title: [Application and evaluation of a pseudotyped virus assay for screening herbs for anti-H5N1 avian influenza virus].
abstract: OBJECTIVE
  To establish a method based on a pseudotyped virus assay for screening drugs for anti-H5N1 avian influenza virus, and then to evaluate anti-virus activity of traditional Chinese herbal compounds with the function of purgation, detoxification, cooling the blood and reinforcing the healthy qi based on seropharmacology.
  
  
  METHODS
  Pseudotyped viruses expressing H5N1 virus hemagglutinin were constructed. Herb-medicated sera were prepared by feeding Wistar rats traditional Chinese herbal compounds. Then inhibitory effects of different drug-medicated sera against pseudotyped virus were evaluated based on incubating serum with pseudotyped viruses or target cells such as Madin-Darby canine kidney cells and human embryonic kidney cell line 293T cells, and the expression of hemagglutinin (HA) protein was detected by flow cytometry.
  
  
  RESULTS
  Pseudotyped virus could be neutralized by specific 2B4 monoclonal antibody at doses of 50 μL (500 μg), 25 μL (250 μg) and 12.5 μL (125 μg), which showed that it was a feasible assay to screen drugs. No significant difference was observed in inhibition against H5N1 pseudotyped virus between herb-medicated sera and control serum (P>0.05), meanwhile, the herb-medicated sera also could not inhibit HA expression.
  
  
  CONCLUSIONS
  Pseudotyped virus assay is a safe, effective, and exact method for screening anti-H5N1 avian influenza virus herbs, and herb-medicated sera prepared by administration of herbs with the function of purgation, detoxification, cooling the blood and reinforcing the healthy qi do not contain active ingredient or monomer which can inhibit HA and sialic acid-α 2,3-galatose.
authors.affiliation: Department of Integrated Traditional Chinese and Western Medicine, Shenzhen Third People's Hospital, Shenzhen 518020, Guangdong Province, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Liu
authors.forename: Guo-liang
authors.initials: GL
authors.lastname: Zhang
authors.forename: Li
authors.initials: L
authors.lastname: Shen
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zeng
authors.forename: Bao-luo
authors.initials: BL
authors.lastname: Zhou
authors.forename: Cheng-hai
authors.initials: CH
authors.lastname: Liu
authors.forename: Guang
authors.initials: G
authors.lastname: Nie
mesh_terms: D000818:Animals; D000998:Antiviral Agents / Q000494:pharmacology*; D002460:Cell Line; D004285:Dogs; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D004365:Drugs, Chinese Herbal / Q000494:pharmacology*; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000378:metabolism; D006801:Humans; D053124:Influenza A Virus, H5N1 Subtype / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000998:Antiviral Agents; D004365:Drugs, Chinese Herbal; D019267:Hemagglutinin Glycoproteins, Influenza Virus