Naunyn-Schmiedeberg's Arch. Pharmacol. 273, 401--413 (1972) �9 by Springer-Verlag 1972

Properties of the Histamine Stores Affected in the Anaphylactic and Anaphylatoxin Shock

of the Guinea Pig I . E f fec t of P r e t r e a t m e n t w i t h A n a p h y l a t o x i n *

G. GARB]~ and K. D. FRIWDB]~RG

Institut fiir Pharmakologie und Toxikologie der Universitat G6ttingen

Received February 23, 1972

Summary. 1. Anaphylatoxin lowers the histamine content in the guinea pig lung. This decrease ,can be demonstrated only immediately after the shock. 1 h later the decrease is no longer significant and normal values are being restored within 1 day.

2. The lung hisLamine content cannot be depressed to a larger extent and/or for a longer period of time by repeated anaphylatoxin injections neither during taehyphylaxis (every second hour) nor when tachy-phylaxis is diminished (every day) nor when it has completely disappeared (every second day).

3. Anaphylatoxin pretreatment (3 times every second hour) does not protect from the lethal effect of anaphylactic shock provoked either by a large or by a small dose of antigen. The plasma histamine level is only slightly decreased and a similar effect can be achieved by pretreatment with rat serum. Pretreatment with Ringer solution caused an increase in the plasma histamine level during anaphylactie shock. I t seems unIikely that an anaphylactic shock can be diminished by a pre- ceding depletion of the histamine stores with the aid of anaphylatoxin.

Key words: Histamine -- Anaphylactic Shock -- Anaphylatoxin Shock.

Guinea pig h i s tamine is l i be ra t ed in the anaphy lac t i c shock as well as in t he shock p r o v o k e d b y anaphy la tox in . The quest ion has been ra ised ve ry ear ly whe ther anaphy lac t i e reac t ions migh t be m e d i a t e d b y endogenous a n a p h y l a t o x i n genera t ion and whe ther an a n a p h y l a c t i c shock can t ake place dur ing a n a p h y l a t o x i n t a chyphy lax i s . I n i so la ted organs a n a p h y l a t o x i n fai led to reduce a following anaphy lac t i c reac t ion b u t the resul ts Jn the i n t a c t an imal were con t r ad i c to ry (for de ta i l ed references see Gier tz a n d Hahn , 1966). Some au thors r epo r t ed t h a t a desensi t iz ing effect of a n a p h y l a t o x i n aga ins t an t igen does in fact exist . T h e y assumed t h a t i t was based on a dep le t ion of the h i s t amine stores

* A preliminary report of a part of this study was presented during the 30. Ta- gung (25.--28. 9. 1966) of the Deutsche Pharmakologische Gesellschaft in Kiel (Friedberg and Poppe, 1967).

402 G.Garbe and K.D.Friedberg:

(Halpern e ta l . , 1956; Fr ick et al., 1962; Hahn , 1967; H a h n etal . , 1969, 1970). I n experiments of Friedberg e ta l . (1963, 1964, 1965), however, anaphy la tox in p re t r ea tmen t failed to reduce the lethal effect of an- aphylact ie shock. I t is the in t en t ion of this paper Co s tudy the changes of the his tamine stores in the guinea pig lung following the admin is t ra t ion of anaphyla toxin . I n addi t ion i t has been measured to what ex ten t the symptoms of an anaphylact ic shock might be influenced by a deplet ion of the h is tamine stores with the aid of anaphyla tox in .

Methods

Male and female guinea pigs were sensitized against native hen egg white. 3 times (every second day) each animal received 2.5 ml of a 400/0 solution of egg white interperitoneally. Shock was elicited not earlier than 22 days after sensi- tization had been started. The challenging dose usually was 1 ml/kg of a 20 ~ solution of the egg white. This is on the whole equivalent to 20 mg/kg ovalbumin (Speetor, 1956). The challenging dose was injected into the V. jugularis. Blood for the plasma histamine determination was withdrawn from the A. carotis. Both vessels were cannulated with polyethylene catheters under local anesthesia (0.5 ml procain solution 2~ s.c.). In order to prevent the acute anaphylactie shock 100 ~g/kg mepyramine maleate were injected 1 min prior to the antigen via the venous catheter. When plasma histamine levels were determined, aminoguanidine (10 mg/kg) was injected (i.v.) 0.5 rain prior to the antigen. 1.5 rain after the antigen injection 4--5 ml blood were withdrawn. Sodium oxalate (4 mg/ml) and amino- guanidine (2 ~g/ml) were added immediately. The survival time was registered and the lung removed for analysis. Histamine was determined as described by Shore and coworkers (1959). The histamine concentrations arc given as ~tg base per g or ml.

