the passive transfer of immediate type hypersensitivity...

Journal of Clinical InvestigationVol. 44, No. 1, 1965

The Passive Transfer of Immediate Type Hypersensitivityfrom Man to Other Primates *

RoY PATTERSON,t JORDANN. FINK, EDWIN T. NISHIMURA,$ ANDJACOB J. PRUZANSKY

(From the Allergy Unit, Department of Medicine, and the Department of Pathology,Northwestern University Medical School, Chicago, Ill.)

There have been various obstacles to investi-gating the immediate type of spontaneous hyper-sensitivity in man. These have included the lackof in vitro methods of measuring allergic antibodyand a limitation on the types of investigations thatcan be done in man without risk. Models of hy-persensitivity using immunized animals have notinvolved antibody of the type occurring in spon-taneous allergy in man. The passive transfer ofimmediate cutaneous reactivity [Prausnitz-Kustnerreaction ( 1 ) ] to other human subjects has re-mained the classical method of investigation of al-lergic antibody. This technique is limited to thestudy of cutaneous wheal and erythema reactionsand carries with it the risk of transmission ofcertain viral diseases. By this method, allergicantibody has been characterized as a nonprecipi-tating, heat labile, skin-sensitizing antibody thatpersists at the injected site for long periods oftime (2). The nature of the serum fraction con-taining allergic antibody, believed to be a y1A typeof immunoglobulin by several investigators (3),has been the subject of controversy in regard toeither electrophoretic migration or sedimentationcharacteristics as demonstrated by ultracentrifugalanalysis.

The availability of an animal model with the im-mediate type of spontaneous hypersensitivity asfound in dogs with spontaneous pollen hyper-

* Submitted for publication July 13, 1964; acceptedSeptember 24, 1964.

Supported by U. S. Public Health Service researchgrant AI-04199 from the National Institute of Allergyand Infectious Diseases.

t Career Development awardee, U. S. Public HealthService no. 5-K3-AI-19,460-02.

t Career Development awarded, U. S. Public HealthService no. GMK3-244-CS. Present address: Depart-ment of Pathology, School of Medicine, University ofCalifornia, Los Angeles.

sensitivity has provided a means of investigationof the immunopathology resulting from the inter-action of allergic antibody and antigen. Withthis model, the hypersensitivity has been trans-ferred to normal dogs (4) and the canine allergicantibody characterized and compared with humanallergic antibody (5).

The immediate type of spontaneous hypersensi-tivity, a common clinical occurrence, is most fre-quently directed against materials that are mix-tures of poorly characterized antigens such as pol-len or animal epithelial proteins. The occurrenceof immediate wheal and erythema type of hyper-sensitivity in certain patients to animal proteinsthat are available in relatively pure form has pro-vided a means of studying single antigen-allergicantibody systems (6, 7).

It has been reported that allergic antibody (re-agin, skin-sensitizing antibody, atopic antibody)from man will sensitize the skin of other primates(8). This transfer of cutaneous reactivity to aheterologous species has recently been directedtoward the investigation of cutaneous reactions ofvarious antigens and their respective skin-sensi-tizing antibodies in monkeys (9, 10).

Based on the previous observation of passivecutaneous sensitization of monkeys by local in-jection of human allergic sera, the present studiesextend these observations to demonstrate passivesystemic sensitization by human allergic antibody.The antigen-allergic antibody reaction in the pas-sively sensitized primate recipients with resultantrelease of histamine, local or generalized increasein vascular permeability, and smooth muscle re-action is correlated with symptoms of broncho-spasm, conjunctivitis, or anaphylaxis. Micro-scopic abnormalities of lung and myocardium arerecorded that result from the hypersensitive re-sponse. The study provides comparative observa-

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PASSIVE TRANSFEROF HUMANHYPERSENSITIVITY TO OTHERPRIMATES

tions between immediate hypersensitivity reactionsas a result of antigen-allergic antibody combinationin dogs and primates.

MethodsPrimate recipients. Four Macaca irus, three Macaca

mulatta (rhesus), and one Cercopithecus aethiops (Afri-can green) monkey were used in these experiments.All of these had been shown (10) to be suitable cu-taneous recipients for human reaginic serum. All monkeysappeared in good general health and had negative reac-tions to tuberculoprotein skin testing.

