Immunoelectron Microscopic Analysis of Human Cytomegalovirus ( HCMV ) Antigens Reacting with Specific Monoclonal Antibodies.
- Author:
Eung Soo HWANG
;
Chang Yong CHA
;
Hae Kyung PARK
;
Ju Young SEOH
- Publication Type:Original Article
- MeSH:
Antibodies, Monoclonal*;
Capsid;
Cytomegalovirus*;
Fibroblasts;
Glutaral;
Glycoproteins;
Goats;
Humans*;
Immunoglobulin G;
Lung;
Polyethylene Glycols;
Polymers;
Virion
- From:Journal of the Korean Society for Microbiology
1997;32(5):601-610
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
For the purpose of morphologic analysis of human cytomegalovirus (HCMV) antigens reacting with specific monoclonal antibodies, we observed HCMV particles after immunogold staining. HCMV was cultured in human fetal lung fibroblasts to be concentrated by polyethylene glycol 6,000. The HCMV stock was dropped onto Formva-coated grids and was fixed by 2% glutaraldehyde. The grids were reacted with MCMVA57, 93, 135 or with SCMVM1, 6, 14, 49 monoclonal antibodies (MoAbs) follwed by gold (10 nm)-conjugated goat anti-mouse IgG. Then the grids were stained with 2.5% uranyl acetate to be observed under Hitachi 500 or Jeol 1,200 electron microscope. When HCMV was reacted with SCMVM14 and SCMVM49 MoAbs, gold particles were adsorbed to virion envelopes, suggesting that the reactive antigens were envelope proteins. In cases of MCMVA135 and SCMVM6 MoAbs, gold particles were adsorbed to dense bodies as well as to virion envelope. These results, together with the previous results of immunologic and genetic characterization, suggested that the reactive antigens of MCMVA135 and SCMVM6 MoAbs were gB homologue and structural protein, respectively. In case of SCMVM1 MoAb, gold particles were adsorbed to capsids, envelopes, and dense bodies, suggesting that the reactive antigen was structural protein. In case of SCMVM8 MoAb, gold particles were observed between the envelopes and capsids, which space was supposed to be the tegument, suggesting that the reactive antigen was carbohydrate moiety of glycoprotein or its polymer. In cases of MCMVA57 and MCMVA93 MoAbs, gold particles were adsorbed to only dense bodies, suggesting that the reactive antigens were precursors of structural proteins.