Anaphylatoxin (AT) was prepared from rat serum or plasma: rats were injected intraperitoneally with 0.75 ml/kg of a 5~ heparin solution. 15 rain later they were killed by a blow in the head and the carotid artery opened by an incision into the throat. The blood was collected in siliconized glass tubes containing 0.1 ml of the heparin solution. Rat serum was obtained from untreated animals. AT ac- tivation was performed with sephadex G 75 fine (i0 mg/ml) at 37~ (30 min). The lethal dose as determined by Friedberg and coworkers (1964) was 3--4 ml/kg. Subsequent experiments carried out with 3 ml/kg AT serum showed a lethality of 100~ (Friedberg st al., 1969). This result was confirmed with each series of AT serum which had been tested that way. Experiments, using AT plasma, were performed with 6 ml/kg, using AT serum even with 9 ml/kg. To prevent death in AT shock 25 tzg/kg mepyramine maleate were injected 1 min prior to the AT. The experiments according to Hahn etal. (1970) were done with 50 ~g/kg mepyramine maleate. Pretreatment by intravenous injections was done into the V. saphena of the hind leg. The vessel was exposed by a skin incision.

Statistical Evaluation

Each experiment was done at least in 5 animals. The measured parameters were distributed normally. The mean (~) and the standard error of the mean (8~) were calculated. The experiments were compared by the modified t-test respecting E-distribution. All p-values less than 0.2 are given in the tables and figures. A dif- ference was considered significant when p was less than 0.05.

Properties of the Histamine Stores. I 403

The lethality quotient was calculated as follows: The time between the injec- tion of the antigen and the cardiac arrest of the animal had been transformed to 100/min. The normal distribution of this quotient could be documented. The mean (2) and the standard error of the mean (s2) were calculated and the modified t-test was applied.

Results

A. Lung Histamine Content a/ter AT-Application

6 ml/kg AT plasma were injected intravenously into 63 guinea pigs. 21 animals which did not receive antihistamine died within 3- -5 rain. The lung histamine content was reduced significantly by about 25 ~ of the control level~ In those animals which had received antihistamine and were killed in general anestesia 1 h after AT, the lung histamine content was still reduced by 15 ~ After 24 h normal values were again present (Fig. i).

Next we wanted to know whether an intensified AT pret reatment is capable of lowering the lung histamine to a larger extent and for a longer period. As described by Friedberg el al. (1964) AT taehyphylaxis began to vanish 24 h after application and disappeared completely within 40 h. Therefore AT injections were performed under mepyramine cover every 24 h. 1 h after 5 or 10 injections of the same AT dose (6 ml/kg) the animals were killed in general anesthesia. The lung histamine content was unchanged when compared with controls (Table 1).

30,0

His ~glg

20.0

10.0

n= 20 21 21 2~ 0

C l xA r

th 24h

Fig. I. Recovery of the lung histamine content after anaphylatoxin application. C controls, untreated animals; AT anaphylatoxin plasma 6 ml/kg

404 G. Garbe and K.D. Friedberg:

Table 1. Lung histamine content after anaphylatoxin pretreatment. A T = anaphy- latoxin pla~na 6 ml/kg. The five~old and ten/old injections were carried out under mepyramine cover (25 pg/kg); all animals were killed I h after the last injection

Pretreatment Lung histamine content (txg/g)

$ 4 - s ~ n p

untreated animals 21,1 4- 1,9 20 "/. 5• AT in 5 days 18,8 -t- 2,4 20 --

10• AT in 10 days 26,7 -t- 3,4 11 -- 11• AT in 6 days 23,6 4- 1,8 23 --

Table 2. Scheme o/ the pretreatment with anaphylatoxin plasma ( = AT) without mepyramine cover similar to the arrangement described by Halpern et al. (1956)

