المؤلفون: Donna Beer-Stolz, Herbert J. Zeh, Andrew A. Amoscato, Nicole E. Schapiro, Antonio Romo de Vivar Chavez, Michael T. Lotze, Patricia Loughran, Michael E. de Vera, Stephen H. Thorne, Xiaoyan Liang

المصدر: Journal of Leukocyte Biology. 86:599-607

مصطلحات موضوعية: medicine.medical_specialty, Time Factors, Liver tumor, Cell Survival, Injections, Subcutaneous, Genetic Vectors, Immunology, Cell, Poly (ADP-Ribose) Polymerase-1, Apoptosis, Inflammation, Pharmacology, Biology, Transfection, Mice, Random Allocation, Liver Neoplasms, Experimental, Immune system, Genes, Reporter, Transduction, Genetic, Cell Line, Tumor, Internal medicine, medicine, Animals, Immunology and Allergy, HMGB1 Protein, Pyruvates, B cell, Cell Proliferation, Luciferases, Renilla, Dose-Response Relationship, Drug, Interleukin-6, Lentivirus, Antibodies, Monoclonal, Cancer, Cell Biology, medicine.disease, Tumor Burden, Mice, Inbred C57BL, Endocrinology, medicine.anatomical_structure, Fluorescent Antibody Technique, Direct, Cell culture, Female, Poly(ADP-ribose) Polymerases, medicine.symptom, Colorectal Neoplasms, Microtubule-Associated Proteins

الوصف: The first demonstration of ethyl pyruvate inhibition of liver tumor growth associated with induction of tumor apoptosis, diminished HMGB1 release, and decreased inflammation is reported. EP is a potent inhibitor of HMGB1 release that has significant anti–inflammatory activities and exerts a protective effect in animal models of inflammation. As inflammation is linked to cancer growth, we hypothesized that EP would have anti–tumor activity and explored its effects in a liver tumor model. Mice injected intraportally with MC38 colorectal cancer cells led to the growth of visible hepatic tumors within 2 weeks. Pretreatment with EP 30 min prior to infusion of tumor cells and continuing daily for 9 days inhibited tumor growth significantly in a dose–dependent manner, with 80 mg/kg EP achieving >70% reduction in the number of tumor nodules when compared with untreated animals. Delayed treatment with EP also suppressed tumor growth significantly, although to a lesser extent. Tumors had early, marked leukocytic infiltrates, and EP administration decreased innate (NK cells, monocytes) and adaptive (T and B cell lymphocytic) immune cell infiltrates acutely and significantly in the liver. Serum IL–6 and HMGB1 levels, which were elevated following tumor injection, were decreased significantly in EP–treated animals. Tumors showed an increase in apoptosis in EP–treated mice, and tumor cells treated in vitro with EP had marked increases in LC3–II and cleaved PARP, consistent with enhanced autophagic flux and apoptosis. Thus, EP inhibition of tumor growth in the liver was mediated by tumor (induction of apoptosis) and host (decreased inflammation) effects. EP administration may have a therapeutic role in the treatment of cancer in conjunction with other therapeutic agents.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::0f4894d2740f2ec03537e1140ae4e18bTest
https://doi.org/10.1189/jlb.0908578Test

المؤلفون: Michael T. Lotze, Ramin Lotfi

المصدر: Journal of Leukocyte Biology. 83:456-460

مصطلحات موضوعية: Cell Survival, CpG Oligodeoxynucleotide, Immunology, Cell Culture Techniques, Inflammation, Biology, Flow cytometry, Antigens, CD, medicine, Animals, Humans, Immunology and Allergy, Respiratory Burst, Innate immune system, medicine.diagnostic_test, Immunity, Dendritic Cells, Cell Biology, respiratory system, Eosinophil, Flow Cytometry, medicine.disease, Acquired immune system, Transplant rejection, Eosinophils, medicine.anatomical_structure, Major basic protein, biology.protein, medicine.symptom

