FAILURE OF SOMATOSTATIN TO INHIBIT HISTAMINE SECRETION BY MAST CELLS G. W. READ, DONNA J. KOERKER AND DAVID LAGUNOFF
Departments of Pathology, Medicine, and Physiology and Biophysics, School of Medicine, University of Washington, Seattle, Washington 98195, U.S.A.
(Received 1 September 1976) Somatostatin
(somatotrophin release inhibiting factor) antagonizes the secretion of somatotrophin (Brazeau, Vale, Burgus, Ling, Butcher, Rivier & Guillemin, 1973), insulin (Alberti, Christensen, Christensen, Hansen, Iversen, Lundbaeck, Seyer-Hansen & Orskov, 1973), thyrotrophin (Hall, Besser, Schally, Coy, Evered, Goldie, Kastin, McNeilly,Mortimer,Phenekos, Tunbridge & Weightman, 1973), glucagon (Iversen, 1974; Koerker, Ruch, Chideckel, Palmer, Goodner, Ensinck & Gale, 1974; Mortimer, Tunbridge, Carr, Yeomans, Lind, Coy, Bloom, Kastin, Mallinson, Besser, Schally & Hall, 1974), gastrin (Bloom, Mortimer, Thorner, Besser, Hall, Gomez-Pan, Roy, Russell, Coy, Kastin & Schally, 1974), gastric acid (Barros D'Sa, Bloom & Baron, 1975; Gomez-Pan, Reed, Albinus, Shaw, Hall, Besser, Coy, Kastin & Schally, 1975) and pepsin (Gomez-Pan et al. 1975) but appears to have no effect on secretion of follicle\x=req-\ stimulating hormone, luteinizing hormone, prolactin, corticotrophin, corticosteroids (Hall et al. 1973), adrenaline, or noradrenaline (Christensen, Christensen, Hansen & Lundbaeck, 1975). It has been proposed that somatostatin acts by altering a late stage of the granule secretory process, probably at a step which is Ca2+-dependent (Curry & Bennet, 1974). Mast cells store histamine in intracellular granules and can be triggered to secrete the amine by a granule extrusion mechanism (Lagunoff, 1976). It was, therefore, of interest to determine whether somatostatin would inhibit mast cell degranulation and histamine release. Suspensions of mast cells were prepared as previously described (Lagunoff, 1975). The cells were incubated with the secretagogues (10 min at 37 °C) in the presence or absence of 10"9 to "6 M-somatostatin. (In a few cases the somatostatin was added 15 min earlier than the secretagogues, but this did not change the results.) The cells were then centrifuged at 200g, and the histamine released into the medium was assayed by the fluorometric method. Histamine remaining in the cells was extracted by lysis with 2-5 % trichloroacetic acid. No effect on histamine release was observed at any concentration of somatostatin. Table 1 shows the data obtained with 10~6 M-somatostatin. Somatostatin caused little or no histamine release by itself. To ensure that the somatostatin was active, the two preparations used in these experiments were tested for inhibition of insulin secretion (Koerker et al. 1974) and for reaction with specific antibody in the radioimmunological assay of somatostatin (Ensinck, Laschansky, Chideckel, Palmer & Goodner, 1976). Both preparations were active by these two criteria. It therefore appears that somatostatin has no inhibitory effect on histamine secretion by mast cells. * Present address: Pharmacology Department, University of Hawaii, Honolulu, Hawaii 96822, U.S.A. f Address reprint requests to D.L.
Table 1. Effect of somatostatin
on
histamine release produced by six agents (means +S.EM.)
mast
cell
degranulating
Histamine release (%)
Secretagogue t
Concentration
Trials
Saline control
Compound 48/80 Dextran T40 +.