Dose of AT-plasma

Day morning afternoon

1. 2 ml/kg 2 ml/kg 2. 3 ml/kg 3 ml/kg 3. 4 ml/kg 4 ml/kg 4. 5 ml/kg 5 ml/kg 5. 6 ml/kg 6 ml/kg 6. 6 ml/kg --

F ina l l y an expe r imen ta l scheme was adop t ed for AT p r e t r e a t m e n t s imilar to t h a t one descr ibed b y H a l p e r n et al. (1956) (Table 2). A T - p l a s m a was in jec ted twice da i ly for 5 days and once on the 5th d a y wi thou t an t ih i s t amine cover increasing the dose of A T each day . 1 h af te r the l as t in jec t ion the animals were k i l led and the h i s tamine con ten t of the lung was de te rmined . As compared to the lung h is tamine conten t in u n t r e a t e d an imals no change was observed (Table 1).

B. Plasma His tamine Level after AT-App l i ca t ion

AT plasma was injected into 10 guinea pigs. All animals died in acute shock (< 10 rain). The plasma histamine levels were not elevated (0.096 4- 0.024 ~g/ml;

4- s~5; n = 8). When AT-serum was injected instead of AT-plasma, however, high histamine levels were found in 19 guinea pigs (1.54 4- 0.18 ttg/ml; v5 4- s~; p < 0.001). The following experiments showed that the heparin content of the AT plasma released an amount of histaminase from the liver sufficient to destroy all the released histamine as previously reported by Bernauer et al. (1964). This enzyme could be blocked by i.v. injection of aminoguanidine (10 mg/kg) and ad- dition of aminoguanidine to the blood samples (2tzg/ml) (1,23-t-0,04[zg/ml;

4- s~; n = 5). The elevated histamine level after application of AT serum could be reduced by previous injection of heparin (500IU/kg; 0.27 4- 0.09 ttg/ml;

4- s~; n = 5). For these reasons all the following experiments were performed with AT serum.

Properties of the Histamine Stores. I 405

The same rise in plasma histamine following AT application was observed in animals which had received no or only a single AT injection before. Even in animals which had a totM of 5 AT injections (one in- jection every other day) the plasma histamine response did not decrease. All animals died in shock as the antihistamine was omitted before the last AT injection (Table 3).

C. Anaphylaetic Shock in AT.Pretreated Guinea Pigs

Under the same conditions we tested whether a following anaphylactic shock was modified. Actively sensitized animals received AT-serum under mepyramine cover 5 times in 48 h intervals. 24 h after the last AT application shock was elicited by i.v. injection of the antigen

Table 3. Increase in plasma histamine level by anat~hylato:cin serum (9 ml/kg) (= AT) . Injections were per]ormed every second day, the last one without

mepyramine cover

Pretreatment Plasma histamine (~g/ml)

Untreated animals 0,05 =t= 0,005 8 1• AT 1,54 =t= 0,18 19 2x AT 1,36 ~= 0,20 5 5• AT 1,40 ~ 0,18 13

./. < 0,001 < 0,001 < 0,005

His i tJg/g; 20.0

10.0

0

J ! _

n= 20 20

'q ,q

IN C C Ag AT+Ag

His pg/ml i

2.0 - I

1 "1 1

1.0

I

0 n= 11 12 Ag AT+Ag

Fig.2. Anaphylac'~ie shock in anaphylatoxin pretreated animals. Left part: lung histamine content; right part: plasma histamine level. C controls, untreated animals; C sens controls, sensitized animals; Ag guinea pigs treated with antigen; A T ~ Ag guinea pigs treated with anaphytatoxin serum (9 ml/kg; 5 times every second day) and 1 day later with antigen (native hen egg white equivalent to

20 mg/kg ovalbumin)

406 G. Garbe and K.D. Friedberg:

2 His pg /ml

plasma

2 mg/kg antigen

C AT

20 mg/kg antigen

c AT

§ lO0/min

15

I0

0 n= 12

surviving C C AT animals n= (5) (I) (01

AT (5)

N C AT C AT His pg/g 21.8 18.2 15.9 IZI 11.1

lung tissue *-2.3-+1.3 *-1.9 *-1.5 +-1.6 n= tO 12 12 12 12

= 0 2 ~-0.I .=0.I .=0.001 t - t e s t ~ - - ~" .r 0.02

Fig. 3. Plasma histamine level, lethality quotient and lung histamine content in anaphylactic shock after injection of AT (v~ q- s&). C Control animals received antihistamine only in the same time interval; A T antihistamine and AT (9 ml/kg) was injected 15 or 60 rnin prior to the antigen (= 2 or 20 mg/kg ovalbumin);

N lung histamine content in untreated guinea pigs

( ~ 2 0 mg/kg ovalbumin). All animals died in acute shock. The lung histamine content was diminished to the same degree in pretreated or in control animals; histamine release into blood plasma, too, was un- changed (Fig. 2).

