istamine hypersensitivity in mice induced by Bordeteffa pert&s or pharmacologic beta adrener Effects of adrenergic, Yukio
Matsumura,
John
H. Vaughan,
M.D.,** M.D.
cholinergic, Eng
M.
and other Tan,
M.D.,
drugs*
and
La Jolla, Calif.
The effects of prostagla.ndin E,, E,, F,, (PGE,, PGE, PGF,,), isoproterenol, epr’nephrine, norepinephrine, salbntamol, pmctolol, atropine, aminophylline, and corticosterone on the hypersensitivity to anaphylaxti, histamine, and serotonin in Bordetella pertussis-treated mice and propranolol-treated mice uere investigated. Female BLASW (ICR) mice, 27-29 gm, were injected with pertnssis vaccine intravenously 4 days before challenge with antigen, histamine, or seroton;i?z. Alternatively, instead of pertussis vaccine, propranolol was injected intraperitoneally 45 min before histamine challenge. Test arugs were administered intraperitoneally 15 min before challenge. PGE, and PGE, at a narrow range of between 10 and 100 pg and epinephrine at 100 fig protected both pertussisand propranolol-treated mice. Isoproterenol (25 pg) and aminophilline (800 pg) protected P-blocked mice, but did not protect pertussistreated mice even with very high doses (1,000 and 3,200 pg, respectively), although salbutamol (500 pg) did. PGF,,, norepinephrine, and atropine zLlere not protective at all. Practolol, a p,-blocker, given intraperitonea.lly 30 min. before histamine neither sensitized normal mice nor changed the efleot of isoproterenol or salbntamol in pertussis-treated mice. Corticosterone 10 mg/kg rednoed the number of deaths from histamine in P-blocked mice, but not in pertussis-treated mice. The protective eflect is discussed in connection u’ith probable effects of the drugs on intracellular cyclic adenosine monophosphate (CAMP) levels.
Bordetella pertussis-induced histamine hypersensitivity is an interesting animal model for the study of hyperreactivity to anaphylaxis and to chemical mediators in human bronchial asthma. Pertussis treatment altered the responses of mice to the ,8-adrenergic effects of epinephrine.l, 2 Adrenalectomy3 and adrenal demedullation4 of mice produced similar histamine hypersensitivity, which could From the Division of Allergy and Immunology, Scripps Clinic and Research Foundation. Supported by United States Public Health Service grants AI 00214 and AM 12198. Read in part at the Thirtieth Annual Meeting of the American Academy of Allergy, Miami, Fla., 1974. Received for publication Nov. 3, 1975. Accepted for publication Feb. 17, 1976. Reprint requests to: Dr. Eng M. Tan, Scripps Clinic and Research Foundation, 476 Prospect St., La Jolla, Calif. 92037. “Publication No. 55 from the Department of Clinical Research, Scripps Clinic and Research Foundation, La Jolla, Calif. *“Present address: Department of Medicine and Physical Therapy, Faculty of Medicine, University of Tokyo, Tokyo, Japan. 701.
58,
No.
3, pp.
395404
3%
Matsumura,
Tan,
and
Vaughan
J. ALLERGY
CLIN. IMMUNOL. SEPTEMBER 1976
be blocked with small doses of epinephrine, but much higher doses of epinephrine were required to protect pertussis-treated mice.j No histopathologic changes were observed in adrenal medulla of pertussis-treated mice.6 Pharmacologic /3-adrenergic blocking agents enhanced the histamine hypersensitivity in pertussis-treated mice7 and also produced this hypersensitivity in normal mice without pertussis treatment if sufficient amounts of the drugs were given.7-Q On the contrary, a-adrenergic blocking agents did not sensitize normal mice but diminished the pertussis-induced histamine hypersensitivity. 7g8~lo This evidence suggested impairment of the P-receptor-adenyl cyclase-cyclic adenosine monophosphate (CAMP) system of pertussis-treated mice. However, whether pertussis-induced histamine hypersensitivity is derived from ,&blockade or from other mechanisms has not been clarified. Adrenergic p-effects are considered to be mediated by adenyl cyclase and CAMP. In the present study, substances that might affect intracellular levels of cyclic nucleotides at different points in the reaction and through different mechanisms were tested to compare histamine hypersensitivity in pertussis-treated mice and in mice with pharmacologically induced P-adrenergic blockade. MATERlALS Experimental
AND METHODS animals
Female, B-10 week old, HLA-SW 27 to 29 g-m each, were used after arrival.
(ICR) mice (Hilltop Lab Animals, they were kept in our animal room
Inc., Scottsdale, at least a week
Pa.), after
Drugs Propranolol hydrochloride (Inderal injectable, Ayerst Laboratories, Inc., New York, N. Y.), isoproterenol (Sigma Chemical Co., St. Louis, MO., or Isuprel, Winthrop, New York, N. Y.), epinephrine (Adrenalin Chloride solution, Parke, Davis & Co., Detroit, Mich.), norepinephrine, corticosterone (Sigma Chemical Co., St. Louis, MO.) atropine sulfate (Burroughs Wellcome & Co., Research Triangle Park, N. C.), aminophylline (Searle Laboratories, Chicago, Ill.), histamine (histamine diphosphate, Matheson Coleman 6t Bell, Norwood, Ohio), and serotonin (serotonin creatinine sulfate monohydrat,e, Aldrich Chemical Co., Milwaukee, Wis.) were purchased. Prostaglandin E,, E,, and F,, (PGE,, PGE,, mnd PGFz,) were obtained through the courtesy of Dr. J. E. Pike, The Upjohn Co., Kalamazoo, Mich. Salbutamol (Albuterol) and practolol were kindly supplied by Schering Corporation, Kenilworth, N. J. and Ayerst Laboratories, Inc., N. Y., respectively.
Experimental
procedure
Mice were pretreated with either Bordetella pertussis vaccine or propranolol as described.11, 12 Bordetella pertussis vaccine was prepared as beforell: 5 or 10 yg in 0.2 ml phosphate-buffered saline (PBS: 0.15 M NaCl with 0.01 M phosphate, pH 7.4) was injected intravenously 4 days before testing. Propranolol hydrochloride, 40 or 250 pg in 0.2 ml PBS, was given intraperitoneally 45 min before histamine challenge. Histamine was dissolved in PBS, and 0.2 ml solution containing 1 mg histamine base was injected intraperitoneally. In some experiments with pertussis-treated mice, intraperitoneal challenge with 3 mg of serotonin base, or intravenous antigen challenge to induce passive systemic anaphylaxis, was used instead of histamine challenge. Passive systemic anaphylaxis was induced by sensitizing animals with mouse anti-egg albumin antibody and challenging them with 0.5 mg of egg albumin 6 hr later, as described before.11 Prostaglandins were dissolved in PBS immediately before use, and 0.2 ml of the solution was injected intraperitoneally 15 min before histamine. Catecholamines,
VOLUME NUMBER
58 3
Histamine
100 Et
0.5
yg Prostaglandin FIG. tivity 4
days
show
before
the
glandins. mice, higher
challenge
45 min histamine.
before Closed
cumulative PGE,
(10
and PGE, (20 doses of PGE,
with
mortality to
1
histamine. bars show 40
of pg)
and
100
52
1. The effects of prostaglandins, induced by Bordetella pertussis
toneally before
100
0.5
F2rr
PGE,, PGE,, and PGF,a on are shown. Pertussis vaccine mg histamine Prostaglandins the each PGE,
base. were
mortality control (40
of group pg)
hypersensitivity
the histamine was given
Propranolol injected
was given intraperitoneally
prostaglandin injected
significantly
to 100 fig) and PGE, (20 pg) protected and PGE, and any dose of PGE,a offered
hypersensiintravenously
group, with
PBS
protected
and instead
intraperi15 min open of
bars prosta-
pertussis-treated
P-blocked mice. no protection.
