DIPHTHERIA. 303 



against the ten-times fatal dose. Each cubic centimeter 

 of this stronger serum was described as an antitoxic unit, 

 and, of course, contained ten immunizing units. Still 

 later it was shown that the limits of strength were by 

 no means reached, and he succeeded in making serums 

 three hundred times the normal strength, each cubic 

 centimeter of which contained 300 immunizing units, 

 or 30 antitoxic units. 



In the course of the development of strength in the 

 serum the exact meaning of u immunizing unit " grad- 

 ually became obscured, until it is at present an expres- 

 sion of strength rather than one of quantity. 



While it is difficult to define an immunizing unit, it is 

 not at all difficult for one skilled in laboratory technique 

 to determine the number present in a sample of serum. 

 There are three rules of practice: 



1. Determine accurately the least certainly fatal dose 

 of a sterile diphtheria toxin for a standard guinea-pig. 



2. Determine accurately the least quantity of the 

 serum that will protect a guinea-pig against ten times 

 the determined least certainly fatal dose of toxin. 



3. Express the required dose of antitoxic serum as a 

 fraction of a cubic centimeter and multiply it by ten. 

 There will then be as many immunizing units in i c.cm. 

 of the serum as there are parts in the resulting fraction. 



Example: It is found that o.oi c.cm. of toxin kills at 

 least 9 out of 10 guinea-pigs. It is then regarded as the 

 least certainly fatal dose. Guinea-pigs receive ten 

 times this dose (o. i c.cm.) and varying quantities of the 

 serum, measured by dilution, say, y^Vir c.cm., y^nr c.cm., 

 ygVfr c - cm - The first two live. The fraction y^^nr is 

 now multiplied by 10; ^oW X 10 = 2"3T> an( ^ we find that 

 there are as many units per cubic centimeter in the serum 

 as there are parts in the result i. <?., 250. 



The most accurate definition of an immunizing unit is: 

 ten times the least amount of antitoxic serum that will 

 protect a standard (3OO-gram) guinea-pig against ten 

 times the least certainly fatal dose of diphtheria toxin. 