Ionophore A23187 Antigen % f
Concanavalin K%
Polymyxin
*Spontaneous histamine
No somatostatin
10"6 M-somatostatinf
0
3± 2 42 ± 23 ± 40 ± 15 ±
10 5 5 3 3 3 3
Mg/ml mg/ml 10"6 mol/1 0-8 units/ml 10 Mg/ml 2 Mg/ml 1 1
release in untreated mast cells
values.
:
9 20 ± 6 35 ±12 11± 3 15± 3 49 ± 6 44
(average
=
5
±
±
0-5 %)
15±
50± 11 was
subtracted from all the
fSuppliers: dihydrosomatostatin, Bachern Inc. and Salk Institute; compound 48/80, Burroughs Wellcome Co.; dextran T40, Pharmacia; ionophore A23187, gift from Dr R. L. Hamill, Lilly Research Laboratories; pertussis vaccine, Eli Lilly; concanavalin A and polymyxin B, Sigma Chemical Co.; phosphatidylserine, Applied Science Laboratories. % Dextran-, antigen-, and
phosphatidylserine, 10 Mg/ml. ^Pertussis vaccine was used
before
concanavalin as
the
-induced histamine release
antigen.
were
tested in the presence of
Rats were inoculated with 2 units of the vaccine 12-19
use.
Supported by grants from the U.S.P.H.S. (GM-18740, GM-13543, HL-03174). supported in part by the University of Hawaii.
days
G.W.R.
REFERENCES
Alberti, K. G. M. M., Christensen, N. J., Christensen, S. E., Hansen, A. P., Iversen, J., Lundbaeck, K., SeyerHansen, K. & Orskov, H. (1973). Lancet ii, 1299-1301. Barros D'Sa, A. A. J., Bloom, S. R. & Baron, J. H. (1975). Lancet i, 886-887. Bloom, S. R., Mortimer, C. H., Thorner, M. O., Besser, G. M., Hall, R., Gomez-Pan, ., Roy, V. M., Russell, R. C. G., Coy, D. H., Kastin, A. J. & Schally, A. V. (1974). Lancet ii, 1106-1109. Brazeau, P., Vale, W., Burgus, R., Ling, N., Butcher, M., Rivier, J. & Guillemin, R. (1973). Science 179, 77-79.
Christensen, N. J., Christensen, S. E., Hansen, A. P. & Lundbaeck, K. (197'5). Metabolism 24, 1267-1272. Curry, D. L. & Bennet, L. L. (1974). Biochemical and Biophysical Research Communications 60, 1015-1019. Ensinck, J., Laschansky, E., Chideckel, E., Palmer, J. & Goodner, C. (1976). Clinical Research 24, 115a. Gomez-Pan, ., Reed, J. D., Albinus, M., Shaw, B., Hall, R., Besser, G. M., Coy, D. H., Kastin, A. J. & Schally, A. V. (1975). Lancet i, 888-890. Hall, R., Besser, G. M., Schally, A. V., Coy, D. H., Evered, D., Goldie, D. J., Kastin, A. J., McNeilly, A. S., Mortimer, C. H., Phenekos, C, Tunbridge, W. M. G. & Weightman, D. (1973). Lancet ii, 581-584. Iversen, J. (1974). Scandinavian Journal of Clinical and Laboratory Investigation 33,125-129. Koerker, D. J., Ruch, W., Chideckel, E., Palmer, J., Goodner, C. J., Ensinck, J. & Gale.C. C. (1974). Science 184,482-484. Lagunoff, D. (1975). In Techniques of biochemical and biophysical morphology, Vol. 2, pp. 283-305. Eds D. Glick & R. Rosenbaum. New York: John Wiley & Sons. Lagunoff, D. (1976). In Bronchial asthma: mechanisms and therapeutics, pp. 383-407. Eds E. B. Weiss & M. S. Segal. Boston: Little, Brown and Company. Mortimer, C. H., Tunbridge, W. M. G., Carr, D., Yeomans, L., Lind, T., Coy, D. H., Bloom, S. R., Kastin, ., Mallinson, C. N., Besser, G. M., Schally, . V. & Hall, R. (1974). Lancet i, 697-701.