Lower
and
salbutamol, atropine and aminophylline were prepared in PBS, and 0.5 ml solution was injected intraperitoneally 13 min before histamine unless otherwise stated. Practolol, 1800 pg in 0.2 ml PBS, was injected intraperitoneally 30 min before histamine. Cortieosterone was dissolved in PBS containing 40v0 propylene glycol, and 0.05 ml was injected intraperitoneally 30 min before histamine, because corticosterone was not soluble in PBS, and because a greater dose of propylene glyeol itself protected mice from histamine death. A test group consisted of 10 mice. In certain instances, test groups consisted of more than 10 animals, as will be indicated in the text. In each experiment an equal volume of the solvent that had been mixed with the testing drug was injected into one group of 10 animals to serve as controls. The mortality of the group receiving a test drug was compared with the mortality of the respective control group 2 hr after histamine challenge by using either the xs test or Fisher’s testis at 5v0 level of statistical significance.
RESULTS Prostaglandins
The behavior of mice injected with PGE,, PGE,, and PGF,, was unchanged, except that they developed diarrhea 5 to 10 min after injection at all doses tested. As shown in Fig. 1, PGE, and PGE, protected pertussis-treated mice from histamine hypersensitivity at doses of 10 to 40 pg, 10 ,ug; respectively, but lower or higher doses were not protective. PGF,, did not protect them at any dose tested. The effects of postaglandins on propranolol-induced and pertussis-induced histamine hypersensitivity were similar. PGE, (10 pg) and PGE, (10 and 100 pg) also protected pertussis-treated mice from enhanced hypersensitivity to passive systemic anaphylaxis, but PGF,, (14 pg, similar in molarity to 10 pg of PGE, and PGE,) did not (Table I).
3
Matsumura,
TABLE I. anaphylaxis
Tan,
Effect of induced
Pertussis
(+g)
and
prostaglandins by Bordetella 1
PGE, PGE,
10
on the pertussis
Prostaglandin
1: 1:
J. ALLERGY
Vaughan
PGF;.
(pg)
enhanced in mice* 1
hypersensitivity
No. died/No.
0 (PBS) 0 (PBS) 10 0.5
tested
3120 1 o/20 4120 5/10
1;: 14
1: :0
of high doses of pertussis-treated
Isoproterenol Salbutamol
of
21140 24145 20/30
“Pertussis vaccine was injected 4 days before test. min before 1 mg histamine base challenge. tsignificantly lower than PBS control (p < 0.05).
lsoproterenol
and
and
Mortality
(%I
:it 22t 40
salbutamol
were passively sensitized challenge. Prostaglandins
on
13123 8/20 12128 Test
systemic
:+
ig 4110
isoproterenol mice*
passive
15
“Pertussis vaccine was given intravenously 4 days before test. Mice mouse anti-egg albumin 6 hr before 0.5 mg egg albumin intravenous were injected intraperitoneally 15 min before antigen challenge. tSignificantly lower than pertussis-treated PBS group (p < 0.05).
TABLE II. Effects hypersensitivity
to
CLIN. IMMUNOL. SEPTEMBER !976
the
histamine
3110
6120 5/10 l/29?
* drugs
were
given
by
intraperitoneally
15
salbutamol
As shown in Fig. 2, 25 to 100 pg of isoproterenol protected P-blocked mice from histamine hypersensitivity very effectively when 40 pg of propranolol was used to induce ,&adrene?gic blockade. When higher doses of propranolol (250 pg) were used, isoproterenol was less protective, but was still effective at higher doses. Isoproterenol, however, did not protect mice from pertussis-induced histamine hypersensitivity at doses that were effective in propranolol-treated mice. Table II shows that very high doses of isoproterenol (500 and 1,000 pg) were still ineffective in protecting mice from pertussis-induced histamine hypersensitivity. Furthermore, salbutamol at 100 pg was not effective, although high doses such as 500 and 1,000 pg did significantly reduce mortality from histamine in pertussis-treated mice. Neither isoproterenol nor salbutamol by themselves caused any abnormal symptoms in the animals even with the highest doses used. Bractolol
Intraperitoneal injection of 100 pg practolol, a pure ,8,-adrenergic blocking agent, did not produce histamine hypersensitivity in normal mice challenged 30 min later with 1 mg histamine base. The same injection of practolol changed neither the histamine hypersensitivity of pertussis-treated mice nor the effect of isoproterenol (100 pg) given 15 min before histamine challenge in pertussistreated mice.
VOLUME NUMBER
Histamine
58 3
lsoproterenol FIG.
2. The
effects
of
catecholamines
induced by different doses bars show the mortalities challenge. instead
Open of test
bars
Epinephrine and
of of
show
Bordetella the
groups
cumulative
Norepinephrine
Aminophylline
aminophylline
on
pertussis
and test
receiving mortality
of
hypersensitivity
the
histamine
propranolol drugs 15
control
groups,
hypersensitivity are
min
shown. Closed before histamine
which
received
PBS
drugs.
TABLE III. Effect of corticosterone pe’rtussis or by pharmacologic
on the P-blockade
I Pretreatment
Corticosterone 0 (Control)
Bordetella pertussis (10 wg iv) Propranolol(40 pg ip)
histamine in mice*
Mot 1 l/20
hypersensitivity
(mg/kg 5
induced
body
by
B~rde~e~~~
weight) 10
5/10 ;;ti
WO$
“Mice were pretreated either with pertussis vaccine 4 days before histamine or with propranolol 45 min before h&amine. Corticosterone was dissolved in PBS conta,ining 40y0 propylene glyeol, and 0.05 ml solution was injected intraperitoneally 15 min before histamine. Bxme volume of the solvent was given to the control group. Histamine was dissolved in PBS, and 0.02 ml containing 1 mg histamine base was injected intraperitoneally. fIXumber died/Number tested. $Significantly lower than the control (p < 0.05).
Epinephrine
and
norepinephrine
After the intraperitoneal injection of 100 pg epinephrine, but not of 50 yg, mice became lethargic and some of them died quietly within 5 to 10 min with rapid respirations or convulsions, One hundred pg of epinephrine protected mice from both pertussis-induced and propranolol-induced histamine hypersensitivity (Fig. 2), although 50 pg failed to protect them. This was unrelated to the possibility that we had selected animals resistant to histamine, since survivors retested with histamine alone two days later showed the same percent mortality as normal controls. Norepinephrine (25 to 100 pg) was not significantly protective in any instance. Since epinephrine was protective and this agent has both alpha and beta effects, it was conceivable that a relationship between these two properties was re-
480
Matsumura,
Tan,
and
J. ALLERGY
Vaughan
PBS
100
PBS
lsoprotercnol FIG.
