- Email: [email protected]
Effect of beta adrenergic stimulation and blockade on cutaneous reactivity to histamine
Effect of beta adrenergic stimulation and blockade on cutaneous reactivity to histamine Nelson Lamkin, Jr., M.D., Phil Lieberman, M.D., Richard Shereff, M.D., E. William Rosenberg, M.D., and Harry Robinson, Sc.D. Mentphis, Tenn.
It has been previously demonstrated that iontophoresis of beta adrenergic agents will alter the size of immediate hypersensitivity shin tests. It was unclear whether mast cell (inhibition of histamine this alteration was due to an effect on the dermal release) or on the cutaneous vasculatwre (inhibition of capillary permeability). For this rea.son isoproterend, propranolol, diphenhydramine as a positive control, and saline as a negative control were iontophoresed onto the forearm of 10 atopic and 10 n-topic adult subjects. In order to bypass histamine release from mast cells the patients were then challenged direCtly with histamine by the “prick” technique. The size of the resultant wheals was noted. The data obtained allowed the following conclzlsions: (1) The atop&o group ?WpOnded to histamine with greater wheal size of diphyenhydramine effectively T’eduCed than the non&topic group. (8) Iontophoresis the magnitude of the histamine wheal in both groups. (8) Isoproterenol decreased the wheal size in both grozcps. (4) Propranolol increased the wheal size in only the nonatopk group. (5) The successful modzllation of the histamine-induced wheat and flare indio&?d that these drugs, regardless of their eflect on the der?nCLl mast Cell, exert a measltrable eflect on the target organ (vasculature).
The adenyl cyclase-cyclic adenosine monophosphate (CAMP) system is intimately linked to mediator release from sensitized mast cells and basophils in allergic reactions. Beta adrenergic stimulation leads to increased intracellular levels of CAMP and subsequent decrease in mediator re1ease.l This effect of CAMP on mediator release has been demonstrated by various in vitro systems.2-4 Recently iontophoresis was employed to demonstrate a similar effect in viva.’ The iontophoresis of beta adrenergic agents reduced the size of and even abolished the immediate hypersensitivity skin test reaction in atopic patients. It was unclear whether this modulation of skin test reactivity was the result of the effect of CAMP on mediator release from dermal mast cells or the effect of this compound on the cutaneous vasculature.6-9 The present study was designed to clarify this issue. From the Section of Allergy-Immunology, the Division of Dermatology, and the Department of Preventive and Community Medicine, University of Tennessee Department of Medicine. Supported in part by United States Public Health Service Grant No. AI 00320. Presented in part at the Thirty-first Annual Meeting of the American Academy of Allergy, San Diego, Calif., Feb. 19, 1975. Received for publication March 26, 1975. Accepted for publication July 15, 1975. Reprint requests to: Phil Lieberman, M.D., University of Tennessee, Department of Medicine, 951 Court-240 Dobbs, Memphis, Tenn. 38163. Vol. 57, No. 5, pp. 449-453
450
TABLE
Lamkin
J. ALLERGY CLIN.
et al.
1. Concentration,
pH, and
Propranolol hydrochloride
net charge
Iaoproterenol hydrochloride
of drugs
used and
Diphenhydramine hydrochloride
rationale
IMMUNOL. MAY IF76
for their
use
Physiologic saline
1 mg/ml 5 mg/mI Adjusted to 4.0 2.5 with citric acid Positive Positive
10 mg/ml 4.5
0.9% solution 7.0
Positive
Neutral
Positive
Positive
Positive
Rationale for inclusion in study
p-adrenergic blocking agent -
@-adrenergic stimulating agent -
Applied to all negatlve electrodes for electrical conductivity and to one positive electrode as control for exoerimental drugs .