Departments of Pathology, Medicine, and Physiology and Biophysics, School of Medicine, University of Washington, Seattle, Washington 98195, U.S.A.
(Received 1 September 1976) Somatostatin
(somatotrophin release inhibiting factor) antagonizes the secretion of somatotrophin (Brazeau, Vale, Burgus, Ling, Butcher, Rivier & Guillemin, 1973), insulin (Alberti, Christensen, Christensen, Hansen, Iversen, Lundbaeck, Seyer-Hansen & Orskov, 1973), thyrotrophin (Hall, Besser, Schally, Coy, Evered, Goldie, Kastin, McNeilly,Mortimer,Phenekos, Tunbridge & Weightman, 1973), glucagon (Iversen, 1974; Koerker, Ruch, Chideckel, Palmer, Goodner, Ensinck & Gale, 1974; Mortimer, Tunbridge, Carr, Yeomans, Lind, Coy, Bloom, Kastin, Mallinson, Besser, Schally & Hall, 1974), gastrin (Bloom, Mortimer, Thorner, Besser, Hall, Gomez-Pan, Roy, Russell, Coy, Kastin & Schally, 1974), gastric acid (Barros D'Sa, Bloom & Baron, 1975; Gomez-Pan, Reed, Albinus, Shaw, Hall, Besser, Coy, Kastin & Schally, 1975) and pepsin (Gomez-Pan et al. 1975) but appears to have no effect on secretion of follicle\x=req-\ stimulating hormone, luteinizing hormone, prolactin, corticotrophin, corticosteroids (Hall et al. 1973), adrenaline, or noradrenaline (Christensen, Christensen, Hansen & Lundbaeck, 1975). It has been proposed that somatostatin acts by altering a late stage of the granule secretory process, probably at a step which is Ca2+-dependent (Curry & Bennet, 1974). Mast cells store histamine in intracellular granules and can be triggered to secrete the amine by a granule extrusion mechanism (Lagunoff, 1976). It was, therefore, of interest to determine whether somatostatin would inhibit mast cell degranulation and histamine release. Suspensions of mast cells were prepared as previously described (Lagunoff, 1975). The cells were incubated with the secretagogues (10 min at 37 °C) in the presence or absence of 10"9 to "6 M-somatostatin. (In a few cases the somatostatin was added 15 min earlier than the secretagogues, but this did not change the results.) The cells were then centrifuged at 200g, and the histamine released into the medium was assayed by the fluorometric method. Histamine remaining in the cells was extracted by lysis with 2-5 % trichloroacetic acid. No effect on histamine release was observed at any concentration of somatostatin. Table 1 shows the data obtained with 10~6 M-somatostatin. Somatostatin caused little or no histamine release by itself. To ensure that the somatostatin was active, the two preparations used in these experiments were tested for inhibition of insulin secretion (Koerker et al. 1974) and for reaction with specific antibody in the radioimmunological assay of somatostatin (Ensinck, Laschansky, Chideckel, Palmer & Goodner, 1976). Both preparations were active by these two criteria. It therefore appears that somatostatin has no inhibitory effect on histamine secretion by mast cells. * Present address: Pharmacology Department, University of Hawaii, Honolulu, Hawaii 96822, U.S.A. f Address reprint requests to D.L.
Table 1. Effect of somatostatin
on
histamine release produced by six agents (means +S.EM.)
mast
cell
degranulating
Histamine release (%)
Secretagogue t
Concentration
Trials
Saline control
Compound 48/80 Dextran T40 +.
Ionophore A23187 Antigen % f
Concanavalin K%
Polymyxin
*Spontaneous histamine
No somatostatin
10"6 M-somatostatinf
0
3± 2 42 ± 23 ± 40 ± 15 ±
10 5 5 3 3 3 3
Mg/ml mg/ml 10"6 mol/1 0-8 units/ml 10 Mg/ml 2 Mg/ml 1 1
release in untreated mast cells
values.
