title: High-throughput screening of chemical libraries for modulators of gene promoter activity of HLA-B2705: environmental pathogenesis and therapeutics of ankylosing spondylitis.
abstract: OBJECTIVE
  Ankylosing spondylitis (AS) is a highly heritable disease with HLA-B27 being the strongest susceptible gene. In order to survey the environmental triggers for arthritis development, we used a high-throughput technique to screen the effects of 12,264 chemicals on the HLA-B27 gene promoter.
  
  
  METHODS
  Promoter reporter transfectants 293T-HLA-B27 and HeLa-HLA-B27 were tested using robotics with 12,264 chemicals. Chemicals that modulated HLA-B27 promoter activity > 150% or < 40% were selected for further evaluation of IC50/EC50 and cell viability.
  
  
  RESULTS
  The primary screening using the 293T-HLA-B27 promoter reporter cell line yielded 5.1% hits that either suppressed (556 chemicals) or enhanced (68 chemicals) the HLA-B27 promoter activity. A secondary reconfirmation screening was carried out with these 624 candidates using HeLa-HLA-B27 promoter reporter cells under several different culture conditions. The yield of positive candidates was 130, of which 47 were derived from natural products. Based on the bio-information of those positive natural products, 21 chemicals were selected for analysis by dose-response IC50/EC50 experiments. Eight compounds showed potential pharmacological activities. Two suppressors are both derived from an herbal medicine (lei gong teng) that has been used for decades to treat immune diseases. The 6 activators all belonged to a group of chemicals known as flavonoids, widely distributed among dietary fruits and vegetables.
  
  
  CONCLUSIONS
  Several common dietary products that contain certain flavonoids might be environmental risk factors for AS; the Chinese traditional herb lei gong teng might be a potential drug for patients who are HLA-B27-positive. These results provide new research directions for the pathogenesis and therapeutics of AS.
authors: Department of Rheumatology, Beijing Hospital, Beijing, China.
authors: Like
authors: L
authors: Zhao
authors: Chin-Hsiu
authors: CH
authors: Liu
authors: David
authors: D
authors: Yu
mesh_terms: D002460:Cell Line; D004305:Dose-Response Relationship, Drug; D004777:Environment; D005419:Flavonoids / Q000009:adverse effects; D020022:Genetic Predisposition to Disease / Q000235:genetics; D015796:HLA-B27 Antigen / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D008403:Mass Screening / Q000379:methods*; D010936:Plant Extracts / Q000494:pharmacology / Q000627:therapeutic use; D011401:Promoter Regions, Genetic / Q000187:drug effects* / Q000235:genetics; D012307:Risk Factors; D054852:Small Molecule Libraries / Q000494:pharmacology*; D013167:Spondylitis, Ankylosing / Q000188:drug therapy* / Q000453:epidemiology / Q000209:etiology*; D013489:Suppression, Genetic / Q000187:drug effects
chemical_list: D005419:Flavonoids; D015796:HLA-B27 Antigen; D010936:Plant Extracts; D054852:Small Molecule Libraries; C041579:lei gong teng