Mast cells and anti-inflammatory drugs: studies of mediator release and calcium mobilization.

by Grant Richardson Stenton.Thesis (Ph.D.)--Chinese University of Hong Kong, 1996.Includes bibliographical references (leaves 259-287).Abstract --- p.iAcknowledgements --- p.iiiPublications --- p.ivAbbreviations --- p.vContents --- p.viiChapter Chapter 1 --- IntroductionChapter 1 1.1. --- Historical Background --- p.2Chapter 1.2. --- Origin and distribution of mast cells --- p.2Chapter 1.3. --- Mast cell heterogeneity --- p.3Chapter 1.4. --- Mast cell mediators --- p.5Chapter 1.4.1. --- Preformed mast cell mediators --- p.6Chapter 1.4.2. --- Newly synthesised mast cell mediators --- p.7Chapter 1.5. --- Mast cell activation --- p.11Chapter 1.5.1. --- Antigenic pathway of mast cell activation --- p.11Chapter 1.5.1.1. --- Antigen binding and receptor aggregation --- p.12Chapter 1.5.1.2. --- Early events following FcεRI aggregation --- p.13Chapter 1.5.1.3. --- Antigenic induction of mast cell second messenger production --- p.15Chapter 1.5.1.4. --- Phospholipase C activation and mast cells --- p.16Chapter 1.5.1.5. --- Phospholipase A2 activation and mast cells --- p.17Chapter 1.5.1.6. --- Intracellular calcium and mast cells --- p.18Chapter 1.5.1.7. --- Calcium and calmodulin --- p.21Chapter 1.5.1.8. --- Adenylate cyclase activation and mast cells --- p.21Chapter 1.5.2. --- Non-antigenic pathway of mast cell activation --- p.22Chapter 1.6. --- Aims of the study --- p.25Chapter 1.6.1. --- Diuretics --- p.26Chapter 1.6.2. --- Histamine receptor directed compounds --- p.27Chapter 1.6.3. --- Cyclo-oxygenase inhibitors --- p.28Chapter 1.6.4. --- Immunosuppressive compounds --- p.29Chapter Chapter 2 --- Materials and Methods --- p.31Chapter 2.1. --- Materials and methods --- p.32Chapter 2.1.1. --- Secretagogues --- p.32Chapter 2.1.2. --- Anti-allergic compounds --- p.32Chapter 2.1.3. --- Diuretics --- p.32Chapter 2.1.4. --- Immunosuppressants --- p.33Chapter 2.1.5. --- Histamine agonists and antagonists --- p.33Chapter 2.1.6. --- Cyclo-oxygenase inhibitors --- p.33Chapter 2.1.7. --- Materials for buffers --- p.34Chapter 2.1.8. --- Materials for rat sensitization --- p.34Chapter 2.1.9. --- Materials for histamine assay --- p.35Chapter 2.1.10. --- Materials for calcium measurement --- p.35Chapter 2.1.11. --- Materials for prostaglandin D2 measurement --- p.35Chapter 2.1.12. --- Materials for leukotriene C4 measurement --- p.36Chapter 2.1.13. --- Materials for cyclic AMP measurement --- p.36Chapter 2.1.14. --- Miscellaneous --- p.36Chapter 2.2. --- Buffers and stock solutions --- p.37Chapter 2.2.1. --- Buffer ingredients --- p.37Chapter 2.2.2. --- Stock solutions --- p.38Chapter 2.3. --- Animals and cell isolation --- p.39Chapter 2.3.1. --- Animals --- p.39Chapter 2.3.2. --- Sensitization of animals --- p.39Chapter 2.3.3. --- Cell isolation --- p.40Chapter 2.3.4. --- Cell washing and purification --- p.41Chapter 2.3.5. --- Preparation of cells for counting --- p.42Chapter 2.3.6. --- Cell counting on a haemocytometer --- p.42Chapter 2.4. --- General protocol for histamine release and histamine measurement --- p.43Chapter 2.4.1. --- Histamine release --- p.43Chapter 2.4.2. --- Spectroflurometric determination of histamine contents --- p.44Chapter 2.4.3. --- Calculation of histamine levels --- p.45Chapter 2.5. --- Protocol for cellular calcium measurement --- p.47Chapter 2.5.1. --- 45Ca2+ influx measurement --- p.47Chapter 2.5.2. --- Calculation of 45Ca2+ influx --- p.48Chapter 2.5.3. --- Fura-2 fluorescence measurement of intracellular calcium --- p.48Chapter 2.5.4. --- Fura-2 cell loading --- p.48Chapter 2.5.5. --- Fura-2 fluorescence parameters --- p.49Chapter 2.5.6. --- Calculation of basal calcium levels --- p.50Chapter 2.6. --- Protocol for prostaglandin D2 (PGD2) measurement --- p.52Chapter 2.6.1. --- PGD2 production --- p.52Chapter 2.6.2. --- Enzyme Immunosorbent Assay (EIA) method of PGD2 measurement --- p.52Chapter 2.6.3. --- Calculation of (EIA) PGD2 production --- p.53Chapter 2.6.4. --- Radio Immunosorbent Assay (RIA) method of PGD2 measurement --- p.53Chapter 