Serum donors. Serum was obtained from two patientswith ragweed pollinosis and one patient with anaphylac-tic sensitivity to horse serum. Characteristics of serumdonors are shown in Table I. Included for comparisonare the results obtained with canine reaginic serum froma dog with ragweed pollinosis (4).

Antigens. Ragweed pollen antigen was prepared bystandard techniques (11) and expressed as weight of pol-len extracted per volume of final extract. Horse serumalbumin (HSA) and horse gamma globulin (HGG)were obtained commercially.' Nitrogen (N) determina-tions were done by the Markham modification of themicro-Kjeldahl method (2).

Passive cutaneous reactions. One-tenth ml of varioussera was injected intracutaneously. Forty-eight hourslater the monkeys received 4 ml of 0.5% Evans blue dyeintravenously followed in 15 minutes by intracutaneousinjection of 0.05 ml of 0.1% pollen antigens or HSA orHGGcontaining 50 /Ag N per ml. Sites were examinedin 30 minutes, and positive reactions were characterizedby edema and cutaneous blueing (Figure 1).

Systemic sensitization of monkeys. Monkeys weresensitized by intravenous injection of reaginic sera individed doses of 25 or 40 ml per kg body weight. Theinjections were given at daily intervals, and animals werechallenged 48 hours after the last injection. Anaphy-lactic challenge consisted of intravenous injection of 2 mlof 3% ragweed antigen or 2 ml containing 5 mg HSAN preceded by intravenous injection of 4 ml of 0.5%oEvans blue dye. The volume and concentration of rag-weed antigen chosen for intravenous anaphylactic challengewere based on the amount of antigen resulting in canineanaphylaxis in dogs passively sensitized with canine anti-ragweed antiserum. The amount of HSA chosen forchallenge was arbitrarily selected as an amount thatwould presumably be in excess of that required to pro-duce anaphylaxis. That quantities of both antigens werein excess was demonstrated by loss of reactivity to localcutaneous testing in those passively sensitized monkeyssurviving anaphylaxis.

Aerosol exposure of animals. After total body sensi-tization by human allergic serum, unanesthetized monkeyswere placed in an Isolator chamber and aerosolized with3% ragweed delivered by a no. 40 DeVilbiss nebulizerin a manner previously described for similar experiments

1 Pentex Corp., Kankakee, Ill.

using dogs (4). Changes in respiratory rate and char-acter were recorded graphically by a pneumograph andrecording kymograph in the unanesthetized, restrainedmonkeys. One monkey (monkey D, Table II) re-ceived 4 ml of 0.5 % Evans blue dye and 10 mg of pento-barbital per kg body weight, after sensitization with 100ml of human serum. During exposure to ragweed anti-gen in the aerosolization chamber, pneumograph re-cordings were made. After sacrifice by injection of ex-cess pentobarbital, the trachea and bronchi were dissected.A control monkey was treated in similar fashion withthe exception of passive sensitization by human allergicserum.

FIG. 1. MONKEYPASSIVELY SENSITIZED BY LOCAL IN-JECTIONS OF SERIAL FIVEFOLD DILUTIONS OF HUMANORDOG ALLERGIC SERUM. A) Human antiragweed serum(Donor 1), B) human antihorse serum albumin (Donor3), C) heated serum (Donor 1), D) canine antiragweedserum (Donor 4). See Table I. Sites in A and B andfirst dilution of D are positive.

141,

PATTERSON, FINK, NISHIMURA, AND PRUZANSKY

TABLE I

Some characteristics of donors of serum and the serum used for passive sensitization of monkeys

PassivePassive trans- transfer*

Cutaneous reactivity fer* to human to monkeyDonor Clinical history of donor skin skin

1 Ragweed pollinosis with sea- Ragweed + + + + 1:1,250 1:125sonal rhinitis and asthma

2 Ragweed pollinosis with sea- Ragweed ++++ 1:1,250 1:25sonal rhinitis and asthma

3 Rhinitis and asthma after ex- Horse serum albumin,tposure to horses and dogs; 0.08 ,ug N/mi 1:1,250 1:125systemic reaction of asthma,urticaria, and hypotension Horse gammaglobulin,tafter accidental contact of 1.2 sg N/ml 1:10 0horse serum with conjunctiva

4 Dog with spontaneous ragweed Ragweed Positivet 1:5pollinosis with dermatitis,rhinitis, and asthma (seereference 4)

* Recorded as highest serum dilution giving positive titer.t Recorded as minimal amount of antigen giving positive reaction on direct cutaneous testing.t Titer in human skin not done. Positive in transfer to dog skin to a dilution of 1:125.