3. The
effects
of
catecholamine
to histamine, serotonin, venously 4 days before show of
each
the
mortalities control
group
and test. of
the receiving
and
anaphylaxis Test drugs groups
100
PBS
100
PBS 3200 pg
Epincphrine llorcpinephrine aminophylline are were
receiving PBS instead
lminaphylline pertussis-induced
hypersensitivity
shown. Pertussis vaccine was given 15 min before challenge.
test of
on
CLIN. IMMUNQL. SEPTEMBER 1976
drugs, test
and
open
bars
show
given Closed the
intrabars
mortality
drugs.
sponsible for protection. In some experiments, combined isoproterenol (100 pg) and norepinephrine (50 pg) was tested, but it did not prevent death from either histamine or anaphylaxis in pertussis-treated mice. Atropine
Atropine (5 to 30 pug per mouse) did not protect mice either from hypersensitivity to anaphylaxis or from Bordetella pertussis or propranolol-induced histamine hypersensitivity. On the contrary, it appeared to increase death rate from pertussis-induced histamine hypersensitivity. Aminophylline
More tha.n 4 mg of aminophylline made mice hyperreactive to any physical stimuli; they became jumpy and even a sound caused convulsions. Four milligrams of aminophylline was often lethal, but 3.2 mg did not kill them although it made them somewhat overreactive. Aminophylline showed dose-related protection against histamine hypersensitivity induced by low dose (40 ,ug) of propranolol, but did not protect mice from histamine hypersensitivity induced by Bordetella pertussis or by high doses of propranolol (more than 100 pg) (Fig. 2). Corticosterone
Only a high dose (10 mg/kg) of corticosterone protected mice from histamine hypersensitivity induced by 40 pg propranolol, but it did not protect pertussistreated mice (Table III). Effect
on
mortality
caused
by
histamine,
serotonin,
and
systemic
aflaphyiaxi~
A comparison of the effects of different drugs on the percent mortality by histamine, serotonin, and anaphylaxis is shown in Fig. 3. Although
caused 100 pg
VOLUME NUMBER
58 3
Histamine
hypersensitivity
4
of isoproterenol did not protect pertussis-treated mice from hypersensitivity to histamine and anaphylaxis, it prevented death from serotonin (0% instead of 80%). Epinephrine (100 pg) was protective in all cases, but norepinephrine (106 pg) was not, Aminophylline (3.2 mg) did not protect pertussis-treated mice from histamine or serotonin, but did protect them from anaphylactic death (7% instead of 58%). DlSCUSSlON It has been observed that PGE, and PGE, stimulate adenyl cyclase and increase AMP level in humaa leukocytes14 and mouse spleen cells.15 PGF,, is less effectivelj or in some cases works conversely, decreasing the CAMP level.lG In the present study it was demonstrated that certain doses of PGE, and PGE, injected intraperitoneally 15 min before histamine or antigen challenge in passively sensitized animals protected mice from hypersensitivity to histamine and anaphylaxis induced by Bordetella pertussis and pharmacologic p-blockade but that PGF,, did not. These facts suggest some common impairment of pertussis-treated mice and ,&blocked mice, that can be restored by PGE, and PGE, but not by PGF,,, presumably impairment of the normal stimulation of adenyl cyelase through the ,&receptor. The reason high doses of PGEl and PGE, failed to protect mice was not clarified in this study, but a similar phenomenon was observed when pertussis-treated micell and P-blocked mice l2 were protected by exogenous cyclic AMP and dibutyryl CAMP from death by histamine hypersensitivity. Too low or too high a dose of CAMP failed to protect them. In the experiments illustrated in Fig. 1, it appeared that low doses of PGE, and PGE, might even have increased the mortality of mice treated with propranolol. However, the experimental method was constructed in such a way that it did not enable us to determine if this was a significant difference. The results from the experiments with isoproterenol agree with the observations of Fishel, Szentivanyi, and Talmage? and of Townley, Trapani, and Szen tivanyi.* Protection of ,&blocked mice from histamine hypersensitivity by intravenous5 and intraperitonealll epinephrine injection has been reported. Pertussisinduced histamine hypersensitivity, however, was apparently overcome by epinephrine only when the epinephrine was given intravenously 30 to 60 see after histamine5 ; intraperitoneal injection was ineffective.5l 7 This was probably because the dose was not as high as that used in the present study. Norepinephrine is a representative a-stimulant with some p-effect, and was not protective in any of our studies. Isoproterenol, a representative ,&stimulant, protected p-blocked mice effectively at a dose as low as 25 pg, but did not protect pertussis-treated mice even when 1,000 pg was injected. Epinephrine has strong p-effects with some aeffects. The reason only epinephrine protected mice from pertussis-induced hypersensitivity to histamine and anaphylaxis whereas neither isoproterenol nor norepinephrine did is hard to interpret. The possibility that a certain combination of (Y-and p-effect of epinephrine protected mice was unlikely because of two pieces of evidence. First, salbutamol, a pure &-stimulant without a-effect, protected pertussis-treated mice and, second, combinations of isoproterenol and norepinephrine were not protective. A second possibility, that the strong &-effect
2
Matsumura,
Tan,
and
Vaughan
J. ALLERGY
CLIN. IMMUNOL. SEPTEMBER 1976
of isoproterenol interfered with its protective &effect, was not supported since the combination of isoproterenol and practolol was not protective in pertussistreated mice. These results might suggest that &blockade by Bordetek pertmsis was so strong and irreversible that isoproterenol could not overcome it, but high doses of epinephrine and salbutamol could. Polson, Krzanowski, and Szentivanyil* reported that after histamine injection, lungs of pertussis-treated mice showed increased cyclic guanosine monophosphate (cGMP) . Exogenous cGMP did not protect mice from pertussis- or propranololinduced histamine hypersensitivity, although CAMP and some related nucleotides did.ll, I2 It was also noted that cGMP behaved antagonistically to CAMP in some cell functions.lg, 2o Therefore, it is possible that abnormality of another eyelic nucleotide system, cGMP, might be involved in the mechanism of the hypersensitivity of pertussis-treated mice. Lee, Kuo, and GreengardZ1 reported that cholinergic stimulation increased intracellular cGMP, which was inhibited by atropine. In the present study, atropine did not decrease but rather increased the deaths from histamine of pertusis-treated mice. The reason the death rate was increased by atropine was not clarified, but at least a hypercholinergic status as the mechanism of pertussis-induced hypersensitivity appeared to be unlikely. Ball and co-workersz2 reported that a-adrenergic stimulation did not change the CAMP level but raised the cGMP level in human plasma. This possibility of increased a-adrenergic reaction in pertussis-treated mice cannot be denied since a-blockers such as phenoxybenzamine and dibenamine decreased the histamine hypersensitivity,7> 8, lo although norepinephrine did not increase the number of deaths from histamine in the present study. Aminophylline reversed in a dose-related fashion the histamine hypersensitivity induced by pharmacologic P-blockade. Our results agree with the reports by TownlevY and co-workers.8a I7 However, even a high dose (3.2 mg) of aminophylline could not protect pertussis-treated mice from histamine hypersensitivity. Presumably, pertussis-treated mice are not subject to hyperfunction of phosphodiesterase but of some earlier step of the CAMP system. It could be a possibility that the P-receptors become impaired, and the inhibition of CAMP breakdown by the phosphodiesterase inhibitor, aminophylline, does not compensate for this defect. Aminophylline, which failed to prevent deaths from histamine and serotonin of pertussis-treated mice, did reduce the number of deaths from anaphylaxis significantly. One might interpret this in light of the report by Perper, Sanda, and LichtensteinZ3 that theophylline inhibits the immunologic release of histamine in vivo. Logsdon, Middleton, and Coffey”* reported that hydrocortisone not only stimulated adenyl cyclase in leukocytes of asthma patients but also reversed their decreased responsiveness to catecholamines. Corticosterone is the natural steroid hormone in mice. In the present study, 10 mg/kg of eorticosterone given intraperitoneally 30 min before histamine challenge did not protect pertussis-treated mice from histamine hypersensitivity. Townley, Daley and Selenkel? reported that 50 mg/kg of cortisol given intraperitoneally 4 hr before histamine failed to protect /?-blocked mice. In the present study, 10 mg/kg of corticosterone given intraperitoneally 30 min before histamine lowered the rates of death from hista-
VOLUME NUMBER
58 3
Histamine
hypersensitivity
mine of propra,nolol-treated mice, but even this unphysiologically high dose did not show complete protection. It should be noted that all of those substances that protected against pertussisinduced and also propranolol-induced (,&blocked) hypersensitivity of mice were those that have shown to increase intracellular CAMP. Further studies are needed to explain how two representative substances, isoproterenol and aminophylline, which increase intracellular cAMP by different mechanisms, protected mice only from propranolol-induced histamine hypersensitivity but not from that induced by pertussis. The
authors
thank
Mrs.
