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353 Comparison of the effects of anti-allergic drugs on monkey lung and intestinal mast cells
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COMPARISON OF THE EFFECTS OF ANTI-ALLERGIC DRUGS ON MONKEY LUNG ANQ INTESTINAL MAST CELLS. Kim E. Barrett, Ph.D., Eva Srucs, M.D., and Dean D. Metcalfe, M.D., Bethesda, Maryland. Functional heterogeneity clearly exists between mast cells from different tiisues in lower species. To examine to what extent such heterogeneity extends to the mast cell populations of higher animals, we examined the effect of antiallergic drugs on intestinal and lung mast cells (IMC, LMC) simultaneously isolated from individual monkeys. Specimens of small bowel and lung were dispersed in parallel using collagenase. The cell suspensions contained l-3% mast cells and were >90% viable. Both cell types released histamine following challenge with anti-human IgE, with LMC more responsive than IMC (25.3+3.2% vs. 8.9+3.7%, means _+ SEM, n=3). Release was comparably inhibited from LMC and IMC by dibutyryl CAMP (ID50 ~a 0.5 mM), isoprenaline (ID50 z 0.5 PM). and theophylline (ID50 ~a 0.2 mM). Quercetiz also inhibited release, although the drug was more active on IMC (ID50 ca 2 FM) than on LMC (ID50 " 7 @). Treatment wxh diJodium cromoglycate (DSCG) resulted in inhibition of release from IMC, while LMC were minimally affected. This study highlights the inter-species differences among mast cells, since IMC from rats are reportedly refractory to theophylline and DSCG. However, it appears that inter-tissue variations in mast cell function may be less marked in higher animals, and may be minimized if the cells are recovered from the tissues in an identical fashion. In the absence of multiple tissue samples from single human subjects, normal monkeys provide a valuable system for the study of mast cell heterogeneity in higher animals.
355
HISTAMINE RELEASE FROMMUCOSAL-TYPEHUMANLUNG Penny MAST CELLS. A.J. Wardlaw, M.R.C.P.,
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ALTERED MAST CELL THRESHOLDIN EXERCISE-INDUCED ANAPHYLAXIS. G.W. Errington, M.D., Y.A. Nekori, M.D., W-S. Silvers, M.D. and M.D., II A.L. .--,Scltocket, Denver, Colorado Exercise-induced anaphylaxis (:.:IA) is a
recently described syndrome, yet i.ts pathogenic: mechanisms are poorly understood. This study asks whether exercise can alter mist cell thresholds for mediator release. :t- compares the surface area of wheals, measured :&t 15 min. (in square mm) to titrated intradermal iicses of Compound 48/80 (0.05 mg/ml and 0.7 zig/ml) and histamine (0.001 mg/ml and 0.01 mg/ml ) immediately before and after exerr.ise in three groups; normal nonatopic controls (N=5), normal atopic controls (N=S) and patient> with EIA (N=6). There was no statistical difference between the groups in wheal size pre- and postexercise with either doses of Coma-,ound 48/80 or histamine. Among the patients wit?) EIA, however, 2 had larger wheals with 0.5 ma/ml of Compound 48180 post-exercise vs pre-exercire (ratio post/pre 1.68 2 0.75 S.D.) compa,rcd tu the others in the ElA group (0.652 k- is.191). This approaches statistical significanie (P 0.05, Student t test). One of these two patients had symptoms of exercise-induced bronchospasm during the exercise, while all. others d:id not, and the second patient had a predictable onset of cholinergic urticaria as part of I!er syndrome. Therfore a subset of EIA may exist in which exercise enhances the Compound 48!80-induced release of mast cell mediators. Exercise may cause this release in others with Cib, but may require the development of symptoms during the exercise to elicit this enhancec ~‘~?sponsc,
Fitzharris, MD, 0. Cromwell, PhD, J.V. Collins, MD, FRCP and A.B. Kay, MD, PhD, London, U.K. Mast cells in bronchoalveolar lavage (BAL) fluid were studied from 24 patients undergoing The lavage routine fibreoptic bronchoscopy. cells were fixed in Carnoys fluid and stained with Alcian Blue and Safranine. Between 0.04 and 0.59% (mean = 0.18) of the lavage cell population exhibited staining characteristics Cells from 16 typical of mucosal mast cells. subjects were further studied for histamine Histamine was content and releasability. measured using a single isotope radioenzymatic There was a significant (p cO.01) assay. correlation between total histamine content of the lavage cell population and mast cell The mean mast cell histamine content counts. was 6.1 picograms with a range of 2.16 to 10.8 Histamine was released in a dosepicograms. dependent fashion after stimulation with anti-IgE and calcium ionophore with a time course characteristic for mast cells. Factor(s) from cultured, unstimulated, blood mononuclear cells also gave direct histamine release from BAL cells and augmented anti-IgE There was minimal release stimulated release. after incubation with 48/80, phorbol myristate acetate and f-met-leu-phe. These results indicate that BAL cells provide a suitable model for the study of the functional and morphological characteristics of human lung mucosal mast cells.
