The Role of Basophils in Inflammatory Reactions

LAWRENCE M. LICHTENSTEIN, M.D, GIANNI MARONE, M.D., LARRY L. THOMAS, PH.D., AND FLOYD J. MALVEAUX, M.D. Department of Medicine, Division of Clinical Immu.nology, The Johns Hoplzins University, School of M edicine at the Good Samaritan Hospital, Baltimore, Mwy land, U.S.A.

This review demonstrates that basophils reflect skin out an important tissue function of the basophil that may, in and lung mast cell reactivity and show characteristic the end, overshadow the functions of this cell type in the changes in mediator release associated with clinical dis ci.r'culatory system [2,3]. In spite of these uncertainties, the ease. Although the numbers of IgE molecules and IgE basophil remains of vital interest and it may play an important receptors on basophils have been enumerated, these role in both physiological and pathological processes. have, in most instances, little influence on the release of histamine after challenge. There is, rather, a parameter RELATIONSHIP BETWEEN BASOPHIL FUNCTION of "releasability" that may be a major variable in allergic AND SYSTEMIC RESPONSIVENESS disease states. Basophils contain and release histamine, The assumption that the basophil is an important in vivo the eosinophil chemotactic factor of anaphylaxis (ECF component of the inflammatory process is rendered more likely A), a slowly reacting substance of anaphylaxis (SRS-A), by the rather faithful fashion in which basophil function par and a kallikrein. The release process is controlled by allels systemic responsiveness. The basophil and the mast cell hormone-basophil receptor interactions that determine seem to be equally endowed with respect to sUlface represen the cyclic AMP levelj plasma and tissue adenosine levels tation of IgE antibody. There is, therefore, an excellent quan appear prominent in this control. Histamine feeds back titative relationship between the response of basophils to anti to negatively modulate basophil and mast cell release gen, manifested by histamine release, and the response of skin through a specific histamine 2-receptorj it also inhibits mast cells to intradermal antigen administration, manifested by lymphocyte and neutrophil function. Like neutrophils, the wheal and flare reaction [4]. There is a similar quantitative basophils contain beta-glucuronidase while neutrophils correlation between either skin test results or basophil hista contain SRS-A and a low-molecular-weight ECF. The mine release and the response of lung mast cells, as measured stimuli for primary basophil and neutrophil release are, by changes in airway conductance on inhalational challenge however, quite different, although phagocytic sti:t~uli, [5]. Moreover, the response of the basophil to antigen reflects which fail to cause basophil mediator release, potentiate systemic disease. Thus, it can be shown that the quantity of the IgE response. It is concluded that basophils play a antigen required for 50% histamine release varies in patients significant in vivo role in inflammation by acting as an with ragweed hay fever over 4 orders of magnitude. This interface between foreign antigens, the serum cascade sensitivity is correlated in a highly significant fashion with the systems, and other inflammatory cells. degree of symptomatology manifested by these patients during the ragweed season [6]. Another example is that of chronic idiopathic urticaria. In Perhaps more than any other blood-borne cell t~p~, the this disease, the causes of which are unknown, the patients' functional role of the basophil remains enigmatic. ThIS IS not basophils are clearly hyporesponsive, as compared to cells because studies of this cell type are laclOng; it is well known, for obtained from control populations [7] (Fig 1), to immunologic example, that the basophil cell membrane contains receptors stimulation by anti-IgE. The reasons for this decrease in reac for IgE antibody and that granule-associated mediators are tivity are not clear, but we have suggested that it reflects in released on contact with antigen. Rathel', it is because of the vivo release and thus results from functional desensitization; basophils sparse representation, constituting, as it does, less i.e., cells that have released a portion of their histamine become than 1% of the leukocyte population. Because of this low unresponsive to further stimulation. This concept is reinforced percentage and because of the failure, to date, to purify normal by the observation that the basophils of patients with Ul'ticaria basophils, its physical features, biochemical