Blood histamine determinations. Blood histamine lev-els were determined by the method of Shore, Burkhalter,and Cohn (12). During anaphylactic shock histaminelevels were determined on blood samples obtained bycardiac puncture.

Post-mortem examinations were done within 30 min-utes of spontaneous death or sacrifice of monkeys.

Results

Local cutaneous reactivity

Examples of cutaneous reactivity transferredby intracutaneous injections of human sera areshown in Figure 1. Columns A and B demon-strate reactions at transfer sites and the method oftitering passive transfer reactions. Column Cshows the destruction of reaginic antibody byheating reaginic serum to 56° C for 4 hours be-fore passive transfer experiments. In this monkeyeven undiluted canine antiragweed serum gaveonly a weakly positive reaction (column D), butin other experiments the same canine allergic se-rum was positive to a dilution of 1: 5. Table Irecords certain characteristics of serum donorsand their allergic antisera. The comparativeability of the human or canine allergic antisera totransfer reactivity to homologous or heterologousskin is demonstrated by the highest dilution of theantisera that can still successfully transfer cutane-ous reactivity. Transfer of the cutaneous reac-tivity to the heterologous recipient was less suc-

cessful with either the human or canine allergicantisera. Whether this difference in ability tosensitize heterologous skin is a result of variationin response of monkey skin to the antigen-antibodyreaction, inability of human and canine reagin tofix to skin of monkeys to the same degree as inhuman skin, or other explanation, is uncertain.

Systemic sensitization of monkeys by reaginicserum

Monkeys receiving a sufficient amount of re-aginic serum intravenously developed systemicsensitization that could be demonstrated by sev-eral methods.

A) Local cutaneous reactions. The monkeyssensitized with antiragweed or anti-HSA reagindeveloped systemic sensitivity demonstrated bywheal and erythema type reactions after intra-cutaneous challenge with either antigen.

B) Mucosal reactivity. Objectives of this por-tion of the investigation were to demonstratethat immediate type sensitivity in conjunctivalor respiratory mucosa occurs after passive sensi-tization by intravenous administration of suffi-cient amounts of reaginic antiserum.

1) Conjunctival reactivity. After intravenousadministration of 4 ml of 0.5% Evans blue dye,2 drops of 3% ragweed antigen placed in oneconjunctival sac of animals passively sensitizedwith human antiragweed serum resulted in intense

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blueing reaction of the tested conjunctiva. Thecontrol conjunctival sac tested with an extraneous

antigen appeared unchanged after 30 minutes.2) Bronchial reactivity. Monkeys sensitized

by intravenous antiragweed serum received 4 mlof 0.5% Evans blue dye and were exposed byaerosol to 3% ragweed antigen in a closed en-

vironment. Clinical manifestations consisted ofincreased respiratory rate, prolonged expiratoryphase, and intermittent grunting respirations.Flaring of the nasal alae during the acute phaseoccurred in all animals. Expiratory wheezes at

termination of the experiment were present insome monkeys. Exposure of normal monkeysunder similar conditions resulted in no symptoms.

Dissection of bronchi from a passively sensitizedmonkey exposed to ragweed under the above cir-cumstances demonstrated a blueing reaction of thebronchial mucosa. The appearance of the bron-chial mucosa is similar to the appearance of thereactive skin sites in passive cutaneous anaphylaxis,and it is likely that these changes represent ex-

travasation of dye due to an antigen-antibody re-

action in the bronchial mucosa similar to that oc-

curring in the skin sites in a passive cutaneous

anaphylactic reaction. The clinical alterationsoccurring as a result of aerosol exposure to rag-

weed antigen can be observed by graphic record-ings with a pneumograph. The recordings dem-onstrated that tachypnea and some change in thecharacter of respirations occur after exposure toaerosolized ragweed antigen. This model of in-duced respiratory distress should provide a valu-able system for study of acute physiologic altera-tions. The respiratory changes as a result of thepassive sensitization and aerosol exposure to anti-gen may, therefore, be studied by clinical observa-tion, graphic recording of thoracic respiratorymovements, and acute changes in the appearance

of bronchial mucosa demonstrated by the blueingreaction.

