Medical Microbiology and Immunology

, Volume 173, Issue 1, pp 9–17 | Cite as

Hepatitis A-virus in cell culture

V. Neutralizing antibodies against hepatitis A-virus
  • J. Zahn
  • A. Vallbracht
  • B. Flehmig
Article

Abstract

A test system for the detection of neutralizing antibodies against hepatitis A-virus (anti-HAV-Nt) is presented. The anti-HAV-Nt assay is performed with Frhk-4/R cells and the hepatitis A virus (HAV) strain GBM/Frhk-4/R which has been adapted to these cells. Non-neutralized HAV is demonstrated 14 days after infection of Frhk-4/R cells using a radio-immunoassay for detecting newly grown HAV.

The influence of differing amounts of HAV on the anti-HAV-Nt titre and the effect of variations in incubation time of virus-serum mixtures are described. The time course of anti-HAV-Nt is shown in sera from a hepatitis-A patient which were taken at different stages of the disease. Anti-HAV-Nt is compared with anti-HAV and anti-HAV-IgM. It is shown that anti-HAV-Nt correlates closely with anti-HAV and separated anti-HAV-IgG, but only slightly with anti-HAV-IgM.

The test system presented makes possible the demonstration of neutralizing antibodies against HAV in gamma-globulin preparations and during vaccination studies.

Keywords

Cell Culture Hepatitis Incubation Time Vaccination Study 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Bradford M (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedGoogle Scholar
  2. 2.
    Bradley DW, Maynard JE, Hindman SH, Hornbeck CL, Fields HA, McCaustland KA, Cook EH jr. (1977) Serodiagnosis of viral hepatitis A: detection of acute-phase immunoglobulin M anti-hepatitis A virus by radioimmunoassay. J Clin Microbil 5:521–530Google Scholar
  3. 3.
    Casey HL (1970) Experience at the National Communicable Disease Center with micro serological techniques for the diagnosis of viral diseases. Health Lab Sci 76:233–242Google Scholar
  4. 4.
    Flehmig B (1978) Laboratoriumsdiagnose der Hepatitis A-Infektion. Bundesgesundheitsblatt 17:277–287Google Scholar
  5. 5.
    Flehmig B (1980) Hepatitis A-virus in cell culture. I. Propagation of different hepatitis A-virus isolates in a fetal rhesus monkey kidney cell line (Frhk-4). Med Microbiol Immunol 168:239–248PubMedGoogle Scholar
  6. 6.
    Flehmig B (1981) Hepatitis A-virus in cell culture. II. Growth characteristics of hepatitis A-virus in Frhk-4/R cells. Med Immunol 170:73–81Google Scholar
  7. 7.
    Flehmig B, Zahn J (1984) Neuere Ergebnisse in der Hepatitis A-Virus Infektion und Möglichkeiten der Prophylaxe. Ergebnisse aus dem Paul-Ehrlich-Institut, in pressGoogle Scholar
  8. 8.
    Flehmig B, Ranke M, Frank H, Gerth HJ (1978) Application of a solid-phase radioimmunoassay and immune electron microscopy for hepatitis a in diagnosis and research. Med Microbiol Immunol 166:187–194PubMedGoogle Scholar
  9. 9.
    Flehmig B, Ranke M, Berthold H, Gerth HJ (1979) A solid phase radioimmunoassay for detection of IgM antibodies against hepatitis A-virus. J Infect Dis 140:169–175PubMedGoogle Scholar
  10. 10.
    Hollinger FB, Bradley DW, Maynard JE, Dreesman GR, Melnick JL (1975) Detection of hepatitis A viral antigen by radioimmunoassay. J Immunol 115:1464–1466PubMedGoogle Scholar
  11. 11.
    Kärber G (1931) Beitrag zur kollektiven Behandlung pharmakologischer Reihenversuche. Arch Pathol Pharmakol 162:480–483Google Scholar
  12. 12.
    Krugman S, Friedman H, Lattimer C (1975) Viral hepatitis type A. Identification by specific complement fixation and immune adherence tests. N Engl J Med 292:1141–1143PubMedGoogle Scholar
  13. 13.
    Lehmann-Grube F (1978) An improved method for determining neutralizing antibody against lymphocytic choriomeningitis virus in human sera. J Gen Virol 41:377–383PubMedGoogle Scholar
  14. 14.
    Locarini SA, Ferris AA, Lehmann NI, Gust JD (1977) The antibody response following hepatitis A-infection. Intervirology 8:309–318PubMedGoogle Scholar
  15. 15.
    Locarini SA, Coulepis AG, Kaldor J, Gust JD (1980) Copro-antibodies in hepatitis A: detection by enzyme-linked immunosorbent assay and immune electron microscopy. J Clin Microbiol 11:710–716PubMedGoogle Scholar
  16. 16.
    Lodetti E, De Simone F, Nordelli L (1972) Neutralization tests for foot and mouth disease, equine rhino-, porcine entero-Angeszky-, and rhinopneumonitis viruses. Zentralbl Veterinärmed B 19:848–857Google Scholar
  17. 17.
    Loefgren B, Nordenfeldt E, Jonsson S, Kronvall G (1980) New bacterial absorption method for determination of hepatitis A IgM and IgA antibodies. J Med Virol 6:37–44PubMedGoogle Scholar
  18. 18.
    Mathiesen LR, Feinstone SM, Wong DC, Skinhoej P, Purcell RH (1978) Enzyme-linked immunosorbent assay for detection of hepatitis A antigen in stool and antibody to hepatitis A antigen in sera: comparison with solid phase radio-immunoassay, immune electron microscopy and immune adherence hemagglutination assay. J Clin Microbiol 7:184–193PubMedGoogle Scholar
  19. 19.
    Miller WJ, Provost PJ, McAlleer WJ, Ittensohn OL, Villarejos VM, Hilleman MR (1975) Specific immune adherence assay for human hepatitis A antibody. Application to diagnostic and epidemiologic investigations (38783) Proc Soc Exp Biol Med 149:254–261PubMedGoogle Scholar
  20. 20.
    Provost PJ, Ittensohn OL, Villarejos VM, Hilleman MR (1975) A specific complement — fixation test for human hepatitis A employing CR 326 virus antigen. Diagnosis and epidemiology (38669). Proc Soc Exp Biol Med 148:962–969PubMedGoogle Scholar
  21. 21.
    Purcell RH, Wong DC, Moritsugu Y, Dienstag JL, Routenberg JA, Boggs JD (1976) A microtiter solid-phase radioimmunoassay for hepatitis A antigen and antibody. J Immunol 116:349–356PubMedGoogle Scholar
  22. 22.
    Schmidt NJ, Lenette EH, Hanahoe MF (1966) Microneutralization test for the reoviruses. Application to detection and assay of antibodies in sera of laboratory animals. Proc Soc Exp Biol Med 121:1268–1276PubMedGoogle Scholar
  23. 23.
    Vesikari T, Vaheri A (1968) Rubella: a method for rapid diagnosis of a recent infection by demonstration of the IgM antibodies. Br Med J I:221–223Google Scholar

Copyright information

© Springer-Verlag 1984

Authors and Affiliations

  • J. Zahn
    • 1
  • A. Vallbracht
    • 1
  • B. Flehmig
    • 1
  1. 1.Abteilung für Medizinische Virologie und Epidemiologie der ViruskrankheitenHygiene Institut der Universität TübingenTübingenGermany

Personalised recommendations