Amount applied to each electrode
0.75 ml
0.75 ml
Iontophoresis and skin test technique control (decreases white dermographism when iontophoresed into atopic skin)lO 0.75 ml 0.75 ml
Concentration PH
Net ionic charge in solution Polarity of “applicator electrode
MATERIALS AND Patient selection
METHODS
Twenty adult subjects were selected for study. Ten were considered atopie on the basis of a history of allergic rhinitis and/or extrinsic asthma and had at least three positive immediate hypersensitivity skin tests (scratch [1:20 w/v] or intracutaneous CO.02cc of 1:lOOO w/v] ) to a battery of standard aeroallergens (grass, weed, tree, mold, and dust, Greer Laboratories, Lenoir, N. C.). The remaining 10 subjects had a negative allergic history and skin tests. Subjects were not allowed any medication 48 hr prior to testing. No participant exhibited obvious dermographism. lontophoresis
technique
and drugs
The iontophoretic apparatus and technique have been previously described.5 Briefly, three drugs (propranolol hydrochloride, isoproterenol hydrochloride, and diphenhydramine hydrochloride) and a saline control were applied to each subject in a different but random order to filter paper (Whatman No. 1, 12.5 cm circles, W. R. Bakston, Ltd., England) cut to fit on four electrocardiogram electrodes strapped to the volar surface of the forearm for simultaneous iontophoresis. This was done to eliminate any possible difference due to specific forearm location. The threshold current used was 1 ma, which produced a subjective “tingling” sensation in each subject. The concentration, pH, and net charge of the drugs used and the rationale for their use are seen in Table I. Skin testing Immediately following iontophoresis, one drop of histamine phosphate 2.75 mg/cc was applied to each iontophoresed area and the skin pricked with a lancet. In addition a histamine scratch test was performed on the opposite arm as a second control. Fifteen minutes later test sites were simultaneously read for whealing. The wheals were traced onto laboratory film (Parafilm “M, ” American Can Company, Neenah, Wis.) and later enlarged for easier computation with a Polaroid MP-3 Land Camera (Polaroid Corporation, Cambridge, Mass.).
Cutaneous
VOLUME 57 NUMBER 5
TABLE II. Cutaneous response of atopic saline, diphenhydramine, isoproterenol,
No.
Nonatopic: 1
1
8
Diphenhydramine
451
preceded
Propranolol
Age
Saline
::
Iti
49.3 46.1
24.1 20.0
38.7 30.7
58.2 70.7
::
F
45.3 40.4
21.3 30.0
26.0 30.0
52.0 32.3
:i
iti
40.0 31.6
23.9 34.0
36.0 32.7
25.3 79.0
:; 2
iti itlF
44.0 32.0 33.3 29.3
36.0 12.0 26.1 22.7
45.3 12.0 28.0 34.7
40.0 67.3 45.0 57.3
39.2
25.1
31.4
52.1
1 :
ii
M ki
132.2 45.7 69.8
58.8 31.0 18.4
49.0 6i.8
137.1 83.8 36.5
4
:II
F
120.1
53.9
90.3
298.8
2 7
ii 30
ki
41.3 57.7 43.7
36.0 37.0 55.4
26.1 28.2 61.8
67.1 52.0 53.3
9" 10 Mean
30 :i
ki E
47.0 75.8 144.3 74.8
60.3 10.5 75.6 43.7
42.1 59.0 15.6 49.8
82.1 63.2 125.7 100.0
The area contained in each outline was then ascertained exhibited any systemic effects with the drugs used. Analysis
with
by
area (mm21 lsoproterenol
SOX
1; Mean Atopic:
to histamine
and nonatopic subiects to histamine and propranolol iontophoresis Drug and surface
Subject
reactivity
a planimeter.
