUM10P.1050)
Authors: Hedin Karen , Kremer Kimberly , van Wijnen Andre , Westendorf Jennifer 
Published: January 2023

Abstract
Osteoblasts are known to protect acute myeloid leukemia (AML) cells in the bone marrow microenvironment, however, the identity of these protective signals is not known. Because disrupting this protection could help prevent the relapse of AML patients following chemotherapy, we characterized the molecular mechanisms. We previously showed that SDF-1 (CXCL12), a chemokine abundant in the bone marrow, induces apoptosis of AML cells in cocultures unless the leukemic cells are protected via proximity to differentiating osteoblasts. Treating differentiating osteoblasts with either suberoylanilide hydroxamic acid (SAHA) or LBH-589 histone deacetylase (HDAC) inhibitor drugs inhibited their protection of AML cells. RNAseq revealed that SAHA globally inhibits osteoblast differentiation while inducing expression of the scaffold protein, Nherf-1. Remarkably, expressing Nherf-1 in differentiating osteoblasts was both required and sufficient to block osteoblast-mediated protection of AML cells. Together, these results indicate that disrupting the differentiation of osteoprogenitors in the bone marrow inhibits their ability to protect AML cells from apoptosis, and that this can be achieved via either inhibiting HDACs or overexpressing Nherf-1 in osteoblast-lineage cells. HDAC inhibitors or treatments specifically designed to induce Nherf-1 expression either during or immediately prior to chemotherapy may therefore be a useful strategy for reducing the risk of disease relapse in AML patients.
Dissecting early mechanisms of melanoma cell resistance to cytotoxic T lymphocyte attack (TUM10P.1025)
Authors: khazen roxana , Puissegur Marie-pierre , Muller Sabin , Valitutti Salvatore 
Published: January 2023

Abstract
The effector function of naturally occurring Cytotoxic T cells (CTL) or of CTL induced by immunetherapy protocols is often insufficient to achieve clinical remission of melanoma. Consistent with these, we have recently shown that cytotoxic T cells are equally activated upon encountering melanoma cells as compared to conventional targets; yet the apoptotic process seems to be defective or “delayed” in melanoma cells. We are presently interested to investigate mechanisms of melanoma resistance to CTL attack during cell/cell interaction at the immunological synapse, in aiming to improve the effectiveness of melanoma therapies. Our results show that melanoma cell resistance is not mainly related to a deficient intrinsic apoptotic process. Conversely, it seems that melanoma cells have evolved a resistance mechanism which causes a reduced entry and activity of granzyme B, which we showed to be related to a deficiency of perforin binding and polymerization on melanoma cells surface. In addition, melanoma cells exhibited a constitutive lysosome exposure, which increased upon specific contact with CTL, resulting in an intense exocytic activity towards the lytic synapse that contributes to their escape mechanism. Moreover, silencing several key molecules of secretory pathway in melanoma cell increased their susceptibility to cytotoxic attack. Together, our results reveal a new pathway of melanoma cell resistance that might become a potential target of new therapeutic strategies.
Synergistic COX2 induction by IFNγ and TNFα self-limits type-1 immunity in cancer tissues (TUM10P.1027)
Authors: Obermajer Natasa , Wong Jeffrey , Odunsi Kunle , Edwards Robert , Kalinski Pawel 
Published: January 2023