الوصف: There are increased eosinophils in tumors, and they are generally associated with a good prognosis, whereas their presence in rejecting allografts is largely seen as a harbinger of poor outcome. The biologic role of eosinophils in their pathogenesis is more poorly understood than in allergy and asthma. Myeloid conventional dendritic cells (DCs) and conversely, plasmacytoid DCs are similarly associated with a good prognosis in cancer patients. We hypothesize that eosinophils, similar to NK cells, could mature DCs, and that could be responsible for regulating immunity in the setting of necrosis-associated chronic inflammation as occurs in cancer and transplant rejection. We have demonstrated that CpG DNA promotes eosinophil-induced DC maturation. As such, a greater linkage than had previously been considered between innate immune cells such as eosinophils and the adaptive immune response can be considered. Granulocytes were isolated from normal human whole blood by density gradient centrifugation followed by ammonium chloride-potassium lysis of the remaining red cells. Eosinophils were negatively separated using magnetic beads. Immature DCs were generated from CD-14 positively separated monocytes, which were cultured for 6 days with GM-CSF and IL-4. CpG ODN 2395 (CpG-C) as a pathogen-associated molecular pattern surrogate was used to induce eosinophil-based DC maturation. Transwells were used to assess cell–cell interaction between eosinophils and DCs. Eosinophil survival was assessed by flow cytometry; cells, which did not stain with Sytox-Orange, were considered viable. In the presence of CpG-C, eosinophils induced DC maturation. Similar results were obtained when eosinophils were pretreated with CpG for 4 h, washed, and cocultured afterwards with DCs. Eosinophil-induced maturation of DCs directly correlated with the eosinophil:DC ratio. Transwell studies showed that the direct cell–cell interaction between eosinophils and DCs enhances maturation. CpGs did not adversely affect eosinophil survival; thus, we could exclude the possibility that DC maturation was caused by sensing eosinophil cell death. While eosinophil-derived neurotoxin did not contribute to the described effect, DCs took up and internalized major basic protein (MBP), which was released from CpG-stimulated eosinophils, revealed by confocal imaging and flow cytometry. Thus, the DC maturational-inducing effect of eosinophils may be a result of released MBP from eosinophils. CpG-activated eosinophils mature conventional DCs. The role of viral or bacterial products or potentially, host-derived DNA as eosinophil activators with consequent DC maturation should be considered in more detail in the inflammatory settings in which eosinophils have been observed.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::281637e824340def30efe7c2824e830bTest
https://doi.org/10.1189/jlb.0607366Test

المؤلفون: Robbie B. Mailliard, Jie Han, Herbert J. Zeh, Donna B. Stolz, Norimasa Ito, Pawel Kalinski, Richard A. DeMarco, Hannah Rabinowich, Michael T. Lotze

المصدر: Journal of Leukocyte Biology. 81:75-83

مصطلحات موضوعية: T-Lymphocytes, medicine.medical_treatment, Immunology, Active Transport, Cell Nucleus, chemical and pharmacologic phenomena, Biology, Granzymes, Amino Acid Chloromethyl Ketones, TNF-Related Apoptosis-Inducing Ligand, Immune system, Cell Line, Tumor, medicine, Humans, Immunology and Allergy, Macrophage, HMGB1 Protein, Killer Cells, Lymphokine-Activated, Melanoma, Cells, Cultured, Cell Nucleus, Tumor microenvironment, Cell Death, Cell Biology, Immunotherapy, Cytotoxicity Tests, Immunologic, Flow Cytometry, Cell biology, Granzyme B, Cytolysis, Granzyme, Cell culture, biology.protein

