Foreign Serum Research Articles

STANDARDIZATION OF SCARLET FEVER ANTISTREPTOCOCCUS SERUMS

At present, the antitoxic titer of scarlet fever antistreptococcus serums is determined by the blanching of the rash in scarlet fever patients, following the intracutaneous injection of serum, as developed first by Dochez 1 and later also by Blake and Trask, 2 or by the reactions induced in susceptible persons by mixtures of toxin and serum injected intracutaneously, as developed by the Dicks. 3 Attempts to devise a satisfactory in vitro method have not been successful. While the tests on human subjects have been essential in establishing definite standards for measuring the antitoxic value of the serums, they are proving impracticable as routine procedures for standardization. Toxin-antitoxin tests on normal persons are impracticable for the following reasons: the difficulty of securing subjects susceptible to the toxin but not hypersensitive to horse serum; the objection to the repeated injection of foreign serum; the large number of injections that should be made

ON THE ANTIGENS OF RED BLOOD CORPUSCLES

Erythrocytes contain more than one substance responsible for the production of lysins and agglutinins. By injections of alcoholic extracts of horse erythrocytes mixed with a foreign serum, hemolysins and agglutinins can be more readily obtained than by the same quantities of extract alone. The immune sera reacted on horse and donkey blood but not on the blood of other species. The antibodies obtained differ from those prepared in the usual way with unchanged red blood corpuscles. The differences involve the ratio lysin: agglutinin, the specificity, the inhibition of hemolysis by alcoholic extracts, and the flocculation of these extracts. Ordinary anti-horse sera give flocculation reactions with emulsions of alcoholic extracts of corpuscles but to a lesser degree and not so regularly as those produced by means of extracts. The specific substances present in alcoholic blood extracts are probably not proteins. According to this view, species specificity in animals depends not alone on proteins but also on another group of substances.

Further studies on the potency of botulinus toxin.

Repeating titration of toxin from day to day, we have observed that determination of the potency of dilutions of toxin in physiological salt solution has often given inconsistent results. Inquiry into the cause of this inconsistency brought out the fact that NaCl, as well as many other salts of mono- and polyvalent metals, have direct deteriorating effect upon toxin. This effect is more marked the higher the dilution at which the toxin is exposed to the salt. At any given concentration of toxin in a salt solution, deterioration is greater the longer the exposure and the higher the concentration of salts. An attempt to neutralize the effect of a single salt by balancing with other salts having antagonistic physiological action yielded no definite result. If, instead of using salt solution, the toxin is diluted in distilled water, the results of titration become more regular, and solutions of toxin keep fairly well. On the other hand, in the presence of a low concentration of salts, addition of serum or broth seems to protect the toxin from deterioration. Moreover, if the toxin is diluted not in salt solution or distilled water but directly in normal horse serum or even in ordinary broth, its potency is increased as compared with that of the same toxin diluted in distilled water. In view of the tendency of foreign serum to increase the toxic action of botulinus toxin, it is particularly important to determine carefully the type of toxin involved before administering antitoxin. Actual experiments have shown that at least the animals receiving heterdogous antitoxin (type B) together with botulinus toxin (type A) die more promptly than the controls receiving the toxin alone.

Studies on Endothelial Permeability. V

Abstract During the course of the studies on endothelial permeability made with thoracic duct fistulas we have had occasion to study the effect of varying doses of peptone injected intravenously. The effect of peptone has been studied intensively; usually relatively large doses have been injected and it has been found that there undoubtedly follows an increase in the rate at which colloids (hemolysins, agglutinins, antitoxins, foreign serum, etc.) pass through from the capillary bed to the lymph stream. Yet there seems some doubt whether we are dealing with an actual alteration in the permeability of the endothelium or are dealing purely with an increased intraportal pressure, as Abe (1) suggests. Starling inclines to the view that both an increase in intraportal pressure and an increased permeability of the endothelium must take place (2).

