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Reduction of the inflammatory response in rats immunized against prostaglandins
PROSTAGLANDINS
REDUCTION OF
THE
IMMUNIZEO
To
the
INFLAMMATORY AGAINST
RESPONSE
IN
RATS
PROSTAGLANOINS
Editor,
The notion that proataglandins contribute to the pathogenesis of the inflammatory response is supported by three Firstly, prostaglandins are generated in main areuments. carrageenin inflammany types of tissue damage, for-example mation t l-31, thermal injury to the skin (4-61, arthritis uveitis (101 and U.V. induced inflam(7,81 anaphylaxis (91, aspirin-like drugs inhibit the biomation t 111. Secondly, svnthesis of orostaelandins (12-151 and reduce the inflammatThirdly, prostaglandins themselves, in conory response. centrations which occur in inflammatory exudates, reproduce the cardinal signs of inflammation such as erythema (16-191, oedema (20,211 and hyperalgesia (16, 17, 19, 211, either by a direct effect or by potentiation of the inflammatory effect! of other mediators (19211. We have recently immunized rats against prostaglandins and have observed that the inflammatory response to carrageenin was significantly less than that in control animals. 3 weeks old, were divided into two Male Wistar rats, One group was injected subdermally at groups of animals. monthly intervals with 500 ug of bovine serum albumin-PGE, The conjugate emulsified in Freund’s complete adjuvant. other group was injected with Freund’s adjuvant only. After three months two animals from each group were killed and the blood removed for antibody determinations. The antibody titre was estimated by the ability of the plasma to bind tritiated PGEl. Heparinised plasma was added to tritiated prostaglandin E and incubated at 4’C for 24 hJ the protein was then prec i pitated with polyetheleneglycol (211 and the unbound radioactivity measured in a scintillatPlasma ion counter using conventional counting techniques. from the control animals did not bind prostaglandin El. However, plasma from the immunized animals bound 31% of the added radioactivity. In two rats there was cross-reactivity of the antibody to PGE2 but not to PGF20. Inflammation was induced in the paw of rats starved overnight and the next day, injected (subplantarl with 0.1 ml of 0.5% carrageenin. The contralateral (control) paw was injected with 0.1 ml of saline. The increase in the
DECEMBER
10, 1974
VOL. 8 NO. 5
433
PROSTAGLANDINS
rat paw volume was measured by the mercury displacement method and calculated by subtracting the volume of the injected with saline from the volume of the paw injected with carrageenin.
paw
The oedema produced by carrageenin was significantly The reless in animals immunized against prostaglandins. duction was already present 60 min after carrageenin inject[SEE TABLE ion and persisted over the period of 4 hours. BELOW).
CARRAGEENIN PAW OEOEMA IN RATS ImlNIZEO
GROUP
n
C%!k#,
AGAINST PROSTAGLANOIN El
INCREASE0 IN RAT PAW VOLUME (~11 f S.E.M. Oh 4h Zh lh
CaVTROL
10
359 k12
-4.1 *14
346.2 k59.0
566.6 +54.2
705.9 k40.5
ImLNIZEO
11
330 iEi.2
12.4 ki3.6
206.7 k19.0
395.6 A31.1
607.7 k35.0
N.S.
N.S.
(0.05
CO.01 <0.005
P*
*The difference between the ho groups wera analysed by “Student’s ttest” using the method for unpaired samples. N.S. - Not significant.
These
volvement
results provide of prostaglandins
additional evidence in the inflammatory
for the inresponse,
and furthermore suggest that active immunization against sndogenous mediators such as prostaglandins would possibly be an important therapeutic tool, free from the usual side-
Prostaglandins have effects of anti-inflammatory drugs, also been implicated in fever (21,251 and hyperalgesia (16, 17,191 and it would be of interest to determine whether immunization reduced the intensity of the febrile response, or the hyperalgesia associated with inflamed or damaged tissue.
Immunization against prostaglandins can be a powerful experimental tool for assessing the role of prostaglandins Whilst these experiments were in in physiological events. Horton and Poyser progress, trus cycle in guinea-pig is against PGF2e.