الوصف: High mobility group box 1 (HMGB1) is one of the recently defined damage-associated molecular pattern molecules, passively released from necrotic cells and secreted by activated macrophage/monocytes. Whether cytolytic cells induce HMGB1 release from tumor cells is not known. We developed a highly sensitive method for detecting intracellular HMGB1 in tumor cells, allowing analysis of the type of cell death and in particular, necrosis. We induced melanoma cell death with cytolytic lymphokine-activated killing (LAK) cells, tumor-specific cytolytic T lymphocytes, TRAIL, or granzyme B delivery and assessed intracellular HMGB1 retention or release to investigate the mechanism of HMGB1 release by cytolytic cells. HMGB1 release from melanoma cells (451Lu, WM9) was detected within 4 h and 24 h following incubation with IL-2-activated PBMC (LAK activity). HLA-A2 and MART1 or gp100-specific cytolytic T lymphocytes induced HMGB1 release from HLA-A2-positive and MART1-positive melanoma cells (FEM X) or T2 cell-loaded, gp100-specific peptides. TRAIL treatment, however, induced HMGB1 release, and it is interesting that this extrinsic pathway-mediated cell death was blocked with the pancaspase inhibitor N-benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone. Conversely, granzyme B delivery did not induce HMGB1 release. HMGB1, along with other intracellular factors released from tumor cells induced by cytolysis, may be important components of the disordered tumor microenvironment. This has important implications for the immunotherapy of patients with cancer. Specifically, HMGB1 may promote healing or immune reactivity, depending on the nature of the local inflammatory response and the presence (or absence) of immune effectors.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::3f82c1ff130194209bdd05a507fb7698Test
https://doi.org/10.1189/jlb.0306169Test

المؤلفون: Michael T. Lotze

المصدر: Journal of Leukocyte Biology. 66:195-200

مصطلحات موضوعية: Clinical immunology, Immunology, Immunology and Allergy, Cell Biology, Biology, Antigen-presenting cell

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::cd54934aec1f54c68dc84399dd2ce8afTest
https://doi.org/10.1002/jlb.66.2.195Test

المؤلفون: Paola Grandi, Nduka Amankulor, Xiaoran Zhang, Aparna Rao, Christopher P Deibert, Jason Kim, Paola Sette, Michael T. Lotze

المصدر: Neuro-Oncology. 17:v123-v123

مصطلحات موضوعية: Cancer Research, medicine.medical_treatment, Biology, medicine.disease, NKG2D, Molecular biology, Isotype, Natural killer cell, medicine.anatomical_structure, Cytokine, Oncology, Glioma, Blocking antibody, Gene expression, medicine, biology.protein, Neurology (clinical), Antibody, Abstracts from the 20th Annual Scientific Meeting of the Society for Neuro-Oncology

الوصف: Mutations in isocitrate dehydrogenase 1 and 2 are common in gliomas. 50-80% of WHO II/III gliomas possess IDH1/IDH2 mutation. Methylation-related transcriptional repression is a feature of IDH mutant (IDHmut) tumors. Epigenetic repression of natural killer (NK) cell ligands is a common mechanism of immune escape in cancer, but it is unknown if this occurs in gliomas. We compared IDHmut and IDHwt tumor expression (z-score) and methylation (mean methylation score) data from the TCGA low-grade glioma database, and found two NKG2D ligands (ULBP1, ULBP3) that were downregulated (-0.47, p = 0.02; -1.28, p < 0.01) and hypermethylated (-0.31, p < 0.01; -0.15, p < 0.01) in IDHmut tumors. We validated this finding by measuring ULBP1 and ULBP3 gene expression in cells derived from IDHwt and IDHmut patients and found both genes were repressed in IDHmut cells (-0.85 fold expression, p = 0.01; -0.57 fold expression, p = 0.01). To determine whether repression of NKG2D ligand expression resulted in escape from NK recognition and specific lysis, we these cells with NK cells. We observed significantly (p = 0.03) reduced specific lysis of IDHmut cell (3.62%) as compared to IDHwt cells (24.13%). To determine NK activation, we measured inflammatory cytokine release in co-cultures, no basal release were detected in either tumor or NK monocultures. IDHmut co-cultures has significantly reduced IFN-g (82.08 pg/mL vs 250.1 pg/mL, p = 0.02) and TNF-a (151.2 pg/mL vs 650.2 pg/mL, p < 0.01) release compared to IDHwt co-cultures. To validate that IDHmut immune escape is NKG2D dependent, we blocked NKG2D binding using an antibody. Addition of isotype antibody did not significantly reduce specific lysis in IDHwt co-culture (10.40% vs 10.20%, p = 0.94). Addition of NKG2D blocking antibody to IDHwt significantly reduced specific lysis below isotype (3.34% vs 10.40%, p < 0.01). In conclusion, we were able to demonstrate that IDH mutant gliomas are able to escape from NK immune surveillance through repression of NKG2D ligand expression.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::d7c8377769beccc5141bc0ca6799b6f4Test
https://doi.org/10.1093/neuonc/nov217.44Test

