EXPERIMENTS ON PASSIVE TRANSFER OF ECZEMATO[JS ALLERGY* H. HAXTHAUSEN, IvLD.
In 1949 II. S. Lawrence (1) described a method by which he had been able in man to transmit tuberculin allergy by intracutaneous injection of white blood cells from a positively reacting donor to a previously negative receptor. Lawrence usually employed 60—100 ml. heparinized blood to which a solution of bovine fibrinogen was added. This procedure accelerates the sedimentation of the red blood cells, so that the major part of the white cells may be isolated by centrifugation of the plasma fraction. These results revived the interest in experiments on passive transmission of eczematous allergy, which indeed has several points of resemblance to tuberculin allergy. Thus it is a well-known fact that intracutaneous injection of eczematogenous allergens—e.g., nickel, chromium and mercury salts, as well as sulfonamides—in hypersensitive individuals produces a late intracutaneous reaction which clinically and histologically quite resembles a positive tuberculin reaction. Although previous experiments on passive transmission of eczematous. allergy with suspension of cells from excised lymph nodes and with white blood cells
obtained through centrifugation of the blood had failed to give any definite positive results (Haxthausen (2)), it was still decided to carry out some additional experiments with a technic similar to the one employed by Lawrence. As bovine fibrinogen was not available to us, we used instead an addition of 10 ml. gum-
arabic saline to 100 ml. heparinized blood. This gives a quick sedimentation so that it is practicable to obtain a considerable part of the white blood cells after 1 hour of centrifugation of the plasma fraction. The proportion between granulocytes and lymphocytes in this fraction is about the same as found in a fresh sample of normal blood. As numerous preceding experiments had shown that the serum or plasma is unable to transmit the effect concerned, in the following experiments the cells were not washed—in order to injure them as little as possible. The precipitate plus the small amounts of plasma adherent were suspended in 0.5 ml. saline, and this suspension was injected intracutaneously in amounts of 0.1 ml. on the back of the experimental subjects, who beforehand had been tested with the allergen concerned and found to give a negative reaction. As a control, a corresponding amount of white blood cells from a normal subject or from a patient with eczematous allergy to some other substance was injected into a symmetrical area of the skin.
After 24 hours all the injections gave rise to red papular infiltrations of the skin, sometimes fairly large in size. But the real test for hypersensitiveness was not performed until this non-specific reaction—which, as a matter of fact, appeared also on use of cells from the subject himself—had subsided, i.e., after 2—4 days. * From
the University Clinic of Dermatology, Rigshospitalet, Copenhagen (Chief Professor H. Haxthausen, M.D.). Received for publication April 8, 1952. 293
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The test proper consisted in application of patch tests to the areas of the experimental and control injections. On employment of soluble allergens—e.g., metal salts—additional intracutaneous tests on corresponding areas were performed with solutions of suitable concentration. Finally, a patch test and, when indicated, an intracutaneous injection of allergen were also applied to normal skin. In experiments carried out with cells from patients affected with different forms of allergy, cross-tests were carried out with a corresponding technic. The reactions were examined after 24 and 48 hours; the intracutaneous reactions also 15 and 30 mm. after the injection. In addition to these studies in eczematous allergy, experiments on passive transfer were carried out also in the cases of 9 subjects giving a strong positive tuberculin reaction and 3 patients giving a strongly positive trichophytin reaction. A priori the possibility could not be excluded altogether that the gum-arabic saline might be somewhat injurious to the white blood cells. On this account the experiments were supplemented by a number of tests, each carried out at the start with 200 ml. heparinized blood, from which the white cells were obtained with fractionated centrifuging (1000 r.p.m. for 10 mm.) without any addition whatever. In this way, it proved possible to obtain the white blood cells in
about the same total amount as in the gum-arabic saline experiments. The number of donors with eczematous allergy and the character of the allergen in the respective cases are evident from Table 1. Because of the scanty injection material, as a rule, only one receptor was employed in each of these experiments; and only in 3 cases were two receptors employed. RESULTS
The results of the 66 passive transfer experiments turned out perfectly negative. In 1 case (nickel allergy) both the patch test and the intracutaneous test yielded slightly positive reactions at the sites of the test proper, whereas the controls turned out negative. Just in this case, however, the primary infiltration from the injected cell-suspension was considerably more pronounced at the sites of the
tests (injected cells from hypersensitive donor) than at the sites of control injected white cells from "normal" donor. On this account it seems only reasonable to assume that here it was a matter of a non-specific reaction.