properties, . and have significantly less histamine than cells from normal indi subsequently, its importance in the overall defense mechamsms viduals; about 0.7 ± 0.1 versus 1.3 ± 0.3 pg per basophil [7]. of the body, remain uncertain. One of the curious features of Whatever the mechanism, this is another instance of basophil the basophil is that, although its origins are different from t~lOse function mirroring a systemic disease process. of the tissue mast cell, it has evolved capacities essentially similar to those of the tissue mast cell. Although there are IgE RECEPTORS ON THE BASOPHIL MEMBRANE differences in the functions of the 2 cell types, these fade in importance as one considers the marked similarities. The rela The basophil and the mast cell are the only cell types that tionship between these 2 cell types is reinforced by the fact that have surface receptors for the Fc portion of IgE immunoglob as one surveys the mammalian species there seems to be an ulins. Because of obvious difficulties in obtaining these cells, inverse ratio between the numbers of circulating basophils and the nature of this receptor has not been studied in human tissue-fixed mast cells [1]. Moreover, the recent work of Asken basophils. However, studies with a line ofleukemic rat basophils ase and Dvorak on cutaneous basophil hypersensitivity points and rat mast cells have revealed that the receptor is an asym metric molecule with a molecular size of about 150,000 daltons [8-10). The isolated receptor molecule retains the a~ility to Publication #289, O'Neill Research Laboratories, The Good Samar combine with IgE, and equilibrium constants of approxll11ately itan Hospital. This work was supported by Grants AI ?7290 and AI 10 10 MIL have been noted for this interaction [10]. Preliminary 08270 from the National Institute of Allergy and InfectIOUs DIseases, National Institutes of Health, Bethesda, Maryland. data indicate that the cross-linking of these receptors by anti Reprint requests to: Dr. Lawrence M. Lichtenstein, The Good Sa receptor antibody can trigger the cell to release histamine maritan Hospital, 5601 Lock Raven Boulevard, Baltimore, Maryland [11). It is well known that histamine release requires bivalent 21239. antigen, and there is good evidence that cross-linking of the IgE 65 66 LICHTENSTEIN ET AL Vol. 71, No.1

LU If) 50 understanding of diseases such as asthma or urticaria; a signifi « p <.01 LU (20) cant variable may be the relative ease with which the mast cells ..J Pat,ents • LU ex: 40 Controls 0 (2.3) of these patients release mediators in response to nonspecific \ LUz stimuli. In this hypothesis, of course, we assume that basophil ~ .30 and mast cell releasability vary concordantly; this idea has not ~ pe05 If) been proved, but it seems likely on the basis of the arguments I 20 laid out above. I- Z LU u 10 BASOPHIL AND MAST CELL MEDIATORS ex: LU a.. Another strong statement to support the similarity between \ 10-2 10- 1 basophils and mast cells is that they.contain a relatively similar ANTI-IgE CONCENTRATION()JgAbN/mi.l complement of mediators. Both cell types are largely filled with histamine-containing granules that can be released by appro FIG 1. Anti-IgE-induced histamine release from the basophils of 20 patients with chronic idiopathic urticaria compared to that in 23 priate stimuli. It has been demonstrated recently that both cell controls. The patients and controls had a similar range and mean in types also contain 2 other mediators that may play a role in terms of age and were matched with respect to sex. allergic reactions, the eosinophil chemotactic factor of anaphy laxis (ECF-A) and the slow reacting substance of anaphylaxis antibody by antigen provides the stimulation for this process (SRS-A) [15-18]. The importance of the eosinophil in inflam [12]. It now seems likely that the first event in the basophil matory diseases is a fascinating question that will, of course, be \ release reaction is an approximation of the cell membrane Fc dealt with by others in this volume. Both basophils and mast receptors. cells contain a variety of low-molecular-weight factors that are The number of IgE molecules per basophil, ranging from potent chemotactic agents and highly specific for eosinophils. The number and nature of these factors has not yet been 3,000 to 6~0,000, is quantitatively related to the serum IgE concentratIOn (r = 0.94) [13]. The number of receptors for IgE delineated. Similarly, despite almost 10 years of effort, the exact on human basophils has a similar range (15,000 to about chemical nature of