THE RELATION OF ANTIBODY TO THE RATE OF DISAPPEARANCE OF CIRCULATING ANTIGEN

1. In rabbits previously immunized to horse serum the rate of disappearance of horse serum after reinjection is somewhat more rapid than in control animals not previously immunized. But when the average duration of the persistence of antigen in ten previously immunized rabbits is compared with the average duration of persistence in ten normal controls, the difference is not impressive. This holds true for animals previously immunized by repeated injections but having no free circulating antibody, as well as for animals having a high titer of antibody at the time of reinjection. The same animal may dispose of circulating antigen as rapidly at a time when the blood contains no precipitin as when there is a high titer of circulating precipitin. 2. The intravenous injection of large amounts of potent antihorse rabbit serum immediately after an injection of horse serum does not materially accelerate the rate of disappearance of the horse serum from the circulation. While the number of days that circulating antigen was demonstrable in two experiments of this type was less than the average of controls the duration of persistence was within the limits of variation of normal controls. 3. Under the condition of the experiments intravascular union of antigen and antibody is an unimportant factor in the mechanism for disposal of antigen. By exclusion it therefore seems probable that the rate of disappearance of antigen is largely dependent upon cell avidity for the antigen. 4. Rabbits show a wide range of individual variation in the rapidity with which foreign serum is appropriated by the tissue cells, and in their ability to form precipitins. There does not appear to be any close relation between the amount of precipitin set free in the circulation and the rate of disappearance of precipitinogen. 5. The type of interrelation between precipitin and precipitinogen-demonstrable not infrequently in patients with serum disease-in which the precipitinogen diminishes rapidly when the precipitin rises to the crest of its curve is sometimes reproduced in rabbits. The type of curve which has been observed in patients with little or no serum disease in which the precipitinogen persists steadily in the circulation for many weeks and little or no antibody is formed, has not been observed in the present series of rabbits.

The dominant reacting tissues in anaphylaotic, peptone and histamine shock

Canine anaphylactic shock, peptone shock and histamine shock are currently assumed to be physiologically identical reactions. In each shock there is a sudden, pronounced fall in arterial blood pressure, the carotid pressure being reduced to about 25 mm. Hg. by the end of two minutes. Recovery usually begins about the tenth minute, the arterial pressure being restored to normal in from 30 minutes to 90 minutes, depending upon the severity of the reaction. In each shock, fatal results may be produced by the injection of large doses or by the use of highly sensitized animals. In each shock there is a pronounced splanchnic engorgement and cyanosis, the production of hemorrhagic lesions in the intestinal mucosa, and a reduction in blood coagulability. In order to test the assumed physiological identity of the three shocks, we have endeavored to determine the topographical distribution of the dominant reacting tissues in each shock. 1. Canine anaphylactic shock. Anaphylactic shock (fall in arterial blood pressure) does not take place in dehepatized (Eckfistula) dogs. This is not only true for the mildly sensitized dogs previously reported, but is equally true for highly sensitized dogs giving the fatal type of the reaction. Practically no change in arterial blood pressure is produced in these highly sensitized animals, even on the intravenous injection of as large a dose as 30 c.c. of specific foreign (horse) serum. The liver is therefore not only the dominant but the essential reacting organ in canine anaphylactic shock. 2. Canine peptone slzfock. The severity of the peptone reaction is reduced in dehepatized dogs, and is further reduced in completely eviscerated dogs. There are therefore important though not dominant hepatic and intestinal factors in this shock. In dehepatized dogs, recovery from the shock does not take place, the animals dying in about 60 minutes.

On the mechanism by which antigen is removed from the circulation

In studying the rate of disappearance of horse serum and the curves for circulating precipitin in a group of serum-treated patients it was noted that individuals who have severe serum disease are good precipitin formers and that at the time the precipitin in the circulation reaches the crest of its curve or soon thereafter the precipitinogen rapidly disappears from the blood stream. On the other hand in those individuals who after a first administration of foreign serum, show very mild or no symptoms of serum disease little or no precipitin is demonstrable in the patient's serum and the precipitinogen persists in the circulation for a long period. Intermediate types were also encountered. From these results it seemed at least plausible to assume that an important factor in determining the rate of disappearance of the foreign serum from the circulation was an intravascular union of antibody and antigen. That such an assumption is erroneous seems probable from the following experiments:In one series of expe...

LOCAL DESENSITIZATION IN HYPERSENSITIVE INDIVIDUALS AND ITS BEARING ON THE PREVENTION OF HAY-FEVER

Cutaneous reactions in hypersensitive individuals are of two quite distinct types. The reactions frequently observed in patients with hay-fever or bronchial asthma and those obtained after sensitization by foreign serum consist essentially of an urticaria-like lesion with a central wheal and a surrounding zone of erythema. Such reactions appear a few minutes after application of the test substance, usually reach their full development in half an hour or less, and fade out completely in one or two hours. The skin, at the site of the test then appears normal. There is no visible evidence of cell destruction. It appears to be essentially a vascular phenomenon with localized edema. Such reactions may often be obtained in hypersensitive individuals with extracts of pollens, animal hair, dandruff or feathers, with foreign serum, with many food proteins and occasionally with bacterial or other proteins. These immediate urticaria-like reactions have little in common with the

RELATION OF ANTIBODY AND ANTIGEN TO SERUM DISEASE SUSCEPTIBILITY

1. Studies on nineteen patients to whom foreign serum had been administered for therapeutic purposes are reported. Analysis of the results obtained by following the precipitin and precipitinogen in the circulation and comparing these factors with the time of appearance, intensity, and duration of the symptoms shows that the nineteen patients fall into three groups. 2. The first group includes eleven patients. These were good precipitin formers, they had severe serum disease, and the precipitinogen disappeared from the circulation near the time that the symptoms subsided. 3. The second group includes four patients who had little or no serum disease, in whose circulation little or no precipitin was demonstrable, and in whom the precipitinogen persisted in the circulation as long as the patients could be kept under observation-from 52 to 67 days. 4. The remaining four patients form a more or less distinctly intermediate group. 5. The results lend further support to the conception of serum disease as an antigen-antibody reaction. 6. The possibility that our results indicate a factor which may be important in the mechanism of natural immunity is discussed.