المؤلفون: Michael Brancho, Aofei Li, Xioran Zhang, Paola Grandi, Hideho Okada, Yigang Chang, Michael T. Lotze, Nduka Amankulor

المصدر: Neuro-Oncology. 16:v107-v107

مصطلحات موضوعية: Cancer Research, Mutation, medicine.medical_treatment, Mutant, Biology, medicine.disease, NKG2D, medicine.disease_cause, Abstracts, Cytolysis, Cytokine, Oncology, Epigenetic Repression, Glioma, Immunology, medicine, Cancer research, Neurology (clinical), Psychological repression

الوصف: Mutations in isocitrate dehydrogenase 1 and 2 (IDH1 and IDH2) are common in gliomas. 50-80% of WHO II/III gliomas possess IDH1/IDH2 mutation. Methylation-related transcriptional repression is a feature of IDH mutant (IDHmut) tumors. Epigenetic repression of natural killer (NK) cell ligands is a common occurrence in cancer, but it is unknown if this occurs in primary brain tumors. Here, we demonstrate transcriptional repression of NKG2D activating ligands (NKG2DLs) in IDHmut gliomas, and correlate this with decreased susceptibility of IDHmut cells to natural killer (NK) cell-mediated cytolysis in vitro. Our data suggest that NK cell-astrocyte contact is required for production of Th1 cytokines IFN-γ and TNF-α, and that production of these cytokines is limited in IDHmut astrocytes. We also show that NKG2DL-specific antibody blockade can result in significant reduction in NK-cell mediated cytolysis in vitro. Furthermore, we infect IDHmut cells with JDNI7 virus containing ULBP3 plasmid DNA to induce overexpression of this ligand. Upon ULBP3 overexpression, these cells are significantly more sensitive to NK-cell mediated lysis. Taken together, our data suggest a potential immune surveillance role for the NKG2DL, ULBP3, during IDHmut gliomagenesis.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::4762e564aa03b3a89bfa5e655582f660Test
https://doi.org/10.1093/neuonc/nou257.3Test

المؤلفون: Anthony A. Rayner, Michael T. Lotze, Elizabeth A. Grimm, Steven A. Rosenberg, Debra J. Wilson

المصدر: JNCI: Journal of the National Cancer Institute.

مصطلحات موضوعية: Cancer Research, education.field_of_study, Cellular immunity, Lymphokine-activated killer cell, Population, Cell, Lymphokine, hemic and immune systems, chemical and pharmacologic phenomena, Biology, Molecular biology, Cytolysis, medicine.anatomical_structure, Oncology, Cell culture, Aldesleukin, Immunology, medicine, education

الوصف: Activated killer cells are generated by the incubation of peripheral blood mononuclear leukocytes (PBL) in the lymphokine interleukin 2 (IL-2). Unseparated populations of these lymphokine-activated killer (LAK) cells lyse a variety of fresh noncultured human tumor targets, but they do not kill normal PBL. This study analyzed the generation and lytic specificity of LAK cell clones. Of 49 (84%) clones isolated by limiting-dilution techniques from a whole population of LAK cells, 41 manifested significant LAK cell activity. LAK cell clones had varied cell surface phenotypes. Clones with high and intermediate LAK cell activity were Leu 2+3-4+7-DR+Tac+ and Leu 2-3+4+7-DR+Tac+, respectively. Single LAK cell clones lysed multiple fresh human tumor targets including autologous sarcoma, 5 allogeneic sarcomas, and a colon cancer in addition to the cultured cell line K562. Autologous PBL were not lysed. Tumor targets were each lysed by multiple LAK cell clones. Sixteen subclones were derived from 5 of these LAK cell clones. These subclones had 99% or greater probability of being derived from a single cell. These subclones also exhibited lysis of multiple tumor targets. These findings suggest the existence of a shared determinant, expressed by multiple human tumors, which is recognized in common by multiple LAK cell clones.

الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_________::c1f69037080ce702d8cfb6fdc2dcafacTest
https://doi.org/10.1093/jnci/75.1.67Test