SRS-A remains undefined, although the 600,000), and the degree ofreceptor saturation is also a function studies of Orange et al suggest that it is a low-molecular-weight, of the serum IgE level (r = 0.84) [43]. More surprisingly, the sulfur-containing lipid [19]. Whether there aTe more than 1 total number of IgE receptors is also closely related to the SRS-A and whether some or all of these mediators are products serum IgE level (r = 0.92) [43]. The basis for this association of arachidonic acid metabolism also remains to be established. between the serum IgE level and its receptor on basophils, i.e., One of the most interesting mediators recently identified as whet~er .it consists of receptor modulation by serum IgE or a being represented in-the basophil, and probably in mast cells genetiC hnkage between the 2 parameters is unknown. The is a kallikrein [20]. This high-molecular-weight enzyme is usu: equilibrium constant for the reaction betw~en human IgE and ally assayed by means of its ability to hydrolize a synthetic \ 9 the. basoph~ re~eptor ranges from about 10 - 1OIoM/ L [44]. substrate, tosyl arginine methyl ester (TAMe). Recent studies ThIS VarlatlOn m receptor affinity for IgE is unrelated to the by Newball and Talamo indicate, however, that the esterase s:ruJ? IgE co ncentr~tion and appears to be rather normally activity and the ability to split a kinin-like molecule from serum dlstnbuted on both sldes of the mean of approximately 7 X 109 kininogen are subserved by the same molecule. The nature of MIL. Thu.s, there is heterogeneity with respect to the affinity the kinin generated has not been studied, but it is identified by of basophil receptors for IgE. Preliminary data also indicate its ability to react with an antibody that specifically recogniz~s that. there is h:terogeneity in the IgE antibody molecules the nonapeptide bradykinin. The basophil kallikrein represents obtamed from different patients; there is a two- to threefold the only immunologic mechanism for the activation of the difference in the ability of different sera (normalized to contain kallikreinkinin system and thus places this system, which ha the sa~e IgE antibody and total IgE) to passively sensitize been elegantly described in chemical terms, in a somewhat \ basophils from a normal donor for histamine release. Both mOl:e biolo~ical perspective [21]. A related feature of anaphy eleme.nts of heterogeneity may well be important to an under lactIC reactIOns that has not yet been fully studied is the ability standmg of the allergic responses of man. of IgE-mediated reactions to activate the coagulation sequenc ~ .

BASOPHIL "RELEASABILITY" We have been particularly interested in the relationship between the number of IgE molecules per basophil and the w \ r~sP.onse ofth~se c~lls to immunologic challenge. The surprising (f) 60

Clotting changes have been observed in rabbit anaphylaxis by have isoproterenol or histamine receptors and other subclasses Pinkard, Tanigawa, and Halonen [22], and our own studies of lack them [27]. Nevertheless, the early data led to the hypoth human anaphylaxis have uncovered similar changes [23]. It esis that the antigen-IgE interaction leads to a transient fall in seems conceivable that the basophil kallikrein interacts not cyclic AMP that constitutes an early step in the release process. only with the high-molecular-weight kininogen, but also with In fact, an appropriate change in cyclic AMP has been dem Hageman factor, for which the high-molecular-weight kinino onstrated in purified rat mast cells subjected to a variety of gen is a cofactor. If this were the case, we would have a ready stimuli, but only very recently have we been fortunate enough explanation for the changes in coagulation ability noted during to obtain a highJy purified preparation of basophils (from a anaphylactic reactions. patient with chronic myelocytic leukemia) with which we could make direct measurements. In this individual's basophils (Fig THE BIOCHEMICAL MECHANISM(S) OF 3) there was an unanticipated, very transient rise in cyclic AMP MEDIATOR RELEASE about 30 sec after antigenic stimulation; by 1 min, however, Having dermed some of the immunologic parameters and well before the initiation of histamine release, the anticipated described some of the basophil mediators, we think it appro fall in cyclic AMP level occurred. priate to discuss what is known about the biochemical mecha "Recent studies by Sullivan et al with rat mast cells have nism of mediator release. The process by which basophil gran shown the same transient rise and fall and