ovalbumin and Liver-Neoplasms

Topics: Aged; Animals; Base Sequence; Carcinoma, Hepatocellular; Cell Nucleus; Cytoplasm; Humans; Kinetics; Liver; Liver Neoplasms; Models, Biological; Neoplasms, Experimental; Ovalbumin; Oxidation-Reduction; Polyribosomes; Protein Biosynthesis; Rats; RNA, Messenger; S-Adenosylhomocysteine; S-Adenosylmethionine

Seleno-ovalbumin (Se-OVA) was a selenium conjugating protein synthesized by the combination of ovalbumin (OVA) and inorganic selenium. In this paper, the structure of Se-OVA was characterized, and the anticancer effect of Se-OVA on hepatocellular carcinoma HepG2 cells was investigated. Through FT-IR, UV, endogenous fluorescence and XRD assays, it was found that the structural characterization of Se-OVA changed after seleno-modification. In addition, the cell assays showed that Se-OVA could induce apoptosis of HepG2 cells by arresting cell cycle in S phase, generating intracellular reactive oxygen species, reducing the mitochondrial transmembrane potential, and triggering the Bax- and Bcl-2-mediated mitochondria apoptosis pathway. These findings revealed that Se-OVA might serve as a novel anticancer drug for cancer adjuvant therapy.

Topics: Antineoplastic Agents; Apoptosis; Carcinoma, Hepatocellular; Cell Cycle; Cell Line, Tumor; Cell Survival; Dose-Response Relationship, Drug; Hep G2 Cells; Humans; Liver Neoplasms; Membrane Potential, Mitochondrial; Mitochondria; Organoselenium Compounds; Ovalbumin; Reactive Oxygen Species; Signal Transduction; Spectroscopy, Fourier Transform Infrared

Immunostimulatory monoclonal antibodies (ISmAb) that unleash antitumor immune responses are showing efficacy in cancer clinical trials. Anti-B7-H1 (PD-L1) monoclonal antibodies (mAb) block a critical inhibitory pathway in T cells, whereas anti-CD137 and OX40 mAbs provide T-cell costimulation. A combination of these ISmAbs (anti-CD137 + anti-OX40 + anti-B7-H1) was tested using a transgenic mouse model of multifocal and rapidly progressing hepatocellular carcinoma, in which c-myc drives transformation and cytosolic ovalbumin (OVA) is expressed in tumor cells as a model antigen.. Flow-cytometry and immunohistochemistry were used to quantify tumor-infiltrating lymphocytes (TIL) elicited by treatment and assess their activation status and cytolytic potential. Tolerance induction and its prevention/reversal by treatment with the combination of ISmAbs were revealed by in vivo killing assays.. The triple combination of ISmAbs extended survival of mice bearing hepatocellular carcinomas in a CD8-dependent fashion and synergized with adoptive T-cell therapy using activated OVA-specific TCR-transgenic OT-1 and OT-2 lymphocytes. Mice undergoing therapy showed clear increases in tumor infiltration by activated and blastic CD8(+) and CD4(+) T lymphocytes containing perforin/granzyme B and expressing the ISmAb-targeted receptors on their surface. The triple combination of ISmAbs did not result in enhanced OVA-specific cytotoxic T lymphocyte (CTL) activity but other antigens expressed by cell lines derived from such hepatocellular carcinomas were recognized by endogenous TILs. Adoptively transferred OVA-specific OT-1 lymphocytes into tumor-bearing mice were rendered tolerant, unless given the triple mAb therapy.. Extension of survival and dense T-cell infiltrates emphasize the translational potential of combinational immunotherapy strategies for hepatocellular carcinoma. Clin Cancer Res; 19(22); 6151-62. ©2013 AACR.

