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Journal of Chinese Integrative Medicine ›› 2010, Vol. 8 ›› Issue (11): 1036-1040.doi: 10.3736/jcim20101106

• Original Expreimental Research • Previous Articles     Next Articles

Application and evaluation of a pseudotyped virus assay for screening herbs for anti-H5Nl avian influenza virus

Hong Liu a,Guo-liang Zhanga,Li Shenb,Zhen Zengb,Bao-luo Zhou c,Cheng-hai Liub,Guang Niea   

  1. Department of Integrated Traditional Chinese and Western Medicine, Shenzhen Third Peopled Hospital, Shenzhen 518020, Guangdong Province, China
    b Institute of Liver Diseases, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China
    c Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200025, China
  • Received:2010-03-12 Accepted:2010-05-20 Online:2010-11-20 Published:2010-11-15
  • Contact: Guang Nie E-mail:fqngl008@163.com

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.

Conclusion

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.

Key words: Pseudotyped virus, H5N1 avian influenza, Drugs,Chinese herbal

Figure 1

Effects of different concentrations of herb-medicated sera on RLA of H5N1 pseudotyped virus Data were represented as $\overline{x}$±s, n=3."

Figure 2

Protective effects of 20% herb-medicated sera on RLA of H5N1 pseudotyped virusData were represented as $\overline{x}$±s, n=3."

Figure 3

Effects of 4 different herb-medicated sera on HA expression of H5N1 pseudotyped virus A: Blank control group; B: Positive control group; C: GXR group; D: JDR group; E: LXR group; F: FZR group; G: Negative control group. The upper right corner values represent the proportion of positive cells."

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