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Production of human monoclonal antibodies neutralizing rabies virus: An alternative to rabies immune globulin of human blood donation
113 IMMUNIZATION WITH EXTERNAL AND/OR INTERNAL STRUCTURAL PROTEINS PROTECTS MICE FROM INFECTION WITH THE PARANYXOVIRUS SV5 R E RANDALL AND D F YOUNG. Department of Biochemistry & Microbiology, University of St. Andrew, Fife, U.K.
Honoclonal antibodies attached to solid matrices were used to purify external and internal The resulting solid matrixstructural proteins of SV5 from infected tissue culture cells. antibody-antigen (SMAA) complexes were used to immunize mice and evidence will be presented that Mice immunized such complexes induce both vigorous humoral and cell mediated immune response. with the nucleo- or matrix proteins showed the greatest degree of protection from subsequent There was no correlation between the degree of protection and the level of challenge with SV5. Immunization with multiple virus neutralizing antibody in the sera of infected animals. proteins was achieved by attaching monoclonal antibodies with different specificities to the same solid matrix.
114 PRODUCTION OF HUMAN MONOCLONAL ANTIBODIES NEUTRALIZING RABIES VIRUS: AN ALTERNATIVE TO RABIES IMMUNE GLOBULIN OF HUMAN BLOOD DONATION. M. LAFON (I), L. EDELMAN (21, E. MONCHATRE (21, M. LAFAGE (1) and P. SUREAU (1) Unit6 Rage, Institut Pasteur, 75724 Paris Cddex 15. (2) Unit6 d'kmnunohdmatologie et d'Irmnunopathologie, Institut Pasteur.
(1).
After exposure to rabies, immune globulin of human origin are of proven effectiveness as an addition to vaccine treatment. The immune globuline are supposed to neutralize the virus by targeting the surface antigen: the glycoprotein. Until now, the rabies insrune globulins are collected from blood of vaccinated donors. Human monoclonal antibodies prepared in vitro with specific neutralising activity to the rabies virus could become equally effective alternative to the lack human blood donation products. Human hybridomas secreting anti-rabies glycoprotein immunoglobulin, were established by EBV infection and fusion of human peripheral blood lymphocytes with an heteromyeloma (SPM4-0). Peripheral blood lymphocytes were collected from a pre-exposure immunized donor with a neutralizing titer serum of 63 IU per ml. Specificity of hybridoma supernatants was tested by enzyme immunoassay on rabies virus infected cells which expressed glycoprotein on their surface as rabies antigen. IgG subclasses and IgG concentrations were determined by modified Elisa procedure. Neutralization activity of hybridoma supernatants against different rabies serotypes and isolates, was tested by a standard in vitro neutralization test using a constant amount of antibody. Three stable hybridoma lines were capable to neutralize rabies virus.
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Honoclonal antibodies attached to solid matrices were used to purify external and internal The resulting solid matrixstructural proteins of SV5 from infected tissue culture cells. antibody-antigen (SMAA) complexes were used to immunize mice and evidence will be presented that Mice immunized such complexes induce both vigorous humoral and cell mediated immune response. with the nucleo- or matrix proteins showed the greatest degree of protection from subsequent There was no correlation between the degree of protection and the level of challenge with SV5. Immunization with multiple virus neutralizing antibody in the sera of infected animals. proteins was achieved by attaching monoclonal antibodies with different specificities to the same solid matrix.
114 PRODUCTION OF HUMAN MONOCLONAL ANTIBODIES NEUTRALIZING RABIES VIRUS: AN ALTERNATIVE TO RABIES IMMUNE GLOBULIN OF HUMAN BLOOD DONATION. M. LAFON (I), L. EDELMAN (21, E. MONCHATRE (21, M. LAFAGE (1) and P. SUREAU (1) Unit6 Rage, Institut Pasteur, 75724 Paris Cddex 15. (2) Unit6 d'kmnunohdmatologie et d'Irmnunopathologie, Institut Pasteur.
(1).
After exposure to rabies, immune globulin of human origin are of proven effectiveness as an addition to vaccine treatment. The immune globuline are supposed to neutralize the virus by targeting the surface antigen: the glycoprotein. Until now, the rabies insrune globulins are collected from blood of vaccinated donors. Human monoclonal antibodies prepared in vitro with specific neutralising activity to the rabies virus could become equally effective alternative to the lack human blood donation products. Human hybridomas secreting anti-rabies glycoprotein immunoglobulin, were established by EBV infection and fusion of human peripheral blood lymphocytes with an heteromyeloma (SPM4-0). Peripheral blood lymphocytes were collected from a pre-exposure immunized donor with a neutralizing titer serum of 63 IU per ml. Specificity of hybridoma supernatants was tested by enzyme immunoassay on rabies virus infected cells which expressed glycoprotein on their surface as rabies antigen. IgG subclasses and IgG concentrations were determined by modified Elisa procedure. Neutralization activity of hybridoma supernatants against different rabies serotypes and isolates, was tested by a standard in vitro neutralization test using a constant amount of antibody. Three stable hybridoma lines were capable to neutralize rabies virus.
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