434
(261 demonstrated that the oesprolonged in animals immunized
DECEMBER
10, 1974
VOL. 8 NO. 5
PROSTAGLANDINS
It will be interesting to find out whether immunization is effective in other species as well as in other systems in which prostaglandins are suspected to play a part, e.g. the control of parturition, renal auto-regulation, gastric secretion and tissue lipolysis. We wish to thank Gr. S. Wilkinson who prepared the bovine serum albumin-El conjugate. S.H.Ferreira, R.J.Flower, Madeleine F.Parsons and J.R.Vane. Dept. of Pharmacology, Wellcome Research Laboratories, Langley Court, Beckenham, Kent, ENGLAND.
REFERENCES 1.
Willis, A.L.: Prostaglandins, Peptides and Amines. (P. Mantegazza and E.W. Horton, Editors). Academic Press, London, 1969, p 31.
2.
Di Rosa, M., Giroud, J.P. and Willoughby, D.A.: Studies of the mediators of the acute inflammatory response induced in rats in different sites by carrageenin and turpentine. J. Path., 104:15, 1971.
3.
Moncada, S., Ferreira, S.H. and Vane, J.R.: Sensitization of pain receptors of the dog knee joint by prostaglandins. In the press., 1974.
4.
Jonsson, C.E.: Smooth muscle stimulating lipids in peripheral lymph after experimental burn injury. Stand. J. Plast. Reconstr. Surgery.. S:l, 1971.
6.
Jonsson, C.E. and Hamberg, M.: Advances in the Biosciences 9 (S. Bergstrf5mand S. Bernard, Editors). Pergamon Press, Vieweg, Braunschweig, 1973, p. 441.
6.
Arturson, G., Hamberg, M. and Jonsson, C.E.: Prostaglandins in human burn blister fluid. Acta. Physiol. Stand., 07:270. 1973.
DECEMBER
10, 1974
VOL. 8 NO. 5
435
PROSTAGLANDINS
7.
Blackham, A., Farmer, J.B.. Radziwonik, H. and J. : Rabbit monarticular arthritis and synovial glandins. Br. J. Pharmacol., 48:343, 1973.
Westwick, prosta-
0.
Higgs, G.A., Vane, J.R., Hart, F.D. And Wojtulewski, J.A.: The effects of anti-inflammatory drugs on prostaglandin concentrations in synovial fluid from patients with rheumatoid arthritis, In the press.
9.
Piper, P.J. and Vane, factors in anaphylaxis inflammatory drugs.
J.R.: Release of additional and its antagonism by antiNature, London, 223:20, 1969.
10.
Eakins, K.E., Whitelock. R.A.F., Bennett, A.C. and Martenet, A.C.: Prostaglandin-like activity in ocular inflammation, Br. J. Med. J., 3:452, 1973.
11.
Greaves, M.W., Sdndergaard, J.: Pharmacological agents released in ultraviolet inflammation studied by continuous skin perfusion. J. Invest. Dermatol., 54:365, 1970.
12.
Vane, J.R.: a mechanism New Biol.,
13.
Flower, R.J., Gryglewski, R., Herbaczynska-Cedro, The effects of anti-inflammatory and Vane, J.R.: on prostaglandin biosynthesis. Nature New Biol., 104, 1972.
14.
Ferreira, acin and spleen.
15.
Smith, hibits Nature
16.
Solomon, L.M., Juhlin, L. Prostaglandin on cutaneous 51:280, 196A. Dermatol.,
Inhibition of prostaglandin of action for aspirin-like 231:232, 1971.
S.H., aspirin Nature
K. drugs 238:
Moncada, S. and Vane, J.R.: Indomethabolish prostaglandin release from the New Biol., P. 231, 237, 1971.
Aspirin selectively J.R. and Willis, A.L.: prostaglandin production in human platelets. New Biol., 235, 1971. -231,
17.
Juhlin. S., Michaelson, ions in healthy subjects and atopic dermatitis. 49:251, 1969.
18.
Crunkhorn, Pearl to prostaglandins 41:49. 1971.
436
synthesis as drugs. Nature
and in
and Kirschenbaum, vasculature.
G.: and Acta.
Willis, rat and
J.
in-
M.B.: Invest.