:
9 20 ± 6 35 ±12 11± 3 15± 3 49 ± 6 44
(average
=
5
±
±
0-5 %)
15±
50± 11 was
subtracted from all the
fSuppliers: dihydrosomatostatin, Bachern Inc. and Salk Institute; compound 48/80, Burroughs Wellcome Co.; dextran T40, Pharmacia; ionophore A23187, gift from Dr R. L. Hamill, Lilly Research Laboratories; pertussis vaccine, Eli Lilly; concanavalin A and polymyxin B, Sigma Chemical Co.; phosphatidylserine, Applied Science Laboratories. % Dextran-, antigen-, and
phosphatidylserine, 10 Mg/ml. ^Pertussis vaccine was used
before
concanavalin as
the
-induced histamine release
antigen.
were
tested in the presence of
Rats were inoculated with 2 units of the vaccine 12-19
use.
Supported by grants from the U.S.P.H.S. (GM-18740, GM-13543, HL-03174). supported in part by the University of Hawaii.
days
G.W.R.
REFERENCES
Alberti, K. G. M. M., Christensen, N. J., Christensen, S. E., Hansen, A. P., Iversen, J., Lundbaeck, K., SeyerHansen, K. & Orskov, H. (1973). Lancet ii, 1299-1301. Barros D'Sa, A. A. J., Bloom, S. R. & Baron, J. H. (1975). Lancet i, 886-887. Bloom, S. R., Mortimer, C. H., Thorner, M. O., Besser, G. M., Hall, R., Gomez-Pan, ., Roy, V. M., Russell, R. C. G., Coy, D. H., Kastin, A. J. & Schally, A. V. (1974). Lancet ii, 1106-1109. Brazeau, P., Vale, W., Burgus, R., Ling, N., Butcher, M., Rivier, J. & Guillemin, R. (1973). Science 179, 77-79.
Christensen, N. J., Christensen, S. E., Hansen, A. P. & Lundbaeck, K. (197'5). Metabolism 24, 1267-1272. Curry, D. L. & Bennet, L. L. (1974). Biochemical and Biophysical Research Communications 60, 1015-1019. Ensinck, J., Laschansky, E., Chideckel, E., Palmer, J. & Goodner, C. (1976). Clinical Research 24, 115a. Gomez-Pan, ., Reed, J. D., Albinus, M., Shaw, B., Hall, R., Besser, G. M., Coy, D. H., Kastin, A. J. & Schally, A. V. (1975). Lancet i, 888-890. Hall, R., Besser, G. M., Schally, A. V., Coy, D. H., Evered, D., Goldie, D. J., Kastin, A. J., McNeilly, A. S., Mortimer, C. H., Phenekos, C, Tunbridge, W. M. G. & Weightman, D. (1973). Lancet ii, 581-584. Iversen, J. (1974). Scandinavian Journal of Clinical and Laboratory Investigation 33,125-129. Koerker, D. J., Ruch, W., Chideckel, E., Palmer, J., Goodner, C. J., Ensinck, J. & Gale.C. C. (1974). Science 184,482-484. Lagunoff, D. (1975). In Techniques of biochemical and biophysical morphology, Vol. 2, pp. 283-305. Eds D. Glick & R. Rosenbaum. New York: John Wiley & Sons. Lagunoff, D. (1976). In Bronchial asthma: mechanisms and therapeutics, pp. 383-407. Eds E. B. Weiss & M. S. Segal. Boston: Little, Brown and Company. Mortimer, C. H., Tunbridge, W. M. G., Carr, D., Yeomans, L., Lind, T., Coy, D. H., Bloom, S. R., Kastin, ., Mallinson, C. N., Besser, G. M., Schally, . V. & Hall, R. (1974). Lancet i, 697-701.