No
subjects
of data
Testing with the paired t test showed no significant differences between the wheal areas produced by histamine on skin pretreated with saline iontophoresis and untreated skin in atopic and nonatopic subjects. Therefore, in analyzing the drug effects, the iontophoresed saline area was used aa the control value. Differences between the saline and drug wheal sizes within both atopic and nonatopic groups were evaluated by the paired t test. RESULTS
Data for each subject are seen in Table II. As a group, atopic individuals demonstrated significantly (p < 0.02) greater skin reactivity to histamine, as judged by wheal area, than nonatopic subjects. The whealing produced by histamine was modified by the iontophoresed agents. Diphenhydramine significantly decreased the wheal response in atopics (p < 0.005) and nonatopics (p < 0.02). Similar results were obtained when the beta adrenergic agonist, isoproterenol, was used. Both groups responded with a significantly decreased wheal area (P <
0.02).
When the beta adrenergic blocking agent-propranolol-was used, no significant difference in wheal size as compared to the control was seen in the atopic group. However, for the nonatopic subjects, propranolol did result in larger wheal areas than the control (p < 0.05). Statistical data are summarized in Table III.
452
Lamkin
TABLE
III.
J. ALLERGY CLIN.
et al.
Comparison
of
saline
control
and
drug
responses
for
atopic
and
IMMUNOL. MAY 1976
nonatopic
subjects Subjects
Diphenhydramine Isoproterenol Propranolol
Atopic Nonatopic Atopic Nonatopic Atopic Nonatopic
I
Paired “t” value
3.19 3.12
2.86 2.84 1.46 2.31
I
p value <0.005 <0.02 <0.02 <0.02
DISCUSSION
The purpose of this study was to determine to what degree, if any, the effects of agents which modify the CAMP-beta adrenergic system are directed toward the cutaneous vascular system as opposed to the dermal mast cell population. The modification of exogenous histamine whealing by these agents was evaluated in this regard. The technique of iontophoresis was used because of its previously demonstrated applicability and reproducibility as an in vivo technique.5 Also an attempt was made to determine if there were differences in response to these agents between atopic and nonatopic individuals. The present study demonstrates that the cutaneous response to histamine is subject to the effects of drugs which alter the metabolism of CAMP. Thus the previously demonstrated effect of these drugs on the immediate skin test5 may well have been due to an effect on the target organ (vasculature) rather than on the effector organ (mast cell). These results are contrary to those obtained by Perper, Sanda, and Lichtenstein.” In a previous study employing rhesus monkeys, these investigators noted that theophylline and salbutamol significantly decreased wheal formation following Ascclris challenge, but had no effect on whealing produced by the intradermal injection of histamine. However, the multiple differences in the studies make valid comparisons impossible. The studies differed with respect to drugs employed, their doses and route of administration, and the species involved. Of interest in our study is the difference in response between the atopic and nonatopic populations. The atopic patients responded more intensely to histamine. The responses to isoproterenol and propranolol are also worthy of note. As can be seen from the results in Table III, the beta agonist isoproterenol produced equally significant decrements in wheal size in both groups. This effect was anticipated and parallels the action of this agent in cellular preparations involving mediator release2e4and in a previous in vivo study involving immediate hypersensitivity skin tests.5 In contrast, propranolol failed to significantly alter wheal size in the atopic population but effectively increased wheal size in the nonatopic group. The results obtained with propranolol suggest the presence of a pre-existing cutaneous beta adrenergic blockade which partially inhibited the action of this
