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Home / Equine Research Bank / Respir Res / Passive immunotherapy for influenza A H5N1 virus infection w...
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

                  References

                  This article includes 39 references

                  Citations

                  This article has been cited 34 times.
                  1. Barker D, Han X, Wang E, Dagley A, Anderson DM, Jha A, Weaver SC, Julander J, Nykiforuk C, Kodihalli S. Equine Polyclonal Antibodies Prevent Acute Chikungunya Virus Infection in Mice.. Viruses 2023 Jun 29;15(7).
                    doi: 10.3390/v15071479pubmed: 37515166google scholar: lookup
                  2. Tharmalingam T, Han X, Wozniak A, Saward L. Polyclonal hyper immunoglobulin: A proven treatment and prophylaxis platform for passive immunization to address existing and emerging diseases.. Hum Vaccin Immunother 2022 Apr 29;18(2):1886560.
                    doi: 10.1080/21645515.2021.1886560pubmed: 34010089google scholar: lookup
                  3. Pan X, Wu Y, Wang W, Zhang L, Xiao G. Development of horse neutralizing immunoglobulin and immunoglobulin fragments against Junín virus.. Antiviral Res 2020 Feb;174:104666.
                    doi: 10.1016/j.antiviral.2019.104666pubmed: 31760108google scholar: lookup
                  4. Beigel JH, Aga E, Elie-Turenne MC, Cho J, Tebas P, Clark CL, Metcalf JP, Ozment C, Raviprakash K, Beeler J, Holley HP Jr, Warner S, Chorley C, Lane HC, Hughes MD, Davey RT Jr. Anti-influenza immune plasma for the treatment of patients with severe influenza A: a randomised, double-blind, phase 3 trial.. Lancet Respir Med 2019 Nov;7(11):941-950.
                    doi: 10.1016/S2213-2600(19)30199-7pubmed: 31582360google scholar: lookup
                  5. Kotey E, Lukosaityte D, Quaye O, Ampofo W, Awandare G, Iqbal M. Current and Novel Approaches in Influenza Management.. Vaccines (Basel) 2019 Jun 18;7(2).
                    doi: 10.3390/vaccines7020053pubmed: 31216759google scholar: lookup
                  6. Pan W, Xie H, Li X, Guan W, Chen P, Zhang B, Zhang M, Dong J, Wang Q, Li Z, Li S, Yang Z, Li C, Zhong N, Huang J, Chen L. Patient-derived avian influenza A (H5N6) virus is highly pathogenic in mice but can be effectively treated by anti-influenza polyclonal antibodies.. Emerg Microbes Infect 2018 Jun 13;7(1):107.
                    doi: 10.1038/s41426-018-0113-2pubmed: 29899428google scholar: lookup
                  7. Nasiri H, Valedkarimi Z, Aghebati-Maleki L, Abdolalizadeh J, Kazemi T, Esparvarinha M, Majidi J. Production and purification of polyclonal antibody against F(ab')2 fragment of human immunoglobulin G.. Vet Res Forum 2017;8(4):307-312.
                    pubmed: 29326789
                  8. Hui DS, Lee N, Chan PK, Beigel JH. The role of adjuvant immunomodulatory agents for treatment of severe influenza.. Antiviral Res 2018 Feb;150:202-216.
                    doi: 10.1016/j.antiviral.2018.01.002pubmed: 29325970google scholar: lookup
                  9. Zhao Y, Wang C, Qiu B, Li C, Wang H, Jin H, Gai W, Zheng X, Wang T, Sun W, Yan F, Gao Y, Wang Q, Yan J, Chen L, Perlman S, Zhong N, Zhao J, Yang S, Xia X. Passive immunotherapy for Middle East Respiratory Syndrome coronavirus infection with equine immunoglobulin or immunoglobulin fragments in a mouse model.. Antiviral Res 2017 Jan;137:125-130.
                    doi: 10.1016/j.antiviral.2016.11.016pubmed: 27890674google scholar: lookup
                  10. Terryn S, Francart A, Lamoral S, Hultberg A, Rommelaere H, Wittelsberger A, Callewaert F, Stohr T, Meerschaert K, Ottevaere I, Stortelers C, Vanlandschoot P, Kalai M, Van Gucht S. Protective effect of different anti-rabies virus VHH constructs against rabies disease in mice.. PLoS One 2014;9(10):e109367.
                    doi: 10.1371/journal.pone.0109367pubmed: 25347556google scholar: lookup
                  11. Hui DS, Lee N, Chan PK. Adjunctive therapies and immunomodulatory agents in the management of severe influenza.. Antiviral Res 2013 Jun;98(3):410-6.
