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The Journal of Immunology, 1976, 117: 1440-1446.
Copyright © 1976 by The American Association of Immunologists, Inc.
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Activation of the Complement Attack Mechanism in the Fluid Phase and its Control by C567-INH: Lysis of Normal Erythrocytes Initiated by Zymosan, Endotoxin, and Immune Complexes1,2,

Thomas F. Lint3, Carole L. Behrends, Patricia J. Baker4 and Henry Gewurz5

From the Departments of Immunology, Rush Medical College, and Microbiology, University of Illinois at the Medical Center, Chicago, Illinois 60612

Abstract

Addition of zymosan-serum complexes to guinea pig erythrocytes in guinea pig complement-EDTA was found to result in substantial lysis of the bystander cells in the presence of polycations such as poly-L-lysine of 178,000 daltons. Involvement of the alternative C pathway was shown, and the optimum time, temperature, and eruthrocyte and polycation concentrations were defined; a surprising efficiency was observed at low temperatures and high cell concentrations. Several lines of evidence indicated that this hemolysis was mediated via the
Figure 1
complex of the C system and modulated by serum inhibitors of
Figure 2
(
Figure 3
-INH): lysis was observed only with zymosan-serum complexes possessing C-consuming activity; it was not observed in C5-depleted guinea pig serum but was restored upon addition of purified C5; the addition of partially purified
Figure 4
-INH substantially depressed hemolysis; and poly-L-lysine which is known to neutralize
Figure 5
-INH in solution resulted in substantial enhancement of hemolysis. We also sought to determine whether the addition of complement activators directly to erythrocyte-serum mixtures could result in the hemolysis of bystander erythrocytes. It was found that zymosan, endotoxin, antigen-antibody complexes, and aggregated human {gamma}-globulin each could initiate such bystander lysis under appropriate conditions. Lysis again was favored by increased erythrocyte concentrations, low temperatures, and the presence of polycations such as poly-L-lysine, and was found to be mediated via the C system.
Figure 6
-INH blocked cytolysis whereas poly-L-lysine potentiated hemolysis by neutralization of
Figure 7
-INH. These experiments emphasize the propensity for
Figure 8
formation and lysis of bystander erythrocytes during C activation generally, the role of
Figure 9
-INH in the control of this lysis, and the susceptibility of these interactions to modulation by highly charged macromolecules.

Footnotes

1 This work was presented in part to the American Society for Experimental Pathology, April 16, 1975 (1).

2 This work was supported by grants from the Leukemia Research Foundation, Inc., and the Hunter Trust.

3 Dr. Lint was supported by National Institutes of Health Postdoctoral Fellowship 1 F32 A105188-01.

4 Presented in partial fulfillment of the requirements for the Doctor of Philosophy Degree in the Graduate College, University of Illinois at the Medical Center, Chicago, Illinois.

5 H. G. holds the Thomas J. Coogan, Sr., Chair in Immunology established by Marjorie Lindheimer Everett.






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