ALTERATIONS IN GRANULOCYTEFWNCTEON WRENHUMAN BLOODLEUCOCYTESARE INCUBATEDWTH A&TX-IgE (REVERSED-TYPE ANAPHYLAXIS), G.El. @al&, FIMLS, R. Moqbrl, PhD, A.J. MacDonald, BSc and A.B. Kay, MD, PhD, London, UK. When whole blood human leucocytes were incubated with a F(ab’)2 anti-humnn IgE kCE) or control F(ab’)2 prepared from normal rabbit IgG @EC), this was associated with ia) increased cytotoxicity of eosinophils and neutrophils for complementor antibody-coated schisrosomula and (b) enhanced expression of complement (C3b) and IgG (Fc) receptors. These effects were dependent both on the time of incubation, and the concentration of ME. The changes were elicited by&E-stimulated mononuclear cells (also containing basophils) but not from NE-stimulated eosinophils or neucrophils. These effects were not observed airer incubation with o(Ec. The IgE-medi ated alterations in schistosomular killing and receptor expression were inhibitable, in a dose-dependent fashion, by disodium cromoglycate (DSCG). Virtually complete reversal of enhancement by this c;mpound was obtained by concentrations of lo-- mol/L. The results support the view that components of the allergic response may play a role both in the amplification of hypersensitivty reactions and the enhancement of the effector function of inflammatory cells in immunity t<> helminths. there is additional evidence that Furthermore, DSCG has a direct effect on human granulocytes as well as mast cells.
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COMPARISON OF THE EFFECTS OF ANTI-ALLERGIC DRUGS ON MONKEY LUNG ANQ INTESTINAL MAST CELLS. Kim E. Barrett, Ph.D., Eva Srucs, M.D., and Dean D. Metcalfe, M.D., Bethesda, Maryland. Functional heterogeneity clearly exists between mast cells from different tiisues in lower species. To examine to what extent such heterogeneity extends to the mast cell populations of higher animals, we examined the effect of antiallergic drugs on intestinal and lung mast cells (IMC, LMC) simultaneously isolated from individual monkeys. Specimens of small bowel and lung were dispersed in parallel using collagenase. The cell suspensions contained l-3% mast cells and were >90% viable. Both cell types released histamine following challenge with anti-human IgE, with LMC more responsive than IMC (25.3+3.2% vs. 8.9+3.7%, means _+ SEM, n=3). Release was comparably inhibited from LMC and IMC by dibutyryl CAMP (ID50 ~a 0.5 mM), isoprenaline (ID50 z 0.5 PM). and theophylline (ID50 ~a 0.2 mM). Quercetiz also inhibited release, although the drug was more active on IMC (ID50 ca 2 FM) than on LMC (ID50 " 7 @). Treatment wxh diJodium cromoglycate (DSCG) resulted in inhibition of release from IMC, while LMC were minimally affected. This study highlights the inter-species differences among mast cells, since IMC from rats are reportedly refractory to theophylline and DSCG. However, it appears that inter-tissue variations in mast cell function may be less marked in higher animals, and may be minimized if the cells are recovered from the tissues in an identical fashion. In the absence of multiple tissue samples from single human subjects, normal monkeys provide a valuable system for the study of mast cell heterogeneity in higher animals.
