Several epitopes on native human complement C9 are involved in interaction with the C5b-8 complex and other C9 molecules.
Ten monoclonal antibodies (mAb) against native human C9 exhibiting various inhibitory effects on the hemolytic activity of C9 (Bausback, J., Kontermann, R. and Rauterberg, E. W., Immunobiology 1988. 178: 58) were further analyzed regarding their reactivities with monomeric C9 (mC9), polymerized C9 (pC9), and the non-lytic SC5b-9 complex in enzyme-linked immunosorbent assay and with the membrane attack complex (MAC) generated on rabbit erythrocytes analyzed by flow cytometry. In addition, the inhibitory effects of mAb on zinc-induced C9 polymerization were investigated. One epitope of the C-terminal half of C9b exposed on the surface of pC9 and the MAC seems not to participate directly in lytic function or polymerization since no inhibitory effect of the respective mAb was observed. The nine other mAb directed against epitopes of the C9a part exhibit various inhibitory potentials. The mAb inhibit either hemolysis or polymerization, or both processes. Due to the reactivity with the tested antigens the mAb can be divided into two groups. mAb of the first group bind with nearly the same affinity to all four antigens, whereas mAb of the second group react preferentially with mC9 while their affinity to pC9, SC5b-9 and the MAC is reduced. Comparison of reaction patterns and inhibitory effects strongly suggest that different epitopes on the surface of native C9 are involved in interaction of C9 with C5b-8 and/or in C9-C9 interaction. The finding that mAb inhibiting polymerization of C9 in vitro have no inhibitory effect on hemolysis confirms that C9 polymers are no prerequisite for lysis.
Institut für Immunologie, Universität Heidelberg, FRG.
R
R
Kontermann
R
R
Deppisch
E W
EW
Rauterberg
D000911:Antibodies, Monoclonal / Q000276:immunology; D003186:Complement C9 / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D015938:Complement Membrane Attack Complex / Q000276:immunology / Q000378:metabolism; D003165:Complement System Proteins / Q000276:immunology / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes; D006801:Humans; D011485:Protein Binding; D015032:Zinc / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D003186:Complement C9; D015938:Complement Membrane Attack Complex; D000939:Epitopes; C042295:complement C5b-8 complex; D003165:Complement System Proteins; D015032:Zinc