ovalbumin and Stomach-Neoplasms

Coupling of multiplex isobaric tags for relative and absolute quantitation (iTRAQ) to a sensitive linear ion trap (LTQ) mass spectrometer (MS) is a challenging, but highly promising approach for quantitative high-throughput proteomic profiling. Integration of the advantages of pulsed-Q dissociation (PQD) and collision-activated dissociation (CAD) fragmentation methods into a PQD-CAD hybrid mode, together with PQD optimization and data manipulation with a bioinformatics algorithm, resulted in a robust, sensitive and accurate iTRAQ quantitative proteomic workflow. The workflow was superior to the default PQD setting when profiling the proteome of a gastric cancer cell line, SNU5. Taken together, we established an optimized PQD-CAD hybrid workflow in LTQ-MS for iTRAQ quantitative proteomic profiling that may have wide applications in biological and biomedical research.

Topics: Algorithms; Animals; Caseins; Cattle; Cell Line, Tumor; Chickens; Computational Biology; Cytochromes c; Horses; Humans; Mass Spectrometry; Muramidase; Myoglobin; Ovalbumin; Proteomics; Sensitivity and Specificity; Serum Albumin, Bovine; Stomach Neoplasms

Priming of CTLs at mucosal sites, where various tumors are originated, seems critical for controlling tumors. In the present study, the effect of the oral administration of OVA plus adjuvant cholera toxin (CT) on the induction of Ag-specific mucosal CTLs as well as their effect on tumor regression was investigated. Although OVA-specific TCRs expressing lymphocytes requiring in vitro restimulation to gain specific cytotoxicity could be detected by OVA peptide-bearing tetramers in both freshly isolated intraepithelial lymphocytes and spleen cells when OVA was orally administered CT, those showing direct cytotoxic activity without requiring in vitro restimulation were dominantly observed in intraepithelial lymphocytes. The magnitude of such direct cytotoxicity at mucosal sites was drastically enhanced after the second oral administration of OVA with intact whole CT but not with its subcomponent, an A subunit (CTA) or a B subunit (CTB). When OVA plus CT were orally administrated to C57BL/6 mice bearing OVA-expressing syngeneic tumor cells, E.G7-OVA, in either gastric tissue or the dermis, tumor growth was significantly suppressed after the second oral treatment; however, s.c. or i.p. injection of OVA plus CT did not show any remarkable suppression. Those mucosal OVA-specific CTLs having direct cytotoxicity expressed CD8alphabeta but not CD8alphaalpha, suggesting that they originated from thymus-educated cells. Moreover, the infiltration of such OVA-specific CD8(+) CTLs was observed in suppressed tumor tissues. These results indicate that the growth of ongoing tumor cells can be suppressed by activated CD8alphabeta CTLs with tumor-specific cytotoxicity via an orally administered tumor Ag with a suitable mucosal adjuvant.

Topics: Animals; Antigens, Neoplasm; CD8 Antigens; Cell Line, Tumor; Cholera Toxin; Cytotoxicity, Immunologic; Female; Intestinal Mucosa; Lymphocyte Activation; Lymphocyte Subsets; Mice; Mice, Inbred C57BL; Mouth Mucosa; Ovalbumin; Skin Neoplasms; Stomach Neoplasms; T-Lymphocytes, Cytotoxic

A combined method has been developed for selective cytolysis in vitro as well as in vivo using a photosensitizer haematoporphyrin-protein conjugate as the targeting molecule and low-power He-Ne laser (632.8 nm) irradiation in order to activate the sensitizer to its excited, toxic triplet energy state. The specificity of the procedure was demonstrated in vitro by purging a mixed cell population from one component, and in vivo in an animal (nude mice) xenograft tumour model, where human cancer cells were destroyed by the immunotargeting method using monoclonal-antibody-haematoporphyrin (mAb-HP) conjugate (a-PNAr-I mAbs, which bind to the cell surface antigens of gastric cancer cells) and soft laser irradiation. The cell destruction was dependent on the doses of both mAb-HP and He-Ne laser light energy, and occurred only in target cell populations.

Topics: Adenocarcinoma; Animals; Hematoporphyrin Photoradiation; Humans; Hybridomas; Immunotoxins; Lasers; Mice; Mice, Inbred BALB C; Mice, Nude; Neoplasm Transplantation; Ovalbumin; Receptors, Mitogen; Stomach Neoplasms

Sera from 111 patients with various gastro-intestinal (GI) diseases were studies for the presence of antibodies to human serum albumin (HSA), bovine serum albumin (BSA) and ovalbumin (OA) by passive haemagglutination assay. The antibody titre to BSA was higher than that to HSA or OA. The anti-BSA antibody was demonstrated in upper GI diseases i.e. esophageal cancer, gastric ulcer, gastric cancer and duodenal ulcer, and not in lower GI disease i.e. Crohn's disease, ulcerative colitis and colon cancer. Both the mean titre and the incidence of the anti-BSA antibody tended to be higher in women than in men, and the titre was in a positive correlation with serum gamma-globulin levels. Sephadex G-200 column chromatography revealed that the anti-BSA antibody was widely distributed between void volume and 7S fraction.

Topics: Adolescent; Adult; Antibodies; Antibodies, Neoplasm; Child; Colonic Neoplasms; Esophageal Neoplasms; Female; Gastric Juice; Gastrointestinal Diseases; Humans; Inflammation; Intestinal Diseases; Male; Middle Aged; Ovalbumin; Peptic Ulcer; Serum Albumin; Serum Albumin, Bovine; Stomach Neoplasms