Abstract
Maintenance of CTL-, Th1-, and NK cell-mediated type-1 immunity is essential for effective anti-tumor responses. Unexpectedly, we observed that IFNγ and TNFα, the critical soluble mediators produced by type-1 immune effector cells, synergize in the induction of cyclooxygenase 2 (COX2), the key enzyme in prostaglandin (PG)E2 synthesis, and the hyper-activation of myeloid-derived suppressor cells (MDSCs) within the tumor microenvironment (TME) of ovarian cancer patients. This MDSC hyper-activation by type-1 immune cells and the subsequent over-expression of indoleamine 2,3-dioxygenase (IDO), inducible nitric oxide synthase (iNOS/NOS2), IL-10, and additional COX2 result in strong feedback suppression of type-1 immune responses. Importantly, this self-limiting feedback could be eliminated by COX2 blockade, allowing amplification of type-1 immunity and prolonged survival of tumor bearing mice. Our data demonstrate a new mechanism underlying the self-limiting nature of type-1 immunity in the human TME and provide rationale for targeting the COX2-PGE2 axis to enhance the effectiveness of cancer immunotherapies.
Generation and characterization of neutralizing human recombinant antibodies against antigenic site II of rabies virus glycoprotein (VAC11P.1106)
Authors: Sun Lina , Wei Jingshuang , Qu Jing , Li Dexin , Liang Mifang 
Published: January 2023

Abstract
Currently recommended treatment for individuals exposed to rabies virus (RV) is post-exposure prophylaxis (PEP) through the combined administration of rabies vaccine and rabies immune globulin (RIG). Human monoclonal antibodies (mAbs) that neutralize RV offer an opportunity to replace RIG for rabies PEP. Here a combinatorial human Fab library was constructed using antibody genes derived from the blood of RV-vaccinated donors. Selections of this library against purified RV virions resulted in the identification of 11 unique Fab antibodies specific for RV glycoprotein. Of the Fab antibodies, five were converted to full human IgG1 format. The human IgG antibodies revealed high binding affinity and neutralizing activities against RV fixed strains through a rapid fluorescent focus inhibition test (RFFIT) in vitro as well as the early stage protective function after exposure to RV infection in vivo. Furthermore, epitope mapping and binding competition analysis showed that all of obtained human neutralizing and protective antibodies were directed to the antigenic site Ⅱ of RV glycoprotein. Our results provide not only important insight into the protective immune response to RV in humans, but also more candidates eligible for use in a mAb cocktail aimed at replacing RIG for rabies post-exposure prophylaxis.
Bacterial lysates activates B cell differentiation and IL-10 production via Toll-Like Receptor-2 recognition (VAC11P.1064)
Authors: Jimenez-Martinez Maria , Salazar Alberto , Galicia-Carreon Jorge , Castillo Axel , Andaluz Maria , Nieto Jane 
Published: January 2023

Abstract
Bacterial lysates (BL) are used to improve immune response in patients with respiratory infections. BL are constituted by a mixture of bacterial antigens derived from different bacterial species; species included in these lysates are 14 different bacteria: S. pneumoniae, K. pneumoniae, B. catarrahalis, S. aureus, H. influenza, S. alpha, and beta, S. faecalis, S. epidermidis, B. pertussis, Proteus sp., Pseudomonas sp., E. coli, and C. pseudodiphteriae (BL-14) It has been suggested that BL activate innate response through TLR, however if BL-14 are able to induce B cell activation is not known and this was the aim of our study. Peripheral mononuclear blood cells (PBMC) were obtained from healthy donors and cultured with/without BL-14 (IPI ASAC Pharma) at different concentrations, after stimulation with BL-14, supernatant were collected and IL-2, IL-4, IL-6, IL-10, TNF, IFN-γ, and IL-17A (BD Biosciences), were measured with cytometric bead arrays. PBMC were harvested at 24, 48 and 72 h, labelled against anti- CD19, CD38, CD69 or TLR2 (BD Biosciences), and analysed by flow cytometry. We observed dose-dependent increased activation of CD19+. After 72 h we observed a diminished frequency of B cells precursors and expansion of transitional B cells, these cells were TLR2+ cells. A significant increased concentration of IL-10 and IL-6 was observed at 24h, and 48 h after BL-14 stimulation. BL induces expansion of transitional B cells and IL-10 production through TLR2 recognition
Different alpha fetoprotein epitope-specific CD8 T cells possess distinct antitumor function (VAC13P.1130)
Authors: He Yukai , Wu Sha , Peng Yibing , Wang Lan , Hong Yuan , Celis Esteban , Wu Juan 
Published: January 2023