1. I •

Hahn et al. (1969) reported tha t the plasma histamine level in anaphylaxis could be :reduced by three preceding AT-injections. Since these findings are opposite to our results we tested whether this effect could be confirmed with a single AT-injection immediately preceding the injection of antigen. The t ime interval was 15 and 60 rain, according to the recovery of the lung histamine content reported above. Since no

Properties of the Histamine Stores. Z 407

2 His tJg/ml

plasma

C

I cs ,:5 [ ~

Ri Set A T

20, ] 4:lO0/rnin

'~ 0 n= surviv/ng C

animals n= ( I )

i

Ri Ser (I) (2)

i AT ( I )

N C Ri Set" AT His pg /g 21,8 19.0 17.0 18.0 17.7

lung tissue +-2.3 +-3.1 +-2,5 +2.7 +1.6 n= 10 6 12 12 12

t - tesl "/" -- .;0.2 ' - .~ 0.2

Fig.4. Plasma histamine level, lethality quotient and lung histamine content in anaphylactie shock (= 2 mg/kg ovalbumin) 15 and 60 rain after a threefold i.v. injection of 9 ml/kg Ringer (= Ri), rat serum (= Set) or anaphylatoxin (~ AT) (2 =t= s&}. C anaphylac~ic shock in untreated animals; AT lung histamine content

in untreated guinea pigs

difference could be observed the results were gathered up. In addition we used a large and a small dose of antigen equivalent to 20 and 2 mg ovalbumin/kg. The control animals received antihistamine only (me- pyramine 50 ~zg/kg i.v.). The plasma histamine level in anaphylactie shock using the small antigen dosage is similar to tha t one reported by Hahn et al. (1969).

a) Plasma His tamine Level. The plasma histamine concentrations were not altered significantly by the AT-pretreatment, bu t the increased lethality quotient observed ff the high dose of antigen was administered seemed to bee accompanied by a slight increase in the histamine level (Fig. 3).

408 G. Garbe and K.D.Friedberg:

r

His p g / m l ! p l asma

20- aclOO/min

I0 -

./.

.0.-,=e_. surv iv ing C

an imals n= (o)

I 2~

1

iH Se.r AT

]

i

I I : l 26 12 12 Ri Set AT (2) (0) ( I )

N C Ri 5er AT His pg/g 21.8 18.0 15.0 12.1 16.8

lung tissue +-2.3 -+4.1 +-1.5 +-2.1 +-2.3 n= 10 6 26 12 12

t - tes t "/" .~. ,:0.025_ ,:0.01_ .c0.2_

Fig.5. Plasma histamine level, lethality quotient and lung histamine content in anaphylactie shock (= 20 mg/kg ovalbumin) 15 and 60 min after a threefold i.v. injection of 9 ml/kg Ringer (= ~i), rat serum (= Set) or anaphylatoxin (=AT) (4 ~ s~). C anaphylactie shock in untreated animals; N lung histamine content

in untreated guinea pigs

b) Lethality. The overall lethality was not altered. But when the large antigen dose was used the lethality quotient did increase signifi- cantly, indicating tha t the animals died earlier (Fig. 3).

c) Lung Histamine Content. The loss of histamine from the lung was more pronounced in AT-pretreated animals especially when the ]arge antigen dose was used (Fig. 3).