Pamela
Sweeney
for
her
skillful
assistance.
REFERENCES I Fishel, C. W., and Szentivanyi, A.: The absence of adrenaline-induced hyperglycemia in pertussis-sensitized mice and its relation to histamine and serotonin hypersensitivity, J. ALLERGY 34: 439, 1963. 2 Reed, C. W., Cohen, M., and Enta, T.: Reduced effect of epinephrine on circulating eosinophils in asthma and after beta.-adrenergic blockade or Bordetella per&s& vaeeine, J. ALLERGY 46: 90, 1970. 3 Kind, L. S.: The altered reactivity of mice after inoculation with Bordetella pertussis vaccine, Bacterial. Rev. 22: 173, 1958. 4 Bergman, R. K., and Munoz, J.: Adrenal medullary hormones and sensitization of mice to histamine by the histamine sensitizing factor of Bordetella pertmsis, Nature 205: 910, 1965. 5 Bergman, R. K., and Munoz, J.: Protection against histamine shock by catecholamines in Bordetella pertzwlsis-treated, adrenalectomized, or adrenergic blocked mice, Proc. Sot. Exp. Biol. Med. 122: 428, 1966. 6 Malkiel, 8.: Anaphylactic shock in the mouse vaccinated with Eemophilus pertwsis. IV. Studies on the adrenal gland, J. ALLERGY 2'7: 445, 1956. Sensitization and desensitization of 7 Fishel, C. W., Szentivanyi, A., and Talmage, D. W.: mice to histamine and serotonin by neurohumors, J. Immunol. 89: 8, 1962. 8 Townley, R. G., Trapani, I. L., and Szentivanyi, A.: Sensitization to anaphylaxis and to some of its pharmacological mediators by blockade of the beta adrenergic receptors, J. ALLERGY 39: 177, 1967. 9 Pieroni, R. E., and Levine, L. : Properties of the immunogenic and sensitizing activities of Bordetella pertussis in mice. I. Enhancement of peptone shock, J. ALLERGY 39: 25, 1967. 10 Kind, L. 8.: Inhibition of histamine death in pertussis-inoculated mice by dibenzgline and adrenergic blocking agent, J. ALLERGY 25: 33, 1954. 11 Matsumura, Y., Tan, E. M., and Vaughan, J. H.: The protective effects of nucleotides on the histamine hypersensitivity of Borcletella pertussis-treated mice, J. ALLERGY CLIN. IMMUNOL. 54: 191, 1974. 12 Matsumura, Y., Tan, E. M., and Vaughan, J. H.: Hypersensitivity to histamine and systemic anaphylaxis in mice with pharmacologic beta adrenergic blockade: Protection by nucleotides, J. ALLERGY CLIN. IYMUNOL. 58: 387,1976. 13 Fisher, R. A.: Statistical methods for research workers, ed. 6, London, 1936, Oliver & Boyd, Ltd., pp. 100-102. 14 Bourne, H. R., Lichtenstein, L. M., and Melmon, K. L.: Pharmacologic control of allergic histamine release in vitro: Evidence for an inhibitory role of 3’, 5’-adenosine monophosphate in human leukocytes, J. Immunol. 108: 685, 1972. 15 Matsumura, Y., Tan, E. M., and Vaughan, J. H.: Nucleotide uptake and interconversion into cyclic AMP by mouse spleen cells. (In preparation.) 16 Tauber, A. I., Kaliner, M., Stechsehulte, D. J., and Austen, K. F.: Immunologic release of histamine and slow reacting substance of anaphylaxis from human lung. V. Elects of prostaglandins on release of histamine, J. Immunol. 111: 27, 1973. 17 Townley, R. G., Daley, D., and Selenke, W.: The effect of agents used in the t,reatment of bronchial asthma on carbohydrate metabolism and histamine sensitivity after beta-adrenergic blockade,J. ALLERGY 45: 71, 1970.
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and
Vaughan
J. ALLERGY
cw IMMUNOL. SEPTEMBER 1976
18 Poison, J. B., Krzanomski, J. J., and Szentivanyi, A.: Effect of histamine (H) on the pulmonary levels of cyclic nucleotides in normal mice and under conditions of pharmacologic or bacterial sensitization, J. ALLERGY CLIN. IMMUNOL. 53: 100, 1974. (Abst.) 19 Kaliner, M., Orange, R. P., and Austen, K. F.: Immunological release of histamine and slow reacting substance of anaphylaxis from human lung. IV. Enhancement by cholinergic and alpha adrenergic stimulation, J. Exp. Med. 136: 556, 1972. 20 Zurier, R. B., Weissmann, G., Hoffstein, S., Kammerman, S. and Tai, H. H.: Mechanisms of lysosomal enzyme release from human leukocytes. II. Effects CAMP and cGMP, autonomic agonists, and agents which affeet microtubule function, J. Clin. Invest. 53: 297, 1974. 21 Lee, T.-P., Kuo, J. F., and Greengard, P.: Role of muscarinic cholinergic receptors in regulation of guanosine 3’ :5’-cyclic monophosphate content in mammalian brain, heart muscle, and intestinal smooth muscle, Proc. Natl. Acad. Sci. U.S.A. 69: 3287, 1972. 22 Ball, J. H., Kaminsky, N. I., Hardman, J. G., Broadus, A E., Sutherland E. W. and Liddle G. W.: Effects of catecholamines and adrenergic-blocking agents on plasma and urinary cyclic nucleotides in man. J. Clin. Invest. 51: 2124, 1972. 23 Perper, R. J., Sanda, M., and Lichtenstein, L. M.: The relationship of in vitro and in. utio allergic histamine release : Inhibition in primates by CAMP active agents, Int. Arch. Allergy 43: 837, 1972. 24 Logsdon, P. J., Middleton, E., Jr., and Coffey, R. G.: Stimulation of leukocyte adenyl cyclase by hydrocortisone and isoproterenol in asthmatic and non-asthmatic subjects, J. ALLERGY CLIN. IMMUNOL. 50: 45, 1972.