355
HISTAMINE RELEASE FROMMUCOSAL-TYPEHUMANLUNG Penny MAST CELLS. A.J. Wardlaw, M.R.C.P.,
399
ALTERED MAST CELL THRESHOLDIN EXERCISE-INDUCED ANAPHYLAXIS. G.W. Errington, M.D., Y.A. Nekori, M.D., W-S. Silvers, M.D. and M.D., II A.L. .--,Scltocket, Denver, Colorado Exercise-induced anaphylaxis (:.:IA) is a
recently described syndrome, yet i.ts pathogenic: mechanisms are poorly understood. This study asks whether exercise can alter mist cell thresholds for mediator release. :t- compares the surface area of wheals, measured :&t 15 min. (in square mm) to titrated intradermal iicses of Compound 48/80 (0.05 mg/ml and 0.7 zig/ml) and histamine (0.001 mg/ml and 0.01 mg/ml ) immediately before and after exerr.ise in three groups; normal nonatopic controls (N=5), normal atopic controls (N=S) and patient> with EIA (N=6). There was no statistical difference between the groups in wheal size pre- and postexercise with either doses of Coma-,ound 48/80 or histamine. Among the patients wit?) EIA, however, 2 had larger wheals with 0.5 ma/ml of Compound 48180 post-exercise vs pre-exercire (ratio post/pre 1.68 2 0.75 S.D.) compa,rcd tu the others in the ElA group (0.652 k- is.191). This approaches statistical significanie (P 0.05, Student t test). One of these two patients had symptoms of exercise-induced bronchospasm during the exercise, while all. others d:id not, and the second patient had a predictable onset of cholinergic urticaria as part of I!er syndrome. Therfore a subset of EIA may exist in which exercise enhances the Compound 48!80-induced release of mast cell mediators. Exercise may cause this release in others with Cib, but may require the development of symptoms during the exercise to elicit this enhancec ~‘~?sponsc,
Fitzharris, MD, 0. Cromwell, PhD, J.V. Collins, MD, FRCP and A.B. Kay, MD, PhD, London, U.K. Mast cells in bronchoalveolar lavage (BAL) fluid were studied from 24 patients undergoing The lavage routine fibreoptic bronchoscopy. cells were fixed in Carnoys fluid and stained with Alcian Blue and Safranine. Between 0.04 and 0.59% (mean = 0.18) of the lavage cell population exhibited staining characteristics Cells from 16 typical of mucosal mast cells. subjects were further studied for histamine Histamine was content and releasability. measured using a single isotope radioenzymatic There was a significant (p cO.01) assay. correlation between total histamine content of the lavage cell population and mast cell The mean mast cell histamine content counts. was 6.1 picograms with a range of 2.16 to 10.8 Histamine was released in a dosepicograms. dependent fashion after stimulation with anti-IgE and calcium ionophore with a time course characteristic for mast cells. Factor(s) from cultured, unstimulated, blood mononuclear cells also gave direct histamine release from BAL cells and augmented anti-IgE There was minimal release stimulated release. after incubation with 48/80, phorbol myristate acetate and f-met-leu-phe. These results indicate that BAL cells provide a suitable model for the study of the functional and morphological characteristics of human lung mucosal mast cells.
ALTERATIONS IN GRANULOCYTEFWNCTEON WRENHUMAN BLOODLEUCOCYTESARE INCUBATEDWTH A&TX-IgE (REVERSED-TYPE ANAPHYLAXIS), G.El. @al&, FIMLS, R. Moqbrl, PhD, A.J. MacDonald, BSc and A.B. Kay, MD, PhD, London, UK. When whole blood human leucocytes were incubated with a F(ab’)2 anti-humnn IgE kCE) or control F(ab’)2 prepared from normal rabbit IgG @EC), this was associated with ia) increased cytotoxicity of eosinophils and neutrophils for complementor antibody-coated schisrosomula and (b) enhanced expression of complement (C3b) and IgG (Fc) receptors. These effects were dependent both on the time of incubation, and the concentration of ME. The changes were elicited by&E-stimulated mononuclear cells (also containing basophils) but not from NE-stimulated eosinophils or neucrophils. These effects were not observed airer incubation with o(Ec. The IgE-medi ated alterations in schistosomular killing and receptor expression were inhibitable, in a dose-dependent fashion, by disodium cromoglycate (DSCG). Virtually complete reversal of enhancement by this c;mpound was obtained by concentrations of lo-- mol/L. The results support the view that components of the allergic response may play a role both in the amplification of hypersensitivty reactions and the enhancement of the effector function of inflammatory cells in immunity t<> helminths. there is additional evidence that Furthermore, DSCG has a direct effect on human granulocytes as well as mast cells.
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