Passive immunotherapy for influenza A H5N1 virus infection with equine hyperimmune globulin F(ab')2 in mice

doi:10.1186/1465-9921-7-43

Respiratory Research

Volume 7

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Research

Passive immunotherapy for influenza A H5N1 virus infection with equine hyperimmune globulin F(ab')₂in mice

Jiahai Lu¹^* , Zhongmin Guo¹^* , Xinghua Pan²^* , Guoling Wang¹^* , Dingmei Zhang¹^* , Yanbin Li³^* , Bingyan Tan¹ , Liping Ouyang¹ and Xinbing Yu¹

¹Sun Yat-sen University, Guangzhou 510080, China

²Kunming General Hospital of Chengdu Military Area, Kunming, 650000, China

³Haerbin Veterinary research institute, Haerbin, 150000, China

author email corresponding author email* Contributed equally

Respiratory Research 2006, 7:43doi:10.1186/1465-9921-7-43


Published:	23 March 2006

Abstract

Background

Avian influenza virus H5N1 has demonstrated considerable pandemic potential. Currently, no effective vaccines for H5N1 infection are available, so passive immunotherapy may be an alternative strategy. To investigate the possible therapeutic effect of antibody against highly pathogenic H5N1 virus on a mammal host, we prepared specific equine anti-H5N1 IgGs from horses vaccinated with inactivated H5N1 virus, and then obtained the F(ab')₂fragments by pepsin digestion of IgGs.

Methods

The horses were vaccinated with inactivated H5N1 vaccine to prepare anti-H5N1 IgGs. The F(ab')₂fragments were purified from anti-H5N1 hyperimmune sera by a protocol for 'enhanced pepsin digestion'. The protective effect of the F(ab')₂fragments against H5N1 virus infection was determined in cultured MDCK cells by cytopathic effect (CPE) assay and in a BALB/c mouse model by survival rate assay.

Results

By the protocol for 'enhanced pepsin digestion', total 16 g F(ab')₂fragments were finally obtained from one liter equine antisera with the purity of over 90%. The H5N1-specific F(ab')₂fragments had a HI titer of 1:1024, and the neutralization titre of F(ab')₂reached 1: 2048. The in vivo assay showed that 100 μg of the F(ab')₂fragments could protect BALB/c mice infected with a lethal dose of influenza H5N1 virus.

Conclusion

The availability of highly purified H5N1-specific F(ab')₂fragments may be promising for treatment of influenza H5N1 infection. Our work has provided experimental support for the application of the therapeutic equine immunoglobulin in future large primate or human trials.