Cutaneous vascular reactpatients with urticaria Derm. Venereal. Stockholm,
A.L.: man.
Cutaneous reactions Br. J. Pharmacol.,
DECEMBER
10, 1974
VOL. 8 NO. 5
PROSTAGLANDINS
19.
aspirin-like Prostaglandins, Ferreira, S.H.: Nature New Biol., 24O:ZOO. 1972. analgesia,
20.
Pro-inflamGlenn, E.M., Bowman, 8.3. and Rohloff, N.A.: Prostaglanmatory effects of certain prostaglandins.: dins Cellular Biology (P.W. Rawell and 8.8. Pharris, Plenon Press, New York, London, 1972, Editors). p. 329.
21.
ProstaMoncada, S.. Ferreira. S.H. and Vane, J.R.: glandins, aspirin-like drugs and the oedema of inflammation, Nature, 246:217, 1973.
22.
ProstaFerreira, S.H., Moncada, S. and Vane, J.R.: glandin and the mechanism of analgesia produced by aspirin-like drugs. Dr. J. Pharmacol., 49:86, 1973.
23.
Prostaglandin F2a Van Orden, D.A. and Farley, D.B.: radioimmunoassay utilising polyethyleneglycol separatProstaglandins, 4:215, 1973. ion technique.
24.
Feldberg, W., Gupta, K.P. Milton, A.S. and Wendlandt, s * Effects of pyrogen and antipyretics on prostaglindin activity in cisternal C.S.F. of unanaesthetized J. Physiol., 234:279, 1973. cats.
25.
Prostaglandin release in the central Milton, A.S.: nervous system during endotoxin induced fever.: SuPPlement to Advances in Biosciences 9 (S. Bergstrbm and Pergamon Press, Vieweg, BraunS. Bernard, Editors]. schweig, 1973, p. 495.
26.
Elongation of oestrous Horton, E.W. and Poyser, N.L.: cycle in the guinea-pig following active immunisation Prostaglandins, 5:349, against prostaglandin FZa. 1974.
DECEMBER
10, 1974
VOL. 8 NO. 5
drugs
437
REDUCTION OF
THE
IMMUNIZEO
To
the
INFLAMMATORY AGAINST
RESPONSE
IN
RATS
PROSTAGLANOINS
Editor,
The notion that proataglandins contribute to the pathogenesis of the inflammatory response is supported by three Firstly, prostaglandins are generated in main areuments. carrageenin inflammany types of tissue damage, for-example mation t l-31, thermal injury to the skin (4-61, arthritis uveitis (101 and U.V. induced inflam(7,81 anaphylaxis (91, aspirin-like drugs inhibit the biomation t 111. Secondly, svnthesis of orostaelandins (12-151 and reduce the inflammatThirdly, prostaglandins themselves, in conory response. centrations which occur in inflammatory exudates, reproduce the cardinal signs of inflammation such as erythema (16-191, oedema (20,211 and hyperalgesia (16, 17, 19, 211, either by a direct effect or by potentiation of the inflammatory effect! of other mediators (19211. We have recently immunized rats against prostaglandins and have observed that the inflammatory response to carrageenin was significantly less than that in control animals. 3 weeks old, were divided into two Male Wistar rats, One group was injected subdermally at groups of animals. monthly intervals with 500 ug of bovine serum albumin-PGE, The conjugate emulsified in Freund’s complete adjuvant. other group was injected with Freund’s adjuvant only. After three months two animals from each group were killed and the blood removed for antibody determinations. The antibody titre was estimated by the ability of the plasma to bind tritiated PGEl. Heparinised plasma was added to tritiated prostaglandin E and incubated at 4’C for 24 hJ the protein was then prec i pitated with polyetheleneglycol (211 and the unbound radioactivity measured in a scintillatPlasma ion counter using conventional counting techniques. from the control animals did not bind prostaglandin El. However, plasma from the immunized animals bound 31% of the added radioactivity. In two rats there was cross-reactivity of the antibody to PGE2 but not to PGF20. Inflammation was induced in the paw of rats starved overnight and the next day, injected (subplantarl with 0.1 ml of 0.5% carrageenin. The contralateral (control) paw was injected with 0.1 ml of saline. The increase in the
DECEMBER
10, 1974
VOL. 8 NO. 5
433
PROSTAGLANDINS
rat paw volume was measured by the mercury displacement method and calculated by subtracting the volume of the injected with saline from the volume of the paw injected with carrageenin.
paw
The oedema produced by carrageenin was significantly The reless in animals immunized against prostaglandins. duction was already present 60 min after carrageenin inject[SEE TABLE ion and persisted over the period of 4 hours. BELOW).
