Dabur Research Foundation is an Indian Contract Research Organization

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

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

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

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.

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

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

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

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

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

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

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