Intravenous Injection Of Antigen Research Articles

Enzyme-Potentiated Hyposensitisation

The ability of preparations of hyaluronidase and <i>β</i>-glucuronidase to potentiate the hyposensitising effect of small doses of antigen injected subcutaneously into previously sensitised animals has been investigated in three different models of experimental anaphylaxis. Guinea-pigs were sensitised by egg albumen and repeatedly challenged by exposure to an aerosol of the antigen at weekly intervals. A small dose of antigen by injection increased their pre-convulsion times on subsequent challenge, and this was potentiated by the addition of hyaluronidase and <i>β</i>-glucuronidase. Rats were sensitised by horse serum and challenged 18 days later with intravenous antigen, deaths being noted within 24 h. Ten days after sensitisation, subcutaneous injections of antigen and <i>β</i>-glucuronidase greatly reduced the mortality on subsequent challenge although antigen injected alone had no effect. In mice sensitised by horse serum and challenged by pinnal anaphylaxis, the anamnestic increase in sensitivity produced by a subcutaneous dose of antigen was prevented by <i>β</i>-glucuronidase. Using this model, tolerance was induced by the intravenous injection of antigen and this was prevented by the addition of hyaluronidase.

Ultrastructural alterations in guinea pig mast cells during anaphylaxis.

Guinea pigs passively sensitized with rabbit antisera were subjected to lethal anaphylaxis by an intravenous injection of antigen (bovine serum albumin or horse spleen ferritin). Specimens of lung were obtained for ultrastructural examination. During anaphylaxis pulmonary mast cells exhibit degranulative phenomena associated with intracellular fragmentation of cytoplasmic granule constituents. No apparent extracellular expulsion of granules was observed. It is suggested that histamine release under these circumstances occurs initially intracellularly, followed by amine liberation from the cell.

Anaphylactic reaction in the cat following intraventricular and intravenous injections of antigen.

Laγ-globuline bovine, en injections intraveineuses et dans le ventricule lateral du cerveau, a produit, chez le chat, l'apparition d'anticorps specifiques dans la circulation et la sensibilisation determinant une reaction anaphylactique d'intensite variable.

Hemmung der anaphylaktischen histaminasefreisetzung durch protamin beim meerschweinchen

Previous experiments had shown that in guinea pigs the liver is the source of the plasma histaminase during the anaphylactic shock as well as after injections of heparin. Furthermore, the liver of guinea pigs is also involved in the anaphylactic release of heparin-like substances which have a histaminase releasing activity. The present study indicates that the release of anaphylactic histaminase into the plasma is mediated by heparin. In sensitized guinea pigs the plasma histaminase activity after intravenous injection of antigen (1 mg ovalbumin/kg) is strongly reduced by simultaneous injection of protamine (1 or 2 mg/kg) into a different vein, although higher doses (5 mg/kg or more) of protamine have like heparin a strong histaminase releasing effect. The effect of protamine on the anaphylactic histaminase release is in accordance with its effect on the histaminase release produced by heparin injection. Heparin doses which provoke the same plasma histaminase activity as the anaphylactic shock are also neutralized by 1 mg protamine/kg. In vitro, protamine has no inhibiting action on the activity of plasma histaminase. Given in different doses, heparin revealed no inhibitory effect on the anaphylactic histaminase release. Protamine is ineffective against the anaphylactic histamine release into the plasma of guinea pigs and does not alter the acute or subacute death rate in shock.

In-vitro detection of reaginic antibody.

Atopic disease in man is distinguished by the presence of a special type of antibody, variously termed reaginic, skin sensitizing, or Prausnitz-Kustner antibody. While universal agreement niay not exist on the exact relationship of the atopic reagin to symptoms in atopic disease, it is nonetheless the most reliable indicator of the naturally occurring sensitized state that is peculiar to atopy in man, and appears unrelated to levels of blocking, hemagglutinating, or precipitating antibodies, which at times may be detected in the sera of atopic individuals. It is also known that the level of reaginic antibody is influenced by repeated injections of antigen, being initially increased, and later dirninishing to the point of disappearance following continuous, long term hyposcnsit1zation.l This influence on reaginic antibody appears to bear a closer relationship to improvement in the clinical state than do the increascs in blocking and hemagglutinating antibodies developed during hyposensitization. The basic method for the detection and study of the rcaginic antibody for more than 40 years has been the passive local sensitization of normal human skin by atopic serum, the Prausnitz-Kustner phcnomenon.2 Since the first recognition of reagin, investigators have atteinptcd to characterize the antibody by methods which would eliminate thc technical and biologic problems involved in using human skin as an indicator of its presence. Among the earliest characteristics of reagin recognized were its inability to form precipitins, fix complement, or passively sensitize the guinea pig to While the usual laboratory animals are also not susceptible to passive sensitization of skin with human sera, the typical P-K phenomenon can be demonstrated in the skin of the macaca irus monkey when intravenous injection of Evans blue dyc is used to stain the lesions as in passive cutaneous anaphylaxis. Intravenous injection of antigen simultaneously with the dye is more successful in visualizing lesions, and is recommended over direct intradermal injection of antigen into the tcst site.4 Other properties of reagin which have been helpful in defining its character and in separating it from other antibodies are its therniolability anti inability to pass the placental barrier.j Since it is destroyed by heating at 56 for periods of approximately 4 hours, it may be readily separated from blocking antibody.6 Its failure to cross the placental harrier has been useful in relating it to certain of the immunoglobulins.7 Among the in-vitro tests for the demonstration of antibodies in allergic sera,

