Beatrice Carrier Seegal

Immunofluorescence Demonstration of a Muscle Binding, Complement-Fixing Serum Globulin Fraction in Myasthenia Gravis.

Summary 1. A muscle binding, complement fixing component has been demonstrated in the crude globulin fraction of a pool of serum from 10 patients with myasthenia gravis of recent onset and progressive character, by means of the immunofluorescence technic. This component could not be demonstrated in a similarly prepared normal serum globulin pool. Untagged myasthenic globulin was shown to block competitively adherence of fluorescein tagged myasthenic globulin to skeletal muscle striations; whereas prior treatment of muscle sections with normal serum globulin intensified staining with fluorescein tagged myasthenic globulin. Individual myasthenia gravis serums included in the pool also blocked staining with fluorescein conjugated myasthenic globulin. Normal serums did not block adherence of the fluorescein tagged myasthenic globulin to skeletal muscle. 2. The myasthenia gravis globulin fraction was shown to fix guinea pig complement to human skeletal muscle, by successive treatment of muscle sections with myasthenic globulin, guinea pig complement, and fluorescein conjugated rabbit anti-guinea pig complement. Normal serum globulin failed to fix complement by this technic. 3. Thirty-one myasthenia gravis serums were screened, with 11 normal serums, and 5 serums from patients with other generalized myopathies, for their ability to fix guinea pig complement to skeletal muscle. Thirteen myasthenic serums gave unequivocal evidences of complement fixation, using the immunofluorescence technic. No normal serums fixed complement. Serum from one patient with paroxysmal myoglobinuria also fixed complement. Serum from one patient with acute dermatomyositis gave rise to fluorescence in the sarcolemma when layered onto skeletal muscle followed successively by guinea pig complement and fluorescein conjugated rabbit anti-guinea pig complement.

The Development of Hypertension and Nephritis in Normal and Adrenalectomized Rats Treated With Cortisone.

Summary 1. The administration of cortisone did not prevent the development of experimental cytotoxic serum nephritis in the rat. 2. Moderate hypertension developed in nephritic rats with intact adrenal glands when injected with cortisone, whereas striking hypertension appeared in adrenalectomized nephritic rats similarly treated. 3. The nephritic rats rendered hypertensive by the administration of cortisone presented no greater histological evidence of renal damage than did normotensive control nephritic animals.

THE ASSOCIATION OF TYPE SPECIFIC HEMOLYTIC STREPTOCOCCI WITH ACUTE GLOMERULONEPHRITIS : AT THE PRESBYTERIAN AND BABIES HOSPITALS, NEW YORK, N. Y., IN THE YEARS 1936-1942

The mechanism by which acute glomerulonephritis develops is unknown, but there is general agreement that an infection initiates the process. This infection commonly occurs in the upper respiratory tract but classical instances of acute glomerulonephritis have followed infections of the skin, subcutaneous tissues, lungs and peritoneal cavity. An antecedent hemolytic streptococcal pharyngitis has not been excluded in all these instances. Clinical, bacteriological and immunological evidence indicates that the hemolytic streptococcus is the precipitating agent in the great majority of the infections that precede acute glomerulonephritis (1-7). It should be noted, however, that acute glomerulonephritis has followed pneumococcic pneumonia (8), subacute bacterial endocarditis (9), and gastrointestinal infections due to various enteric organisms (1). Recently Rammelkamp, Weaver, and Dingle (10) presented evidence indicating that the attack rate of acute glomerulonephritis following Group A hemolytic streptococcus infections varied considerably. In contrast, the incidence of acute rheumatic fever following such infections appeared to be relatively constant. Rammelkamp and his associates postulated that this difference in attack rate was due to the episodic appearance of certain strains of hemolytic streptococci which were more nephritogenic than others. Their collected data