CARRAGEENIN PAW OEOEMA IN RATS ImlNIZEO
GROUP
n
C%!k#,
AGAINST PROSTAGLANOIN El
INCREASE0 IN RAT PAW VOLUME (~11 f S.E.M. Oh 4h Zh lh
CaVTROL
10
359 k12
-4.1 *14
346.2 k59.0
566.6 +54.2
705.9 k40.5
ImLNIZEO
11
330 iEi.2
12.4 ki3.6
206.7 k19.0
395.6 A31.1
607.7 k35.0
N.S.
N.S.
(0.05
CO.01 <0.005
P*
*The difference between the ho groups wera analysed by “Student’s ttest” using the method for unpaired samples. N.S. - Not significant.
These
volvement
results provide of prostaglandins
additional evidence in the inflammatory
for the inresponse,
and furthermore suggest that active immunization against sndogenous mediators such as prostaglandins would possibly be an important therapeutic tool, free from the usual side-
Prostaglandins have effects of anti-inflammatory drugs, also been implicated in fever (21,251 and hyperalgesia (16, 17,191 and it would be of interest to determine whether immunization reduced the intensity of the febrile response, or the hyperalgesia associated with inflamed or damaged tissue.
Immunization against prostaglandins can be a powerful experimental tool for assessing the role of prostaglandins Whilst these experiments were in in physiological events. Horton and Poyser progress, trus cycle in guinea-pig is against PGF2e.
434
(261 demonstrated that the oesprolonged in animals immunized
DECEMBER
10, 1974
VOL. 8 NO. 5
PROSTAGLANDINS
It will be interesting to find out whether immunization is effective in other species as well as in other systems in which prostaglandins are suspected to play a part, e.g. the control of parturition, renal auto-regulation, gastric secretion and tissue lipolysis. We wish to thank Gr. S. Wilkinson who prepared the bovine serum albumin-El conjugate. S.H.Ferreira, R.J.Flower, Madeleine F.Parsons and J.R.Vane. Dept. of Pharmacology, Wellcome Research Laboratories, Langley Court, Beckenham, Kent, ENGLAND.
REFERENCES 1.
Willis, A.L.: Prostaglandins, Peptides and Amines. (P. Mantegazza and E.W. Horton, Editors). Academic Press, London, 1969, p 31.
2.
Di Rosa, M., Giroud, J.P. and Willoughby, D.A.: Studies of the mediators of the acute inflammatory response induced in rats in different sites by carrageenin and turpentine. J. Path., 104:15, 1971.
3.
Moncada, S., Ferreira, S.H. and Vane, J.R.: Sensitization of pain receptors of the dog knee joint by prostaglandins. In the press., 1974.
4.
Jonsson, C.E.: Smooth muscle stimulating lipids in peripheral lymph after experimental burn injury. Stand. J. Plast. Reconstr. Surgery.. S:l, 1971.
6.
Jonsson, C.E. and Hamberg, M.: Advances in the Biosciences 9 (S. Bergstrf5mand S. Bernard, Editors). Pergamon Press, Vieweg, Braunschweig, 1973, p. 441.
6.
Arturson, G., Hamberg, M. and Jonsson, C.E.: Prostaglandins in human burn blister fluid. Acta. Physiol. Stand., 07:270. 1973.
DECEMBER
10, 1974
VOL. 8 NO. 5
435
PROSTAGLANDINS
7.
Blackham, A., Farmer, J.B.. Radziwonik, H. and J. : Rabbit monarticular arthritis and synovial glandins. Br. J. Pharmacol., 48:343, 1973.
Westwick, prosta-
0.
