Hemolysis of Human Red Cells by Saponin Following Viral Action

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Abstract
Red blood cells were altered by Shope''s strain 15 of swine influenza virus (SW). To remove the virus receptors, allan-toic fluid of the chick embryo infected with SW was added to packed human red cells of Group O and the mixture kept at 37 C for 2 hrs. The cells were washed with saline, and after removing the supernate, anti-SW ferret serum was added, and the mixture kept at room temp. for 15 min. The cells were washed with saline and made up to a 4% suspension in saline. The virus receptors were removed from the cells. Two controls were used: (1) cells treated similarly with unin-fected allantoic fluid, and (2) cells whose receptors were coated with SW. Hemolysis from addition of 0.5 ml. of a 1:15,000 dilution of saponin to 0.5 ml. of the treated 4% cell suspension was detd. with a photoelectric colorimeter. Cells with virus receptors removed were significantly more resistant to hemolysis than cells with receptors intact or coated by virus. The effect of saponin on cells treated with filtrates of the 4Z strain of Vibrio cholera was tested. Virus receptors were removed by adding the filtrate in borate buffer at pH 7.6 containing 0.1% CaCl2 to packed cells at 37 C for 2 hrs. The cells were washed with saline and made up to a 4% suspension in saline. The cells were placed in contact with a 1:18,000 dilution of saponin, and hemolysis detd. as above. Cells with virus receptors removed were significantly more resistant to hemolysis than cells with intact receptors. Cells with unaltered virus reached the 50% hemolytic end-point in 20 min., whereas cells without receptors did not reach the same point until 50 min. Cells with virus receptors removed were not modified in their reactions to hemolysis by hypotonic solns. The mechanism of hemolysis is unknown.

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