Merrill W. Chase

Inhibition of experimental drug allergy by prior feeding of the sensitizing agent.

Summary Through the feeding of certain allergenic compounds to the non-sensitive subject, a state of resistance may be established against subsequent experimental sensitization of the skin by the same substance.

The Cellular Transfer of Cutaneous Hypersensitivity to Tuberculin

In studies on experimental drug allergy, it has been found that specific hypersensitiveness of the “delayed type” is transferable to normal guinea pigs by means of cells in exudates recovered from sensitized guinea pigs. 1 Resemblances between the delayed type of reaction to drugs and the classical tuberculin reaction have prompted investigation as to whether cells from tuberculin-sensitive animals may likewise transfer tuberculin sensitivity. The experiments show that guinea pigs receiving such cells acquire for a limited time a skin hypersensitivity that exhibits the essential features of the typical tuberculin reaction. Guinea pigs were rendered hypersensitive to tuberculin by subcutaneous injection of killed human tubercle bacilli suspended in paraffin oil,2,3 usually mixed with vaseline; each animal received 0.5 to 2.5 mg of dried tubercle bacilli in a total inoculum of 1 cc. Between 5 and 9 weeks later, the cutaneous reactivity to tuberculin then being pronounced, exudates were induced by the intraperitoneal injection of about 28 cc paraffin oil into each of a group of guinea pigs so sensitized. After 48 hours, the peritoneal cavities were washed out with heparinized Tyrode solution containing gelatin or normal guinea pig serum. The washings were combined and the cells recovered from the aqueous layer by minimal centrifugation. The sedimented cells were resuspended in fresh washing fluid by gentle pipetting, and again spun down. A similar washing was made, using Tyrode solution mixed with 1/10 volume of normal guinea pig serum. The washed cells were then suspended in serum-Tyrode and immediately injected into male albino guinea pigs. The yield of cells amounted to 0.1 to 0.15 cc per donor, and recipients were usually given the cells of between 2 and 10 donors. The cells were comprised of 15 to 30% of polymorphonuclear leucocytes, 20 to 35% of lymphocytes, and 50 to 65% of large mononuclear cells.

Experiments on Transfer of Cutaneous Sensitivity to Simple Compounds

In the course of experiments to passively transfer skin sensitiveness to simple compounds an attempt was made to induce sensitivity by injecting peritoneal exudates from sensitized animals. Guinea pigs were rendered sensitive to picryl chloride in the manner previously described, 1 using conjugates (guinea pig stromata treated with picryl chloride) in conjunction with intraperitoneal injections of killed tubercle bacilli. To produce exudates, killed tubercle bacilli (or tuberculin) were injected intraperitoneally about 3 weeks from the beginning of the treatment, when substantial tuberculin hyper-sensitivity was established. On injecting such exudates intraperitoneally into normal animals, the recipients in most of the experiments were seen to develop sensitivity to picryl chloride; when then a drop of an oil solution of the substance was put on the skin, erythematous reactions, mostly of high color, were apparent on the next day. The phenomenon was found to be due not to the clarified fluid but to the sediment obtained upon centrifugation. Among the possible explanations, an active sensitization through residual antigenic material in the peritoneal exudates seems rather improbable because a transfer was possible also with the exudate of animals in which the injection of dead tubercle bacilli and picryl stromata was made under the skin of the neck (using as a vehicle “Aquaphor” (Duke Laboratories) and paraffin oil, according to a method devised by Freund 2 ). This is further supported by the appearance of the sensitivity after a short interval, namely, 2 days after injection of sufficient material, and fading of the reactivity within a few days. Finally, there is preliminary evidence that moderate heating sufficient to kill the exudate cells abolishes the effect. Consequently one would be inclined to assume that the sensitivity is produced by an activity in the recipient of the surviving cells, if not by antibodies carried by these.

A simplified and efficient system for separating proteins by preparative polyacrylamide electrophoresis: With notes on horse heart myoglobin

Abstract A simplified but effective method for preparative polyacrylamide gel electrophoresis is presented, suitable for single runs of ∼15 mg of protein, which allows: (1) a gel casting shaped and cooled so that proteins do not assume curvature as they migrate down the column; (2) direct contact of the bottom of the gel with buffer; (3) use of reduced electrical field since anode and cathode are spaced only 9.5 cm apart; (4) increased stability of the anodal face of the gel, by employing a dilute running buffer and consequently a still lower current (7 to 8 mA at 150 V); (5) obtaining successive effluents in separate Visking sacs, at appropriate time intervals, and removal of proteins in absence of an electrical field; (6) rinsing of the column between collections; (7) advance planning of the actual times of collection, calculated from the rates of migration of proteins of interest relative to the time of passage of (yellow) “myoglobin standard” determined in analytical columns run under the same conditions. Working conditions are designed to eliminate gel deformations during electrophoresis. Individual effluents and rinsings are dialyzed and concentrated by negative pressure in a simple device.

Studies on the Sensitization of Animals with Simple Chemical Compounds

(1) Following the onset of sensitivity to picric acid (PA) by means of a ‘split-adjuvant’ technique, an anamnestic rise in the degree of sensitivity occurs accompanied by a qualitative cha

Irreverent recollections from Cooke and Coca, 1928–1978

The story commences with two strong personalities, Arthur F. Coca (1875-1959) and Robert A. Cooke ( 18801959). Both had had childhood allergies and as physicians they proceeded to specialize in this area whit h concerned them intimately. And both were active with the teaching affiliation between Cornell University Medical College located at 26th St. and First Avenue in Manhattan (across from Bellevue Hospitalj and the New York Hospital on West 16th Street just west of Fifth Avenue. Coca was more the solitary laboratory type, interested in immunology and allergenic extracts and in teaching; Cooke was the physician and organizer of allergy clinics. These two individuals, at first working hand in hand, later apart, largely shaped the new discipline in America and afforded training in clinical allergy to many selected individuals who later established their own practices. Several physicians in private practice also helped to establish the discipline, such as Joseph Harkavy and Bret Ratner in New California.