C) Anaphylaxis. Monkeys systemically sensi-tized by the administration of human allergic se-

rum developed anaphylactic shock as a result ofintravenous challenge with specific antigen (Ta-ble II). Manifestations of anaphylaxis includeddiarrhea, vomiting, collapse, dyspnea, and death.All of these symptoms were not present in eachshocked animal. A striking change occurred insome animals receiving Evans blue dye before

anaphylactic challenge. Associated with the oc-currence of clinical manifestations in somemonkeys, there was a generalized cutaneous blue-ing indicating an increase in capillary permeabilityin the skin. Other monkeys in anaphylaxis de-veloped irregular areas of local cutaneous edemawith an appearance similar to urticaria, and amore marked localized blueing of these areas waspresent than occurred in generalized blueing.The generalized cutaneous reaction appeared to beassociated with more severe anaphylaxis than thelocalized urticarial type.

The blood histamine concentration of the nor-mal monkey was in the range of 0.07 ug histamineper ml. (Normal dog blood histamine in thislaboratory ranges from 0.03 to 0.09 jtg of hista-mine per ml). During the period of anaphylacticshock, blood histamine ranged from 0.29 jug hista-mine per ml (monkey A, Table II) to 0.53 jughistamine per ml (monkey C, Table II). Thus,approximately a four- to eightfold increase in bloodhistamine concentration occurred during anaphy-laxis. Lee-White clotting times (13) showed nochange during the period of anaphylaxis in con-trast to the infinite clotting times observed duringacute canine passive anaphylaxis (4).

Post-mortem abnormalities in monkeys afteranaphylactic shock were observed only in theheart and lungs. Microscopic examination ofliver, kidneys, spleen, and large and small intes-tines of monkeys C and F were done and showedno definite abnormalities. Microscopic examina-tion of lungs from monkeys C, D, F, and G showedacute emphysema. Thus, acute emphysema oc-curred whether there was an intravenous challengealone (monkey G), aerosol challenge alone (mon-key D), or both (monkeys C and F). Abnormali-ties were observed in the hearts of all monkeysfollowing anaphylaxis. The microscopic changesobserved in these hearts are recorded in Table III.Figure 2 depicts an example of a myocardial lesionwith a focal infiltrate of lymphocytes and neutro-philes. Although the pericardial adipose tissueshows some inflammatory reaction, the myocar-dium constitutes the primary site of the exudativereaction. Arteritis, focal necrosis, and inflamma-tory cells in the myocardium of monkey D werepresent although challenged only by aerosol ex-posure to antigen. However, the concentrationof aerosolized ragweed antigen used in these

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TABLE III

Summary of pathological findings in myocardium of monkeys after passive sensitizationand aerosol or intravenous challenge

InflammatoryMonkey* Vessels Hyperemia Hemorrhage Necrosis cells

C Moderate arteritis 4+ 4+ focal 3+ Mt 2+Moderately Nt 2 +

diffuseD Slight arteritis None None 4+ focal M 3+

N 4+E None None None None None

(Control)F Focal arteritis None None None NoneG Endarteritis 3+ 2 + focal 2 + focal M 2+

N 2+H None None None None None

(Control)

* See Table II.t M = monocytic cells.

N = neutrophilic granulocytes.

experiments was in great excess of that whichwould be encountered in environmental exposure.