In another case (turpentine allergy) the patch test produced after 4 days a slightly positive reaction; but this was seen not only at the site of the test proper but also at the control site and on untreated skin areas. Most likely, we were here dealing with an early development of sensitization induced by the patch test which was applied prior to the experiment proper (see addendum on recent studies by Baer and Sukberger). In 4 cases—namely, 1 of allergy to chromium, 1 of allergy to mercury and 2 of allergy to lucosil—a quick so-called "immediate"
reaction was seen after intracutaneous injection at the site of the test—i.e., an urticarial lesion surrounded by reflex erythema, distinctly more pronounced
than seen at the control site. In contrast hereto, the reverse condition—i.e., a stronger wheal reaction at the control site—was not observed in any instance. In one of the above-mentioned four cases of allergy (here chromium allergy) 2
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receptors were employed, but only one of them presented the above-described difference mentioned in whealing. Whether we are here meeting with accidental differences in the reactivity of the subject, or whether some specific effect may be asserting itself, cannot be decided on. Although this phenomenon was observed but seldom, I think it still deserves more thorough investigation. Finally, it is noteworthy, that with the technic here employed all attempts at transmission of hypersensitiveness to tuberculin itself turned out to be failures. Addendum
After the conclusion of the present work a paper was published by Baer and Sulzberger (3), reporting their investigations of the same problem with employment of the technic originally given by Lawrence. Out of the total 27 experiments on transmission of the allergy, 21 turned out negative, in 2 the outcome was dubious, and in 4 cases the patch test was found to be positive—but, nota bene, it was positive at the site of control and at uninjected skin sites as well as at the TABLE I Number of Donors and Character of the Allergy The first figures give the numbers of experiments with gum arabic; those in parentheses give the numbers of experiments without employment of gum-arabic saline.
A. Bichromate 6 (2). Mercury 6 (3). Nickel 15 (3). Percaine 1 (0). Nupercaine 0 (2). Quinine 0 (1). Lucosil (a soluble sulfonamide) 0 (3). Dinitrochlorbenzene 1 (0). B. Nitrobenzyl chloride 1 (0). Coal tar 1 (0). Adhesive tape 2 (0). Aetherol. portugalli 1 (0). Turpentine 1 (0). Asparagus 1 (0). Mercaptothiobenzene 1 (0). Leather 1 (0). Peruvian balsam 0 (1). Primula 1 (0). C. Trichophytin 2 (1). Tuberculin 6 (3). Group A comprises the cases on which the patch test was performed as well as the intracutaneous reaction. Group B includes the cases in which only the patch test was performed. Group C comprises 12 cases in which only the intracutaneous test was performed.
site of the test proper. The authors discuss the possibilities of an adequate explanation of these positive reactions. They speculate that it may have been a matter of passive sensitization; but it is also possible that the injected white blood cells from the allergic donor might somehow increase the capacity of the normal skin for active sensitization. Out of these 4 positive cases 3 involved allergy to turpentine. One of my own cases—strange to say, also a case of allergy to turpentine—gave similar positive reactions. So, perhaps turpentine occupies a special position as regards the appearance of this phenomenon. SUMMARY AND CONCLUSION
Attempts at passive transmission of eczematous allergy through intracutaneous injection of white blood cells obtained from heparinized blood after addition of gum-arabic saline, sedimentation and centrifugation have yielded negative re-
sults. Only in two cases was a slightly positive reaction observed and this could readily be explained in some way other than due to passive sensitization.
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Also experiments on transmission of tuberculin allergy turned out negative. In four cases the intracutaneous test injection was followed by an "immediate" wheal reaction, i.e., an urticarial lesion that was more pronounced than at the site of the control, whereas the reverse has not been observed so far. As this phenomenon has been seen in such a few cases it would hardly be safe at present to attach any particular significance to it, even though it deserves more thorough investigation. REFERENCES 1. LAWRENCE, H. S.: The cellular transfer of cutaneous type sensitivity to tuberculin in man. Proc. Soc. Exper. Biol. & Med. 71: 516, 1949. 2. HAXTHAIJSEN, H.: Studies on the role of the lymphocytes as "transmitter" of the hypersensitiveness in allergic eczema. Acta derma. yen. 27: 136, 1947. 3. BAmt, R. L. AND SULZBERGSR, M. B.: Attempts at passive transfer of allergic eczematous
sensitivity in man. J. Invest Dermat. 18: 53, 1952.
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