Abstract
Using lentivector (lv) immunization and epitope-optimization, we have recently identified three CD8 epitopes in the hepatocellular carcinoma (HCC) associated antigen of murine alpha fetoprotein (AFP) that could effectively activate CD8 T cells in mice, resulting in potent antitumor effect that can completely prevent mice from tumor challenge and prevent carcinogen-induce autochthonous HCC. In this report, we studied whether the epitope-specific CD8 responses can be further enhanced by peptide boost and investigate if the newly identified epitope specific T cells can kill AFP+ tumor cells and have equal antitumor activity. We found that the lv-primed AFP-specific CD8 responses can be boosted by AFP peptides even at the peak of primary responses. As much as 20 folds increase of AFP-specific CD8 responses can be achieved by peptide boost. However, the kinetics and the antitumor effect of different epitope-specific CD8 responses are drastically different. While the epitope AFP499-specific CD8 T cells possess potent CTL activity against AFP+ tumor cells and generate strong antitumor effect in vivo, the AFP212-specific CD8 T cells have a very limited CTL activity and antitumor effect though they can produce high level of IFNg. The mechanisms underpinning the functional differences will be reported. This finding demonstrates that cancer vaccines must be able to elicit tumor-killing immune effector cells in order to generate antitumor effect.
Neuronal nitric oxide synthase: a possible inflammatory mediator responsible for host-induced susceptibility to herpes simplex encephalitis (VIR9P.1153)
Authors: Ferraioli Adrianna , Lundberg Patric 
Published: January 2023

Abstract
Herpes Simplex Virus Type-1 (HSV-1) infection can result in a severe disease of the central nervous system termed Herpes Simplex Encephalitis (HSE). In both animals and humans, the immune response is known to play a role in HSE progression. Using resistant and susceptible HSE mouse models, we have characterized late-stage (&gt;day 7 post-infection (p.i.)) inflammation by which susceptible mice succumb to HSE. Previous data have indicated that immune dysregulation between the two strains is evident at day 6 p.i. Thus, we propose that activation of specific pathways before day 6 p.i. result in an inflammatory cascade that contributes to HSE. To demonstrate this, resistant C57BL/6 and susceptible 129S6 male mice were infected with HSV-1 and brainstems were collected days 1-4 p.i. Microarrays, Ingenuity Pathway Analysis (IPA), and QRT-PCR were performed on RNA, and protein levels were determined via westerns. IPA predicated several inflammatory functions, such as migration of immune cells, to be 129S6-biased (p&lt;0.05). Among the dysregulated transcripts identified by the microarray, neuronal nitric oxide synthase (nNOS) was shown to be significantly upregulated in susceptible 129S6 mice on day 3 p.i at both the RNA (p&lt;0.01) and protein level (p&lt;0.001). Our results implicate nNOS as a regulator of HSE pathogenesis and we are currently performing animal studies to better define the role of nNOS in HSE progression.
The application of clustered regularly interspaced short palindromic repeats genome engineering to the study of host antiviral factors (VIR9P.1160)
Authors: Omokehinde Tolu , Soll Steven , Bieniasz Paul 
Published: January 2023