VOLUME 57 NUMBER 5
Cutaneous
reactivity
to histamine
453
drug on the vasculature in atopics. However, the similarity of responses of both groups to isoproterenol is not in keeping with this hypothesis. In summary, iontophoresis of drugs affecting CAMP metabolism effectively alters the cutaneous response to histamine. It is therefore assumed that regardless of the effects these drugs may have on the dermal mast cell after allergen challenge, at least part of their action is directed against the cutaneous vasculature. REFERENCES 1 Reed, C. E.: The pathogenesis of asthma, Med. Clin. North Am. 58: 55,1974. 2 Yamamoto, S., Greaves, M. W., and Fairley, V. M.: Cyclic AMP-induced inhibition of IgE-mediated hypersensitivity in human skin, Immunology 24: 77, 1973. release: In vitro 3 Lichtenstein, L. M., and DeBernardo, R.: IgE mediated histamine separation into two phases, Int. Arch. Allergy Appl. Immunol. 41: 56, 1971. 4 Ishizaka, T., Ishizaka, K., Orange, R. P., and Au&en, K. F.: Pharmacologic inhibition of the antigen-induced release of histamine and slow reacting substance of anaphylaxis (SRS-A) from monkey lung tissues mediated by human IgE, J. Immunol. 106: 1267, 1971. 5 Shereff, R. II., Harwell, W., Lieberman, P. L., Rosenberg, E. W., and Robinson, H.: Effect of beta adrenergic stimulation and blockade on immediate hypersensitivity skin test reactions, J. ALLERGY CLIN. IMMUNOL. 52: 328,1973. 6 Ichikawa, A., Nagasaki, M., Umezu, K., Hayaski, H., and Tomita, K. : Effect of cyclic adenosine 3’5’-monophosphate on edema and granuloma induced by carrageenin, Biochem. Pharmacol. 21: 2615, 1972. 7 Ichikawa, A., Hayashi, H., Minami, M., and Tomita, K.: An acute inflammation induced by inorganic pyrophosphate and adenosine triphosphate, and its inhibition by cyclic 3’5’adenosine monophosphate, Biochem. Pharmacol. 21: 317, 1972. 8 Lugnier, C., Bertrand, Y., and Stoclet, J. C.: Cyclic nucleotide phosphodiesterase inhibition and smooth vascular relaxation, Eur. J. Pharmacol. 19: 134, 1972. 9 Triner, L., Nahas, G. G., Vulliemoz, Y., Overweg, N. I. A., Veresky, M., Habif, D. V., and Ngai, S. H.: Cyclic AMP and smooth muscle function, Ann. N. Y. Acad. Sci. 185: 458, 1971. dermographia, Arch. 10 Kalz, F., Bower, C. M., and Prichard, H.: Delayed and persistent Dermatol. Syphilol. 61: 772, 1950. in vitro and in vivo 11 Perper, R. J., Sanda, M., and Lichtenstein, L. M.: The relationship allergic histamine release : inhibition in primates by CAMP active agents, Int. Arch. Allergy 48: 837, 1972.
It has been previously demonstrated that iontophoresis of beta adrenergic agents will alter the size of immediate hypersensitivity shin tests. It was unclear whether mast cell (inhibition of histamine this alteration was due to an effect on the dermal release) or on the cutaneous vasculatwre (inhibition of capillary permeability). For this rea.son isoproterend, propranolol, diphenhydramine as a positive control, and saline as a negative control were iontophoresed onto the forearm of 10 atopic and 10 n-topic adult subjects. In order to bypass histamine release from mast cells the patients were then challenged direCtly with histamine by the “prick” technique. The size of the resultant wheals was noted. The data obtained allowed the following conclzlsions: (1) The atop&o group ?WpOnded to histamine with greater wheal size of diphyenhydramine effectively T’eduCed than the non&topic group. (8) Iontophoresis the magnitude of the histamine wheal in both groups. (8) Isoproterenol decreased the wheal size in both grozcps. (4) Propranolol increased the wheal size in only the nonatopk group. (5) The successful modzllation of the histamine-induced wheat and flare indio&?d that these drugs, regardless of their eflect on the der?nCLl mast Cell, exert a measltrable eflect on the target organ (vasculature).