Loss of capacity to accept sensitization

One month after the experiment recorded inTable II, monkeys A and B, which had previouslybeen passively sensitized by intravenous injectionof 120 or 75 ml of human allergic sera, re-spectively, were injected with human sera, thistime locally with 0.1 ml of sera from Donors 1,2, and 3 and these sites tested for the presence ofimmediate type sensitivity to ragweed, HSA, orHGG. Reactions were negative, indicating theloss of ability to accept skin sensitivity as a re-sult of prior injection of human serum. The ini-tial injections of human sera that resulted inpassive sensitization of the monkeys had also ac-tively sensitized the monkeys. Thus, after a latentperiod, the injection of additional human reaginicserum failed to sensitize the monkey skin. Seracollected from monkeys A and B were testedfor the presence of precipitating antibody againsthuman serum, human gamma globulin, and hu-man serum albumin by ring precipitin and Ouch-terlony type double gel diffusion precipitin reac-tions (2). No precipitating antibody was detectedby these tests, but the sera of monkeys A and Bdemonstrated the presence of circulating antibodyagainst human serum protein antigens by sensi-tization of normal monkey skin for passive cu-taneous anaphylactic reactions against human se-

rum. In addition, monkeys A and B demon-strated symptoms of anaphylaxis accompanied bythe generalized cutaneous blueing reaction whenchallenged intravenously with 2 ml of human se-rum. This reaction never occurred in monkeysat the time of the first injection of human serum.

DiscussionMethods of studying allergic antibody include

the passive transfer of cutaneous reactivity in man

FIG. 2. A SECTION OF EPICARDIAL FAT ANDMYOCARDIUMSHOWINGA FOCAL INFILTRATE OF LYMPHOCYTESAND NEU-TROPHILES. The primary site of the inflammatory reac-tion is in the myocardium. X 80.

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and the transfer of cutaneous and total sensitivityin dogs. These models provide a means of com-parative study of this antibody. The transfer oflocal cutaneous and total body sensitivity by hu-man allergic serum to monkeys provides a thirdmajor model for the study of allergic antibody.

The events presumed to occur in a passive cu-taneous reaction after sensitization with allergicantibody in man include the attachment of allergicantibody to skin cells, reaction of injected antigenwith antibody, the release of chemical mediatorsfrom cells, and resultant increase in capillarypermeability. The latter is demonstrated by thecharacteristic wheal reaction and made more visibleby extravasation of Evans blue dye. The passivecutaneous anaphylactic reaction of Ovary (2) is asimilar reaction as a result of induced, skin-fixinganimal antibody. The allergic antibody occurringin man will attach to normal human leukocytesin vitro by passive sensitization as detected byhistamine release determination (14).

Canine allergic antibody from dogs with rag-weed pollinosis, asthma, and dermatitis will pas-sively transfer cutaneous, bronchial mucosal, andanaphylactic sensitivity (4). Histamine levelsand pathologic changes during anaphylaxis indogs passively sensitized with this canine reaginicserum can thus be compared with induced anaphyl-axis in the dog in which the antibody has not yetbeen defined. This model of the immediate type ofhypersensitivity provides a useful comparison be-tween the immediate type of hypersensitivity oftwo species, man and dog.

Human allergic antibody has the following im-munologic characteristics: it is a nonprecipitatingantibody, which is heat labile at 56° C, fixes toskin, and persists at the skin site (3). Canineallergic antibody has identical characteristics incompleted studies using presently developed im-munologic techniques (5). Although in vitromethods of detecting antibody in human allergicsera have evolved ( 15 ), it has not been establishedthat these in vitro techniques actually measure al-lergic antibody.

As demonstrated in the experiments described,the injection of sufficient amounts of human al-lergic serum into monkeys results in total bodysensitization. The monkey that has been totallysensitized in this manner can react to antigenicchallenge with the specific antigen-human antibody

reaction in a manner similar to the sensitive hu-man. The use of the monkey for this type ofsensitization is obligatory, since no other non-human species has been found to accept the trans-fer of human allergic antibody. Skin-sensitizingactivity of canine antisera will transfer to man andmonkey, but human allergic antisera will not trans-fer to dog, guinea pig, or rabbit.