                    doi: 10.1016/j.antiviral.2013.03.019pubmed: 23578727google scholar: lookup
                  12. Hu H, Voss J, Zhang G, Buchy P, Zuo T, Wang L, Wang F, Zhou F, Wang G, Tsai C, Calder L, Gamblin SJ, Zhang L, Deubel V, Zhou B, Skehel JJ, Zhou P. A human antibody recognizing a conserved epitope of H5 hemagglutinin broadly neutralizes highly pathogenic avian influenza H5N1 viruses.. J Virol 2012 Mar;86(6):2978-89.
                    doi: 10.1128/JVI.06665-11pubmed: 22238297google scholar: lookup
                  13. Howard MK, Sabarth N, Savidis-Dacho H, Portsmouth D, Kistner O, Kreil TR, Ehrlich HJ, Barrett PN. H5N1 whole-virus vaccine induces neutralizing antibodies in humans which are protective in a mouse passive transfer model.. PLoS One 2011;6(8):e23791.
                    doi: 10.1371/journal.pone.0023791pubmed: 21876771google scholar: lookup
                  14. Howard WA, Peiris M, Hayden FG. Report of the 'mechanisms of lung injury and immunomodulator interventions in influenza' workshop, 21 March 2010, Ventura, California, USA.. Influenza Other Respir Viruses 2011 Nov;5(6):453-4, e458-75.
                    doi: 10.1111/j.1750-2659.2011.00278.xpubmed: 21848616google scholar: lookup
                  15. Beigel JH. ANTIVIRAL COMPOUNDS IN THE PIPELINE TO TACKLE H1N1 INFLUENZA INFECTION.. Drugs Future 2010 May;35(5):385-392.
                    doi: 10.1358/dof.2010.035.05.1487081pubmed: 21738286google scholar: lookup
                  16. Wallach MG, Webby RJ, Islam F, Walkden-Brown S, Emmoth E, Feinstein R, Gronvik KO. Cross-protection of chicken immunoglobulin Y antibodies against H5N1 and H1N1 viruses passively administered in mice.. Clin Vaccine Immunol 2011 Jul;18(7):1083-90.
                    doi: 10.1128/CVI.05075-11pubmed: 21613458google scholar: lookup
                  17. Ng WC, Wong V, Muller B, Rawlin G, Brown LE. Prevention and treatment of influenza with hyperimmune bovine colostrum antibody.. PLoS One 2010 Oct 26;5(10):e13622.
                    doi: 10.1371/journal.pone.0013622pubmed: 21049034google scholar: lookup
                  18. Smith SM, Gums JG. Antivirals for influenza: strategies for use in pediatrics.. Paediatr Drugs 2010 Oct 1;12(5):285-99.
                    doi: 10.2165/11532530-000000000-00000pubmed: 20799758google scholar: lookup
                  19. Oh HL, Akerström S, Shen S, Bereczky S, Karlberg H, Klingström J, Lal SK, Mirazimi A, Tan YJ. An antibody against a novel and conserved epitope in the hemagglutinin 1 subunit neutralizes numerous H5N1 influenza viruses.. J Virol 2010 Aug;84(16):8275-86.
                    doi: 10.1128/JVI.02593-09pubmed: 20519402google scholar: lookup
                  20. White MR, Boland P, Tecle T, Gantz D, Sorenson G, Tornoe I, Holmskov U, McDonald B, Crouch EC, Hartshorn KL. Enhancement of antiviral activity of collectin trimers through cross-linking and mutagenesis of the carbohydrate recognition domain.. J Innate Immun 2010;2(3):267-79.
                    doi: 10.1159/000272313pubmed: 20375547google scholar: lookup
                  21. Leider JP, Brunker PA, Ness PM. Convalescent transfusion for pandemic influenza: preparing blood banks for a new plasma product?. Transfusion 2010 Jun;50(6):1384-98.
                    doi: 10.1111/j.1537-2995.2010.02590.xpubmed: 20158681google scholar: lookup
                  22. Hui DS, Lee N, Chan PK. Clinical management of pandemic 2009 influenza A(H1N1) infection.. Chest 2010 Apr;137(4):916-25.
                    doi: 10.1378/chest.09-2344pubmed: 20022969google scholar: lookup
                  23. Beigel JH, Voell J, Huang CY, Burbelo PD, Lane HC. Safety and immunogenicity of multiple and higher doses of an inactivated influenza A/H5N1 vaccine.. J Infect Dis 2009 Aug 15;200(4):501-9.