The adenyl cyclase-cyclic adenosine monophosphate (CAMP) system is intimately linked to mediator release from sensitized mast cells and basophils in allergic reactions. Beta adrenergic stimulation leads to increased intracellular levels of CAMP and subsequent decrease in mediator re1ease.l This effect of CAMP on mediator release has been demonstrated by various in vitro systems.2-4 Recently iontophoresis was employed to demonstrate a similar effect in viva.’ The iontophoresis of beta adrenergic agents reduced the size of and even abolished the immediate hypersensitivity skin test reaction in atopic patients. It was unclear whether this modulation of skin test reactivity was the result of the effect of CAMP on mediator release from dermal mast cells or the effect of this compound on the cutaneous vasculature.6-9 The present study was designed to clarify this issue. From the Section of Allergy-Immunology, the Division of Dermatology, and the Department of Preventive and Community Medicine, University of Tennessee Department of Medicine. Supported in part by United States Public Health Service Grant No. AI 00320. Presented in part at the Thirty-first Annual Meeting of the American Academy of Allergy, San Diego, Calif., Feb. 19, 1975. Received for publication March 26, 1975. Accepted for publication July 15, 1975. Reprint requests to: Phil Lieberman, M.D., University of Tennessee, Department of Medicine, 951 Court-240 Dobbs, Memphis, Tenn. 38163. Vol. 57, No. 5, pp. 449-453
450
TABLE
Lamkin
J. ALLERGY CLIN.
et al.
1. Concentration,
pH, and
Propranolol hydrochloride
net charge
Iaoproterenol hydrochloride
of drugs
used and
Diphenhydramine hydrochloride
rationale
IMMUNOL. MAY IF76
for their
use
Physiologic saline
1 mg/ml 5 mg/mI Adjusted to 4.0 2.5 with citric acid Positive Positive
10 mg/ml 4.5
0.9% solution 7.0
Positive
Neutral
Positive
Positive
Positive
Rationale for inclusion in study
p-adrenergic blocking agent -
@-adrenergic stimulating agent -
Applied to all negatlve electrodes for electrical conductivity and to one positive electrode as control for exoerimental drugs .
Amount applied to each electrode
0.75 ml
0.75 ml
Iontophoresis and skin test technique control (decreases white dermographism when iontophoresed into atopic skin)lO 0.75 ml 0.75 ml
Concentration PH
Net ionic charge in solution Polarity of “applicator electrode
MATERIALS AND Patient selection
METHODS
Twenty adult subjects were selected for study. Ten were considered atopie on the basis of a history of allergic rhinitis and/or extrinsic asthma and had at least three positive immediate hypersensitivity skin tests (scratch [1:20 w/v] or intracutaneous CO.02cc of 1:lOOO w/v] ) to a battery of standard aeroallergens (grass, weed, tree, mold, and dust, Greer Laboratories, Lenoir, N. C.). The remaining 10 subjects had a negative allergic history and skin tests. Subjects were not allowed any medication 48 hr prior to testing. No participant exhibited obvious dermographism. lontophoresis
technique
and drugs
The iontophoretic apparatus and technique have been previously described.5 Briefly, three drugs (propranolol hydrochloride, isoproterenol hydrochloride, and diphenhydramine hydrochloride) and a saline control were applied to each subject in a different but random order to filter paper (Whatman No. 1, 12.5 cm circles, W. R. Bakston, Ltd., England) cut to fit on four electrocardiogram electrodes strapped to the volar surface of the forearm for simultaneous iontophoresis. This was done to eliminate any possible difference due to specific forearm location. The threshold current used was 1 ma, which produced a subjective “tingling” sensation in each subject. The concentration, pH, and net charge of the drugs used and the rationale for their use are seen in Table I. Skin testing Immediately following iontophoresis, one drop of histamine phosphate 2.75 mg/cc was applied to each iontophoresed area and the skin pricked with a lancet. In addition a histamine scratch test was performed on the opposite arm as a second control. Fifteen minutes later test sites were simultaneously read for whealing. The wheals were traced onto laboratory film (Parafilm “M, ” American Can Company, Neenah, Wis.) and later enlarged for easier computation with a Polaroid MP-3 Land Camera (Polaroid Corporation, Cambridge, Mass.).
Cutaneous
VOLUME 57 NUMBER 5
TABLE II. Cutaneous response of atopic saline, diphenhydramine, isoproterenol,
No.