Matsumura,
John
H. Vaughan,
M.D.,** M.D.
cholinergic, Eng
M.
and other Tan,
M.D.,
drugs*
and
La Jolla, Calif.
The effects of prostagla.ndin E,, E,, F,, (PGE,, PGE, PGF,,), isoproterenol, epr’nephrine, norepinephrine, salbntamol, pmctolol, atropine, aminophylline, and corticosterone on the hypersensitivity to anaphylaxti, histamine, and serotonin in Bordetella pertussis-treated mice and propranolol-treated mice uere investigated. Female BLASW (ICR) mice, 27-29 gm, were injected with pertnssis vaccine intravenously 4 days before challenge with antigen, histamine, or seroton;i?z. Alternatively, instead of pertussis vaccine, propranolol was injected intraperitoneally 45 min before histamine challenge. Test arugs were administered intraperitoneally 15 min before challenge. PGE, and PGE, at a narrow range of between 10 and 100 pg and epinephrine at 100 fig protected both pertussisand propranolol-treated mice. Isoproterenol (25 pg) and aminophilline (800 pg) protected P-blocked mice, but did not protect pertussistreated mice even with very high doses (1,000 and 3,200 pg, respectively), although salbutamol (500 pg) did. PGF,,, norepinephrine, and atropine zLlere not protective at all. Practolol, a p,-blocker, given intraperitonea.lly 30 min. before histamine neither sensitized normal mice nor changed the efleot of isoproterenol or salbntamol in pertussis-treated mice. Corticosterone 10 mg/kg rednoed the number of deaths from histamine in P-blocked mice, but not in pertussis-treated mice. The protective eflect is discussed in connection u’ith probable effects of the drugs on intracellular cyclic adenosine monophosphate (CAMP) levels.
Bordetella pertussis-induced histamine hypersensitivity is an interesting animal model for the study of hyperreactivity to anaphylaxis and to chemical mediators in human bronchial asthma. Pertussis treatment altered the responses of mice to the ,8-adrenergic effects of epinephrine.l, 2 Adrenalectomy3 and adrenal demedullation4 of mice produced similar histamine hypersensitivity, which could From the Division of Allergy and Immunology, Scripps Clinic and Research Foundation. Supported by United States Public Health Service grants AI 00214 and AM 12198. Read in part at the Thirtieth Annual Meeting of the American Academy of Allergy, Miami, Fla., 1974. Received for publication Nov. 3, 1975. Accepted for publication Feb. 17, 1976. Reprint requests to: Dr. Eng M. Tan, Scripps Clinic and Research Foundation, 476 Prospect St., La Jolla, Calif. 92037. “Publication No. 55 from the Department of Clinical Research, Scripps Clinic and Research Foundation, La Jolla, Calif. *“Present address: Department of Medicine and Physical Therapy, Faculty of Medicine, University of Tokyo, Tokyo, Japan. 701.
58,
No.
3, pp.
395404
3%
Matsumura,
Tan,
and
Vaughan
J. ALLERGY
CLIN. IMMUNOL. SEPTEMBER 1976
be blocked with small doses of epinephrine, but much higher doses of epinephrine were required to protect pertussis-treated mice.j No histopathologic changes were observed in adrenal medulla of pertussis-treated mice.6 Pharmacologic /3-adrenergic blocking agents enhanced the histamine hypersensitivity in pertussis-treated mice7 and also produced this hypersensitivity in normal mice without pertussis treatment if sufficient amounts of the drugs were given.7-Q On the contrary, a-adrenergic blocking agents did not sensitize normal mice but diminished the pertussis-induced histamine hypersensitivity. 7g8~lo This evidence suggested impairment of the P-receptor-adenyl cyclase-cyclic adenosine monophosphate (CAMP) system of pertussis-treated mice. However, whether pertussis-induced histamine hypersensitivity is derived from ,&blockade or from other mechanisms has not been clarified. Adrenergic p-effects are considered to be mediated by adenyl cyclase and CAMP. In the present study, substances that might affect intracellular levels of cyclic nucleotides at different points in the reaction and through different mechanisms were tested to compare histamine hypersensitivity in pertussis-treated mice and in mice with pharmacologically induced P-adrenergic blockade. MATERlALS Experimental
AND METHODS animals
Female, B-10 week old, HLA-SW 27 to 29 g-m each, were used after arrival.
(ICR) mice (Hilltop Lab Animals, they were kept in our animal room
Inc., Scottsdale, at least a week
Pa.), after
Drugs Propranolol hydrochloride (Inderal injectable, Ayerst Laboratories, Inc., New York, N. Y.), isoproterenol (Sigma Chemical Co., St. Louis, MO., or Isuprel, Winthrop, New York, N. Y.), epinephrine (Adrenalin Chloride solution, Parke, Davis & Co., Detroit, Mich.), norepinephrine, corticosterone (Sigma Chemical Co., St. Louis, MO.) atropine sulfate (Burroughs Wellcome & Co., Research Triangle Park, N. C.), aminophylline (Searle Laboratories, Chicago, Ill.), histamine (histamine diphosphate, Matheson Coleman 6t Bell, Norwood, Ohio), and serotonin (serotonin creatinine sulfate monohydrat,e, Aldrich Chemical Co., Milwaukee, Wis.) were purchased. Prostaglandin E,, E,, and F,, (PGE,, PGE,, mnd PGFz,) were obtained through the courtesy of Dr. J. E. Pike, The Upjohn Co., Kalamazoo, Mich. Salbutamol (Albuterol) and practolol were kindly supplied by Schering Corporation, Kenilworth, N. J. and Ayerst Laboratories, Inc., N. Y., respectively.
Experimental
procedure
Mice were pretreated with either Bordetella pertussis vaccine or propranolol as described.11, 12 Bordetella pertussis vaccine was prepared as beforell: 5 or 10 yg in 0.2 ml phosphate-buffered saline (PBS: 0.15 M NaCl with 0.01 M phosphate, pH 7.4) was injected intravenously 4 days before testing. Propranolol hydrochloride, 40 or 250 pg in 0.2 ml PBS, was given intraperitoneally 45 min before histamine challenge. Histamine was dissolved in PBS, and 0.2 ml solution containing 1 mg histamine base was injected intraperitoneally. In some experiments with pertussis-treated mice, intraperitoneal challenge with 3 mg of serotonin base, or intravenous antigen challenge to induce passive systemic anaphylaxis, was used instead of histamine challenge. Passive systemic anaphylaxis was induced by sensitizing animals with mouse anti-egg albumin antibody and challenging them with 0.5 mg of egg albumin 6 hr later, as described before.11 Prostaglandins were dissolved in PBS immediately before use, and 0.2 ml of the solution was injected intraperitoneally 15 min before histamine. Catecholamines,
VOLUME NUMBER
58 3
Histamine
100 Et
0.5
yg Prostaglandin FIG. tivity 4
days
show
before
the
glandins. mice, higher
challenge
45 min histamine.
before Closed
cumulative PGE,
(10
and PGE, (20 doses of PGE,
with
mortality to
1
histamine. bars show 40
of pg)
and
100
52
1. The effects of prostaglandins, induced by Bordetella pertussis
toneally before
100
0.5
F2rr
PGE,, PGE,, and PGF,a on are shown. Pertussis vaccine mg histamine Prostaglandins the each PGE,
base. were
mortality control (40
of group pg)
hypersensitivity
the histamine was given
Propranolol injected
was given intraperitoneally
prostaglandin injected
significantly
to 100 fig) and PGE, (20 pg) protected and PGE, and any dose of PGE,a offered
hypersensiintravenously
group, with
PBS
protected
and instead
intraperi15 min open of
bars prosta-
pertussis-treated
P-blocked mice. no protection.