2.6.5. --- Calculation of (RIA) PGD2 concentration --- p.54Chapter 2.7. --- Protocol for leukotriene C4 (LTC4) measurement --- p.54Chapter 2.7.1. --- LTC4 production --- p.54Chapter 2.7.2. --- Enzyme Immunosorbent Assay (EIA) method of LTC4 measurement --- p.55Chapter 2.7.3. --- Calculation of (EIA) LTC4 concentration --- p.55Chapter 2.8. --- Protocol for cyclic adenosine monophosphate (cAMP) measurement --- p.56Chapter 2.8.1. --- cAMP production --- p.56Chapter 2.8.2. --- Radio Immunosorbent Assay (RIA) method of cAMP measurement --- p.56Chapter 2.8.3. --- Calculation of cAMP concentration --- p.57Chapter 2.9. --- Statistical analysis --- p.57Chapter Chapter 3 --- "Frusemide, Bumetanide and DSCG" --- p.58Chapter 3.1. --- Introduction --- p.59Chapter 3.1.1. --- Frusemide and bumetanide as loop diuretics --- p.59Chapter 3.1.2. --- Effects of frusemide and bumetanide on the airways --- p.59Chapter 3.1.3. --- Effects of frusemide on mast cells --- p.60Chapter 3.1.4. --- Experimental aims --- p.61Chapter 3.2. --- Materials and methods --- p.62Chapter 3.3. --- Results --- p.63Chapter 3.3.1 --- "Effects of frusemide, bumetanide and DSCG on immunologically induced histamine release from rat peritoneal mast cells" --- p.63Chapter 3.3.2. --- "Effects of frusemide, bumetanide and DSCG on compound 48/80 induced histamine release from rat peritoneal mast cells" --- p.64Chapter 3.3.3. --- "Effects of frusemide, bumetanide and DSCG on compound 48/80 induced histamine release from rat peritoneal mast cells suspended in calcium free buffer" --- p.65Chapter 3.3.4. --- "Effects of frusemide, bumetanide and DSCG on ionophore A23187 and thapsigargin induced histamine release from rat peritoneal mast cells" --- p.65Chapter 3.3.5. --- Cross-tachyphylaxis effects of frusemide and bumetanide --- p.66Chapter 3.3.6. --- Effects of DSCG on the inhibition of anaphylactic histamine release due to frusemide --- p.67Chapter 3.3.7. --- Effects of frusemide and DSCG on immunologically and non-immunologically induced 45Ca2+ uptake --- p.67Chapter 3.3.8. --- Effects of frusemide and DSCG on immunologically and non-immunologically induced changes in the free intracellular calcium concentration of rat peritoneal mast cells --- p.68Chapter 3.3.9. --- Effects of frusemide and bumetanide on the spontaneous and secretagogue induced PGD2 production from rat peritoneal mast cells --- p.69Chapter 3.3.10. --- Effects of frusemide and DSCG on cellular cAMP levels --- p.70Chapter 3.4. --- Discussion --- p.101Chapter 3.5. --- Summary --- p.111Chapter 3.6. --- Conclusion --- p.114Chapter 3.7. --- Future studies --- p.114Chapter Chapter 4 --- Histamine Receptor Directed Compounds --- p.115Chapter 4.1. --- Introduction --- p.116Chapter 4.1.1. --- Histamine receptor subtypes --- p.116Chapter 4.1.2. --- Histamine effects on the airways --- p.117Chapter 4.2. --- Signal transduction mechanisms --- p.118Chapter 4.2.1. --- H1-receptors --- p.118Chapter 4.2.2. --- H2-receptors --- p.119Chapter 4.2.3. --- H3-receptors --- p.120Chapter 4.3. --- Histamine receptors and mast cells --- p.120Chapter 4.3.1. --- Effects of histamine agonists and antagonists on mast cells --- p.120Chapter 4.3.2. --- Experimental aims --- p.122Chapter 4.3.3. --- Materials and methods --- p.123Chapter 4.4. --- Results --- p.123Chapter 4.4.1. --- Effects of the test compounds on the spontaneous histamine release from rat peritoneal mast cells --- p.123Chapter 4.4.2. --- Effects of the test compounds on anti-IgE induced histamine release from rat peritoneal mast cells --- p.125Chapter 4.4.3. --- Effects of the test compounds on compound 48/80 induced histamine release from rat peritoneal mast cells --- p.126Chapter 4.4.4. --- Effects of the test compounds on anti-IgE and compound 48/80induced histamine release from rat peritoneal mast cells in calcium free buffer --- p.126Chapter 4.4.5. --- Effects of the test compounds on ionophore A23187 induced histamine release from rat peritoneal mast cells --- p.127Chapter 4.4.6. --- "Effects of histamine antagonists on dimaprit, imetit and impromidine induced histamine release from rat peritoneal