The availability of the monkey sensitized withhuman serum provides several potentially usefulareas of investigation. The cutaneous reactionhas been used as a method of antigen evaluation(9, 10) and study of fractions of allergic antisera(16). The use of Evans blue dye as an indicatorin these reactions provides an indication of whealand erythema reactivity, making a reaction morevisible for graphic recording. The extravasationof Evans blue dye at the site of cutaneous reac-tion is an indication of the increased capillarypermeability as a result of the antigen-antibodyreaction and release of mediators. This occurs inIa local mucosal reaction demonstrated in these ex-periments in which antigen was placed in contactwith the conjunctival mucosa. Blueing of thebronchial mucosa occurs in those monkeys withsystemic sensitivity after exposure to specific anti-gen in aerosol form with subsequent developmentof symptoms of respiratory distress. Finally,during systemic anaphylactic shock in the passivelysensitized monkey injected with Evans blue dyebefore antigenic challenge, a cutaneous blueingreaction is consistently observed. This varies indifferent animals from a generalized blueing tocircumscribed areas of cutaneous blueing with anappearance not dissimilar from that of acute urti-carial reactions, as may be seen in anaphylaxis inman. The generalized blueing suggests the oc-currence of an increase in cutaneous capillarypermeability as the result of the systemic reaction.

The events and mechanisms resulting in anaphy-lactic shock in the passively sensitized monkey ap-pear to be consistent with the following patho-physiologic pattern. After intravenous injectionof human allergic serum, reaginic antibody resultsin systemic sensitization with fixation of the anti-body to cells demonstrated by the general cutane-ous and bronchial reactivity to antigen. Afterintravenous challenge, the antigen-allergic antibodyinteraction results in histamine release demon-strated by increase in blood histamine levels. The

146


subsequent increase in vascular permeability dem-onstrated by generalized cutaneous blueing is fol-lowed by clinical symptoms of shock. The con-comitant occurrence of smooth muscle spasm isshown by symptoms of vomiting, micturition, anddiarrhea in varying degree, and the pulmonarysymptoms and pathologic demonstration of pul-monary emphysema may represent acute changesresulting from the smooth muscle spasm in thebronchi. The variations in type or severity ofsymptoms and variation in severity of pathologicchanges represent biologic differences between ex-perimental subjects. The occurrence of the patho-logic changes of acute emphysema in some of themonkeys examined after anaphylactic shock rep-resents pulmonary abnormalities that may havecontributed to the systemic symptoms of anaphy-laxis. Since the symptoms of anaphylactic shockdo not occur in passively sensitized monkeys chal-lenged by aerosol alone, it appears reasonable toconsider the anaphylactic shock primarily the re-sult of the intravenous challenge, with pulmonarymanifestations a secondary phenomenon.

The pathologic abnormalities in the monkeysafter passive anaphylaxis represent changes re-sulting from the specific conditions of method ofpassive sensitization, challenge, duration ofanaphylaxis, and whether death occurs spontane-ously. The acute emphysema demonstrated inthese monkeys after anaphylaxis is an abnormalitynot unexpected in view of the respiratory symp-toms occurring during shock and similar observa-tions in some humans (-17) and some dogs (4) af-ter spontaneous or experimental anaphylaxis.The myocardial abnormalities observed in all sen-sitized monkeys exposed to the respective antigenare striking in terms of consistency and character.These changes appear to represent primary changesas the result of the antigen-allergic antibody inter-action. Similar myocardial changes have not beendescribed in man (17). Such myocardial abnor-malities may represent specific species localizationof pathologic changes in the monkey in anaphy-laxis [comparable to the liver, gall bladder, andintestinal tract abnormalities in the dog (4, 5)],or similar abnormalities may be found when largerseries of post-mortem studies of human anaphy-laxis are available. Further investigation of themyocardial abnormalities described in these ex-periments is in progress. Studies to correlate

the myocardial abnormalities with severity andduration of anaphylactic shock are required.

The duration of passively transferred systemicsensitivity will depend on the antibody contentof the sensitizing serum, amount of serum given,and the half-life of human reaginic antibody (whichhas not yet been determined) in the monkey. Fi-nally the duration of the passive sensitivity in themonkey depends on the development of monkeyantibody against the sensitizing serum and im-mune elimination of the sensitizing antibody fromthe monkey. The present experiments indicatethat monkeys receiving amounts of serum usedin these studies cannot be used for sensitizationafter 1 month; two monkeys developed sensitivityto the human sera used for passive immunization.