Lower
and
salbutamol, atropine and aminophylline were prepared in PBS, and 0.5 ml solution was injected intraperitoneally 13 min before histamine unless otherwise stated. Practolol, 1800 pg in 0.2 ml PBS, was injected intraperitoneally 30 min before histamine. Cortieosterone was dissolved in PBS containing 40v0 propylene glycol, and 0.05 ml was injected intraperitoneally 30 min before histamine, because corticosterone was not soluble in PBS, and because a greater dose of propylene glyeol itself protected mice from histamine death. A test group consisted of 10 mice. In certain instances, test groups consisted of more than 10 animals, as will be indicated in the text. In each experiment an equal volume of the solvent that had been mixed with the testing drug was injected into one group of 10 animals to serve as controls. The mortality of the group receiving a test drug was compared with the mortality of the respective control group 2 hr after histamine challenge by using either the xs test or Fisher’s testis at 5v0 level of statistical significance.
RESULTS Prostaglandins
The behavior of mice injected with PGE,, PGE,, and PGF,, was unchanged, except that they developed diarrhea 5 to 10 min after injection at all doses tested. As shown in Fig. 1, PGE, and PGE, protected pertussis-treated mice from histamine hypersensitivity at doses of 10 to 40 pg, 10 ,ug; respectively, but lower or higher doses were not protective. PGF,, did not protect them at any dose tested. The effects of postaglandins on propranolol-induced and pertussis-induced histamine hypersensitivity were similar. PGE, (10 pg) and PGE, (10 and 100 pg) also protected pertussis-treated mice from enhanced hypersensitivity to passive systemic anaphylaxis, but PGF,, (14 pg, similar in molarity to 10 pg of PGE, and PGE,) did not (Table I).
3
Matsumura,
TABLE I. anaphylaxis
Tan,
Effect of induced
Pertussis
(+g)
and
prostaglandins by Bordetella 1
PGE, PGE,
10
on the pertussis
Prostaglandin
1: 1:
J. ALLERGY
Vaughan
PGF;.
(pg)
enhanced in mice* 1
hypersensitivity
No. died/No.
0 (PBS) 0 (PBS) 10 0.5
tested
3120 1 o/20 4120 5/10
1;: 14
1: :0
of high doses of pertussis-treated
Isoproterenol Salbutamol
of
21140 24145 20/30
“Pertussis vaccine was injected 4 days before test. min before 1 mg histamine base challenge. tsignificantly lower than PBS control (p < 0.05).
lsoproterenol
and
and
Mortality
(%I
:it 22t 40
salbutamol
were passively sensitized challenge. Prostaglandins
on
13123 8/20 12128 Test
systemic
:+
ig 4110
isoproterenol mice*
passive
15
“Pertussis vaccine was given intravenously 4 days before test. Mice mouse anti-egg albumin 6 hr before 0.5 mg egg albumin intravenous were injected intraperitoneally 15 min before antigen challenge. tSignificantly lower than pertussis-treated PBS group (p < 0.05).
TABLE II. Effects hypersensitivity
to
CLIN. IMMUNOL. SEPTEMBER !976
the
histamine
3110
6120 5/10 l/29?
* drugs
were
given
by
intraperitoneally
15
salbutamol
As shown in Fig. 2, 25 to 100 pg of isoproterenol protected P-blocked mice from histamine hypersensitivity very effectively when 40 pg of propranolol was used to induce ,&adrene?gic blockade. When higher doses of propranolol (250 pg) were used, isoproterenol was less protective, but was still effective at higher doses. Isoproterenol, however, did not protect mice from pertussis-induced histamine hypersensitivity at doses that were effective in propranolol-treated mice. Table II shows that very high doses of isoproterenol (500 and 1,000 pg) were still ineffective in protecting mice from pertussis-induced histamine hypersensitivity. Furthermore, salbutamol at 100 pg was not effective, although high doses such as 500 and 1,000 pg did significantly reduce mortality from histamine in pertussis-treated mice. Neither isoproterenol nor salbutamol by themselves caused any abnormal symptoms in the animals even with the highest doses used. Bractolol
Intraperitoneal injection of 100 pg practolol, a pure ,8,-adrenergic blocking agent, did not produce histamine hypersensitivity in normal mice challenged 30 min later with 1 mg histamine base. The same injection of practolol changed neither the histamine hypersensitivity of pertussis-treated mice nor the effect of isoproterenol (100 pg) given 15 min before histamine challenge in pertussistreated mice.
VOLUME NUMBER
Histamine
58 3
lsoproterenol FIG.
2. The
effects
of
catecholamines
induced by different doses bars show the mortalities challenge. instead
Open of test
bars
Epinephrine and
of of
show
Bordetella the
groups
cumulative
Norepinephrine
Aminophylline
aminophylline
on
pertussis
and test
receiving mortality
of
hypersensitivity
the
histamine
propranolol drugs 15
control
groups,
hypersensitivity are
min
shown. Closed before histamine
which
received
PBS
drugs.
TABLE III. Effect of corticosterone pe’rtussis or by pharmacologic
on the P-blockade
I Pretreatment
Corticosterone 0 (Control)
Bordetella pertussis (10 wg iv) Propranolol(40 pg ip)
histamine in mice*
Mot 1 l/20
hypersensitivity
(mg/kg 5
induced
body
by
B~rde~e~~~
weight) 10
5/10 ;;ti
WO$
“Mice were pretreated either with pertussis vaccine 4 days before histamine or with propranolol 45 min before h&amine. Corticosterone was dissolved in PBS conta,ining 40y0 propylene glyeol, and 0.05 ml solution was injected intraperitoneally 15 min before histamine. Bxme volume of the solvent was given to the control group. Histamine was dissolved in PBS, and 0.02 ml containing 1 mg histamine base was injected intraperitoneally. fIXumber died/Number tested. $Significantly lower than the control (p < 0.05).
Epinephrine
and
norepinephrine
After the intraperitoneal injection of 100 pg epinephrine, but not of 50 yg, mice became lethargic and some of them died quietly within 5 to 10 min with rapid respirations or convulsions, One hundred pg of epinephrine protected mice from both pertussis-induced and propranolol-induced histamine hypersensitivity (Fig. 2), although 50 pg failed to protect them. This was unrelated to the possibility that we had selected animals resistant to histamine, since survivors retested with histamine alone two days later showed the same percent mortality as normal controls. Norepinephrine (25 to 100 pg) was not significantly protective in any instance. Since epinephrine was protective and this agent has both alpha and beta effects, it was conceivable that a relationship between these two properties was re-
480
Matsumura,
Tan,
and
J. ALLERGY
Vaughan
PBS
100
PBS
lsoprotercnol FIG.