Observations on the specific exhaustion of cutaneous reactions

Cutaneous reactions in hypersensitive individuals are of two quite distinct types. The reactions observed in patients with hay fever or asthma and after sensitization by foreign serum develop in a few minutes, consist essentially of a wheal and erythema, and fade out completely in one to two hours. The skin then appears normal. There is no visible evidence of cell destruction. Such reactions may be obtained with extracts of pollen, animal dandruff and feathers, with food proteins, with foreign serum, and occasionally with bacterial or other proteins. This type of cutaneous reaction has little in common with the local cutaneous reactions to tuberculin, typhoidin, luetin, and mallein. Here the reaction does not usually develop for 12 to 24 hours; it is characterized by induration and persistent signs of inflammation, requires many days to fade out entirely and clearly involves cell destruction. Zinsser has recently shown that the local tuberculin reaction in the guinea-pig is independent of the development of a state of anaphylaxis and it is highly probable that the same holds true for the reactions of this type produced by other substances of bacterial origin. Although it has commonly been assumed that the immediate skin reactions with urticaria-like lesions are manifestations of anaphylaxis, it has not been demonstrated that the mechanism consists of an antigen-antibody reaction. Similar reactions may sometimes be obtained with non-antigenic substances such as aspirin, salicylates and quinine, and there are a few substances, notably histamine, morphine and pituitrin, which produce this type of reaction in normal individuals. It appears to be essentially a vascular phenomenon with localized edema. In an effort to determine the nature of these urticaria-like skin reactions we have studied their exhaustibility by a simple procedure in five hypersensitive patients, the subjects of hay fever or bronchial asthma.

EFFECT OF INJECTION OF NONSPECIFIC PROTEIN ON DIPHTHERIA VIRULENCE TESTS IN GUINEA-PIGS

CONTROVERSY CONCERNING THE VALUE OF HORSE SERUM Bingel, 1 noting that toxic cases of diphtheria, in which the most marked effect might be anticipated from antitoxin treatment, react only slightly, questioned the possibility of nonspecific reaction due to a foreign serum. He treated alternating cases of diphtheria entering the hospital with normal horse serum and with antitoxin. He was unable to see any appreciable difference between the 471 horse serum treated cases and the 466 antitoxin treated cases. The bacilli disappeared after four weeks in 89.6 per cent, of the antitoxin treated cases and in 94.6 per cent, of the horse serum cases. He concluded that the success of the serum therapy in diphtheria was dependent on the larger amount of horse serum used with the larger doses of antitoxin. Perhaps Bingel's results with horse serum may be due to the fact that only 45.7 per cent, of his antitoxin

THE CALCIUM TREATMENT OF THE SERUM DISEASE.

The peculiar pathologic phenomena, such as urticaria and articular pains, which may occur after the introduction of foreign serum of any kind, have been grouped together under the convenient name of the serum disease. We recently³discussed the theories as to the causation of this remarkable condition. Many efforts have been made to prevent this disease, which constitutes an annoying disadvantage incidental to serum therapy of all kinds. Some ten years ago Wright⁴showed that as the symptoms of the serum disease develop there occurs a diminution in the coagulability of the blood and that calcium salts appeared to have a preventive effect. Hence he suggested that calcium salts should be administered in order to forestall urticaria, etc., and that they should be given also to remove more rapidly these troubles when they have supervened. Recently the good effects of the calcium salts in these directions, especially the

Original Papers Used for Advertising.

Chicago, March 30, 1901. To the Editor: —I notice in the advertising columns ofThe Journalof March 30, an advertisement which states: Official Figures Reported to theAmerican Medical Associationby the Chairman of Section on Pediatrics, and then it goes on to give statistics where 6325 cases of diphtheria were treated with the advertiser's antitoxin, with a mortality of only 4.11 per cent., and 859 cases treated with all other American and foreign serums with a mortality of 7.46 per cent. These figures are taken from an article which is furnished to whoever applies to the manufacturer of the antitoxin referred to, and on the inside of the cover it states that is presented with the compliments of the author. I wish to ask if it is possible that this article was written as an advertisement for this particular antitoxin? If this is a fact, is it