thus have under ules are exteriorized is similar to the secretory mechanism of scored the similarity between these 2 cell types [28]. Therefore, other cells: it is an active, energy-requiring process in which it seems likely that the in vivo secretion of histamine and other calcium acts as the link between stimulation and secretion mediators is partially modulated by hormones that, by inter [24]. One of the more essential elements of the biochemical acting with specific receptors on mast cells and basophils, alter control of the secretory process in basophils is related to cyclic the intracellular level of cyclic AMP. Of the several endogenous nucleotide metabolism. It was observed about a decade ago hormones that may function in this way, the most interesting that agents such as beta-adrenergic agonists (e.g., isoproterenol) is histamine itself. Histamine acts on specific basophil histanline and phosphodiesterase inhibitors (e.g., theophylline) which in 2-receptors to increase the cyclic AMP level and turn off crease intracellular cyclic AMP inhibited the release of hista histamine release [29]. The same phenomenon has been noted mine and other mediators [25]. Subsequent measurements in in mast cells: Chakrin and his colleagues have demonstrated whole leukocyte prepal'ations showed that these drugs, at about that in the presence of histamine 2-antagonists, such as buri the same concentrations required for the inhibition of histamine mamide or metiamide, there is potentiation of the release of release [26], increase the cyclic AMP level. The difficulty, of histamine and other mediators from anti-IgE-challenged, course, is that it was unclear whether the cyclic AMP levels chopped monkey skin or lung [30]. Thus, a histamine feedback being recorded had anything to do with the few basophils mechanism controls the release of basophil and mast cell me present in the leukocyte preparations. At the beginning of this diators in vivo. Histamine operating through histamine 2-recep research effort it seemed reasonable to assume that most of the tors also controls the function of other inflammatory cell types, leukocytes responded similarly to the different agonists, but we and this fact should be kept in mind in evaluating patients on now know this is Dot true. Neutrophils and lymphocytes behave the histamine 2-antagonists [31,32]. quite differently; even among lymphocytes some subclasses Another potentially important element in the control of basophil function has recently been described by Marone in 14 this laboratory. He found that adenosine increases leukocyte cyclic AMP levels and inhibits mediator release. Perhaps more interesting is the parallel observation that human serum, when added together with the adenosine deaminase inhibitor erythl'o 30 12 cAMP HISTAMINE/ 9(2-hydl'oxy-3-monyl) adenine (ENHA), also is a potent inhib itor of mediator release [45]. These observations suggest that physiological levels of adenosine in body fluids or in serum are 10 major elements in the control of basophil and perhaps mast cell w U) mediator release. It is well appreciated that histamine, again w

w en that split fragments of C3 and C5, the C3a and C5a anaphyla Immunology II, vol 3. Edited by L Brent, J Holborow. Amsterdam, North Holland Publishing Company, 1974, pp observation that highly purified human neutrophils were able 171-181 3. Askenase PS: The role of basophils, mast cells and vasoamines in to generate SRS-A when stimulated by a calcium flux induced hypersensitivity reactions with a delayed time co urse, Progress by an ionophore [38]. Neutrophils also released a low-molecu in Allergy. Edited by P Kallos, BH Waksman, Basel, S. Krager, lar-weight eosinophil chemotactic factor similar to ECF-A on 1977, pp 199-320 standard phagocytic stimulation with aggregated IgG or serum 4. Norman PS, Lichtenstein LM, Ishizaka K: Diagnostic tests in treated zymosan particles [39]. Thus, the neutrophils were ragweed hay fever. J Allerg Clin Immunol 52:210-224, 1973 5. Bruce CA, Rosenthal RR, Lichtenstein LM, Norman PS: Diagnos· 1 found to contain some of the same mediators that had until tic tests in ragweed-allergic asthma. J Allerg Clin Immunol that time been associated with basophils. Conversely, using 53:230-239, 1974 1 first the basophils from the patient with chronic myelocytic 6. Lichtenstein LM, Norman PS, Osler AG, Winkenwerder WL: In leukemia and, more recently, from people with normal baso vitro studies of human ragweed allergy: Changes in cellular and humoral activity associated with specific desensitization. J Clin phils, Dr. Thomas