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Respiratory research2006; 7(1); 43; doi: 10.1186/1465-9921-7-43

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

Abstract: 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')2 fragments by pepsin digestion of IgGs. Methods: The horses were vaccinated with inactivated H5N1 vaccine to prepare anti-H5N1 IgGs. The F(ab')2 fragments were purified from anti-H5N1 hyperimmune sera by a protocol for 'enhanced pepsin digestion'. The protective effect of the F(ab')2 fragments 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')2 fragments were finally obtained from one liter equine antisera with the purity of over 90%. The H5N1-specific F(ab')2 fragments had a HI titer of 1:1024, and the neutralization titre of F(ab')2 reached 1: 2048. The in vivo assay showed that 100 microg of the F(ab')2 fragments could protect BALB/c mice infected with a lethal dose of influenza H5N1 virus. Conclusions: The availability of highly purified H5N1-specific F(ab')2 fragments 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.
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Publication Date: 2006-03-23 PubMed ID: 16553963PubMed Central: PMC1459145DOI: 10.1186/1465-9921-7-43Google Scholar: Lookup
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  • Comparative Study
  • Journal Article
  • Research Support
  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

This research article investigates the potential therapeutic effect of equine antibodies against the highly pathogenic H5N1 avian influenza virus, demonstrating that the purified antibody fragments could protect mice from a lethal dose of the virus.

Methodology

  • The researchers initiated the study by preparing specific anti-H5N1 Immunoglobulins G (IgGs) from horses that had been vaccinated with an inactivated H5N1 virus. These IgGs are essentially proteins used by the immune system to fight off foreign substances like the H5N1 virus.
  • They further obtained F(ab’)2 fragments of these IgGs by treating them with pepsin, an enzyme that breaks down proteins. F(ab’)2 fragments are parts of the antibody that contain the antigen-binding sites.
  • The therapeutic effect of the produced F(ab’)2 fragments was then evaluated in test tube experiments using MDCK (Madin-Darby Canine Kidney) cells, and in a BALB/c mouse model. MDCK cells are often used in virology studies, while BALB/c is a type of lab mice.

Results

  • Through the ‘enhanced pepsin digestion’ protocol they used, the researchers successfully extracted 16 grams of F(ab’)2 fragments from one liter of equine sera, with a purity of over 90%.
  • These H5N1-specific F(ab’)2 fragments demonstrated strong antivirus activity, as indicated by a Hemagglutination Inhibition (HI) titer of 1:1024. The HI test measures the ability of antibodies to prevent red blood cells from clumping, which is a direct impact of a virus.
  • The neutralization titre, the highest dilution of antiserum that can neutralize the virus, of F(ab’)2 reached 1: 2048, indicating a very high potential for neutralizing the virus.
  • An in vivo assay showed that 100 micrograms of the F(ab’)2 fragments could protect the BALB/c mice from a lethal dose of the H5N1 Influenza virus.

Conclusions

  • The research showed promising results in using highly purified H5N1-specific F(ab’)2 fragments for the treatment of H5N1 infection.
  • Additionally, this study has provided experimental support for the potential therapeutic use of these equine-derived immunoglobulins in larger primates or humans in future clinical trials.

Cite This Article

APA
Lu J, Guo Z, Pan X, Wang G, Zhang D, Li Y, Tan B, Ouyang L, Yu X. (2006). Passive immunotherapy for influenza A H5N1 virus infection with equine hyperimmune globulin F(ab’)2 in mice. Respir Res, 7(1), 43. https://doi.org/10.1186/1465-9921-7-43

Publication

Respiratory research
ISSN: 1465-993X
NlmUniqueID: 101090633
Country: England
Language: English
Volume: 7
Issue: 1
Pages: 43

Researcher Affiliations

Lu, Jiahai
  • Sun Yat-sen University, Guangzhou 510080, China. jiahailu@yahoo.com.cn
Guo, Zhongmin
    Pan, Xinghua
      Wang, Guoling
        Zhang, Dingmei
          Li, Yanbin
            Tan, Bingyan
              Ouyang, Liping
                Yu, Xinbing

                  MeSH Terms

                  • Animals
                  • Cell Line
                  • Chick Embryo
                  • Dogs
                  • Female
                  • Horses
                  • Immunization, Passive / methods
                  • Immunoglobulin Fab Fragments / immunology
                  • Immunoglobulin Fab Fragments / therapeutic use
                  • Influenza A Virus, H5N1 Subtype / immunology
                  • Influenza Vaccines / immunology
                  • Influenza Vaccines / therapeutic use
                  • Mice
                  • Mice, Inbred BALB C
                  • Orthomyxoviridae Infections / immunology
                  • Orthomyxoviridae Infections / prevention & control

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