Topics: Adoptive Transfer; Animals; Antibodies, Monoclonal; B7-H1 Antigen; Carcinoma, Hepatocellular; CD4-Positive T-Lymphocytes; CD8-Positive T-Lymphocytes; Cell Line, Tumor; Cytotoxicity, Immunologic; Disease Models, Animal; Immunotherapy; Liver Neoplasms; Lymphocyte Activation; Lymphocyte Count; Lymphocytes, Tumor-Infiltrating; Membrane Glycoproteins; Mice; Mice, Inbred C57BL; Mice, Transgenic; Ovalbumin; OX40 Ligand; Survival; T-Lymphocytes, Cytotoxic; Tumor Necrosis Factor Receptor Superfamily, Member 9; Tumor Necrosis Factors

Many diseases are characterized by the changes of either glycan structure or glycosylation site of glycoproteins. The glycan profiling can provide the overview of glycosylation in despite of the absence of the glycosylation sites, which in turn simplifies the complexity of disease diagnosis. Herein, we describe a simple method to profile the N-linked glycans by MALDI-TOF MS with the enrichment using oxidized ordered mesoporous carbon, taking advantages of the size-exclusive effect of mesopore against proteins as well as the interaction between glycans and carbon. Twenty four N-linked glycans derived from ovalbumin could be efficiently detected with high signal-to-noise (S/N) ratios and sufficient peak intensities. In the analysis of complex serum samples, 32 N-linked glycans could be profiled, and 5 (4 core-fucosylated glycans) of them were distinguished from liver cancer and healthy samples.

Topics: Carbon; gamma-Cyclodextrins; Glycosylation; Humans; Liver Neoplasms; Ovalbumin; Oxidation-Reduction; Polysaccharides; Porosity; Signal-To-Noise Ratio; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

The reason the adaptive immune system fails in advanced liver tumors is largely unclear. To address this question, we have developed a novel murine model that combines c-myc-induced autochthonous tumorigenesis with expression of a cognate antigen, ovalbumin (OVA). When c-myc/OVA transgenic mice were crossed with liver-specific inducer mice, multifocal hepatocellular carcinomas co-expressing OVA developed in a tetracycline-dependent manner with a short latency and 100% penetrance. Transferred OVA-specific T cells, although infiltrating the tumor at high numbers, were hyporesponsive, as evidenced by a lack of in vivo cytotoxicity and interferon gamma production. This allowed the tumor to progress even in the presence of large numbers of antigen-specific T cells and even after vaccination (OVA+CpG-DNA). Interestingly, T cell receptor down-modulation was observed, which may explain antigen-specific hyporesponsiveness. This model is helpful in understanding liver cancer-specific mechanisms of T cell tolerance and dissection of antigen-specific and nonspecific mechanisms of immunotherapies in the preclinical phase.

Topics: Adoptive Transfer; Animals; Antigen-Presenting Cells; Crosses, Genetic; Disease Models, Animal; DNA Primers; Flow Cytometry; Genes, myc; Genotype; Humans; Liver Neoplasms; Mice; Mice, Transgenic; Ovalbumin; Receptors, Antigen, T-Cell; Reverse Transcriptase Polymerase Chain Reaction; Survival Analysis

Topics: Animals; Carcinoma, Hepatocellular; Cytotoxicity Tests, Immunologic; Guinea Pigs; Immune Sera; Immunity, Cellular; Liver Neoplasms; Lymphocytes; Lymphotoxin-alpha; Mast-Cell Sarcoma; Mice; Ovalbumin; Rabbits; Spleen

Synthesis of ovalbumin in fragmented oviduct magnum explants of immature, estrogen-stimulated chicks has been studied in the presence of exogenous tRNA. tRAN from Novikoff hepatoma specifically inhibited ovalbumin synthesis, determined by precipitation with antisera. In addition, the major protein(s) synthesized in the presence of hepatoma tRNA had higher electrophoretic mobility than ovalbumin, as shown by sodium dodecyl sulfate polyacrylamide gel electrophoresis. tRNAs from rat liver, rooster liver, and hen oviduct did not affect ovalbumin synthesis, although oviduct tRNA is stimulatory during the earlier stages of estrogen stimulation.

Topics: Animals; Carcinoma, Hepatocellular; Chickens; Dactinomycin; Estradiol; Female; Liver; Liver Neoplasms; Male; Neoplasms, Experimental; Ovalbumin; Oviducts; Rats; Ribonucleases; RNA, Transfer