Nonatopic: 1
1
8
Diphenhydramine
451
preceded
Propranolol
Age
Saline
::
Iti
49.3 46.1
24.1 20.0
38.7 30.7
58.2 70.7
::
F
45.3 40.4
21.3 30.0
26.0 30.0
52.0 32.3
:i
iti
40.0 31.6
23.9 34.0
36.0 32.7
25.3 79.0
:; 2
iti itlF
44.0 32.0 33.3 29.3
36.0 12.0 26.1 22.7
45.3 12.0 28.0 34.7
40.0 67.3 45.0 57.3
39.2
25.1
31.4
52.1
1 :
ii
M ki
132.2 45.7 69.8
58.8 31.0 18.4
49.0 6i.8
137.1 83.8 36.5
4
:II
F
120.1
53.9
90.3
298.8
2 7
ii 30
ki
41.3 57.7 43.7
36.0 37.0 55.4
26.1 28.2 61.8
67.1 52.0 53.3
9" 10 Mean
30 :i
ki E
47.0 75.8 144.3 74.8
60.3 10.5 75.6 43.7
42.1 59.0 15.6 49.8
82.1 63.2 125.7 100.0
The area contained in each outline was then ascertained exhibited any systemic effects with the drugs used. Analysis
with
by
area (mm21 lsoproterenol
SOX
1; Mean Atopic:
to histamine
and nonatopic subiects to histamine and propranolol iontophoresis Drug and surface
Subject
reactivity
a planimeter.
No
subjects
of data
Testing with the paired t test showed no significant differences between the wheal areas produced by histamine on skin pretreated with saline iontophoresis and untreated skin in atopic and nonatopic subjects. Therefore, in analyzing the drug effects, the iontophoresed saline area was used aa the control value. Differences between the saline and drug wheal sizes within both atopic and nonatopic groups were evaluated by the paired t test. RESULTS
Data for each subject are seen in Table II. As a group, atopic individuals demonstrated significantly (p < 0.02) greater skin reactivity to histamine, as judged by wheal area, than nonatopic subjects. The whealing produced by histamine was modified by the iontophoresed agents. Diphenhydramine significantly decreased the wheal response in atopics (p < 0.005) and nonatopics (p < 0.02). Similar results were obtained when the beta adrenergic agonist, isoproterenol, was used. Both groups responded with a significantly decreased wheal area (P <
0.02).
When the beta adrenergic blocking agent-propranolol-was used, no significant difference in wheal size as compared to the control was seen in the atopic group. However, for the nonatopic subjects, propranolol did result in larger wheal areas than the control (p < 0.05). Statistical data are summarized in Table III.
452
Lamkin
TABLE
III.
J. ALLERGY CLIN.
et al.
Comparison
of
saline
control
and
drug
responses
for
atopic
and
IMMUNOL. MAY 1976
nonatopic
subjects Subjects
Diphenhydramine Isoproterenol Propranolol
Atopic Nonatopic Atopic Nonatopic Atopic Nonatopic
I
Paired “t” value
3.19 3.12
2.86 2.84 1.46 2.31
I
p value <0.005 <0.02 <0.02 <0.02
DISCUSSION
The purpose of this study was to determine to what degree, if any, the effects of agents which modify the CAMP-beta adrenergic system are directed toward the cutaneous vascular system as opposed to the dermal mast cell population. The modification of exogenous histamine whealing by these agents was evaluated in this regard. The technique of iontophoresis was used because of its previously demonstrated applicability and reproducibility as an in vivo technique.5 Also an attempt was made to determine if there were differences in response to these agents between atopic and nonatopic individuals. The present study demonstrates that the cutaneous response to histamine is subject to the effects of drugs which alter the metabolism of CAMP. Thus the previously demonstrated effect of these drugs on the immediate skin test5 may well have been due to an effect on the target organ (vasculature) rather than on the effector organ (mast cell). These results are contrary to those obtained by Perper, Sanda, and Lichtenstein.” In a previous study employing rhesus monkeys, these investigators noted that theophylline and salbutamol significantly decreased wheal