2. 3 x A T

I f the hypothesis of I t ahn et al. (1969) was right tha t an anaphylactic shock could be mitigated by preceding AT injections, the effect should be more marked ff the AT t)retreatment was intensified. Since the AT

Properties of the Histamine Stores. I 409

Table 4. Lung histamine content after various pretreatments with anaphylatoxin (= AT) or rat serum (= Ser). AT was injected (f) under mepyramine protection

(50 #g/kg) and the animals killed various times a/terwards

Pretreatment by Scheme of treatment Histamine

[xg/g lung tissue 2 -[-s2 n p

AT

Ser

[ ] 19 ,8 • 2 , 0 6 - -

~" 1" [] 16,0 4- 2,8 6 -- t" 1" "[" [] 27,7 • 9,4 6 -- 1" f r 15'[] 17,34-i,1 6 -- j" f ~- 60' [] 16,7 4- 3,0 5 - -

~" [ ] 2 3 , 1 4 - 5 , 6 6 - - 0 2 4h

~" [] 17,1 4- 3,2 7 - -

~" t [] 13,8 • 2,6 6 < 0,05 0 2 4h

Untreated animals 21,8 4- 2,3 10 "/.

serum was administered in a large volume (three times 9 ml/kg), Ringer solution and also native rat serum, were used for comparison. The anti- histamine (mepyramine maleate 50 ~g/kg) was given 0,5 rain prior to the first Ringer, serum or AT injection.

a) Plasma Histamine Level. When the animals received Ringer solution instead of the AT pre t rea tment the plasma histamine level during anaphylaxis was increased. After serum pretreatment , however, the plasma histamine level was slightly decreased, to about the same degree as after AT pre t rea tment (Fig. 4, 5).

b) Lethality. No obvious changes in the overall lethali ty or significant alterations of the lethali ty quotient could be obtained (Fig. 4, 5).

c) Lung Histamine Content. The lung histamine content was no t significantly changed with one exception (Figs.4,5). I n order to learn whether it could be altered already by the pret reatment , different schedules of AT and rat serum applications were investigated (Table4). Wi th one and fairly doubtful exception no significant decrease could be achieved.

d) Comparison o /Two Dif[erent Control Sera. Our experiments with normal ra t serum had an opposite result as those reported by H a h n et al. (1970). We therefore prepared "act ive serum" as described by these authors by incubat ing fresh serum for 45 rain at 37 ~ C. 15 min after p re t rea tment we challenged the animals with the small antigen dose ( ~ 2 mg/kg ovalbumin). Our results ment ioned above could be con-

28 Naunyn-Schmiedeberg's Arch. Pharmacol., Vol. 273

410 G.Garbe and K.D.Friedberg:

3 His pg /m l

plasma

C o

+ lO0/min l i

20-

!

,o t

O- C

surviving animals n= (2)

15 rain

~5

el Ri 5er Ser

37 "45 '

cN

I I

s el e Oi Ser Ser

37~ '

( I ) (o1 (2)

cu

AT

AT

( I )

N C Ri Se t Ser A T 3 7 0 4 5 ,

His l.lg/g 21.8 184 17.6 17.9 16.0 18.1 lung tissue +-4.0 +-3.7 +2.1 +-4.0 +2.5 +-2.5

n= I0 5 4 6 4 6

t - tes t % - - - ":0.2 - ./. . . . .

Fig. 6. Plasma histamine level, lethality quotient and lung histamine content in anaphylactie shock (= 2 mg/kg ovalbumin) 15 min after a threefold i.v. injection of 9 ml/kg Ringer (= _Ri), normal rat serum (= Net), rat serum incubated 45 rain at 37 ~ (~ Ser, 45' 37 ~ and anaphylatoxin (=AT) (2 • s2). C anaphylaetic shock

in untreated animals; 2g lung histamine content in untreated guinea pigs

firmed. In animals pretreated with Ringer solution plasma histamine levels rose more markedly. When antigen was injected in animals which had received either normal rat serum or rat serum which had been exposed to 37 degrees centigrade for 45 min nearly the same result was obtained as by pre t rea tment with AT (Fig. 6).

Discussion

Our experiments demonstra te t ha t AT does reduce the histamine content of the lung by about 25 ~ of control levels but t ha t this reduct ion does only occur in animMs dying in acute shock. Lung histamine stores

Properties of the Histamine Stores. I 411

are quickly recovering otherwise in surviving animals. Therefore the AT tachyphylaxis as present in isolated tissues and in the intact animals cannot be explained by an exhaustion of the histamine stores since taehyphylaxis outlasts the longer period of t ime than necessary to refill the histamine stores. Because of the quick recovery lung histamine stores cannot be reduced by repeated injections of AT. I t is not of relevance whether the AT injections are made in the presence or absence of the state of taehyphylaxis . The same fact is reflected by the unchanged high plasma histamine levels which were present even in animals which had received four AT injections before. This confirms the findings of Fried- berg et al. (1963, 1964, 1965).