Abstract
Human immunodeficiency virus (HIV), is a retrovirus that infects cells of the human immune system, including CD4­+ T cells, macrophages and dendritic cells. Previously, two of the candidate genes identified showed a capacity to protect cells from HIV-1 infection were Tripartite motif containing 56 (TRIM56) and 2’, 3’-cyclic-nucleotide 3’-phosophodiesterase (CNP). Clustered regularly interspaced short palindromic repeats (CRISPR) and the CRISPR-associated (CAS) protein 9 is a nuclease system involved in defense against invading phages and plasmids in bacteria. This system can be programmed to induce DNA double stranded breaks (DSBs), which when targeting coding regions of genes can create frame shift insertion/deletion (indel) mutations that result in gene knock-out cell lines. Using the CRISPR/Cas9 system, we were able to clone guide sequences targeting TRIM56 and CNP into a LentiCRISPR and LentiGuide plasmid. The cloned plasmids were then used to generate viruses that would deliver the necessary machinery to target and cleave TRIM56 and CNP genes. To test the efficiency of the CRISPR/Cas9 system, guide sequence targeting GFP were cloned. GFP deletion was observed in a 293T-GFP stable cell line and knockout efficiency was determined by analysis. Knockout results of TRIM56 and CNP are still pending, but we expect that TRIM56 and CNP deleted cell lines will demonstrate the contribution of each of these innate immune factors to the interferon response to HIV-1.
HIV infection in humanized mice: role of type I interferon on disease progression (VIR9P.1139)
Authors: Abraham Sojan , Choi Jang Gi , Ye Chunting , Schreiber Gideon , Shankar Premlata , Swamy Manjunath 
Published: January 2023

Abstract
HIV and SIV disease progression is associated with chronic expression of type I IFN and IFN-stimulated genes. A recent study showed that a high-dose injection of IFN-1 early during primary SIV infection in the rhesus macaque model was protective, while neutralization of endogenous IFN-1 was deleterious. In contrast, in the same animal model, prolonged IFN-1 administration accelerated disease development in the chronic stage of the infection. To better understand the role of IFN-1 during HIV infection, we used humanized-PBL mouse model for this study. The advantage of this model is that there is no endogenous interferon production upon HIV infection, allowing analysis of IFN-1 therapy. Since continuous presence of IFN-1 is detrimental in the SIV model, we hypothesized that intermittent treatment with IFN-1 may be protective against HIV infection. To test this hypothesis, we carried out intermittent pegasys (peg-interferon alpha) treatment after HIV infection in human PBL infused NOD-scid IL2rγcnull (hu-PBL) mice. This treatment with pegasys diminished HIV replication and delayed CD4 depletion compared to control mice. To further test the role of IFN, we are currently using humanized BLT mouse model of HIV infection. HIV infection in BLT mice leads to continuous production of high levels of type-I IFNs; and thus, we are studying the effects of continuous or intermittent IFN blockade using type I IFN antagonist. The results will be discussed.
Mechanism of HIV-1 capsid-nucleocapsid polyprotein in genome recognition (VIR9P.1143)
Authors: McCowin Sayo , Keane Sarah , Summers Michael 
Published: January 2023

Abstract
Human Immunodeficiency Virus type-1 (HIV-1) is a retrovirus that is responsible for a widespread epidemic affecting nearly 36 million individuals worldwide. During the late phase of the viral life cycle, the Gag polyprotein specifically binds two copies of the unspliced viral genomic RNA. Gag consists of three structured domains: matrix (MA), capsid (CA), and nucleocapsid (NC). We have shown that mature NC binds to both the monomeric and dimeric 5’-untranslated region (5’-UTR) RNAs, but fails to explain the observation that the dimeric genome is preferentially packaged. Additionally, mature NC does not exist in the cytoplasm, where genome recognition occurs. Thus, this has led to the belief that other domains of Gag may also be involved in the selection of dimeric genomes. We hypothesize that the CA domain contributes to diploid RNA genome selection. In order to test this hypothesis, the polyprotein Capsid-Nucleocapsid (CA-NC) will be purified and titrated into 5’-UTR viral RNA. Previous work has shown that this CA-NC polyprotein is unstable and aggregates in the presence of RNA due to hexameric interactions mediated at the C-terminal domain (CTD). We will utilize the strategy of Gamble et al., by introducing two critical mutations, W184A and M185A, in the CTD domain of CA. These mutations have been shown to block CTD dimerization resulting in the monomeric form of the CA protein, while preserving necessary characteristics of the wild-type CA.