IMMUNOLOGIC AND BIOLOGIC CHARACTERIZATION OF ANTISERA TO BEEF THYROTROPIN PREPARATIONS

Sensitive immunologic techniques and more purified preparations of bovine pituitary thyrotropin have made it possible to re-examine the question of whether thyrotropin acts as an antigen upon injection into heterologous species. This possibility was first indicated by Werner (1, 2) in 1936, to explain the loss of responsiveness to thyrotropin which Collip and Anderson (3) previously had observed to follow repeated injection of crude pituitary extracts. These latter workers, having demonstrated that serum from such refractory animals could neutralize the effect of thyrotropin upon the thyroid in nonrefractory animals, postulated that thyrotropic stimulation had caused an increase in the titer of an antihormone acting to preserve homeostasis by exerting an effect opposite to that of the thyrotropin. In conflict with the antihormone thesis, however, were the observations (4) that a flavianic acid preparation of thyrotropin did not evoke refractoriness, could stimulate the thyroid of the refractory animal, and could not be neutralized by the serum of refractory animals-supporting the view that antibody to a foreign protein, rather than an antihormone, had been induced. To date, however, there has been no in vitro demonstration of a circulating antibody specific for thyrotropin (5), nor is it known whether highly purified thyrotropic preparations evoke antibody formation. The results of the present experiments suggest that in the rabbit, bovine thyrotropin itself, or a closely associated and as yet not separable protein, does induce the formation of precipitating antibody; and that antisera containing such antibodies can neutralize the thyrotropic effect of the hor-

The production of glomerulonephritis by immunologic methods

Abstract 1. 1. Specific antikidney, antiglomerular, or antiplacenta serum, when injected in the rat or dog, produces acute glomerulonephritis, which may be fatal within a few days, may heal, or may progress to chronic nephritis resulting in death from renal failure months or years later. Duck antirabbit-kidney serum injected in rabbits produces a similar result. 2. 2. The clinical course and pathologic lesions of this experimental disease resemble those of human nephritis. 3. 3. In the rat, high-titered specific antisera may produce a nephrotic syndrome characterized by edema, hypercholesterolemia, hypoproteinemia, and massive proteinuria. 4. 4. Between the clinical picture of acute nephritis and the onset of chronic nephritis in the rat, there may be an interval of several months during which time the animal appears normal. 5. 5. Nephritis produced in rats, rabbits, or dogs by the injection of specific duck or chicken antikidney serum often has a latent period of several days. 6. 6. Evidence has been presented that, at least in the rabbit, the onset of nephritis after a latent period is the result of a reaction between the injected antikidney foreign protein (fowl gamma globulin) attached to kidney parenchyma and the antibodies produced by the rabbit to this antigen. 7. 7. The lesions of delayed nephritis in the rabbit, rat, or dog are similar to those which occur in nephritis with no latent period. 8. 8. Evidence has been developed from chemical and histochemical studies of the glomerulus of the dog and rat which indicates that the basement membrane of the glomerulus may be the source of the antigen inciting production of the nephrotoxic antibody. 9. 9. The new immunologic and histochemical techniques which have been used in the study of experimental nephrotoxic nephritis may in the future give data on the mechanism of human nephritis.

Effect of Anti-Placenta Serum on Development of Foetus in the Pregnant Rat

Conclusion (1) Rabbit-anti-rat-placenta serum effectively interferes with the normal development of the placenta and foetus in the pregnant rat. (2) The same result is obtained with much smaller injections of rabbit anti-rat-whole-blood serum. (3) Rabbit anti-rat-serum serum fails to influence pregnancy unless injected in sufficient volume to induce anemia. (4) Anti-hormone serum and normal rabbit serum are without effect on pregnancy in the rat. (5) The immunological factor or factors responsible for the resorption of the foetuses following the injection of anti-placental serum and anti-whole-blood serum are as yet unknown.

Range of Antibiotic Activity of Protoanemonin.

Summary 1. Protoanemonin inhibited the growth of all the aerobic and anaerobic bacteria, the fungi and the protozoa tested. 2. The maximum dilution of protoanemonin which was effective against the bacteria and fungi varied from 1-6000 to 1-300,000. 3. The anti-bacterial activity of protoanemonin was independent of the acidity of the medium, the size of the inoculum and the age of the culture, within the limits tested. 4. The two protozoa were prevented from qrowing in dilutions of protoanemonin ranging from 1-200.000 to 1-1,600,000. 5. No inhibition by protoanemonin of the growth of the two bacteriophages and the influenza virus was demonstrable by the techniques employed. 6. A dilution of 1-1.000,000 protoanemonin was toxic for chicken tissue culture epithelial and fibroblastic cells.