Higgs, G.A., Vane, J.R., Hart, F.D. And Wojtulewski, J.A.: The effects of anti-inflammatory drugs on prostaglandin concentrations in synovial fluid from patients with rheumatoid arthritis, In the press.
9.
Piper, P.J. and Vane, factors in anaphylaxis inflammatory drugs.
J.R.: Release of additional and its antagonism by antiNature, London, 223:20, 1969.
10.
Eakins, K.E., Whitelock. R.A.F., Bennett, A.C. and Martenet, A.C.: Prostaglandin-like activity in ocular inflammation, Br. J. Med. J., 3:452, 1973.
11.
Greaves, M.W., Sdndergaard, J.: Pharmacological agents released in ultraviolet inflammation studied by continuous skin perfusion. J. Invest. Dermatol., 54:365, 1970.
12.
Vane, J.R.: a mechanism New Biol.,
13.
Flower, R.J., Gryglewski, R., Herbaczynska-Cedro, The effects of anti-inflammatory and Vane, J.R.: on prostaglandin biosynthesis. Nature New Biol., 104, 1972.
14.
Ferreira, acin and spleen.
15.
Smith, hibits Nature
16.
Solomon, L.M., Juhlin, L. Prostaglandin on cutaneous 51:280, 196A. Dermatol.,
Inhibition of prostaglandin of action for aspirin-like 231:232, 1971.
S.H., aspirin Nature
K. drugs 238:
Moncada, S. and Vane, J.R.: Indomethabolish prostaglandin release from the New Biol., P. 231, 237, 1971.
Aspirin selectively J.R. and Willis, A.L.: prostaglandin production in human platelets. New Biol., 235, 1971. -231,
17.
Juhlin. S., Michaelson, ions in healthy subjects and atopic dermatitis. 49:251, 1969.
18.
Crunkhorn, Pearl to prostaglandins 41:49. 1971.
436
synthesis as drugs. Nature
and in
and Kirschenbaum, vasculature.
G.: and Acta.
Willis, rat and
J.
in-
M.B.: Invest.
Cutaneous vascular reactpatients with urticaria Derm. Venereal. Stockholm,
A.L.: man.
Cutaneous reactions Br. J. Pharmacol.,
DECEMBER
10, 1974
VOL. 8 NO. 5
PROSTAGLANDINS
19.
aspirin-like Prostaglandins, Ferreira, S.H.: Nature New Biol., 24O:ZOO. 1972. analgesia,
20.
Pro-inflamGlenn, E.M., Bowman, 8.3. and Rohloff, N.A.: Prostaglanmatory effects of certain prostaglandins.: dins Cellular Biology (P.W. Rawell and 8.8. Pharris, Plenon Press, New York, London, 1972, Editors). p. 329.
21.
ProstaMoncada, S.. Ferreira. S.H. and Vane, J.R.: glandins, aspirin-like drugs and the oedema of inflammation, Nature, 246:217, 1973.
22.
ProstaFerreira, S.H., Moncada, S. and Vane, J.R.: glandin and the mechanism of analgesia produced by aspirin-like drugs. Dr. J. Pharmacol., 49:86, 1973.
23.
Prostaglandin F2a Van Orden, D.A. and Farley, D.B.: radioimmunoassay utilising polyethyleneglycol separatProstaglandins, 4:215, 1973. ion technique.
24.
Feldberg, W., Gupta, K.P. Milton, A.S. and Wendlandt, s * Effects of pyrogen and antipyretics on prostaglindin activity in cisternal C.S.F. of unanaesthetized J. Physiol., 234:279, 1973. cats.
25.
Prostaglandin release in the central Milton, A.S.: nervous system during endotoxin induced fever.: SuPPlement to Advances in Biosciences 9 (S. Bergstrbm and Pergamon Press, Vieweg, BraunS. Bernard, Editors]. schweig, 1973, p. 495.
26.
Elongation of oestrous Horton, E.W. and Poyser, N.L.: cycle in the guinea-pig following active immunisation Prostaglandins, 5:349, against prostaglandin FZa. 1974.
DECEMBER
10, 1974
VOL. 8 NO. 5
drugs
437