formation following Ascclris challenge, but had no effect on whealing produced by the intradermal injection of histamine. However, the multiple differences in the studies make valid comparisons impossible. The studies differed with respect to drugs employed, their doses and route of administration, and the species involved. Of interest in our study is the difference in response between the atopic and nonatopic populations. The atopic patients responded more intensely to histamine. The responses to isoproterenol and propranolol are also worthy of note. As can be seen from the results in Table III, the beta agonist isoproterenol produced equally significant decrements in wheal size in both groups. This effect was anticipated and parallels the action of this agent in cellular preparations involving mediator release2e4and in a previous in vivo study involving immediate hypersensitivity skin tests.5 In contrast, propranolol failed to significantly alter wheal size in the atopic population but effectively increased wheal size in the nonatopic group. The results obtained with propranolol suggest the presence of a pre-existing cutaneous beta adrenergic blockade which partially inhibited the action of this
VOLUME 57 NUMBER 5
Cutaneous
reactivity
to histamine
453
drug on the vasculature in atopics. However, the similarity of responses of both groups to isoproterenol is not in keeping with this hypothesis. In summary, iontophoresis of drugs affecting CAMP metabolism effectively alters the cutaneous response to histamine. It is therefore assumed that regardless of the effects these drugs may have on the dermal mast cell after allergen challenge, at least part of their action is directed against the cutaneous vasculature. REFERENCES 1 Reed, C. E.: The pathogenesis of asthma, Med. Clin. North Am. 58: 55,1974. 2 Yamamoto, S., Greaves, M. W., and Fairley, V. M.: Cyclic AMP-induced inhibition of IgE-mediated hypersensitivity in human skin, Immunology 24: 77, 1973. release: In vitro 3 Lichtenstein, L. M., and DeBernardo, R.: IgE mediated histamine separation into two phases, Int. Arch. Allergy Appl. Immunol. 41: 56, 1971. 4 Ishizaka, T., Ishizaka, K., Orange, R. P., and Au&en, K. F.: Pharmacologic inhibition of the antigen-induced release of histamine and slow reacting substance of anaphylaxis (SRS-A) from monkey lung tissues mediated by human IgE, J. Immunol. 106: 1267, 1971. 5 Shereff, R. II., Harwell, W., Lieberman, P. L., Rosenberg, E. W., and Robinson, H.: Effect of beta adrenergic stimulation and blockade on immediate hypersensitivity skin test reactions, J. ALLERGY CLIN. IMMUNOL. 52: 328,1973. 6 Ichikawa, A., Nagasaki, M., Umezu, K., Hayaski, H., and Tomita, K. : Effect of cyclic adenosine 3’5’-monophosphate on edema and granuloma induced by carrageenin, Biochem. Pharmacol. 21: 2615, 1972. 7 Ichikawa, A., Hayashi, H., Minami, M., and Tomita, K.: An acute inflammation induced by inorganic pyrophosphate and adenosine triphosphate, and its inhibition by cyclic 3’5’adenosine monophosphate, Biochem. Pharmacol. 21: 317, 1972. 8 Lugnier, C., Bertrand, Y., and Stoclet, J. C.: Cyclic nucleotide phosphodiesterase inhibition and smooth vascular relaxation, Eur. J. Pharmacol. 19: 134, 1972. 9 Triner, L., Nahas, G. G., Vulliemoz, Y., Overweg, N. I. A., Veresky, M., Habif, D. V., and Ngai, S. H.: Cyclic AMP and smooth muscle function, Ann. N. Y. Acad. Sci. 185: 458, 1971. dermographia, Arch. 10 Kalz, F., Bower, C. M., and Prichard, H.: Delayed and persistent Dermatol. Syphilol. 61: 772, 1950. in vitro and in vivo 11 Perper, R. J., Sanda, M., and Lichtenstein, L. M.: The relationship allergic histamine release : inhibition in primates by CAMP active agents, Int. Arch. Allergy 48: 837, 1972.