Construction of fusogenic vesicles bearing specific antibodies. Targeting of reconstituted Sendai virus envelopes towards neuraminidase-treated human erythrocytes.
The cross-linking reagents succinimidyl-4-(p-maleimidophenyl)-butyrate and N-succinimidyl-3-(2-pyridyldithio)-propionate were used to covalently attach antibodies against human erythrocytes to the thiol-containing paraffin, dodecanethiol. The complex formed, dodecanethiol-maleimidophenylbutyrate (or pyridyldithiopropionate)-antibody was inserted into the membranes of reconstituted Sendai virus envelopes. This was achieved by addition of the dodecanethiol-maleimidophenylbutyrate-antibody to a detergent solution (Triton X-100) containing the viral envelope phospholipids and glycoproteins. Removal of the detergent led to the formation of vesicles containing the viral glycoprotein and the dodecanethiol-maleimidophenylbutyrate (or pyridyldithiopropionate)-antibody complexes within the same membrane. Reconstituted Sendai virus envelope-bearing antibodies against human erythrocytes were able to fuse with human erythrocytes (as was reflected by reconstituted Sendai virus envelope-induced hemolysis) from which the natural virus receptors were removed by treatment with neuraminidase. Thus, it appears that anti-human erythrocyte antibodies could substitute for the viral binding protein (hemagglutinin/neuraminidase glycoprotein) in mediating functional binding of the virus particles to the cell plasma membranes. Furthermore, from the results of the present work, it may be inferred that in addition to being the viral-binding protein, hemagglutinin/neuraminidase glycoprotein actively participates in the process of virus-cell fusion.
D000906:Antibodies / Q000276:immunology*; D002472:Cell Transformation, Viral; D003432:Cross-Linking Reagents / Q000378:metabolism; D004229:Dithiothreitol / Q000494:pharmacology; D004912:Erythrocytes / Q000276:immunology*; D006389:Hemagglutinins, Viral / Q000502:physiology; D006801:Humans; D008854:Microscopy, Electron; D009439:Neuraminidase / Q000378:metabolism*; D010222:Parainfluenza Virus 1, Human / Q000502:physiology*; D013388:Succinimides / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000276:immunology; D014759:Viral Envelope Proteins / Q000502:physiology*
D000906:Antibodies; D003432:Cross-Linking Reagents; D006389:Hemagglutinins, Viral; D013388:Succinimides; D013438:Sulfhydryl Compounds; D014759:Viral Envelope Proteins; C013976:dodecylmercaptan; C018151:N-succinimidyl 3-(2-pyridyldithio)propionate; C042424:succinimidyl 4-(4-maleimidophenyl)butyrate; D009439:Neuraminidase; D004229:Dithiothreitol