The effects on the liver of sensitised rats of intravenous injection of antigen.

The effects on the liver of sensitised rats of intravenous injection of antigen.

Prevention by intravenous injection of antigen and antibody of passive Arthus reaction to unrelated immune system.

Summary1. Previous intravenous injection of antigen and antibody prevents development of passive Arthus reaction to an unrelated immune system in guinea pigs. 2. This suppressive effect does not seem to depend on C'depletion but rather on deficiency of platelets in circulating blood.

A Quantitative Study of the Passive Arthus Reaction in the Rabbit Eye

Summary A typical Arthus reaction may be induced in the rabbit uveal tract by the simultaneous injection of adequate amounts of antigen (crystalline egg albumin or bovine γ globulin) into the anterior chamber or the vitreous and of homologous rabbit antibody intravenously. The reaction is an acute, fibrinous, hemorrhagic uveitis which reaches its peak in less than 24 hr and subsides rapidly. Maximal and minimal reactions required respectively 12 mg and about 1 mg of antibody N (when both eyes reacted simultaneously). Reversed reactions (maximum and minimum) were produced with 0.24 and 0.03 mg approximately of antibody N given in the eye at the same time as intravenous antigen. It was possible to produce both direct and reversed reactions with diphtheria toxoid and horse antitoxin at comparable levels. When antigen or antibody was injected into the vitreous, it persisted for several days. These substances, however, disappeared rapidly from the anterior chamber. Direct reactions elicited one or more days after antigen injection into the vitreous were increased in intensity and occurred with unusually low levels of intravenous antibody. This did not happen with the reversed reaction. Evidence is presented to show that the enhancement might be due to localization or fixation of antigen in a more reactive site. Attempted direct and reversed reactions in the cornea failed completely to produce either a typical Arthus response or a delayed opacity comparable to that resulting from local tuberculin injection in a tuberculin-sensitive rabbit. Instead there occurred an iridocyclitis comparable to that which occurred with vitreous or anterior chamber injections. Arthus reactions were readily produced in the conjunctiva and were accompanied by the formation of a plasmoid aqueous comparable to that seen following the subconjunctival injection of glycerine or hypertonic saline solution. It did not prove possible to produce a reaction of the wheal and flare type by local sensitization with small amounts of antibody and the later intravenous injection of antigen. These findings in the rabbit are contrasted with the results of similar experiments in the guinea pig, in which neither an Arthus nor a wheal and flare type of response could be produced by antigen and homologous rabbit antibody injected in various doses and sites and at various intervals in time. Histamine in the anterior chamber of the guinea pig eye gave a well defined reaction.

Inhibitory Effect of Heparin upon Histamine Release by Trypsin, Antigen, and Proteose.

Anaphylactic, trypsin, and peptone shock are closely related phenomena. The prominent symptoms are similar and the intravenous injection of antigen, trypsin, or proteose into intact animals has been shown to result in the liberation of significant amounts of histamine.1-3 It has also been shown that the addition of antigen, trypsin, or proteose to rabbits' blood (rendered incoagulable with heparin) leads to the release of histamine from cells to plasma.4-9 It is possible that the mechanisms of the release are the same in each case. A study of the effect of various agents in inhibiting or augmenting the release of histamine by the above substances suggests itself as a means of determining whether similar mechanisms are involved. Inasmuch as rabbits' blood can be divided into samples which can be considered identical in every respect, this tissue seems to provide an ideal medium for such studies.It has been shown that heparin is capable of inhibiting the proteolysis of various substrates by trypsin,10, 11 a...

