title: In vitro studies on immunotoxic potential of Orange II in splenocytes.
abstract: Orange II, an azo dye, is not permitted in food preparations, but high levels of the dye have been detected in different food commodities. Though there are reports on the toxicity of Orange II but knowledge based on the immunomodulatory properties of Orange II is scanty. The present investigation was undertaken to study the in vitro immunotoxic potential of Orange II in splenocytes. Splenocytes were isolated, cultured and subjected to immunophenotypic analysis, mixed lymphocyte reaction (MLR) assay or stimulated with lipopolysaccharide (LPS) or concanavalin A (Con A) for 72 h. The supernatant was collected for cytokine assays. Orange II showed cytotoxic effects at 100-1000μg/ml concentrations and 50μg/ml was determined as the highest non-cytotoxic dose. Orange II at the non-cytotoxic dose (50μg/ml) significantly altered the relative distribution of T and B-cells, MLR response and the mitogen induced proliferative response of T-cells and B-cells. Consistent with the hypo-responsiveness of the T and B-lymphocytes, Orange II induced a concomitant decline in the secretion of cytokines IL-2, IL-4, IL-6, IFN-γ, TNF-α and IL-17. On the contrary, there was an increase in the production of IL-10, an anti-inflammatory regulatory cytokine, which may be one of the causative factor for immunosuppressive property of Orange II. These results suggest that non-cytotoxic dose of Orange II may have immunomodulatory effects.
authors.affiliation: Food, Drugs and Chemical Toxicology Group, CSIR-Indian Institute of Toxicology Research (IITR), Council of Scientific and Industrial Research, P.O. Box No. 80, Mahatma Gandhi Marg, Lucknow 226 001, India.
authors.forename: Ashish
authors.initials: A
authors.lastname: Yadav
authors.forename: Arvind
authors.initials: A
authors.lastname: Kumar
authors.forename: Premendra Dhar
authors.initials: PD
authors.lastname: Dwivedi
authors.forename: Anurag
authors.initials: A
authors.lastname: Tripathi
authors.forename: Mukul
authors.initials: M
authors.lastname: Das
mesh_terms: D056704:Adaptive Immunity / Q000187:drug effects; D000818:Animals; D001391:Azo Compounds / Q000276:immunology / Q000633:toxicity*; D001557:Benzenesulfonates / Q000276:immunology / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D004396:Coloring Agents / Q000633:toxicity; D016207:Cytokines / Q000032:analysis; D005260:Female; D005434:Flow Cytometry; D007113:Immunity, Innate / Q000187:drug effects; D016130:Immunophenotyping / Q000379:methods; D007959:Lymphocyte Culture Test, Mixed; D051379:Mice; D008807:Mice, Inbred BALB C; D013154:Spleen / Q000166:cytology / Q000187:drug effects* / Q000276:immunology*; D013601:T-Lymphocytes / Q000166:cytology / Q000187:drug effects* / Q000276:immunology*
chemical_list: D001391:Azo Compounds; D001557:Benzenesulfonates; D004396:Coloring Agents; D016207:Cytokines; C014722:2-naphthol orange