has been able to demonstrate that basophils Invest 45:1126-1136, 1966 contain beta-glucuronidase, an enzyme usually associated with 7. Kern F, Lichtenstein LM: Defective histamine release in clu-onic the neutrophil granules. In spite of the similarity in ontogeny urticaria. J Clin Invest 57:1369-1377, 1976 and in the granule associated mediators, the stimuli that induce 8. Kulcyzycki A Jr., Metzger H: The interaction of IgE with rat basophilic leukemia cells. II. Quantitative aspects of binding secretory function in the 2 cell types are completely different. reaction. J Exp Med 140:1676-1695, 1974 The polymorphonuclear cell cannot respond to antigenic stim 9. Conrad DH, Froese A: Characterization of the target cell receptor ulation and basophils cannot respond to phagocytic stimuli. for IgE. II. Polyacrylamide gel analysis of the surface IgE receptor Surprisingly, however, the same phagocytic stimuli that fail to from normal rat mast cells and from rat basophilic leukemia cells. J Immunol 116:319-326, 1976 induce basophil mediator release are able to markedly poten 10. Isersky C, Mendoza GR, Metzger H: The binding of antibodies to tiate the response of the cell to antigen or anti-IgE [45]. Thus, the solubilized and membrane-integrated mouse and rat receptor although zymosan and aggregated IgG by themselves have no for IgE. J Immunol 119:123-130, 1977 effect, they can more than double the response of basophil to 11. Ishiza ka T, Chang TH, Taggart M, Ishizaka K: Histamine release from rat mast cells by antibodies against rat basophilic leukemic 1 antigen or anti-IgE stimulation (Fig 4). This ability does not cell membrane. J Immunol 119:1589-1596, 1977 seem to be the result of phagocytosis, but of effects on the cell 12. Levine BB, Redmond AP: The nature of the antigen-antibody membrane that occur as a result of occupancy of Fc or C complexes initiating the specific wheal and flare reaction in \ receptors. Although it has been known that basophils contain sensitized man. J Clin Invest 47:556-557, 1968 13. Conroy MC, Adkinson NF Jr., Lichtenstein LM: Measurement of receptors for IgG, it has not been appreciated that they have a IgE on human basophils: Relation to serum IgE and anti-IgE· complement receptor [40]. This seems likely, however, since induced histamine release. J ImmunoI118:1317-1321, 1977 the preparation of zymosan particles without serum or with 14. Lichtenstein LM, Conroy MC: The "releasability" of mediators EDTA-treated serum completely abolishes theil' effect [45]. C3- from human basophils and granulocytes. Ninth International \ Congress of Allergology, Buenos Aires. Edited by E Mathov. deficient serum also fails to prepare zymosan particles for Amsterdam, Excerpta Medica, 1977, pp 109-115 enhancement of IgE -mediated histamine release. Thus, it may 15. Kay AB, Stechschulte DJ, Austen KF: An eosinophil leukocyte \ be suggested that the basophils contain a C3b receptor. Not chemotactic factor of anaphylaxis. J Exp Med 133:602-619, 1971 surprisingly, a similar receptor has recently been described on 16. Czarnetzki BM, Konig W, Lichtenstein LM: Antigen-induced eosin. ophil chemotactic factor (ECF) release by human leukocyte rat mast cells [41]. Inflammation 1:201-215, 1975-1976 17. Orange RP The formation and release of slow reacting substance CONCLUSION of anaphylaxis in human lung tissues, Progress in Immunology II, vol. 4. Edited by L Brent, J Holborow. Amsterdam, Nortb The in vivo importance of the basophils in defense or in Holland Publishing Company, 1974, pp 29-39 pathogenetic mechanisms is unclear. We have attempted to 18. Grant JA, Lichtenstein LM: Release of slow reacting substance of I discuss some elements of theil' function and to indicate som e anaphylaxis from human leukocytes. J Immunol 112:897-904, 1974 areas in which they may playa significant role. The observation 19. Orange RP, Murphy RC, Karnovsky MD, Austen KF: The physi· by Dvorak and Askenase that basophils can enter tissue spaces ochemical characteristics and purification of slow reacting sub· expands the potential range of action for these cells. Perhaps stance of anaphylaxis. J Immunol 110:760-770, 1973 the best way to appreciate the function of basophils is to view 20. Newball HH, Talamo RC, Lichtenstein LM: Release of l e llko cyt ~ I them as an interface between foreign antigens and a variety of kallikrein mediated by IgE. Nature 254:635-636, 1975 2l. Colman RW: Formation of human plasma kinin. N Engl J Med inflammatory processes. Thus, basophils serve as the only 291:509-515, 1974 \ known link between immune stimuli and some of the serum 22. Pinckard RN, Tanigawa C, Halonen M: IgE-induced blood coagu· systems t hat participate in inflammation, such as activation of lation alterations in the rabbit: co nsumption of coagulation fac· the kallikreinkinin system. On the other hand, activated serum tors XII, XI, and IX in. vivo. J Immunol 115:525-532, 1975 23. Kaplan AP, Hunt KJ, Sobotka AK, Smith P, Horakova Z, Grainick systems such as complement may interact with basophils; this H, Lichtenstein LM: Human anaphylaxis: A study of mediator possibility is perhaps best illustrated by the recent observation systems. Clin Res 25:361A, 1977 July 1978 THE ROLE OF BASOPHILS IN INFLAMMATORY REACTIONS 69