But this result is contradictory to the finding of Frick et al. (1962). These authors reported, that after 6 days of twice-daily intravenous injections of AT the lung histamine content is reduced by about 75 ~ This low level persisted for nearly 3 weeks. Two possible explanations for this discrepancy can be made: The normal lung histamine content reported by these authors might be too high (mean: 38.6 ~g/g tissue; range 32.0--49.6; n = 16). Our histamine levels (21.i :~ 1.9 ~tg/g; 2 :k s2; n = 20) agree very well with those of Hongar and 8ohild (1953) (mean: 19.0 ~g/g) and giertz and Hahn (1955) (5.0--22.8 ~g/g). The second possible ex- planation may be: the intense contact of the animals with anaphylatoxin-prepara- tions might have led in spite of the short time to a sensitization under the conditions used by Halpern et al. (1956) and Frick etal . (1962). The last AT-injections might have caused a mild anaphylaetic shock, too. An anaphylaetic shock decreases the lung histamine content to about 50 ~ One week is necessary to reestablish normal values (Garbe and Friedberg, 1972).

Another finding of I-Ialpern and Frick can be confirmed: The lung tissue cannot be depleted totally of histamine. As will be shown (Garbe and Friedberg, 1972) the lung of the guinea pig seems to be able to take up histamine from the circulation. Therefore it might be impossible to completely deplete histamine from the lung. Because of these results Frick et al. (1962) assumed that there might be histamine pools in different cell types as already suggested by Hera (1959). This assumption is supported by Garbe and Friedberg (1972).

Since the lung histamine content after an AT mediated histamine release recovers very quickly it is unlikely tha t the following anaphylact ie shock can be weakened. But since H a h n and coworkers (1969, 1970) observed a decreased plasma histamine level and reduced lethali ty rate in anaphylaxis by previous administrat ion of AT, we repeated the ex- periments and modified the method used by t I a h n for the following reasons : Vigorous AT pre t rea tment has a densensitizing effect. This effect will be more pronounced when the t ime interval between the last AT injection and challenge is shortened (15 and 60 rain instead of 120 rain).

First we tried whether this desensitizing effect against anaphylaxis develops already after a single AT injection. Regarding the lung histamine content we see, t ha t there is an additional loss in histamine in AT pretreated animals. Using the low antigen dose we find a small reduct ion of the plasma histamine level. These findings correspond to the results

28*

412 G. Garbe and K. D. Friedberg:

of Hahn. But when the high antigen dose is administered we find a slight increase in the plasma histamine levels by AT pretreatment . I t is surprising tha t this is accompanied by an highly significant increase in the lethali ty quotient, indicating tha t the animals now die very quickly in the anaphylact ic shock.

Our experiments with a threefold AT-pre t rea tment again could only part ial ly confirm the results of H a h n et al. (1969, 1970). I n contrast to the findings of H a h n and coworkers the death rate in anaphylaxis was not decreased by any pre t rea tment and the lethali ty quotient showed no obvious alterations.

I f we regard the anaphylaet io plasma histamine level we see a reduc- t ion by the AT-pre t rea tment as reported by H a h n and coworkers (1969, 1970). But the same effect results f rom a pre t rea tment with normal ra t serum and socatled "act ive serum" (45 rain 37 ~ too. Therefore, the reduced release of histamine cannot be due to a specific effect of AT. This is in accordance with the lung histamine levels. I n contrast to a single AT injection repeated injections failed to intensify the loss of histamine from the lungs.

Surprisingly the pretreatment with I~inger solution did increase plasma hista- mine levels in anphylaxis. The same effect was observed by Hahn et al. (1970) after administration of active serum. As Hahn et al. we have not yet a satisfactory explanation for this phenomenon. We assume that unknown changes in the struc- ture of the histamine stores are responsible for these effects.

We conclude tha t AT itself is not able to modify the anaphylact ic shock by previous histamine liberation.

Acknowledgement. This work was supported by grants of the Deutsche For- schungsgemeinschaft.