The antibody developed by the monkeys againsthuman serum was demonstrated only by activeanaphylaxis or passive cutaneous anaphylaxis andnot by precipitation reactions. Monkeys im-munized with human serum in Freund's adjuvantdevelop precipitating antibodies (18). The dif-ference between these immunologic responses ofmonkeys is probably dependent upon the adjuvantadministered with the antigenic serum.

Summary

Total antibody sensitization of monkeys occurswith sufficient passively transferred human allergicserum. Cutaneous, respiratory, and anaphylacticreactions can be studied with this model of humanhypersensitivity. The symptoms, blood histaminelevels, and pathologic changes in the passivelysensitized monkey can be compared with a similartype of reaction in the dog. Microscopic patho-logic changes occurring in anaphylaxis in monkeyssensitized by human allergic sera were acute pul-monary emphysema and arteritis, focal necrosis,and focal hemorrhages in the myocardium.

References1. Prausnitz, C., and H. - Kfistner. Studien fiber die

Ueberempfindlichkeit. Zbl. Bakt. I. Abt. Orig.1921, 86, 160.

2. Kabat, E. A., and M. M. Mayer. Experimental Im-munochemistry. Springfield, Ill., Charles CThomas, 1961.

3. Stanworth, R. A. Reaginic antibodies in Advances inImmunology, F. J. Dixon, Jr., and J. H. Humphrey,Eds. New York and London, Academic Press,1963, vol 3.

147


4. Patterson, R., and D. B. Sparks. The passive trans-fer to normal dogs of skin reactivity, asthma andanaphylaxis from a dog with spontaneous rag-weed pollen hypersensitivity. J. Immunol. 1962,88, 262.

5. Patterson, R., J. J. Pruzansky, and W. W. Y.Chang. Spontaneous canine hypersensitivity toragweed. Characterization of the serum factortransferring skin, bronchial and anaphylacticsensitivity. J. Immunol. 1963, 90, 35.

6. Patterson, R., J. J. Pruzansky, and S. M. Feinberg.Studies on reactions of human allergic serum withserum protein antigens. I. Methods of passiveimmune elimination and gel diffusion autoradiog-raphy. J. Allergy 1962, 33, 236.

7. Pruzansky, J. J., R. Patterson, and S. M. Feinberg.Studies on reactions of human allergic serumwith serum protein antigens. II. Method ofquantitative demonstration by a coprecipitationtechnique. J. Allergy 1962, 33, 381.

8. Straus, H. W. Studies in experimental hypersensi-tiveness in the rhesus monkey. II. Passive localcutaneous sensitization with human reaginic sera.J. Immunol. 1937, 32, 251.

9. Layton, L. L., and E. Yamanaka. Demonstration ofhuman reagins to foods, cat dander, an insect, andragweed and grass pollens. J. Allergy 1962, 33,271.

10. Layton, L. L., W. E. Greer, F. C. Greene, and E.Yamanaka. Passive transfer of human atopicallergies to catarrhine and platyrrhine primates ofsuborder anthropoidea. Int. Arch. Allergy 1963,23, 176.

11. Hepler, 0. E. Manual of Clinical Laboratory Meth-ods. Springfield, Ill., Charles C Thomas, 1949.

12. Shore, P. A., A. Burkhalter, and V. A. Cohn, Jr.A method for the fluorometric assay of histaminein tissues. J. Pharmacol. exp. Ther. 1959, 127,182.

13. Todd, J. C., and A. H. Sanford. Clinical Diagnosisby Laboratory Methods. Philadelphia and London,W. B. Saunders, 1948.

14. VanArsdel, P. P., Jr., and C. J. Sells. Antigenichistamine release from passively sensitized hu-man leukocytes. Science 1963, 141, 1190.

15. Pruzansky, J. J., and R. Patterson. Binding ofI'-labeled ragweed antigen by sera of ragweed-sensitive individuals. J. Allergy 1964, 35, 1.

16. Buckley, R. H., and R. H. Metzgar. The use of non-human primates for studies of reagin. Fed. Proc.1964, 23, 402.

17. James, L. P., and K. F. Austen. Fatal systemicanaphylaxis in man. New Engl. J. Med. 1964,270, 597.

18. Lichter, E. A., and S. Dray. Immunoelectrophoreticcharacterization of human serum proteins withprimate antisera. J. Immunol. 1964, 92, 91.

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