Skin Sensitization to a Simple Compound by Injections of Conjugates

In view of a recent paper 1 touching upon the subject, we wish to make a preliminary communication of a study under way for some time on the possibility of producing in animals skin sensitivity to drugs by immunizing with conjugates. We have in fact been able to render guinea pigs sensitive to superficial application of picryl chloride by intraperitoneal injections of a conjugate resulting from the treatment of guinea pig erythrocyte stromata with picryl chloride in alkaline solution, killed tubercle bacilli as in previous work having been injected beforehand. Since even minute quantities of the simple substance can sensitize under certain conditions and must be avoided, the chief concern in these experiments was to guard against the inclusion of unchanged picry! chloride in the injection material. This was carried out by adding an excess of glycine which removed any possible remainder of the substance through chemical combination, and by washing with aqueous alcohol. The large majority of animals treated in this way have shown upon subsequent testing with the simple substance typical reactions of the contact dermatitis type.

THE IMMUNOLOGICAL ENIGMA OF TRANSFER FACTOR

Publisher Summary This chapter discusses the development of the knowledge of biology of transfer factor (TF). The extracts of mononuclear cells, predominantly lymphocytes, release many intracellular products, of which some have special biological activities in in vitro systems. But the concern of the immunologist has to do with the biological basis for specificity of transfer, that is, the ability to confer, on a new individual, the DH+ reactivity of the cell donor. This problem has remained a conundrum, which seems closer to be solved now. The chapter discusses the specific and the nonspecific components of leukocyte lysates. TF from highly reactive donors can produce lasting reactivity. Positive skin transfer is the sole criterion of TF. TF is not provided by nonsensitive individuals. TF dialyzes through Visking tubing. Biologically active materials besides TF are diffusible. TFD passes 12,000D membranes and not 3500D membranes. The removal of 3500D diffusate (>90% of weight) raises peptide:RNA ratio.

REPORT OF WORKSHOP III: CHEMICAL NATURE OF AGENTS AND AIRWAY RESPONSES

The search for insights into mechanisms of toxicity of airborne organic dusts, the goal of the workships, was pursued with varying results. The audience fortunately included experienced individuals who made substantial contributions to the Workshop. Dr. Robert M. Adams, dermatologist from Palo Alto and author of OCCUpational Contact Dermatitis,’ reviewed the possibilities for involvement of delayed hypersensitivity in determining airway responses. The skin, with its desmosomes and tonofibrils bridging cells, presents a unique situs for observing reactions of delayed hypersensitivity such as spongiosis and vesiculation. Other tissues would not afford equivalent opportunities for observation. For induction of sensitization, a mild rather than a strong insult to the skin appears to be important; certainly irritation is an assist. Among volatile or dusty chemicals that could be considered as perhaps capable of inducing delayed-type hypersensitivity of lung tissue were these suggestions : formaldehyde, epoxy resins and its catalysts, chromates, acrylics, pyrethrum, chloramine-T, and p-phenylenediamine. The possibility of sensitization effected by one compound and cross-reactivity with chemically related chemicals was pointed out. Dr. John Rankin described a respiratory ailment affecting many rubber workers when a new rubber formulation (a resorcinal-HCHO-resin) was introduced into the plant, among those persons dealing with its application both when hot and during curing. Some showed deep airway distress but skin testing could not distinguish between the affected and nonaffected workers. A quite dilute solution was sprayed on two successive days, after which reactions developed involving the nasal mucosa and lung disease. The latent period was suggestive of a delayed-type hypersensitivity, but no proof for this could be assembled. Toluene diisocyanate, a volatile member of the diisocyanates, is used in preparing polyurethane foam that is widely used for insulation, cushioning, and surface coatings. TDI is reacted with a polyol resin in this process, but not all becomes combined. Dr. John M. Peters described an asthma-like disease among plant workers; those most affected separated themselves from the industry. It is estimated that 50,000 workers are at risk in the U.S.A. Further, “polyurethane kits” are available at hardware stores, these can generate more vapor than in occupational exposures. High dosages in industry, usually from accidental spillage, tends to cause acute bronchitis; low doses provoke asthma in the “sensitized” worker. Many workers sustain acute falls of vital capacity during the workshift. These falls develop on Monday and remain low for the

Purification and Characterization of Allergens

Publisher Summary This chapter provides details of a workshop held to analyze results of experiments conducted to study purification and characterization of allergens. New immunological control methods, useful during separation and standardization of extracts were presented, such as the laiirell immunoelectrophoretic technique with its newer modification and the radioallergosorbent test (RAST test). Studies on nondialyzable constituents of allergenic materials were also presented. Several studies were reported on dialyzable constituents of pollens and of peas. It was observed that delayed-type skin reactions in streptococcal hypersensitivity are provoked by the preparation of streptococcal enzymes called streptokinase/streptodornase (SK/SD). The search for strain-specific antigens of the Mycobacterium tuberculosis group is proceeding. This project is different from the objectives of others currently directed toward further purifications of skin-test materials for the diagnosis of clinical tuberculosis. A proposal was submitted in advance regarding adoption of nomenclature which would assign a codifying symbol to isolated allergens as these are reported.