In Vitro Pharmacology

Inflammation


  • Dendritic cell based ex vivo models
    in vitro Model for identification of anti-inflammatories
    Model Chemokine & cytokine profiling of potential anti inflammatory molecules using Dendritic cells
    Test system
    • Murine bone marrow derived dendritic cell cultures
    Methods
    • Generation of Dendritic cells in vitro
    • Treatment of DCs with LPS and test compounds
    • Effect on secreted levels of key pro-inflammatory markers
    End points
    • Quantitation of TNF-a, IL-1-ß, IL-6, IL-8, MIP-1-a, RANTES, CCL-22, IP-10
    X
  • Target Based Screens for arthritis
    in vitro Model for identification of anti-arthritic compounds
    Model in vitro Model for identification of anti-arthritic compounds
    Test system
    • Human synovial- sarcoma cell line (SW982)
    • Human chondro-sarcoma cell line (SW1353)
    Methods
    • Treatment of cells with test compounds
    • Effect on cellular proliferation Effect on cytokines/chemokines activity that play important role in pathogenesis of arthritis) secreted by LPS/TNF-a/IL-1- stimulated cells - -TNF- a, IL-1- , IL-6, MIP-1- a and MMPs.
    End points Anti-inflammatory effect on secretion of cytokines and matrix metalloproteinases
    X
  • Target Based Screens for Gout
    Model for Xanthine Oxidase inhibitory activity - management of Gout
    Model Evaluation of anti-inflammatory activity in respiratory diseases such as asthma
    Test system
    • Human lung epithelial cell line (A549)
    • Bronchial Smooth Muscle Cell line (BSES-2B)
    Methods
    • Treatment of cells with test compounds
    • Effect on the secretion of the inflammatory mediators secreted by TNF-a stimulated cells treated with test compounds : IL-6, IL-8,IL-1ß, MCP-1, TNF-a, RANTES and GMCSF
    End points Quantitation of inflammatory cytokines
    X
  • Target Based Screens for IBD
    in-vitro Model for protective activity against Inflammatory Bowel Disease (IBD)
    Model in-vitro Model for protective activity against Inflammatory Bowel Disease (IBD)
    Test system
    • Human intestinal epithelial cells (Caco-2)
    • Human colonic epithelial cell line (HT-29)
    Methods
    • Identification of non-cytotoxic concentrations
    • Treatment of cells with test compound
    • Stimulation of cell line with appropriate stimulus (TNF-a or IL-1-ß)
    • Effect on pro-inflammatory markers in culture supernatants by ELISA - IL-8, IL-1ß, IP-10 and MIG
    End points Inhibitory effect on secretion of pro-inflammatory cytokines from intestinal and colonic epithelial cells.
    X
  • Immune cells based model
    In Vitro Model for identification of anti-inflammatories
    Model Chemokine & cytokine profiling of potential anti inflammatory molecules using Immune cells
    Test system
    • Murine macrophage cell line (RAW264.7)
    • Murine splenocytes
    • Human monocytic cell line (THP-1)
    Methods
    • Treatment of cells with LPS and test compounds
    • Effect on secreted levels of key pro-inflammatory markers
    End points Quantitation of TNF-a, IL-1-ß, IL-6, IL-8, MIP-1-a, RANTES, CCL-22, IP-10
    X
  • Target based screens for respiratory inflammation
    Model for identification of anti-inflammatories for respiratory inflammation
    Model Evaluation of anti-inflammatory activity in respiratory diseases such as asthma
    Test system
    • Human lung epithelial cell line (A549)
    • Bronchial Smooth Muscle Cell line (BSES-2B)
    Methods
    • Treatment of cells with test compounds
    • Effect on the secretion of the inflammatory mediators secreted by TNF-a stimulated cells treated with test compounds : IL-6, IL-8,IL-1ß, MCP-1, TNF-a, RANTES and GMCSF
    End points Quantitation of inflammatory cytokines
    X
  • Target Based Screens for Ocular inflammation
    in vitro Model for screening ocular inflammation
    Model Evaluation of anti-inflammatory activity in ocular inflammation
    Test system
    • Conjunctival epithelial cell
    End Point
    Effect on the secretion of the inflammatory mediators; secreted by TNF-a stimulated cells treated with test compounds - IL-12p40, GM-CSF, MCP-1, IL-6, IL-7, IL-8, IFN-?, IL-1a, IL-1ß, IL-3, and IP-10 and RANTES
    X
  • Target Based Screens fo Dermal Inflamation
    in vitro Model for screening ocular inflammation
    Model Evaluation of inflammatory/anti-inflammatory activity in dermal cells
    Test system
    • Human skin fibroblast cell line
    End points
    • Morphology of fibroblasts
    • Effect on levels of key pro-inflammatory markers secreted by PMA (phobol-12-myristate-13-acetate)-stimulated skin fibroblasts treated with test compounds–GM-CSF, IL-1-b
    • Effect on Nitric Oxide levels
    • Effect on levels of COX, Prostaglandins and Arachidonic acid
    X
  • Target Based Screens fo Neuronal inflammation
    in vitro Model for screening ocular inflammation
    Model Evaluation of inflammatory/anti-inflammatory activity in dermal cells
    Test system
    • Human skin fibroblast cell line
    End points
    • Morphology of fibroblasts
    • Effect on levels of key pro-inflammatory markers secreted by PMA (phobol-12-myristate-13-acetate)-stimulated skin fibroblasts treated with test compounds–GM-CSF, IL-1-b
    • Effect on Nitric Oxide levels
    • Effect on levels of COX, Prostaglandins and Arachidonic acid
    X
  • Invitro Model for pain
    Evaluation of analgesic/antinociceptive activity
    Model Evaluation of analgesic/antinociceptive activity
    Test system
    • Human neuroblastoma/glioblastoma cell lines expressing µ and dopioid receptors Induction of pain response by VIP/Forskolin
    End Points
    • Effecton following markers secreted by Forskolin/IL-1-ß stimulated cell streated with test compounds:
    • Effect on PGE2 synthesis
    • Effect on COX-2 activity
    • Effect on cAMP levels
    • Effect on calcium signaling pathway
    X
  • Systemic inflammation
    Model Evaluation of inflammatory/anti-inflammatory activity
    Test system
    • Murine Macrophage cell line (RAW264.7)
    • Human Monocyte cell line (THP-1)
    End Points
    Morphology of macrophages/monocytes Effect on levels of key pro-inflammatory markers secreted by LPS-stimulated cells treated with test compounds – TNF-., IL-1-., MIP-1-,, IL-6, RANTES, IL-12, IFN-., CCL-22, IP-10, IL-4, IL-10, TGF-b Effect on Nitric Oxide levels
    Test system Mouse splenocytes
    Stimulatory agent for IL-17secretion
    Concanavalin A (5 µg/ml)
    Experimental Designe
    • Splenocytes were treated with test item at non-cytotoxic concentrations (1:100 to 1:5) along-with the inflammatory stimulus (ConA-5 µg/ml)
    • After 24 h, culture supernatants were collected from each well
    • Secreted levels of IL-17 were estimated by ELISA
    X