3. The
effects
of
catecholamine
to histamine, serotonin, venously 4 days before show of
each
the
mortalities control
group
and test. of
the receiving
and
anaphylaxis Test drugs groups
100
PBS
100
PBS 3200 pg
Epincphrine llorcpinephrine aminophylline are were
receiving PBS instead
lminaphylline pertussis-induced
hypersensitivity
shown. Pertussis vaccine was given 15 min before challenge.
test of
on
CLIN. IMMUNQL. SEPTEMBER 1976
drugs, test
and
open
bars
show
given Closed the
intrabars
mortality
drugs.
sponsible for protection. In some experiments, combined isoproterenol (100 pg) and norepinephrine (50 pg) was tested, but it did not prevent death from either histamine or anaphylaxis in pertussis-treated mice. Atropine
Atropine (5 to 30 pug per mouse) did not protect mice either from hypersensitivity to anaphylaxis or from Bordetella pertussis or propranolol-induced histamine hypersensitivity. On the contrary, it appeared to increase death rate from pertussis-induced histamine hypersensitivity. Aminophylline
More tha.n 4 mg of aminophylline made mice hyperreactive to any physical stimuli; they became jumpy and even a sound caused convulsions. Four milligrams of aminophylline was often lethal, but 3.2 mg did not kill them although it made them somewhat overreactive. Aminophylline showed dose-related protection against histamine hypersensitivity induced by low dose (40 ,ug) of propranolol, but did not protect mice from histamine hypersensitivity induced by Bordetella pertussis or by high doses of propranolol (more than 100 pg) (Fig. 2). Corticosterone
Only a high dose (10 mg/kg) of corticosterone protected mice from histamine hypersensitivity induced by 40 pg propranolol, but it did not protect pertussistreated mice (Table III). Effect
on
mortality
caused
by
histamine,
serotonin,
and
systemic
aflaphyiaxi~
A comparison of the effects of different drugs on the percent mortality by histamine, serotonin, and anaphylaxis is shown in Fig. 3. Although
caused 100 pg
VOLUME NUMBER
58 3
Histamine
hypersensitivity
4
of isoproterenol did not protect pertussis-treated mice from hypersensitivity to histamine and anaphylaxis, it prevented death from serotonin (0% instead of 80%). Epinephrine (100 pg) was protective in all cases, but norepinephrine (106 pg) was not, Aminophylline (3.2 mg) did not protect pertussis-treated mice from histamine or serotonin, but did protect them from anaphylactic death (7% instead of 58%). DlSCUSSlON It has been observed that PGE, and PGE, stimulate adenyl cyclase and increase AMP level in humaa leukocytes14 and mouse spleen cells.15 PGF,, is less effectivelj or in some cases works conversely, decreasing the CAMP level.lG In the present study it was demonstrated that certain doses of PGE, and PGE, injected intraperitoneally 15 min before histamine or antigen challenge in passively sensitized animals protected mice from hypersensitivity to histamine and anaphylaxis induced by Bordetella pertussis and pharmacologic p-blockade but that PGF,, did not. These facts suggest some common impairment of pertussis-treated mice and ,&blocked mice, that can be restored by PGE, and PGE, but not by PGF,,, presumably impairment of the normal stimulation of adenyl cyelase through the ,&receptor. The reason high doses of PGEl and PGE, failed to protect mice was not clarified in this study, but a similar phenomenon was observed when pertussis-treated micell and P-blocked mice l2 were protected by exogenous cyclic AMP and dibutyryl CAMP from death by histamine hypersensitivity. Too low or too high a dose of CAMP failed to protect them. In the experiments illustrated in Fig. 1, it appeared that low doses of PGE, and PGE, might even have increased the mortality of mice treated with propranolol. However, the experimental method was constructed in such a way that it did not enable us to determine if this was a significant difference. The results from the experiments with isoproterenol agree with the observations of Fishel, Szentivanyi, and Talmage? and of Townley, Trapani, and Szen tivanyi.* Protection of ,&blocked mice from histamine hypersensitivity by intravenous5 and intraperitonealll epinephrine injection has been reported. Pertussisinduced histamine hypersensitivity, however, was apparently overcome by epinephrine only when the epinephrine was given intravenously 30 to 60 see after histamine5 ; intraperitoneal injection was ineffective.5l 7 This was probably because the dose was not as high as that used in the present study. Norepinephrine is a representative a-stimulant with some p-effect, and was not protective in any of our studies. Isoproterenol, a representative ,&stimulant, protected p-blocked mice effectively at a dose as low as 25 pg, but did not protect pertussis-treated mice even when 1,000 pg was injected. Epinephrine has strong p-effects with some aeffects. The reason only epinephrine protected mice from pertussis-induced hypersensitivity to histamine and anaphylaxis whereas neither isoproterenol nor norepinephrine did is hard to interpret. The possibility that a certain combination of (Y-and p-effect of epinephrine protected mice was unlikely because of two pieces of evidence. First, salbutamol, a pure &-stimulant without a-effect, protected pertussis-treated mice and, second, combinations of isoproterenol and norepinephrine were not protective. A second possibility, that the strong &-effect
2
Matsumura,
Tan,
and
Vaughan
J. ALLERGY
CLIN. IMMUNOL. SEPTEMBER 1976
of isoproterenol interfered with its protective &effect, was not supported since the combination of isoproterenol and practolol was not protective in pertussistreated mice. These results might suggest that &blockade by Bordetek pertmsis was so strong and irreversible that isoproterenol could not overcome it, but high doses of epinephrine and salbutamol could. Polson, Krzanowski, and Szentivanyil* reported that after histamine injection, lungs of pertussis-treated mice showed increased cyclic guanosine monophosphate (cGMP) . Exogenous cGMP did not protect mice from pertussis- or propranololinduced histamine hypersensitivity, although CAMP and some related nucleotides did.ll, I2 It was also noted that cGMP behaved antagonistically to CAMP in some cell functions.lg, 2o Therefore, it is possible that abnormality of another eyelic nucleotide system, cGMP, might be involved in the mechanism of the hypersensitivity of pertussis-treated mice. Lee, Kuo, and GreengardZ1 reported that cholinergic stimulation increased intracellular cGMP, which was inhibited by atropine. In the present study, atropine did not decrease but rather increased the deaths from histamine of pertusis-treated mice. The reason the death rate was increased by atropine was not clarified, but at least a hypercholinergic status as the mechanism of pertussis-induced hypersensitivity appeared to be unlikely. Ball and co-workersz2 reported that a-adrenergic stimulation did not change the CAMP level but raised the cGMP level in human plasma. This possibility of increased a-adrenergic reaction in pertussis-treated mice cannot be denied since a-blockers such as phenoxybenzamine and dibenamine decreased the histamine hypersensitivity,7> 8, lo although norepinephrine did not increase the number of deaths from histamine in the present study. Aminophylline reversed in a dose-related fashion the histamine hypersensitivity induced by pharmacologic P-blockade. Our results agree with the reports by TownlevY and co-workers.8a I7 However, even a high dose (3.2 mg) of aminophylline could not protect pertussis-treated mice from histamine hypersensitivity. Presumably, pertussis-treated mice are not subject to hyperfunction of phosphodiesterase but of some earlier step of the CAMP system. It could be a possibility that the P-receptors become impaired, and the inhibition of CAMP breakdown by the phosphodiesterase inhibitor, aminophylline, does not compensate for this defect. Aminophylline, which failed to prevent deaths from histamine and serotonin of pertussis-treated mice, did reduce the number of deaths from anaphylaxis significantly. One might interpret this in light of the report by Perper, Sanda, and LichtensteinZ3 that theophylline inhibits the immunologic release of histamine in vivo. Logsdon, Middleton, and Coffey”* reported that hydrocortisone not only stimulated adenyl cyclase in leukocytes of asthma patients but also reversed their decreased responsiveness to catecholamines. Corticosterone is the natural steroid hormone in mice. In the present study, 10 mg/kg of eorticosterone given intraperitoneally 30 min before histamine challenge did not protect pertussis-treated mice from histamine hypersensitivity. Townley, Daley and Selenkel? reported that 50 mg/kg of cortisol given intraperitoneally 4 hr before histamine failed to protect /?-blocked mice. In the present study, 10 mg/kg of corticosterone given intraperitoneally 30 min before histamine lowered the rates of death from hista-
VOLUME NUMBER
58 3
Histamine
hypersensitivity
mine of propra,nolol-treated mice, but even this unphysiologically high dose did not show complete protection. It should be noted that all of those substances that protected against pertussisinduced and also propranolol-induced (,&blocked) hypersensitivity of mice were those that have shown to increase intracellular CAMP. Further studies are needed to explain how two representative substances, isoproterenol and aminophylline, which increase intracellular cAMP by different mechanisms, protected mice only from propranolol-induced histamine hypersensitivity but not from that induced by pertussis. The
authors
thank
Mrs.
