212 CHEMISTRY OF THE IMMUNITY REACTIONS 



sorbed hemoglobin, which process can be dupHcated experimentally 

 with a combination consisting of a corresponding solid hydrophilic 

 colloid, fibrin, and a hydrophobic colloid dye, carmin^e; this artificial 

 combination behaves exactly like a corpuscle to simple hemol^'tic 

 agents. ^^ 



Repeated alternate freezing and thawing is another physical means 

 of bringing on hemolysis. Heating to 62°-64° C. causes hemolysis 

 of mammalian corpuscles; in cold-blooded animals this seems to occur 

 at a slightly lower temperature. Hypertonic solutions produce hemo- 

 lysis, and it may be that freezing and desiccating cause hemolysis 

 through the resulting hypertonicity.^- 



Some chemical agents are capable of liberating hemoglobin, even 

 when the corpuscles are in isotonic solutions. The ordinar}' salts 

 of serum, of course, do not have this property, but ammonium salts 

 are strongly hemolytic. The chemical agents that dissolve red cor- 

 puscles seem to be those that have the power of penetrating the 

 stroma. Ammonium salts and urea penetrate the corpuscles freel}^ 

 and cause hemolysis. Sugar and NaCl seem not to penetrate the 

 corpuscles, and therefore do not produce hemolysis. Of the perme- 

 ating substances, there seem to be two types: one, like urea, does not 

 produce hemolysis when in a solution of NaCl isotonic with the serum; 

 the other, like ammonium chloride, is not prevented from producing 

 hemolysis by the presence of NaCl.^^ 



All these agents seem to effect hemolysis by acting on the stroma, 

 for when the stroma of corpuscles hardened in formalin has its leci- 

 thin and cholesterol removed with ether, saponin, a powerfully hemo- 

 lytic substance, seems to have no effect. The action of saponin and 

 of many other hemolytic agents can be prevented b}' the presence of 

 cholesterol in excess, suggesting that it is this constituent of the stroma 

 that is affected.^* By studying hemolysis under dark field illumination 

 Dietrich^^ found that in water hemolysis a diffusion of hemoglobin 



^^ Concerning the influence of H-ion concentration on hemolysis see Walbum, 

 Biochem. Zeit., 1914 (63), 221. 



^2 Guthrie, Jour. Lab. Clin. Med., 1917 (3), 87. 



^^ Hamburger, in his book, "Osmotischer Druck und lonenlehre," reviews ex- 

 haustively the physical chemistry of hemolysis. The following is his summary 

 of the permeability of red corpuscles by various substances: 



Organic Substances. — (a) Impermeable for sugars; namely, cane-sugar, dextrose, 

 lactose, also arabit and mannit. (6) Permeable for alcohols, in inverse proportion 

 to the number of hydroxyl groups that they contain; also for aldohj'des (except 

 paraldehyde), ketones, ethers, esters, antipyrin, amides, urea, urethan, l)ile acids 

 and their salts, (c) Slightly permeable for neutral amino-acids (glycocoll, aspar- 

 agin, etc.). 



Inorganic substances, not including the salts of the fixed alkalies, (a) Com- 

 pletely impermeable for the cations Ca, Sr, Ba, Mg. {b) Pcnncable for NHj ions, 

 for free acids and alkalies. 



^■•Ransom, Deut. med. Woch., 1901 (27), 194; Koliert, "Saponinsubstanzen'' 

 Stuttgart, 1904; Abdcrhalden and Le Count, Zeit. exp. Path. u. Ther., 1905 (2), 

 199. Noguchi (Univ. of Penn. Med. Bull., 1902 (15), 327) found lecithin without 

 this proi)erty. 



"Verb. Deul. Patli. (icscll., 190S (12), 202. 



