Further studies on the affinity of sheep-corpuscles for anti-sheep hemolysin

Abstract

In previous studies on the rate of absorption of anti-sheep hemolysin by sheep corpuscles, it was shown that 0.05 c.c. of packed sheep-cells added to a saline solution containing 400 units of hemolysin, will absorb as many as 390 units after 10 minutes extraction at room temperature. The hemolysin was obtained by immunizing rabbits with sheep-cells in the usual manner and a unit was taken to be the smallest quantity which hemolyzed 0.1 c.c. of a 5 per cent. suspension of packed sheep-cells in the presence of 0.1 c.c. (2 units) of pooled guinea-pig complement after 15 minutes incubation in the water-bath. It was observed also that extraction periods of 5, 10, 15 and 20 minutes did not show any marked differences in the number of hemolysin units extracted by the corpuscles. A study of the effect of temperature on the rapidity of hemolysin extraction, indicated but a small increase in the number of units extracted at water-bath as compared with ice-box temperature. With this data on hand, a quantitative study was undertaken of the hemolysin absorption capacity of 0.05 C.C. of packed sheep-cells when employing different concentrations of hemolysin. Accordingly 0.05 C.C. quantities of centrifugalized cells were added to a series of tubes each containing different quantities of hemolysin in I C.C. quantities of saline. The extractions were carried out for 10 minutes at room-temperature, after which period the cells were thrown down by rapid centrifugation and the number of unabsorbed hemolysin units remaining in the supernatant fluid, determined. The following table gives the results obtained in one of a series of such experiments. The marked affinity of sheep-cells for anti-sheep hemolysin is well illustrated in this table. In similar studies by Morgenroth and Arrheniusz an hour extraction period was employed, while the quantity of sheep-cells used for extraction is not stated.