MOUSE BRAIN ANTIGEN

Hellerstrom 1 in 1931 first reported that the intravenous injection of Frei antigen prepared with human pus was followed by a typical febrile response in patients with lymphogranuloma venereum. A similar reaction was not produced in control subjects. Ravaut, Levaditi and Maisler 2 injected monkey brain antigen intravenously. They stated that patients with lymphogranuloma venereum react to intravenous injection of antigen while the intradermal Frei test is still negative. Gay Prieto and Egea Bueno 3 and Flandin and Turiaf 4 reported in favor of its diagnostic value. Findlay 5 expressed the belief that the intravenous test is dangerous. Wlassics 6 found that 30 per cent of his control subjects, chiefly patients with tuberculosis of the skin, gave a positive reaction. The difference in the reports of some of the investigators may be accounted for by differences in potency and amount of antigen used. The question of patients with mixed infections

Late Deaths occurring in Active Anaphylaxis

Conditions are described under which late death from active anaphylaxis occurred in the guinea-pig. The post-mortem appearances were those of gastrointestinal congestion and haemorrhage resembling the changes seen in dogs dying of anaphylactic shock.Late deaths can be produced by the intravenous injection of antigen but more easily by intraperitoneal injections.

The Relationship of Circulating Antibody to the Local Inflammatory Reaction to Antigen (the Arthus Phenomenon)

Summary The results shown in this paper indicate that the presence of circulating precipitin is necessary for local inflammatory reactions of the type described by Arthus. The severity of the Arthus reaction in the skin is directly correlated with the amount of antibody in the blood. When the circulating precipitin disappears with time or is eliminated by an intravenous injection of antigen from an actively sensitized animal known to be capable of giving an Arthus reaction, that animal is no longer sensitive to the skin test. The injection of an amount of antigen just sufficient to remove all the circulating antibody results immediately in the loss of skin sensitivity. All of the evidence obtained indicates that tissue hypersensitiveness in an actively sensitized animal is dependent upon the formation of antibody by the animal and is reflected by the presence of precipitin in the circulation. The importance of antibody in the Arthus test is further brought out in the experiments upon passive transfer of hypersensitiveness. When the precipitin is absorbed from an antiserum, that antiserum loses its power to sensitize passively a normal animal. Such a serum is also unable to elicit a reaction on injection into the skin of a normal animal which has been injected intravenously with the specific antigen. The tissue sensitizing substance in an antiserum appears to be inseparable from and, probably, identical with the precipitin. Several of these observations are contrary to those reported by Kahn in his numerous publications upon the subject and cast doubt upon the conclusions to which Kahn has been led. The existence in the skin or other organs of any sensitizing agency other than the antibody appears not to be essential for an explanation of the mechanism of the Arthus reaction. Such an agency has, furthermore, thus far not been demonstrated. The data here offered supports entirely the contention of Opie that the union of antibody with antigen is responsible for the Arthus reaction.

LOCAL ORGAN HYPERSENSITIVENESS

1. Rabbit eyes sensitized with guinea pig red blood cells or fresh egg white respond with an inflammatory reaction following the intravenous injection of the homologous antigen, but not the heterologous. 2. Two-tenths of 1.0 cc. of a multiple antigen containing ten separate ingredients, or in other words, 0.02 cc. of each foreign protein, when introduced into the rabbit's anterior chamber, is sufficient to produce an altered ocular reactivity such that when 1 cc. of one of the ten antigens is introduced intravenously the eye shows hyperemia of the iris and conjunctiva with more or less edema and lacrimation during the next 24 hours. 3. For as long as 8 months after sensitization eyes will respond with an inflammatory reaction following the intravenous injection of fractions of the total antigen. 4. Repeated daily intravenous injections of a single antigen usually produce no reaction in the sensitized eye after the first few days. Injection of different antigens intravenously on succeeding days produces a continued sterile inflammatory process in the sensitized eye. After the total number of single antigens has been injected, repetition of these injections now fails to produce a similar response. Instead, the eye reaction is at a much lower level and the inflammatory response is manifested only to a few of the antigens injected intravenously. 5. Unless massive doses of antigen are used to desensitize, permanent desensitization of the eye has not occurred in animals which have been followed for at least 8 months. Animals may develop maximal eye responses following repeated intravenous injections of the same antigen, if sufficient time has elapsed between injections. Nevertheless, the eye reactions which can be elicited 6 or 7 months after sensitization are less intense than the initial responses. 6. The ability of the eye inflammation to light up following the intravenous injection of homologous antigen is not due to an initial tissue injury as proven by the fact that the reaction is specific and anterior chambers injured with typhoid vaccine, iodine, saline, glycerine, or albolene will not respond subsequently when the various proteins used for sensitization of the other eyes are injected intravenously. 7. It has been impossible to demonstrate sensitivity in the eye by the intravenous shocking route until at least the 5th day following introduction of the antigen into the anterior chamber. 8. The eye reaction can be produced by the subcutaneous as well as the intravenous injection of antigen. 9. Rabbits vary considerably in the intensity of the eye reaction which can be elicited in them, but only rarely was an animal found which failed completely to give a reaction.