mast cells" --- p.128Chapter 4.4.7. --- "Effects of anti-IgE, dimaprit and imetit on PGD2 production from rat peritoneal mast cells" --- p.128Chapter 4.4.8. --- "Effects of benzalkonium chloride (BAC) on dimaprit, imetit, compound 48/80 and anti-IgE induced histamine release from rat peritoneal mast cells" --- p.129Chapter 4.4.9. --- "Effects of pertussis toxin on dimaprit, imetit, compound 48/80and anti-IgE induced histamine release from rat peritoneal mast cells" --- p.129Chapter 4.4.10. --- "Effects of dimaprit, imetit, compound 48/80 and anti-IgE on the free intracellular calcium concentration of rat peritoneal mast cells" --- p.130Chapter 4.5. --- Discussion --- p.171Chapter 4.5.1. --- The possible existence of histamine receptors on rat peritoneal mast cells --- p.171Chapter 4.5.2. --- "Possible mechanism of action for the histamine releasing actions of dimaprit, imetit and impromidine on rat peritoneal mast cells" --- p.174Chapter 4.6. --- Conclusion --- p.181Chapter 4.7. --- Future studies --- p.182Chapter Chapter 5 --- Cyclo-oxygenase Inhibitors --- p.184Chapter 5.1. --- Introduction --- p.185Chapter 5.1.1. --- Cyclo-oxygenase isozymes --- p.185Chapter 5.1.2. --- Cyclo-oxygenase inhibitors and mast cells --- p.186Chapter 5.1.3. --- Experimental aims --- p.190Chapter 5.2. --- Materials and methods --- p.190Chapter 5.3. --- Results --- p.191Chapter 5.3.1. --- Effects of cyclo-oxygenase inhibitors on immunologically and non-immunologically induced histamine release from rat peritoneal mast cells - --- p.191Chapter 5.3.2. --- Effects of cyclo-oxygenase inhibitors on immunologically and non-immunologically induced PGD2 production from rat peritoneal mast cells --- p.192Chapter 5.3.3. --- Effects of cyclo-oxygenase inhibitors on immunologically and non-immunologically induced LTC4 production from rat peritoneal mast cells --- p.192Chapter 5.3.4. --- Effects of cyclo-oxygenase inhibitors on immunologically and non-immunologically induced 45Ca uptake by rat peritoneal mast cells --- p.193Chapter 5.4. --- Discussion --- p.221Chapter 5.5. --- Summary and Conclusion --- p.225Chapter Chapter 6 --- Immunosuppressive Drugs --- p.228Chapter 6.1. --- Introduction --- p.229Chapter 6.1.1. --- CsA and FK506 binding proteins --- p.230Chapter 6.1.2. --- Distribution of CyPA and FKBP12 --- p.231Chapter 6.1.3 --- Mechanism of immunosuppression --- p.232Chapter 6.1.4. --- The role of calcineurin in IL-2 promoter induction --- p.233Chapter 6.2. --- Immunosuppressive agents and mast cells --- p.234Chapter 6.2.1. --- Introduction --- p.234Chapter 6.2.2. --- CsA and FK506 inhibit mast cell cytokine production --- p.235Chapter 6.2.3. --- "CsA mediated inhibition of mediator release from, and calcium uptake by mast cells and basophils" --- p.236Chapter 6.2.4. --- Inhibition of mediator release from mast cells and basophils by FK506 --- p.239Chapter 6.2.5. --- Aim of this study --- p.240Chapter 6.2.6. --- Materials and methods --- p.241Chapter 6.3. --- Results --- p.241Chapter 6.3.1. --- Effects of CsA and FK506 on immunologically and non-immunologically induced histamine release from rat peritoneal mast cells --- p.241Chapter 6.3.2. --- Effects of CsA and FK506 on immunologically and non-immunologically induced PGD2 production from rat peritoneal mast cells --- p.242Chapter 6.3.3. --- Effects of CsA and FK506 on immunologically and non-immunologically induced 45Ca uptake by rat peritoneal mast cells --- p.243Chapter 6.4. --- Discussion --- p.254Chapter 6.4.1. --- "Effects of CsA on histamine release from, and 45Ca uptake by rat peritoneal mast cells, following immunological and non-immunological activation" --- p.254Chapter 6.4.2. --- Effects of CsA on PGD2 production from rat peritoneal mast cells --- p.256Chapter 6.4.3. --- "Effects of FK506 on histamine release from, and 45Ca uptake by rat peritoneal mast cells, following immunological and non-immunological activation" --- p.256Chapter 6.4.4. --- Effects of FK506 on immunological PGD2 production from rat peritoneal mast cells --- p.257Chapter 6.5. --- Summary --- p.257'Chapter 6.6. --- Future work --- p.258Reference