References

Bernauer, W., Gicrtz, H., Hahn, F., Schmutzler, ~u Seseke, G., Sievers, B. U. : Zum Mechanismus der Histaminaseaktivierung im anaphylaktischen Schock. Naturwissenschaften 51, 412--413 (1964).

Frick, O. L., Halpern, B. N., Liacopoulos, 1 ). : The mechanism of protection from anaphylactic shock in the guinea-pig by pretreatment with anaphylatoxin. J. Physiol. (Lond.) 168, 191--199 (1962).

Friedberg, K. D., Bauer, U. : Anaphylaktischer Schock nach passiver Sensibilisie- rung am anaphylatoxinvorbehandelten Meerschweinchen. Naunyn-Schmiede- bergs Arch. exp. 1)ath. 1)harmak. ~50, 171--172 (1965).

--Engelhardt, G., Meineke, F.: ~ber die Tachyphylaxie der Anaphylatoxin- reaktion und ihre Bedeutung ffir die Anaphylaxie. Int. Arch. Allergy 22, 166--169 (1963).

-- -- -- Untersuchungen fiber die Anaphylatoxin-Tachyphylaxie und ihre Be- deutung for den Ablauf echter anaphylaktischer Reaktionen. Int. Arch. Allergy 25, 154--181 (1964).

Properties of the Histamine Stores. I 413

Friedberg, K.D., Garbe, G., Griitzmaeher, J. : Untersuchungen fiber die anti- anaphylaktisehe Wirkung des Chlorophyllins. Naunyn-Schmiedebergs Arch. Pharmak. 265, 287--300 (1969).

-- Poppe, I. : ~rber die Histaminfreisetzung aus der Meersehweinchenlunge durch Anaphylatoxin. Naunyn-Schmiedebergs Arch. Pharmak. exp. Path. 257, 19 (1967).

Garbe, G., Friedberg, K. D. : Properties of the histamine stores affected in the anaphylaetie and anaphylatoxin shock of the guinea pig. IL Effect of pretreatment with antigen. Naunyn-Sehmiedeberg's Arch. Pharmaeol. 278, 414--421 (1972).

Giertz, H., tIahn, F.: Die inverse Anaphylaxie vom Standpunkt der Histamin- theorie der Anaphylaxie. Int. Arch. Allergy 6, 23--44 (1955).

- - -- Makromolekulare Histaminliberatoren. In: M. Rocha e Silva: Histamine and Antihistaminics. Handbook of exp. Pharmaeol., Vol. XVIII/1, pp. 543--544. Berlin-Heidelberg-New York: Springer 1966.

Hahn, F. : Anaphylatoxin and anaphylaxis. Exerpta Medica International Congress Series Nr. 162, pp. 145--159. Allergology, Proceedings Sixth Congress of the International Association of Allergology. Montreal Nov. 5--11, 1967.

-- Ebner, C., Giertz, H. : Anaphylatoxin tachyphylaxis and anaphylaxis. Int. Arch. Allergy 85, 434--444 (1969).

-- Maack, R., Miiller, G., Mitze, 1~., Ebner, C. : Anaphylatoxin tachyphylaxis and anaphylaetie histamine release. Int. Arch. Allergy 87, 596--606 (1970).

Halpern, B.N., Liacopoulos, P., Briot, M.: Mecanism de protection du eobaye eontre le choe anaphylactique mortel par l'anaphylatoxin. Acta allerg. (Kbh.) 10, 9--14 (1956).

3{ongar, J. L., Sehild, H. O. : Quantitative measurement of the histamine-releasing activity of a series of monoalkylamines using minced guinea-pig lung. Brit. J. Pharmacol. 8, 103--109 (1953).

3/iota, J. : The mecihanism of action of anaphylatoxin. Its effect on guinea pig mast cells. Immunology 2, 403--413 (1959).

Shore, P. A., Burkhalter, A., Cohn, V. H. : A method for the fluorometrie assay of histamine in tissues. J. Pharmacol. exp. Ther. 127, 182--186 (1959).

Speetor, W. S.: Handbook of biological data, p. 190. Philadelphia-London: W. B. Saunders Comp. 1956.

K. D. Friedberg Institut ftir Pharmakologie und Toxikologie der Universiti~t GSttingen D-3400 GSttingen, Geiststr. 9 Germany