                    doi: 10.1086/599992pubmed: 19569973google scholar: lookup
                  24. Prabakaran M, Prabhu N, He F, Hongliang Q, Ho HT, Qiang J, Meng T, Goutama M, Kwang J. Combination therapy using chimeric monoclonal antibodies protects mice from lethal H5N1 infection and prevents formation of escape mutants.. PLoS One 2009 May 22;4(5):e5672.
                    doi: 10.1371/journal.pone.0005672pubmed: 19478856google scholar: lookup
                  25. Virelizier JL. Rationale for treating human influenza infections by passive transfer of specific antibodies.. Influenza Other Respir Viruses 2007 Mar;1(2):61-4.
                    doi: 10.1111/j.1750-2659.2007.00012.xpubmed: 19432636google scholar: lookup
                  26. Sun L, Lu X, Li C, Wang M, Liu Q, Li Z, Hu X, Li J, Liu F, Li Q, Belser JA, Hancock K, Shu Y, Katz JM, Liang M, Li D. Generation, characterization and epitope mapping of two neutralizing and protective human recombinant antibodies against influenza A H5N1 viruses.. PLoS One 2009;4(5):e5476.
                    doi: 10.1371/journal.pone.0005476pubmed: 19421326google scholar: lookup
                  27. Shahzad MI, Naeem K, Mukhtar M, Khanum A. Passive immunization against highly pathogenic Avian Influenza Virus (AIV) strain H7N3 with antiserum generated from viral polypeptides protect poultry birds from lethal viral infection.. Virol J 2008 Nov 28;5:144.
                    doi: 10.1186/1743-422X-5-144pubmed: 19040734google scholar: lookup
                  28. Lim AP, Chan CE, Wong SK, Chan AH, Ooi EE, Hanson BJ. Neutralizing human monoclonal antibody against H5N1 influenza HA selected from a Fab-phage display library.. Virol J 2008 Oct 28;5:130.
                    doi: 10.1186/1743-422X-5-130pubmed: 18957074google scholar: lookup
                  29. Yu H, Gao Z, Feng Z, Shu Y, Xiang N, Zhou L, Huai Y, Feng L, Peng Z, Li Z, Xu C, Li J, Hu C, Li Q, Xu X, Liu X, Liu Z, Xu L, Chen Y, Luo H, Wei L, Zhang X, Xin J, Guo J, Wang Q, Yuan Z, Zhou L, Zhang K, Zhang W, Yang J, Zhong X, Xia S, Li L, Cheng J, Ma E, He P, Lee SS, Wang Y, Uyeki TM, Yang W. Clinical characteristics of 26 human cases of highly pathogenic avian influenza A (H5N1) virus infection in China.. PLoS One 2008 Aug 21;3(8):e2985.
                    doi: 10.1371/journal.pone.0002985pubmed: 18716658google scholar: lookup
                  30. Lim AP, Wong SK, Chan AH, Chan CE, Ooi EE, Hanson BJ. Epitope characterization of the protective monoclonal antibody VN04-2 shows broadly neutralizing activity against highly pathogenic H5N1.. Virol J 2008 Jul 11;5:80.
                    doi: 10.1186/1743-422X-5-80pubmed: 18616831google scholar: lookup
                  31. Law M, Hangartner L. Antibodies against viruses: passive and active immunization.. Curr Opin Immunol 2008 Aug;20(4):486-92.
                    doi: 10.1016/j.coi.2008.06.005pubmed: 18577455google scholar: lookup
                  32. Beigel J, Bray M. Current and future antiviral therapy of severe seasonal and avian influenza.. Antiviral Res 2008 Apr;78(1):91-102.
                    doi: 10.1016/j.antiviral.2008.01.003pubmed: 18328578google scholar: lookup
                  33. Simmons CP, Bernasconi NL, Suguitan AL, Mills K, Ward JM, Chau NV, Hien TT, Sallusto F, Ha do Q, Farrar J, de Jong MD, Lanzavecchia A, Subbarao K. Prophylactic and therapeutic efficacy of human monoclonal antibodies against H5N1 influenza.. PLoS Med 2007 May;4(5):e178.
                    doi: 10.1371/journal.pmed.0040178pubmed: 17535101google scholar: lookup
                  34. Hanson BJ, Boon AC, Lim AP, Webb A, Ooi EE, Webby RJ. Passive immunoprophylaxis and therapy with humanized monoclonal antibody specific for influenza A H5 hemagglutinin in mice.. Respir Res 2006 Oct 14;7(1):126.
                    doi: 10.1186/1465-9921-7-126pubmed: 17040574google scholar: lookup
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