title: Induction of immunostimulatory cytokine genes expression in human PBMCs by a novel semiquinone glucoside derivative (SQGD) isolated from a Bacillus sp. INM-1.
abstract: In the present study, a semiquinone glucoside derivative (SQGD) isolated from a radioresistant bacterium Bacillus sp. INM-1 was evaluated for its immunostimulatory activities. Human peripheral blood mononuclear cells (PBMCs) were stimulated by different doses (30-90 microg/ml) of SQGD for different time (3-12h) intervals at 37°C, and IL-12p40, IL-23p19, IL-10, RelA and c-Jun gene expression analysis was carried out by qRT-PCR method. SQGD dose dependent cytokines protein expression kinetic analysis was carried out using western blotting. As the results of SQGD (30μg/ml) stimulation for 3h at 37°C, significant induction in IL-12p40, IL-23p19 and RelA gene expression was observed in PBMCs compared to unstimulated control cells. However, no such induction in IL-10 and c-Jun gene expression was observed. Time dependent protein expression study indicated significant increase in IL-12p40, IL-12p35, IL-23p19 and RelA protein expression at 3-6h, which was found decrease at 12h upon SQGD treatment. In contrast, IL-10 protein expression was found to enhance significantly at 12h after SQGD treatment to the PBMCs. SQGD dose dependent study showed approximately similar level of induction in IL-12p40, IL-12p35, IL-23p19 and RelA proteins expression at all tested concentration (30-90 microg/ml) compared to control. However, no significant change in the IL-10 and c-Jun protein expression was observed at any SQGD concentration. SQGD treatment (0.25mg/kgbwt.) was also found to enhance anti-keyhole Limpet Hemocynin (KLH) IgM antibodies significantly in the mice immunized by KLH. Thus, SQGD fraction stimulates cellular immunity by inducing immunostimulatory cytokines and humoral immunity by enhancing IgM antibodies and could be a promising immunostimulant. Further studies related to molecular mechanisms offering immunostimulation is underway, will certainly helpful to unravel its mode of action in the biological system.
authors.affiliation: Institute of Nuclear Medicine and Allied Sciences, Delhi, India. rajkumar790@yahoo.com
authors.forename: Raj
authors.initials: R
authors.lastname: Kumar
authors.forename: Rakesh Kumar
authors.initials: RK
authors.lastname: Sharma
authors.forename: Deen Dayal
authors.initials: DD
authors.lastname: Bansal
authors.forename: Dev Dutt
authors.initials: DD
authors.lastname: Patel
authors.forename: Saurabh
authors.initials: S
authors.lastname: Mishra
authors.forename: Lyuba
authors.initials: L
authors.lastname: Miteva
authors.forename: Zlatka
authors.initials: Z
authors.lastname: Dobreva
authors.forename: Veselina
authors.initials: V
authors.lastname: Gadjeva
authors.forename: Spaska
authors.initials: S
authors.lastname: Stanilova
mesh_terms: D000818:Animals; D001407:Bacillus / Q000737:chemistry / Q000276:immunology*; D016227:Benzoquinones / Q000737:chemistry*; D002478:Cells, Cultured; D016207:Cytokines / Q000235:genetics / Q000276:immunology*; D005786:Gene Expression Regulation; D005960:Glucosides / Q000737:chemistry / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D053765:Interleukin-12 Subunit p35 / Q000235:genetics / Q000276:immunology*; D053762:Interleukin-12 Subunit p40 / Q000235:genetics / Q000276:immunology*; D053760:Interleukin-23 Subunit p19 / Q000235:genetics / Q000276:immunology*; D007700:Kinetics; D007962:Leukocytes / Q000276:immunology*; D008297:Male; D051379:Mice; D051996:Transcription Factor RelA / Q000235:genetics
chemical_list: D016227:Benzoquinones; D016207:Cytokines; D005960:Glucosides; C508840:IL23A protein, human; D053765:Interleukin-12 Subunit p35; D053762:Interleukin-12 Subunit p40; D053760:Interleukin-23 Subunit p19; C498143:RELA protein, human; D051996:Transcription Factor RelA; C004532:quinone