Abstract
Mayer and Levine(l) described a reaction step in the lysis of sheep erythrocytes by antibody and complement which follows the action of all of the complement components on erythrocytes sensitized with antibody and takes place in the absence of these components from the fluid phase. They called this reaction step the terminal transformation and referred to erythrocytes which have reacted with antibody and all the complement components, but which have not yet lysed, as E∗. They also observed that an increase in the NaCl concentration in the medium from 0.143 m to 0.19 m does not affect the reaction.
In several recent studies(2-4) on the effect of ionic strength (I) on immune hemolysis and on some of the individual reaction steps which comprise this process, the ionic strength of the reaction medium has been varied over a wide range while maintaining constant osmolarity by substituting K2SO4, or uncharged substances such as sucrose, mannitol or inositol for the NaCl usually employed. None of these investigators, however, appears to have studied the possibility that these changes might also affect the terminal transformation. The present communication records an experiment which shows that this step in immune hemolysis is markedly inhibited in buffers of low ionic strength and that the degree of inhibition depends on the nature of the substance used as a substitute for NaCl.
A veronal-buffered NaCl solution (I = 0.147) was prepared according to the “Alternative Procedure” of Kabat and Mayer (5), except that bivalent cations were omitted. Veronal-buffered solutions of glucose, mannitol and sucrose∗ (each 0.284 m) were prepared by dissolving appropriate amounts of these substances and 1.019 g/1 of sodium 5,5'-diethylbarbiturate in water and adjusting to pH 7.4-7.5 by addition of n HC1 before adjusting to volume.
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