24. Osler AG, Lichtenstein LM, Levy DA: In vitro studies of human 36. Lichtenstein LM: The immediate allergic response: ill vitro sepa reaginic allergy. Adv Immunol 8: 183-231, 1968 ration of antigen activation, decay and histamine release. J 25. Lichtenstein LM, Margolis S: Histamine release in vitro: Inhibition Immunol 107:1122-1130, 1971 by catecholamines and methylxanLhines. Science 161:902-903, 37. Kaliner M. Austen KF: A sequence of biochemical events in the 1968 . antigen-induced release of chemical mediat.ors from sensitized 26. Bourne HR, Lichtenstein LM, Melmon KL: Pharmacologic control human lung tissue. J Exp Med 138:1077-1094, 1973 of allergic histamine release in vitro: evid ence for an inhibitory 38. Conroy MC, Orange RP, Lichtenstein LM: Release of slow reacting role of 3',5'-adenosine monophosphate in human leukocytes. J substance of anaphylaxis (SRS-A) from human leukocytes by the Immunol 108:695-705, 1972 calcium ionophore A23187. J Immunol 116:1677-1681, 1976 27. Roszkowski W, Plaut M, Lichtenstein LM: Selective display of 39. Konig W, Czarnetski BM, Lichtenstein LM: Eosinophil chemotactic histamine receptors on lymphocytes. Science 195:683-685, 1977 factor (ECF). II. Release from human polymorphonuclear leu 28. Sullivan TJ, Parker KL, Kulczycki A Jr., Parker CW: Modulation kocytes during phagocytosis. J Immunol 117:235-241, 1976 of cyclic AMP in purified rat mast cells. III. Studies on the effects 40. Grant JA, Lichtenstein LM: Reversed in vitro anaphylaxis induced of concanavalin A and anti-IgE on cyclic AMP concentrations by anti-IgG: specificity of the reaction and compru'ison with dUTing histamine release. J Immunol 117:713-716, 1976 antigen-induced histamine release. J Immunol 109:20-25, 1972 29. Lichtenstein LM, Gillespie E: The effects of the HI and H2 anti 41. Sher A, McIntyr.e SL: Receptors for C3 on rat peritoneal mast cells. histamines on "allergic" histamine release and its inhibition by J Immunol 119:722-725, 1977 histamin e. J Pharmacol Exp Ther 192:441-450, 1975 42. Hook WA, Siraganian RP, Wahl SM: Complement-induced hista 30. ChaJu'in LW, Krell RD, Mengel J, Young D, Zaher C, Wardell JR: mine release from human basophils. 1. Generation of activity in Effect of a histamine H2-receptor antagonist on immunologically human serum. J Immunol 114:1185-1190, 1975 induced mediator release in vitro. Agents Actions 4:297-303, 1974 43. Malveaux FJ, Conroy MC, Adkinson NF Jr, Lichtenstein LM: IgE 31. Busse WW, Sosman J: Histamine inhibi tion of neutrophil lysosomal receptors on human basophils: Relationship to Serum IgE co n enzyme release: An H2 histamine receptor response. Science centration. J Clin Invest, in press 194:737-738, 1976 44. Marone G, Lichtenstein LM: Adenosine-adenosine deaminase 32. Plaut M, Lichtenstein LM, Henney CS: Properties of a subpopu modulation of histamine release. J Allerg Clin Immunol 61 :131, lation of T Cells bea.ring histamine receptors. J Clin Invest 1978 55:856-874, 1975 45. Thomas LL, Lichtenstein LM: Augmentation of IgE-medicated 33. Plaut M. Lichtenstein LM: Modulation of inflammation by hista histamine release from human basophils. J Allerg Clin Immunol mine receptor-bearing cells, Histamine Receptors. Edited by TO 61: 152, 1978 Yell in. Holliswood, New York, Spectrum Publication, in press 46. Dolovich J, Hargreave FE, Chalmers R, Shier KJ, Gauldie J, 34. Hirschhorn R, Sela E: Adenosine deaminase and immunodefi Bienenstock J: Late cutaneous alLergic responses in isoLated IgE ciency: an in vitro model. Ce ll Immunol 32:350-360, 1977 dependent reactions. J Allerg Clin Immunol 52:33-46, 1973 35. Gillespie E, Lichtenstein LM: H istamine release from human leu 47. Solley GO, GLeich GJ, Jordan RE, Schroeter AI: The Late phase of kocytes: Studies with deuterium ox ide colchicine and cytochal the immediate wheal and flare skin reaction. J CLin Invest asin B. J Clin Invest 51:2941-2947, 1972 58:403-420, 1976