Local Organ Hypersensitiveness

Although there is a considerable and significant literature on the problems of general hypersensitiveness the known facts concerning local hypersensitiveness are few. The early work of Koch, 1 Calmette and Guerin, 2 and v. Pirquet 3 on the tuberculin reaction, the demonstration by Arthus 4 of skin necroses after repeated injection of foreign proteins, the work of Gay 5 on the typhoidin reaction and finally the work of Swift. 6 Dochez and Stevens, 7 and MacKenzie and Hanger, 8 and many others have helped our understanding of local changes in the reactivity of the skin. Furthermore, clinical observations have attested to the fact that apparently arrested areas of inflammation in various parts of the body have been relit by the introduction of the homologous noxious substance into distant sites. Experimental work along similar lines attempting to produce altered reactivity in other organs than the skin has not yielded uniformly positive results. From a review of previous attempts to produce local hypersensitiveness it seemed that the critical requirement was the maintenance of a set of experimental conditions which would keep antigen in juxtaposition to body cells for a relatively long period of time. The structure of the anterior chamber of the eye presents a feature which probably allows this condition to be fulfilled. We had observed that heterologous erythrocytes when injected into the anterior chamber of the rabbits eye would persist for several days. On this account the anterior chamber of the eye was the initial area chosen to test for the production of local hypersensitivity. A number of workers in ophthalmology have attempted to explain certain types of conjunctivitis and keratitis on the basis of “ocular anaphylaxis”.

Development of Hypertension in the Adrenalectomized Nephritic Rat Maintained on NaCl.

Summary In response to cytotoxic serum nephritis, bilaterally adrenalectomized rats develop hypertension as frequently as do animals with intact adrenals. In order for hypertension to appear among such adrenalectomized rats, it is necessary that a satisfactory state of nutrition be maintained over a period of weeks. In our laboratory, the difficulties encountered in fulfilling this provision have been considerable in spite of the employment of a high NaCl regimen. This experience has led us to wonder whether the conflicting results in the literature relating to the question of hypertension in adrenalectomized animals may not be due, in part, to similar difficulties.

Incidence of Hemolytic Streptococci and Pneumococci in the Pharyngeal Flora of Normal Rhesus Monkeys

The normal bacteriologic flora of the pharynx of chimpanzees has been described by Dochez, Shibley and Mills 1 in connection with their experiments on the transmission of the common cold. The bacteria occurring in the throats of apes were similar, for the most part, to those found in human throats, although the percentile incidence varied for some organisms. Apparently, no such complete study of the pharyngeal flora of lower primates has been reported. The few reports of throat-cultures in the monkey are primarily concerned with the presence of C. diphtheriœ in the nasopharynx. Dold and Weigmann, 2 and Ramon and Erber 3 found diphtheria-like organisms in some monkeys, but Jungeblut 4 in 50 normal rhesus monkeys found no C. diphtheriœ. Dold and Weigmann reported the presence of staphylococci and streptococci but did not specify the reaction of these organisms in blood-media. During studies of experimentally induced hemolytic streptococcal infection of rhesus monkeys, a survey of the incidence of hemolytic streptococci in the normal throats of 49 animals∗ was undertaken. The cultures were taken during the 2nd week of January, 1936, and within 9 days of receiving the animals in the laboratory from the dealer. A swab was rubbed firmly over the faucial surface, streaked on sheep-blood agar, and after 24 hours incubation, the plates were searched for hemolytic colonies. From 28 of the 49 cultures a β-streptococcus 5 was isolated. On one plate a minute streptococcus of the type described by Bliss and Long 6 was also isolated. These organisms were tested for production of a soluble hemolysin, lysis of human and rhesus plasma-clot (Tillett and Garner 7 ) and fermentation of lactose, mannite, and salicin. The antigenic group to which the organisms belonged was determined by the method of Lancefield. 8 Six of the strains were tested for the production of a toxin reactive in the skin of silver-fox rabbits.