Pamela
Sweeney
for
her
skillful
assistance.
REFERENCES I Fishel, C. W., and Szentivanyi, A.: The absence of adrenaline-induced hyperglycemia in pertussis-sensitized mice and its relation to histamine and serotonin hypersensitivity, J. ALLERGY 34: 439, 1963. 2 Reed, C. W., Cohen, M., and Enta, T.: Reduced effect of epinephrine on circulating eosinophils in asthma and after beta.-adrenergic blockade or Bordetella per&s& vaeeine, J. ALLERGY 46: 90, 1970. 3 Kind, L. S.: The altered reactivity of mice after inoculation with Bordetella pertussis vaccine, Bacterial. Rev. 22: 173, 1958. 4 Bergman, R. K., and Munoz, J.: Adrenal medullary hormones and sensitization of mice to histamine by the histamine sensitizing factor of Bordetella pertmsis, Nature 205: 910, 1965. 5 Bergman, R. K., and Munoz, J.: Protection against histamine shock by catecholamines in Bordetella pertzwlsis-treated, adrenalectomized, or adrenergic blocked mice, Proc. Sot. Exp. Biol. Med. 122: 428, 1966. 6 Malkiel, 8.: Anaphylactic shock in the mouse vaccinated with Eemophilus pertwsis. IV. Studies on the adrenal gland, J. ALLERGY 2'7: 445, 1956. Sensitization and desensitization of 7 Fishel, C. W., Szentivanyi, A., and Talmage, D. W.: mice to histamine and serotonin by neurohumors, J. Immunol. 89: 8, 1962. 8 Townley, R. G., Trapani, I. L., and Szentivanyi, A.: Sensitization to anaphylaxis and to some of its pharmacological mediators by blockade of the beta adrenergic receptors, J. ALLERGY 39: 177, 1967. 9 Pieroni, R. E., and Levine, L. : Properties of the immunogenic and sensitizing activities of Bordetella pertussis in mice. I. Enhancement of peptone shock, J. ALLERGY 39: 25, 1967. 10 Kind, L. 8.: Inhibition of histamine death in pertussis-inoculated mice by dibenzgline and adrenergic blocking agent, J. ALLERGY 25: 33, 1954. 11 Matsumura, Y., Tan, E. M., and Vaughan, J. H.: The protective effects of nucleotides on the histamine hypersensitivity of Borcletella pertussis-treated mice, J. ALLERGY CLIN. IMMUNOL. 54: 191, 1974. 12 Matsumura, Y., Tan, E. M., and Vaughan, J. H.: Hypersensitivity to histamine and systemic anaphylaxis in mice with pharmacologic beta adrenergic blockade: Protection by nucleotides, J. ALLERGY CLIN. IYMUNOL. 58: 387,1976. 13 Fisher, R. A.: Statistical methods for research workers, ed. 6, London, 1936, Oliver & Boyd, Ltd., pp. 100-102. 14 Bourne, H. R., Lichtenstein, L. M., and Melmon, K. L.: Pharmacologic control of allergic histamine release in vitro: Evidence for an inhibitory role of 3’, 5’-adenosine monophosphate in human leukocytes, J. Immunol. 108: 685, 1972. 15 Matsumura, Y., Tan, E. M., and Vaughan, J. H.: Nucleotide uptake and interconversion into cyclic AMP by mouse spleen cells. (In preparation.) 16 Tauber, A. I., Kaliner, M., Stechsehulte, D. J., and Austen, K. F.: Immunologic release of histamine and slow reacting substance of anaphylaxis from human lung. V. Elects of prostaglandins on release of histamine, J. Immunol. 111: 27, 1973. 17 Townley, R. G., Daley, D., and Selenke, W.: The effect of agents used in the t,reatment of bronchial asthma on carbohydrate metabolism and histamine sensitivity after beta-adrenergic blockade,J. ALLERGY 45: 71, 1970.
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cw IMMUNOL. SEPTEMBER 1976
18 Poison, J. B., Krzanomski, J. J., and Szentivanyi, A.: Effect of histamine (H) on the pulmonary levels of cyclic nucleotides in normal mice and under conditions of pharmacologic or bacterial sensitization, J. ALLERGY CLIN. IMMUNOL. 53: 100, 1974. (Abst.) 19 Kaliner, M., Orange, R. P., and Austen, K. F.: Immunological release of histamine and slow reacting substance of anaphylaxis from human lung. IV. Enhancement by cholinergic and alpha adrenergic stimulation, J. Exp. Med. 136: 556, 1972. 20 Zurier, R. B., Weissmann, G., Hoffstein, S., Kammerman, S. and Tai, H. H.: Mechanisms of lysosomal enzyme release from human leukocytes. II. Effects CAMP and cGMP, autonomic agonists, and agents which affeet microtubule function, J. Clin. Invest. 53: 297, 1974. 21 Lee, T.-P., Kuo, J. F., and Greengard, P.: Role of muscarinic cholinergic receptors in regulation of guanosine 3’ :5’-cyclic monophosphate content in mammalian brain, heart muscle, and intestinal smooth muscle, Proc. Natl. Acad. Sci. U.S.A. 69: 3287, 1972. 22 Ball, J. H., Kaminsky, N. I., Hardman, J. G., Broadus, A E., Sutherland E. W. and Liddle G. W.: Effects of catecholamines and adrenergic-blocking agents on plasma and urinary cyclic nucleotides in man. J. Clin. Invest. 51: 2124, 1972. 23 Perper, R. J., Sanda, M., and Lichtenstein, L. M.: The relationship of in vitro and in. utio allergic histamine release : Inhibition in primates by CAMP active agents, Int. Arch. Allergy 43: 837, 1972. 24 Logsdon, P. J., Middleton, E., Jr., and Coffey, R. G.: Stimulation of leukocyte adenyl cyclase by hydrocortisone and isoproterenol in asthmatic and non-asthmatic subjects, J. ALLERGY CLIN. IMMUNOL. 50: 45, 1972.
